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Sample records for reduviidae em nicotiana

  1. <em>In Vitroem> Antioxidant Properties of Flavonoids and Polysaccharides Extract from Tobacco (<em>Nicotiana> em>tabacum> L. Leaves

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    Jing-Lu Wang

    2012-09-01

    Full Text Available In the present study, antioxidant properties of flavonoids and polysaccharides from tobacco (<em>Nicotiana> tabacumem> L. leaves were evaluated in several <em>in vitroem> systems, e.g., scavenging activities on hydroxyl, superoxide anion, 1,1-diphenyl-2-picrylhydrazyl (DPPH and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS radicals, and reducing power. Flavonoids showed much better activity than polysaccharides in scavenging activities on free radicals. When compared to the positive control, ascorbic acid, both showed weaker antioxidant potential. However, flavonoids possessed comparable superoxide anion, DPPH and ABTS radical scavenging abilities to ascorbic acid at high concentration (600 μg/mL. Meanwhile, it was found that flavonoids had prominent effects on the reducing power, which was equivalent to ascorbic acid, and was significantly higher than polysaccharides. These results clearly indicate that flavonoids are effective in scavenging free radicals and have the potential to be powerful antioxidants. Thus, tobacco leaves could be considered as a potential source of natural antioxidants for food, pharmaceutical, cosmetics or nutraceutical industries.

  2. The role of Nicotiana gluca Graham (paraguayan herbs as an adjuvant in immunomodulation of Newcastle disease vaccine for broilers Estudo da ação de Nicotiana glauca Graham (erva paraguaia como coadjuvante em vacina contra a doença de Newcastle em frangos de corte

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    Fabiane Pereira Gentilini

    2008-07-01

    Full Text Available The <em>Nicotiana glaucaem> is a native plant species from Argentina, but found all over South América, being used against headaches, rheumatism, injuries, ulcers, and so on. Researchers have considered it as having immunomodulation effect. This study was conducted to investigate the use of a aqueous extract of <em>Nicotiana glauca em>Graham as an immunomodulator (adjuvant of a Newcastle disease vaccine.. A total of 56 broilers were distributed into 4 experimental groups. Each one of them received 3 dosages of this vaccine with or without the addition of different concentrations of the extract Using hemmoaglutination inhibition techniques , the results have shown differences (P<0.05 in the third sera collection. An increase in the antibody titer with the inclusion of 5 mg/dosage of the extract (Treatment 3 as compared to 1 mg/dosage (Treatment 2 and 10 mg/dosage of the extract (Treatment 4 was observed, However, birds from Treatment 3 did not differ (P> 0.05 from Treatment 1. These results indicated that further investigations are required, including the use of cytotoxicity tests in vitro, to evaluate the immunomodulation effect of this extract.

     

    KEY WORDS: Immunomodulation effect, <em>Nicotiana glaucaem> Graham, vaccine.

    A <em>Nicotiana glaucaem> Graham é uma espécie nativa da Argentina, bem distribuída na América do Sul, sendo empregada, popularmente, contra dores de cabeça, dores reumáticas, cicatrização de feridas e úlceras, entre outros. Pesquisas têm avaliado a sua ação na potencialização da resposta imune. Assim, com este estudo, buscou-se avaliar a ação de um extrato aquoso de <em>Nicotiana glauca em>Graham como coadjuvante imunológico em uma vacina contra a doença de Newcastle (DNC. Utilizaram-se 56 frangos de corte, distribuídos em quatro grupos experimentais, que receberam tr

  3. Tumores em certos híbridos do gênero nicotiana

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    F. G. Brieger

    1942-01-01

    Full Text Available The present paper gives some observations on the tumour formation and other abnormalities observed in the interspecific hybrids, Nicotiana glauca x N. Langsdorffii and N. glauca x N. Sanderae. Without entering into a detailed discussion of the literature and the various speculations proposed by Kostoff and Whitaker about the causes of tumour formations or on the relations between "spontaneous" and pathogenic tumours (Kostoff, Levine and others, two new observations are described. 1 The abnormalities in the two above mentioned hybrids are quite different. As already reported by other authors, the hybrids N. glauca x N. Langsdorffii are initially quite normal and vigorous, while tumours and other proliferations, on the stem, the stem base and the root appear more or less frequently in older plants. Tumours on leaf are rare and flowers are normal (fig. 1,2,7 and 10. The hybrids N. glauca x N. Sanderae are very different from the former. These remain either weak and rachitic, forming early large tumours at the stem base and on the root (fig. 5 and 6 right half. Numerous shoots with more or less normal leaves may arise from these tumours (fig. 11 and 12. Or the plants show an extremely vigorous and abnormal ramification with a number of serial axial shoots developing in each leaf axis (fig. 3 and 4. The leaves are frequently deformed and the flowers exhibit abnormally coloured patches. When crossing N. glauca with F1 (N. Langsdorffii x N. Sanderae the offspring could clearly be divided into two groups : plants exactly as described for N. glauca x N. Langsdorffii and others which correspond to the types of N. glauca x N. Sanderae. Thus it seems that we are dealing with a monogenic differences, each species, N. Langsdorffii and N. Sanderae, being homozygous for one allele each. These alleles have no phenotypic effect in either pure species, but only in hybrids when combined with the genom of N. glauca. The situation is somewhat similar to the sublethal

  4. Fauna epigeica em sistemas de produção de Nicotiana tabacum L.

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    Marta Sandra Drescher

    2011-10-01

    Full Text Available O Rio Grande do Sul detém grande relevância na produção de tabaco. Adotam-se três sistemas de manejo: convencional, mínimo e plantio direto. Para analisar o impacto desses sistemas sobre a população da fauna do solo, efetuou-se uma amostragem da fauna epigeica. Foram avaliados preparo convencional (PC, cultivo mínimo (CM e plantio direto (PD, além de área de reconversão da fumicultura para a vitivinicultura (RV e de mata nativa (MN. Na amostragem, foram utilizadas dez armadilhas Provid em cada área. Os atributos avaliados foram: abundância de organismos, riqueza, índice de diversidade (H' e equitabilidade de Shannon (J. Collembola (Arthropoda: Hexapoda foi identificada até o nível de família, devido à sua sensibilidade às modificações do ambiente. Efetuaram-se a análise multivariada de Agrupamento Hierárquico e a Análise de Componentes Principais, e os índices H' e J foram comparados pelo teste t, de Student. O PD apresentou maior número total de organismos, isolando-se das demais áreas pela análise de agrupamento hierárquico. Os valores de riqueza foram muito semelhantes entre as áreas, destacando-se a aproximação entre RV e MN. Os tratamentos PC, PD e RV não diferiram estatisticamente quanto ao H', apresentando valores de 1,58; 1,60; e 1,52, respectivamente. CM apresentou menor valor de H' (1,18 e MN, o valor intermediário (1,33. O comportamento estatístico de J foi muito semelhante ao encontrado em H'. Em relação à Classe Collembola, foram identificadas as famílias Entomobrydae, Sminthuridae, Poduridae e Hipogasturidae. PD apresentou exemplares das quatro famílias identificadas, enquanto nas demais áreas foram identificadas apenas Entomobrydae e Sminthuridae. Pela Análise de Componentes Principais (PCA, as diferenças entre as áreas explicaram 32 % da variabilidade encontrada; desse percentual, 59,3 % foram explicados pelos eixos 1 e 2. A presença ou ausência de revolvimento do solo e a rota

  5. ANORMALIDADES EN FLORES DE NICOTIANA

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    Raúl Martínez Crovetto

    2010-11-01

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    En este trabajo se describen las siguientes anormalidades florales: la adherencia de tallos florales, <em>spirody, dialysepaly, dialypetaly, staminodiody, petalodyem> de estambre y la supresión de estilo en <em>Nicotiana rusticaem>, y <em>dialysepaly, dialypetaly, dialycarpely,em> supresión de estilo y <em>staminody> del sépalo en <em>Nicotiana tabacumem>. Algunos de estos términos se proponen por primera vez por los fenómenos teratológicos.

  6. Avaliação de óleos essenciais de plantas aromáticas com atividade inseticida em Triatoma infestans (Klug, 1834 (Hemiptera: Reduviidae = Evaluation of essential oils with insecticidal activity in Triatoma infestans (Klug, 1834 (Hemiptera: Reduviidae

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    Suellem Petilim Gomes

    2011-07-01

    Full Text Available A doenca de Chagas e um dos maiores problemas de saude publica na America Latina. Neste trabalho, foi avaliada a acao inseticida de oleos essenciais de Anacardium humile Cymbopogon nardus, Ocimum basilicum e Eucalyptus urograndis em ninfas do 3‹ e 4‹ estadios dedesenvolvimento de Triatoma infestans por acao topica e pressao a vapor (fumigacao. Para o teste topico foram realizadas duas repeticoes cada uma com cinco ninfas de 3‹ e 4‹ estadios de desenvolvimento de T. infestans. Foi aplicado 1ƒÊL de cada concentracao na regiao dorsal de cada individuo. Para o teste de fumigacao foram utilizados potes de 2 L com tampa vedante. Os oleos essenciais de O. gratissimum e E. urograndis apresentaram atividadeinseticida para T. infestans, e A. humile e C. nardus apresentaram em media 5 e 15% de mortalidade, respectivamente. Para fumigacao, somente o oleo de E. urograndis teve efeito, com 100% de mortalidade sobre T. infestans , sendo que para os outros oleos nao foiobservado efeito de fumigacao. Esse trabalho relata pela primeira vez a acao inseticida de E. urograndis em potes fumigenos para o controle de triatomineos. Os resultados sao importantes devido a busca por novos modelos moleculares com acao inseticida em triatomineos resistentes aos inseticidas sinteticos.Chagas disease is a major public health problem in Latin America. This work aimed to evaluate the insecticide action of essential oils of Anacardium humile, Cymbopogon nardus, Ocimum basilicum and Eucalyptus urograndis on Triatoma infestans nymphs of third and fourth stages of development by topic action and steam pressure (fumigation. Two repetitions were performed at the topic test with five nymphs of third and fourth stages of development of T. infestans. It was applied 1ƒÊL of each concentration on the dorsal region of each nymph. Two-liter pots with sealing covers were used at the fumigation test. The essential oils of O. gratissimum and E. urograndis showed insecticidal

  7. Avaliação de óleos essenciais de plantas aromáticas com atividade inseticida em Triatoma infestans (Klug, 1834 (Hemiptera: Reduviidae Evaluation of essential oils with insecticidal activity in Triatoma infestans (Klug, 1834 (Hemiptera: Reduviidae

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    Silvio Favero

    2011-09-01

    Full Text Available A doença de Chagas é um dos maiores problemas de saúde pública na América Latina. Neste trabalho, foi avaliada a ação inseticida de óleos essenciais de Anacardium humile Cymbopogon nardus, Ocimum basilicum e Eucalyptus urograndis em ninfas do 3° e 4° estádios de desenvolvimento de Triatoma infestans por ação tópica e pressão a vapor (fumigação. Para o teste tópico foram realizadas duas repetições cada uma com cinco ninfas de 3° e 4° estádios de desenvolvimento de T. infestans. Foi aplicado 1µL de cada concentração na região dorsal de cada indivíduo. Para o teste de fumigação foram utilizados potes de 2 L com tampa vedante. Os óleos essenciais de O. gratissimum e E. urograndis apresentaram atividade inseticida para T. infestans, e A. humile e C. nardus apresentaram em média 5 e 15% de mortalidade, respectivamente. Para fumigação, somente o óleo de E. urograndis teve efeito, com 100% de mortalidade sobre T. infestans, sendo que para os outros óleos não foi observado efeito de fumigação. Esse trabalho relata pela primeira vez a ação inseticida de E. urograndis em potes fumígenos para o controle de triatomíneos. Os resultados são importantes devido à busca por novos modelos moleculares com ação inseticida em triatomíneos resistentes aos inseticidas sintéticos.Chagas disease is a major public health problem in Latin America. This work aimed to evaluate the insecticide action of essential oils of Anacardium humile, Cymbopogon nardus, Ocimum basilicum and Eucalyptus urograndis on Triatoma infestans nymphs of third and fourth stages of development by topic action and steam pressure (fumigation. Two repetitions were performed at the topic test with five nymphs of third and fourth stages of development of T. infestans. It was applied 1µL of each concentration on the dorsal region of each nymph. Two-liter pots with sealing covers were used at the fumigation test. The essential oils of O. gratissimum and E

  8. Sobrevivência e ocorrência de muda em Triatoma infestans Klug (Hemiptera, Reduviidae após choque de temperatura Survival and molting occurrence in Triatoma infestans Klug (Hemiptera, Reduviidae after temperature shocks

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    Vera Lúcia C.C. Rodrigues

    1991-12-01

    Full Text Available A sobrevivência e a ocorrência de mudas em espécimes de Triatoma infestans foram estudados num período de 30 dias após choques de temperatura. Foi demonstrado que choques hipertérmico e hipotérmico interferem nesses processos, na dependência da temperatura do choque, tempo de sua duração, fase de desenvolvimento e sexo dos espécimes. Dentre as situações experimentais utilizadas, o choque a 0°C por 12 h pareceu produzir a ação mais deletéria, porém choques a 40°C e 0°C mesmo por uma hora são admitidos como afetando o desenvolvimento hormonal que controla a muda. Casos de aumento de sobrevivência pós-choque são sugeridos como tendo sido favorecidos por ação de proteínas de choque térmico.Survival and molting occurrence were studied in specimens of Triatoma infestans over 30 days after temperature shocks. Hyperthermal and hypotermal shocks could be found to affect both survival and molting incidence as a function of temperature and period of the developement phase and sex of the specimens. Considering the various test conditions, the shock at 0°C for 12 h was found to elicit the most deleterious effect, whereas shocks at 40°C and 0°C even for 1 h are interpreted as affecting the hormonal balance which controls molting. Cases of a rise in post-shock survival are suggested to have been favored by heat-shock protein action.

  9. Resistência ao jejum de Rhodnius stali Lent, Jurberg & Galvão, 1993 (Hemiptera, Reduviidae, Triatominae) em condições de laboratório

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    Cailleaux, Solange Ribeiro Peixoto; Cunha,Vanda; Verly, Sergio; Lamas Junior, Valdir Dias; Jurberg, José

    2011-01-01

    O objetivo deste estudo é avaliar a capacidade de resistência à privação alimentar de Rhodnius stali Lent, Jurberg, & Galvão, 1993, em condições de laboratório sob temperatura e umidade média ambiental registrada. Ovos recémcolocados, de casais previamente separados, foram transferidos para tubos de polipropileno com tampa de rosca, com capacidade para 50 mL, forrados com papel filtro e, em seu interior, foi colocada uma tira do mesmo papel, dobrada, para aumentar a área de contat...

  10. Fauna Triatominae do Estado de Bahia, Brasil.V - Microtriatoma pratai sp. n. Hemiptera, Reduviidae, Triatominae, Bolboderini Triatomidae fauna of the State of Bahia, Brazil. V - Microtriatoma pratai sp. n. - Hemiptera,Reduviidae, Triatominae, Bolderini

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    Ítalo A. Sherlock

    1982-06-01

    Full Text Available Microtriatoma pratai sp. n. de Hemíptera, Reduviidae, Triatominae, Bolboderini, é descrita no presente trabalho com base em um exemplar coletado em domicílio da cidade do Salvador, Estado da Bahia, Brasil. A espécie se distingue facilmente das outras conhecidas do gênero, principalmente por apresentar asas sem manchas e conexivo com largas manchas claras e escuras alternadas.Microtriatoma pratai n. sp. (Hemíptera, reduviidae, Triatominae, Bolboderini is described from a single specimen collected inside a house in the city of Salvador, State of Bahia, Brazil. This species is easily distinguished from the others belonging to the genus principally by the existence of alternating broad light and dark coloured spots on the connexivum and the absence of spots on the hemelytra.

  11. Sobre a variação intra-específica em Triatoma dimidiata (Latreille e Triatoma infestans (Klug (Hemiptera, Reduviidae

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    Herman Lent

    1985-09-01

    Full Text Available A observação de oito exemplares de Triatoma dimidiata (Latreille, 1811 de procedência diversas, com vistas a procurar caracterizar as estruturas da genitália externa dos machos (processo do endosoma, suporte do falosoma, falosoma e visica levou-nos a constatar a mesma variabilidade que a espécie demonstra em relação aos seus caracteres externos de colorido e relações da cabeça e do rosto. Por associação de idéias, ampliamos um estudo anterior sobre a genitália masculina de Triatoma infestans (Klug, 1834, também espécie de larga distribuição geográfica, porém com caracteres externos estáveis. Encontramos assim mesmo, embora em percentagem pequena, que os espinhos do processo do endosoma, habitualmente em número reduzido, podiam não existir. Desta espécie observamos quinze exemplares, de procedências diversas.The analysis of eight speciemens of Triatoma dimidiata (Latreille, 1811 of diverse origins, in order to characterize the structure of the male external genitalia (endosoma process, struts, phallosoma, vesica led us to point out that the same variability is demonstrate by the species in relation to its external colour characters and the relation between the head and rostrum. Triatoma dimidiata is a vector of Chagas' disease in several American countries, from Mexico (extreme north of its distribution to the north of South America, including Colombia, Venezuela, Ecuador and Peru. Similarly we have carried out a large scale study on male genitalia of Triatoma infestans (Klug, 1834 which also has a wide geographic distribution, but with stable external characters. The spines of the endosoma process which generally are few in number, might not exist in a small percentage of cases. We analysed fifteen specimens of diverse origins.

  12. Alguns aspectos da biologia de Triatoma pseudomaculata Corrêa & Espínola, 1964, em condições de laboratório (Hemiptera:Reduviidae:Triatominae

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    Teresa Cristina M Gonçalves

    1997-03-01

    Full Text Available Biology of Triatoma pseudomaculata Corrêa & Espínola, 1964, under Laboratory Conditions (Hemiptera:Reduviidae:Triatominae - Observations were made on the evolutive cycle of Triatoma pseudomaculata, held under laboratory conditions, fed weekly on bird (pigeon. Of 60 eggs obtained, only 34 nymphs reached the adult stage in a period of X(S=398±76 days. The following parameters were observed: the time immature stages took to develop from egg to adult emergence; the occurrence of the first meal; the time-lapse between the presenting of the blood-meal and the begining of feeding; time of feeding; amount of blood ingested; variation of weight 24 hr after the blood-meal and until the next blood-meal; and the defecation pattern. The experiment was carried out for 20 months, held in BOD incubator with the average of temperature and humidity of 28±1ºC and 80±5% RU, respectively

  13. Análise das relações taxonômicas e sistemáticas entre espécies de triatomíneos (Hemiptera: Reduviidae) de colônias mantidas pelo Serviço Especial de Saúde de Araraquara, inferida de seqüências de genes mitocondriais

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    Walter Ceretti Junior

    2008-01-01

    Os insetos da subfamília Triatominae Jeannel, 1919 de Reduviidae (Hemiptera: Heteroptera: Reduviidae), também conhecidos como barbeiros, constituem um grupo amplamente distribuído pela região Neotropical que comporta hoje, 142 espécies ocorrentes e uma fóssil, distribuídas em 18 gêneros, são insetos hematófagos estritos em todas as fases da vida e reconhecidos vetores da Doença de Chagas. Essa enfermidade é considerada um dos mais importantes problemas de saúde na América Latina, com cerca de...

  14. Triatoma brasiliensis (Neiva,1911 (Hemiptera, Reduviidae: observações sobre seu comportamento em relação à fonte alimentar em galinheiro experimental Triatoma brasiliensis Neiva, 1911 (Hemiptera, Reduviidae: observations on dynamics of feeding behavior in an artificial ecotope

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    Vera Lúcia C. C. Rodrigues

    1995-06-01

    Full Text Available O presente trabalho tem a finalidade de estudar o comportamento de triatomíneos no interior de ecótopos artificiais. Para tanto, construiu-se galinheiro experimental com paredes internas formadas por blocos de barro desmontáveis, abrigando aves como fonte alimentar. O galinheiro foi subdividido em 4 áreas, duas destas separadas por telas e tijolos furados, sendo a menor o local de soltura dos triatomíneos e a maior o galinheiro propriamente dito. Dessa maneira buscou-se a coleta de informaç��es a respeito do deslocamento, sobrevida e locais de abrigo de ninfas do 4º estádio de Triatoma brasiliensis, na certeza de que os mesmos iriam se acomodar nas proximidades da fonte alimentar. As paredes do galinheiro foram desmontadas e pesquisadas mensalmente durante 6 meses e as ninfas encontradas, marcadas com diferentes cores de acordo com o mês da leitura e a posição das mesmas nas áreas. Os resultados alcançados mostraram que 88,5% das ninfas estavam engorgitadas e foram encontradas próximas ao local de repouso (poleiro da fonte alimentar. Após a primeira alimentação, não abandonavam as proximidades da fonte. Os resultados alcançados apresentam, não apenas interesse epidemiológico, como também interesse na área de controle dos vetores da doença de Chagas, tendo em vista as possíveis implicações quanto aos procedimentos na aplicação do inseticida.In order to study the behavior of triatomine bugs under artificial ecotopic conditions, an experimental, double-walled poultry coop was built. Its inside walls were made of sun-dried clay blocks which could be broken apart for better access to the insects hiding in their crevices. The coop was subdivided into two main areas of different sizes using wire netting and a wall made of perforated bricks. The chickens, the only food source available for the triatomines, were kept in the larger, rectangular area, further subdivided diagonally. The triatomines were released in the

  15. Estrutura de populações de Rhodnius neglectus Lent e Psammolestes tertius Lent & Jurberg (Hemiptera, Reduviidae em ninhos de pássaros (Furnariidae presentes na palmeira Mauritia flexuosa no Distrito Federal, Brasil Population structure of Rhodnius neglectus Lent and Psammolestes tertius Lent & Jurberg (Hemiptera, Reduviidae in bird nests (Furnariidae on Mauritia flexuosa palm trees in Federal District of Brazil

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    Rodrigo Gurgel-Gonçalves

    2007-03-01

    Full Text Available Rhodnius neglectus Lent, 1954 {Rn}e Psammolestes tertius Lent & Jurberg, 1965 {Pt} são triatomíneos que ocorrem em ninhos de aves, principalmente da família Furnariidae. O ciclo biológico dessas espécies é conhecido em condições de laboratório, sendo poucos estudos em ecótopos silvestres. Para analisar a infestação e estrutura de populações de Rn e Pt em ninhos de aves presentes na palmeira Mauritia flexuosa Linnaeus, em duas estações climáticas do Brasil Central, foram amostradas 41 palmeiras com evidências de nidificação de Phacellodomus ruber Vieillot, 1817 (22 na estação chuvosa e 19 na estação seca em quatro áreas do Distrito Federal. Os insetos foram capturados usando-se coleta manual na copa da palmeira, identificados morfologicamente, separados por sexo e estádio ninfal. Fezes e glândulas salivares de Rn foram examinadas para verificar infecção por Trypanosoma cruzi Chagas, 1909 e/ou T. rangeli Tejera, 1920. Trinta e cinco palmeiras com ninhos de P. ruber estavam infestadas por Rn (85% e 22 por Pt (53%. 442 indivíduos foram coletados na estação seca (200 Rn e 242 Pt e 267 na estação chuvosa (136 Rn e 131 Pt. O único fator relacionado significativamente com a densidade de triatomíneos nas palmeiras foi a área. A estrutura etária das populações mostrou: a maior abundância de adultos nas populações de Pt, b maior abundância de machos em ambas as espécies e c presença de fêmeas ovipondo em ambas as estações. Nenhum dos 177 triatomíneos examinados estava infectado por T. cruzi ou T. rangeli. A estrutura etária das populações de Rn e Pt não diferiu significativamente entre as estações amostradas, indicando ausência de marcada sazonalidade para essas espécies.Rhodnius neglectus Lent, 1954 {Rn}and Psammolestes tertius Lent & Jurberg, 1965 {Pt} are triatomines that occur in bird nests, mainly furnarid nests. Their biological cycles are known in laboratory conditions and few studies were

  16. Comparative biology of the two sister species of Triatominae (Hemiptera: Reduviidae Biologia comparativa de duas espécies irmãs de Triatominae (Hemiptera: Reduviidae

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    Ana Laura Carbajal de la Fuente

    2010-02-01

    Full Text Available INTRODUCTION: Triatoma pseudomaculata and T. wygodzinskyi (Hemiptera: Reduviidae: Triatominae are two Brazilian vectors of Chagas disease. The first is an arboricolous species in sylvatic environment and considered a vector of T. cruzi in peridomestic structures; the second, a rupicolous species in the wild environment of no epidemiological importance. In order to test the assumption that sister species share biological traits, comparative studies of their development cycle and blood ingestion were conducted. METHODS: Eggs laid by five field females of each species were randomly selected. The nymphs were observed daily and fed on mice weekly. The time required to pass through the different stages to adulthood was recorded in days. The triatomines were weighed individually before and after feeding. The mortality rate according to each nymphal stage was calculated. RESULTS AND CONCLUSIONS: Analysis of the results shows that they display only minor biological differences even though they exhibit a distinct ecology. This suggests that the biological traits are important criteria to determine the relationship between species.INTRODUÇÃO: Triatoma pseudomaculata e T. wygodzinskyi (Hemiptera: Reduviidae: Triatominae são dois vetores Brasileiros da doença de Chagas. A primeira é uma espécie arborícola em ambiente silvestre e considerada vetor do T. cruzi em estruturas peridomesticas. A segunda é rupícola em ambientes silvestres e sem importância epidemiológica. Com o objetivo de testar a hipótese que espécies irmãs compartilham características semelhantes, realizamos um estudo comparativo do ciclo biológico e ingesta alimentar. MÉTODOS: Ovos pertencentes a cinco fêmeas de cada espécie provenientes do campo foram selecionados aleatoriamente. As ninfas foram observadas diariamente e alimentadas com camundongos semanalmente. O tempo requerido para passar até o estágio adulto foi registrado em dias. Os triatomíneos foram pesados

  17. Biologia do Triatoma vitticeps (Stal, 1859 em condições de laboratório (Hemiptera: Reduviidae: Triatominae: II. Resistência ao jejum Biology of Triatoma vitticeps (Stal, 1859 under laboratory conditions (Hemiptera: Reduviidae: Triatominae: II. Resistance to starvation

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    Teresa Cristina M. Gonçalves

    1989-03-01

    Full Text Available Em prosseguimento ao estudo da biologia do Triatoma vitticeps (Gonçalves et al., 1988, foram feitas observações sobre a sua resistência ao jejum. Dos 286 ovos obtidos, apenas 201 eclodiram e atingiram o estádio pretendido para as observações. Os demais não eclodiram, não completaram a muda ou morreram sem motivo aparente. As ninfas foram acondicionadas, individualmente em frascos de Borrel, devidamente registrados. Para a alimentação foram utilizados camundongos e a medida que as ninfas atingiam o estágio pretendido a alimentação era suspensa até ocorrer a morte. A avaliação da resistência ao jejum foi feita da seguinte forma: o intervalo de dias entre o último repasto e a morte e entre a muda e a morte. Verificou-se que a resistência está diretamente relacionada com a fase de desenvolvimento. Para os parâmetros último repasto/morte e muda/morte, ambos os sexos foram menos resistentes do que as ninfas de 3º e 2º estádios, respectivamente. O experimento teve duração de 15 meses e neste período as temperaturas máxima e mínima e a umidade relativa do ar variaram em média de 25 ± 2ºC e 81 ± 3% UR, respectivamente. O material foi proveniente da criação de triatomíneos mantida no Departamento de Entomologia do Instituto Oswaldo Cruz.Following the study on the biology of Triatoma vitticeps (Gonçalves et al. 1988 observations have been made on its resistance to starvation. Of the 286 eggs obtained only 201 hatched and reached the intended stage for observation. The others did not eclode, neither reached the ecdisis nor died, without explanation. The nymphs were kept, separately, in Borrel flasks and properly listed. The blood-meal was performed in mice, although the insects were kept without feeding as soon as moulted. The starvation was evaluated in two ways: the time-lapse in days between the last meal/death and between mult/death. The starvation was directly related with the developmental stage. In relation to

  18. Distribuição espacial de populações de triatomíneos (Hemiptera: Reduviidae em palmeiras da espécie Mauritia flexuosa no Distrito Federal, Brasil

    Directory of Open Access Journals (Sweden)

    Gurgel-Gonçalves Rodrigo

    2004-01-01

    Full Text Available Para determinar e analisar a distribuição espacial de populações silvestres de triatomíneos no Distrito Federal, Brasil, foram amostradas 150 palmeiras da espécie Mauritia flexuosa em seis veredas de diferentes paisagens (silvestre, rural e periurbana na estação chuvosa. Os triatomíneos foram identificados morfologicamente, separados por sexo e estadio ninfal e examinados para verificar infecção por Trypanosoma cruzi e Trypanosoma rangeli. Vinte e oito (18,6% palmeiras estavam infestadas por Rhodnius neglectus e 14 (9,5% por Psammolestes tertius. A freqüência dessas espécies em palmeiras com e sem ninhos foi significativamente diferente, sendo superior nas palmeiras com ninhos de aves e mamíferos na copa. O maior número médio de insetos/palmeira foi observado nas áreas rurais, com estimativas de até 838 insetos/hectare. A composição etária das espécies apresentou um padrão diferente, Rhodnius neglectus com predomínio de ninfas e Psammolestes tertius com predomínio de adultos, sendo que muitos ovos de Rhodnius neglectus foram coletados indicando um evento reprodutivo em fevereiro de 2003. Entre os ninhos encontrados nas palmeiras, o do pássaro Phacellodomus ruber (Furnariidae apresentou as maiores abundâncias de triatomíneos, ocorrendo em 42% das palmeiras. A abundância relativa de Rhodnius neglectus e Psammolestes tertius foi maior em áreas rurais que possuíram maior número de ninhos nas palmeiras e menor densidade de palmeiras por hectare. Nenhum dos 96 triatomíneos examinados estava infectado por Trypanosoma cruzi ou Trypanosoma rangeli, apesar de encontrarmos infecção de Rhodnius neglectus por esses parasitas em estudos anteriores.

  19. An overview on the ecology of Triatominae (Hemiptera:Reduviidae).

    Science.gov (United States)

    Galvão, Cleber; Justi, Silvia A

    2015-11-01

    Chagas disease, the American trypanosomiasis, is an important neglected tropical illness caused by the flagellate protozoan Trypanosoma cruzi (Kinetoplastida, Trypanosomatidae) and transmitted by insects of the subfamily Triatominae (Hemiptera: Reduviidae). Here we provide an overview on the current knowledge about Triatominae ecology, its association with human, T. cruzi infection and the immediate consequences of habitat fragmentation. We also discuss the geographic distribution of the species and the importance of predicting their distributions to control programs.

  20. Recombinant protein expression in Nicotiana.

    Science.gov (United States)

    Matoba, Nobuyuki; Davis, Keith R; Palmer, Kenneth E

    2011-01-01

    Recombinant protein pharmaceuticals are now widely used in treatment of chronic diseases, and several recombinant protein subunit vaccines are approved for human and veterinary use. With growing demand for complex protein pharmaceuticals, such as monoclonal antibodies, manufacturing capacity is becoming limited. There is increasing need for safe, scalable, and economical alternatives to mammalian cell culture-based manufacturing systems, which require substantial capital investment for new manufacturing facilities. Since a seminal paper reporting immunoglobulin expression in transgenic plants was published in 1989, there have been many technological advances in plant expression systems to the present time where production of proteins in leaf tissues of nonfood crops such as Nicotiana species is considered a viable alternative. In particular, transient expression systems derived from recombinant plant viral vectors offer opportunities for rapid expression screening, construct optimization, and expression scale-up. Extraction of recombinant proteins from Nicotiana leaf tissues can be achieved by collection of secreted protein fractions, or from a total protein extract after grinding the leaves with buffer. After separation from solids, the major purification challenge is contamination with elements of the photosynthetic complex, which can be solved by application of a variety of facile and proven strategies. In conclusion, the technologies required for safe, efficient, scalable manufacture of recombinant proteins in Nicotiana leaf tissues have matured to the point where several products have already been tested in phase I clinical trials and will soon be followed by a rich pipeline of recombinant vaccines, microbicides, and therapeutic proteins.

  1. Nicotiana glauca poisoning in ostriches (Struthio camelus)

    CSIR Research Space (South Africa)

    Botha, CJ

    2011-01-01

    Full Text Available Putative Nicotiana glauca (wild tobacco) poisoning was diagnosed in a flock of ostriches near Oudtshoorn, South Africa. Post mortem examinations (n = 7) were performed on ostriches (Struthio camelus) that had died. Suspicious leaf remnants (weighing...

  2. Citroenzuur- en nicotinegehalte in enkele variëteiten van Nicotiana tabacum en Nicotiana rustica

    NARCIS (Netherlands)

    Fatton, Numa Edouard

    1939-01-01

    Van technisch rijpe bladen van verschillende varieteiten van Nicotiana rustica en Nicotiana Tabacum werden het citroenzuuren het nicotinegehalte bepaald, uitgedrukt in % op de droge stof. Hetzelfde werd gedaan van bladmoes en hoofdnerf afzonderlijk, verder van de wortel, de stengel, de bloemen en de

  3. Behavior of triatomines (Hemiptera: Reduviidae vectors of Chaga's disease: IV. Fecundity, fertility and longevity of Panstrongylus megistus (Burm., 1835 pairs and virgin females starved under laboratory conditions Comportamento de triatomíneos Hemiptera: Reduviidae vectores da doença de Chagas. IV. Fecundidade, fertilidade e longevidade de casais e fêmeas adultas de Panstrongylus megistus Burm, 1835, privados de alimento, em condições de laboratório

    Directory of Open Access Journals (Sweden)

    Marli Maria Lima

    1987-12-01

    Full Text Available A laboratory study was conducted on the fecundity, fertility and life span of Panstrongylus megistus pairs and on the fecundity and life span of P. megistus virgin females submitted to starvation after the last moulting. Of the mated females, 22.2% laid eggs, 4.4% of which were fertile. Females resisted starvation more than males. Of the starved virgin females, only 10% laid eggs, with a low egg-laying rate (0.47 per female. Resistance to starvation was lower in virgin than in mated females.Efetuou-se, em laboratório, um estudo sobre a fecundidade, fertilidade e duração de vida de casais de Panstrongylus megistus e da fecundidade e duração de vida de fêmeas virgens dessa espécie, submetidos a jejum, após a última ecdise. Das fêmeas acasaladas, 22,2% puseram ovos, dos quais 4,4% estavam férteis. As fêmeas tiveram maior resistência ao jejum que os machos. Das fêmeas virgens em jejum, apenas 10% puseram ovos, com baixa de ovipostura por fêmeas (0,47. A resistência ao jejum foi mais baixa nas fêmeas virgens do que naquelas que permaneceram sempre com os machos.

  4. Genetic variation in oriental tobacco (Nicotiana tabacum L. by agro-morphological traits and simple sequence repeat markers Variação genética em tabaco oriental (Nicotiana tabacum L. por marcadores agro-morfológicos e traços simples de repetição de sequência

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    Reza Darvishzadeh

    2013-06-01

    tabaco simples. As análises de variância mostarram que há pouca alto nível de diversidade genética nos tabacos to tipo oriental baseado nos tratamentos morfológicos incluindo número de folhas, dias para os 50% do florescimento, comprimento da folha, largura da folha, peso fresco da folha, peso seco da folha e maior comprimento e espessura do caule. A classificação dos genótipos usando o método de dados morfológicos por médias de grupos pareados não ponderados usando média aritmética (UPGMA algoritmo baseado nas distâncias Euclidianas quadradas padronizadas resultou quatro grupos distintos de acordo co a própria distribuição geográfica. Nas investigações dos marcadores moleculares, um total de 13 repetições de sequências simples (RSS de iniciadores pares foram utilizados para determinar o polimorfismo do germoplasma de teste. Trinta e cinco alelos foram marcados em 13 locos RSS. O numero médio de alelos por loco (na e o número de alelo efetivo (Ae forma 2,69 e 2,34, respectivamente. Utilizando os dados RSS, coeficientes de similaridade de Jaccard foram produzidos. O agrupamento de genótipos através dos coeficientes de similaridade de Jaccard e usando o método de agrupamento UPGMA que considera três grupos que não tiveram algum alojamento com suas próprias origens. Os resultados revelaram que revelaram que não estão completamente de acordo para classificação baseada em agro-morfológicas por loco e RSS em tabacos do tipo oriental. Por causa da não influencia dos efeitos ambientais no marcador molecular, grupos heteróticos com base em marcadores SSR poderiam estar mais próximos da realidade.

  5. Cladistics and biogeography of the assassin bug genus Rasahus Amyot & Serville (Heteroptera: Reduviidae: Peiratinae)

    NARCIS (Netherlands)

    Morrone, J.J.; Coscarón, M. del C.

    1998-01-01

    The assassin bug genus Rasahus Amyot & Serville (Heteroptera: Reduviidae: Peiratinae) comprises 26 Neotropical species. A cladistic analysis of the genus was carried out using 63 characters from external morphology, body vestiture, and male and female genitalia, with the species considered as termin

  6. Exocrine secretion of wheel bugs (Heteroptera: Reduviidae: Arilus spp): clarification and chemistry

    Science.gov (United States)

    Assassin bugs (Heteroptera: Reduviidae) in the genus Arilus are among the largest of the New World reduviids, and can inflect extremely painful bites. The most common North American species, A. cristatus (the “wheel bug”), possesses eversible osmeterial glands that reportedly produce an obnoxious od...

  7. Cladistics and biogeography of the assassin bug genus Rasahus Amyot & Serville (Heteroptera: Reduviidae: Peiratinae)

    NARCIS (Netherlands)

    Morrone, J.J.; Coscarón, M. del C.

    1998-01-01

    The assassin bug genus Rasahus Amyot & Serville (Heteroptera: Reduviidae: Peiratinae) comprises 26 Neotropical species. A cladistic analysis of the genus was carried out using 63 characters from external morphology, body vestiture, and male and female genitalia, with the species considered as

  8. Systematics of Mepraia (Hemiptera-Reduviidae): cytogenetic and molecular variation.

    Science.gov (United States)

    Calleros, L; Panzera, F; Bargues, M D; Monteiro, F A; Klisiowicz, D R; Zuriaga, M A; Mas-Coma, S; Pérez, R

    2010-03-01

    The haematophagous insects of the subfamily Triatominae (Hemiptera-Reduviidae) have great epidemiological importance as vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Mepraia was originally described as a monotypic genus comprised of Mepraia spinolai, distributed along coastal areas of northern Chile (from Region I to the Metropolitan Region). Recently, some M. spinolai populations have been ranked as a new species named Mepraia gajardoi. Several populations along the distribution range of the genus were sampled, and genetic differentiation was studied based upon the analysis of three molecular markers: cytogenetics (karyotype and chromosome behaviour during meiosis using the C-banding technique), mitochondrial DNA (a cytochrome oxidase I gene fragment), and nuclear ribosomal DNA (intergenic region including the two internal transcribed spacers ITS-1 and ITS-2 and the 5.8S rRNA gene). The data here presented indicate that populations within the Mepraia genus (excluding Region II specimens) can be divided into two separate lineages. One lineage is comprised of specimens from the northernmost Region I and represents M. gajardoi. The other includes samples from the southern III, IV and the Metropolitan Regions, and represents M. spinolai. Region II individuals deserve particular attention as their relationship to the two identified lineages is not clear-cut. While they appear to belong to M. spinolai based on cytogenetics and rDNA markers, COI results indicate a closer relationship to M. gajardoi. This disagreement can be due to mitochondrial DNA introgression or the retention of ancestral polymorphisms.

  9. Cytogenetics and genome evolution in the subfamily Triatominae (Hemiptera, Reduviidae).

    Science.gov (United States)

    Panzera, F; Pérez, R; Panzera, Y; Ferrandis, I; Ferreiro, M J; Calleros, L

    2010-01-01

    The subfamily Triatominae (Hemiptera, Reduviidae), vectors of Chagas disease, includes over 140 species. Karyotypic information is currently available for 80 of these species. This paper summarizes the chromosomal variability of the subfamily and how it may reveal aspects of genome evolution in this group. The Triatominae present a highly conserved chromosome number. All species, except 3, present 20 autosomes. The differences in chromosome number are mainly caused by variation in the number of sex chromosomes, due to the existence of 3 sex systems in males (XY, X(1)X(2)Y and X(1)X(2)X(3)Y). However, inter- and intraspecific differences in the position, quantity and meiotic behavior of constitutive heterochromatin, in the total genome size, and in the location of ribosomal 45S rRNA clusters, have revealed considerable cytogenetic variability within the subfamily. This cytogenetic diversity offers the opportunity to perform cytotaxonomic and phylogenetic studies, as well as structural, evolutionary, and functional analyses of the genome. The imminent availability of the complete genome of Rhodnius prolixus also opens new perspectives for understanding the evolution and genome expression of triatomines. The application of fluorescence in situ hybridization for the mapping of genes and sequences, as well as comparative analyses of genome homology by comparative genomic hybridization will be useful tools for understanding the genomic changes in relation to evolutionary processes such as speciation and adaptation to different environments.

  10. Application of RNA interference in triatomine(Hemiptera:Reduviidae)studies

    Institute of Scientific and Technical Information of China (English)

    Rafaela M.M.Paim; Ricardo N.Araujo; Michael J.Lehane; Nelder F.Gontijo; Marcos H.Pereira

    2013-01-01

    Triatomines(Hemiptera: Reduviidae)are obligate hematophagous insects.They are of medical importance because they are vectors of Trypanosoma cruzi,the causative agent of Chagas disease in the Americas.In recent years,the RNA interference (RNAi)technology has emerged as a practical and useful alternative means of studying gene function in insects,including triatomine bugs.RNAi research in triatomines is still in its early stages,several issues still need to be elucidated,including the description of the molecules involved in the RNAi machinery and aspects related to phenotype evaluation and persistence of the knockdown in different tissues and organs.This review considers recent applications of RNAi to triatomine research,describing the major methods that have been applied during the knockdown process such as the double-stranded RNA delivery mechanism(injection,microinjection,or ingestion)and the phenotype characterization (mRNA and target protein levels)in studies conducted with the intent to provide greater insights into the biology of these insects.In addition to the characterization of insect biomolecules,some with biopharmacological potential,RNAi may provide a new view of the interaction between triatomine and trypanosomatids,enabling the development of new measures for vector control and transmission of the parasite.

  11. [Development of Rhodnius pictipes Stal, 1872 fed on mice and through a silicone membrane (Hemiptera, Reduviidae, Triatominae)].

    Science.gov (United States)

    Rocha, D da S; da Fonseca, A H; Costa, F A; Jurberg, J; Galvão, C

    1997-01-01

    Rhodnius pictipes (Hemiptera, Reduviidae) from Serra Norte, State of Pará, Brazil, acclimatized in an insectary at the Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos, Departamento de Entomologia, Instituto Oswaldo Cruz, were fed through a silicone membrane. In order to know the viability and the efficiency of this membrane compared with insects fed on mice, the number of bloodmeals taken, period of development of the five nymphal instars, longevity of adults, average amount of blood intake in each meal and percent of mortality were observed. A total of 310 insects, were used, comprising 50 nymphs of each instar, as well as 30 male and 30 female adults. Insects fed artificially had reduced minimal and maximal periods of development than the group fed on mice. The largest relative increase of body weight was observed in the 2nd instar followed by the 1st, and the amount of blood ingested increased during the development, to the 5th instar for both groups. There were no significant differences between the groups fed artificially and in vivo according to Tukey's test for p > 0.05. The percent of mortality in the 1st instar was 18% for artificially fed and 16% for the group fed on mice; these percentages decreased as insects developed until the 4th instar, without mortality, returning to increase in the 5th instar. R. pictipes was shown to be easily adaptable to artificial feeding, and could be considered as an important and viable experimental model.

  12. Distribution of solanesol in Nicotiana tabacum

    Institute of Scientific and Technical Information of China (English)

    ZHAO Chun-jian; ZU Yuan-gang; LI Chun-ying; TIAN Cheng-yu

    2007-01-01

    Solanesol is an important secondary metabolite in Nicotiana tabacum. Distribution of solanesol in Nicotiana tabacum was investigated by High Performance Liquid Chromatography (HPLC) method. The quantitative distribution of solanesol in various organs and tissues of N. Tabacum showed that solanesol content, obviously different in all organs, was 6.8, 18.3, 27.5, 45.8, and 68.0 times higher in leaves than that in the stalks, flowers, seeds, fruits and roots, respectively. The contents of solanesol in various parts of leaf, stalk and flower were determined. The content of solanesol in top leaf, middle leaf and bottom leaf gradually decreased (6.124, 5.813 and 5.687 mg·g-1, respectively) and the content of solanesol in various leaf-parts (leaf apex, leaf middle and leaf base) also gradually decreased. The content of solanesol in top stalk was 1.19 times and 1.92 times higher than that in the middle stalk and the bottom stalk, respectively. The content of solanesol in various tissues of stalk (epidermis, cortex and stele) dramatically decreased. The sepal contained higher concentration of solanesol (1.192 mg·g-1) compared to any other parts in flower. The study will provide the base data for the regulation and control of solanesol,moreover, it will provide the scientific evidences for the rational development and utilization of N. Tabacum resources.

  13. Cinco especies nuevas de Pselliopus (Hemiptera: Heteroptera: Reduviidae: Harpactorinae: Harpactorini para México Five new species of Pselliopus (Hemiptera: Heteroptera: Reduviidae: Harpactorinae from Mexico

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    Harry Brailovsky

    2007-06-01

    Full Text Available Se describen 5 especies nuevas de Pselliopus Bergroth (Reduviidae: Harpactorinae recolectadas en México (Guerrero, Hidalgo, Jalisco, Michoacán, Nayarit, Nuevo León, Oaxaca, Puebla, Querétaro, San Luis Potosí, Sinaloa, Sonora y Tamaulipas y en los Estados Unidos de América (Arizona. Pselliopus karlenae Hussey se registra por primera vez para la República Mexicana. Se ilustran caracteres diagnósticos del pronoto, del borde posterior del segmento abdominal VII, de la cápsula genital del macho (pigóforo y parámeros. Se incluye una clave para la identificación de las especies basada principalmente en los genitales masculinos.Five new species of Pselliopus Bergroth (Reduviidae: Harpactorinae from Mexico (Guerrero, Hidalgo, Jalisco,Michoacán, Nayarit, Nuevo León, Oaxaca, Puebla, Querétaro, San Luis Potosí, Sinaloa, Sonora and Tamaulipas and from the United States of America (Arizona are described. Pselliopus karlenae Hussey is a new record for Mexico. Diagnostic taxonomic characters of pronotum, posterior border of abdominal segment VII, male genital capsule (pygophore and parameres are illustrated. An identification key based mainly on the male genitalia is included.

  14. E-beam irradiation for the control of Phytophthora nicotianae var. nicotianae in stonewool cubes

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    Ptaszek Magdalena

    2015-09-01

    Full Text Available Effectiveness of electron beam irradiation was evaluated against Phytophthora nicotianae var. nicotianae, the causal agent of stem base and root rot of tomato. In laboratory trials, irradiation of 7-day-old Phytophthora cultures growing on potato-dextrose-agar (PDA medium with 1 kGy resulted in the disintegration of the pathogen’s hyphae. Increasing the irradiation dose to 3 kGy caused decay of the hyphae. Irradiation of infested stonewool with 5 kGy caused decrease of the pathogen population about 5 times. Application of 20 kGy completely eliminated the pathogen from stonewool. Irradiation of substratum resulted in significant increase of tomato seedlings healthiness, especially when the dose 20 kGy was applied.

  15. Geological Changes of the Americas and their Influence on the Diversification of the Neotropical Kissing Bugs (Hemiptera: Reduviidae: Triatominae.

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    Silvia A Justi

    2016-04-01

    Full Text Available The family Reduviidae (Hemiptera: Heteroptera, or assassin bugs, is among the most diverse families of the true bugs, with more than 6,000 species. The subfamily Triatominae (kissing bugs is noteworthy not simply because it is the only subfamily of the Reduviidae whose members feed on vertebrate blood but particularly because all 147 known members of the subfamily are potential Chagas disease vectors. Due to the epidemiological relevance of these species and the lack of an efficient treatment and vaccine for Chagas disease, it is more common to find evolutionary studies focusing on the most relevant vectors than it is to find studies aiming to understand the evolution of the group as a whole. We present the first comprehensive phylogenetic study aiming to understand the events that led to the diversification of the Triatominae.We gathered the most diverse samples of Reduviidae and Triatominae (a total of 229 Reduviidae samples, including 70 Triatominae species and reconstructed a robust dated phylogeny with several fossil (Reduviidae and Triatominae calibrations. Based on this information, the possible role of geological events in several of the major cladogenetic events within Triatominae was tested for the first time. We were able to not only correlate the geological changes in the Neotropics with Triatominae evolution but also add to an old discussion: Triatominae monophyly vs. paraphyly.We found that most of the diversification events observed within the Rhodniini and Triatomini tribes are closely linked to the climatic and geological changes caused by the Andean uplift in South America and that variations in sea levels in North America also played a role in the diversification of the species of Triatoma in that region.

  16. Evolutionary history of assassin bugs (insecta: hemiptera: Reduviidae: insights from divergence dating and ancestral state reconstruction.

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    Wei Song Hwang

    Full Text Available Assassin bugs are one of the most successful clades of predatory animals based on their species numbers (∼6,800 spp. and wide distribution in terrestrial ecosystems. Various novel prey capture strategies and remarkable prey specializations contribute to their appeal as a model to study evolutionary pathways involved in predation. Here, we reconstruct the most comprehensive reduviid phylogeny (178 taxa, 18 subfamilies to date based on molecular data (5 markers. This phylogeny tests current hypotheses on reduviid relationships emphasizing the polyphyletic Reduviinae and the blood-feeding, disease-vectoring Triatominae, and allows us, for the first time in assassin bugs, to reconstruct ancestral states of prey associations and microhabitats. Using a fossil-calibrated molecular tree, we estimated divergence times for key events in the evolutionary history of Reduviidae. Our results indicate that the polyphyletic Reduviinae fall into 11-14 separate clades. Triatominae are paraphyletic with respect to the reduviine genus Opisthacidius in the maximum likelihood analyses; this result is in contrast to prior hypotheses that found Triatominae to be monophyletic or polyphyletic and may be due to the more comprehensive taxon and character sampling in this study. The evolution of blood-feeding may thus have occurred once or twice independently among predatory assassin bugs. All prey specialists evolved from generalist ancestors, with multiple evolutionary origins of termite and ant specializations. A bark-associated life style on tree trunks is ancestral for most of the lineages of Higher Reduviidae; living on foliage has evolved at least six times independently. Reduviidae originated in the Middle Jurassic (178 Ma, but significant lineage diversification only began in the Late Cretaceous (97 Ma. The integration of molecular phylogenetics with fossil and life history data as presented in this paper provides insights into the evolutionary history of

  17. Lethality of triatomines (Hemiptera: Reduviiae, vectors of Chagas' disease, feeding on blood baits containing synthetic insecticides, under laboratory conditions Letalidade de triatomíneos (Hemiptera: Reduviidae, vetores da doença de Chagas, alimentados sobre iscas de sangue contendo inseticidas sintéticos, em condições de laboratório

    Directory of Open Access Journals (Sweden)

    Marli Maria Lima

    1991-12-01

    Full Text Available A laboratory study was conducted to test the toxicity of synthetic insecticides added to defibrinated sheep blood kept at room temperature and offered as food to the following triatomine species: Triatoma infestans, Panstrongylus megistus, Triatoma vitticeps, Triatoma pseudomaculata, Triatoma brasiliensis and Rhodnius prolixus. The insecticides used, at a concentration of 1g/l, were: HCH, DDT, Malathion and Trichlorfon, and the lethalithy observed at the end of a 7-day period varied according to the active principle of each. HCH was the most effective by the oral route, killing 100% of the insects, except P. megistus (95.7% and T. pseudomaculata (94.1%. Trichlorfon killed the insects at rates ranging from 71.8% (T. vitticeps to 98% (R. prolixus. Malathion was slightly less efficient, killing the insects at rates from 56.8% (T. vitticeps to 97% (T.brasiliensis. DDT was the least effective, with a killing rate of 10% (T. vitticeps to 75% (T.brasiliensis. Since the tests were performed at room temperature, we suggest that baits of this type should be tried for the control of triatomines in the field.Ensaiou-se, em laboratório, o efeito tóxico de inseticidas sintéticos, adicionados ao sangue de carneiro desfibrinado, mantidos em temperatura ambiente, oferecidos como fonte alimentar às seguintes espécies de triatomíneos: Triatoma infestans, Panstrongylus megistus, Triatoma vitticeps, Triatoma pseudomaculata, Triatoma brasiliensis e Rhodnius prolixus. Os inseticidas usados, na concentração de 1g/l, foram: HCH, DDT, Malathion e Triclorfon e a letalidade observada, ao fim de 7 dias variou, de acordo com o princípio ativo. HCH foi o mais efetivo por via oral, matando 100% dos insetos, exceto P. megistus (95,7% e T. pseudomaculata (94,1%. Triclorfon eliminou entre 71,8% (T. vitticeps e 98% (R. prolixus. Malathion ficou ligeiramente abaixo, matando entre 56,8% (T. vitticeps e 97% (T. brasiliensis. DDT foi o menos efetivo, matando entre 10% (T

  18. Fungal flora of the digestive tract of Triatoma infestans (Hemiptera: Reduviidae from Argentina Flora fúngica de tractos digestivos en Triatoma infestans (Hemiptera: Reduviidae en Argentina

    Directory of Open Access Journals (Sweden)

    G. A. Marti

    Full Text Available A survey of the fungal microbiota of the digestive tract of Triatoma infestans (Klug (Hemiptera: Reduviidae adults was carried out. Insects captured in the field from different provinces in Argentina, as well as individuals reared in artificial colonies, were used for dissection. Axenic cultures of the fungal species were identified and were deposited with mycological collections at La Plata , Argentina. A total of 33 fungal species, with the exception of three that were mycelia sterilia, belonging to 11 genera were identified. Thirty two species belonged to Ascomycota (Eurotiomycetes and Sordariomycetes and one to Zygomycota (Zygomycetes. The genera with the greatest number of species were Penicillium (15, Aspergillus (5, and Cladosporium (2. Among the isolated fungi, some of the species were entomopathogenic or pathogens of humans and other animals.En el presente estudio se realizó un relevamiento de la flora fúngica microbiana en tractos digestivos de adultos de Triatoma infestans (Klug (Hemiptera: Reduviidae. Se disecaron insectos capturados del campo en diferentes provincias Argentinas, así como también se utilizaron individuos de una colonia artificial. Fueron realizados cultivos axénicos de las especies fúngicas aisladas, los que fueron identificados y luego depositados en las colecciones de hongos entomopatógenos del CEPAVE La Plata , Argentina. Fueron identificadas 33 especies fúngicas perteneciente a 11 géneros. Treinta y dos especies pertenecen a Ascomycota (Eurotiomycetes y Sordariomycetes y una a Zygomycota (Zygomycetes. Los géneros con mayor número de especies fueron Penicillium (15, Aspergillus (5, y Cladosporium (2. Entre los aislamientos fúngicos, algunas de las especies encontradas son entomopatogénicas o patógenas de humanos y otros animales.

  19. Genetic variation in alkaloid accumulation in leaves of Nicotiana

    Institute of Scientific and Technical Information of China (English)

    Bo SUN; Fen ZHANG; Guo-jun ZHOU; Guo-hai CHU; Fang-fang HUANG; Qiao-mei WANG; Li-feng JIN; Fu-cheng LIN; Jun YANG

    2013-01-01

    Alkaloids are plant secondary metabolites that are widely distributed in Nicotiana species and contribute greatly to the quality of tobacco leaves. Some alkaloids, such as nornicotine and myosmine, have adverse effects on human health. To reduce the content of harmful alkaloids in tobacco leaves through conventional breeding, a genetic study of the alkaloid variation among different genotypes is required. In this study, alkaloid profiles in leaves of five Nicotiana tabacum cultivars and Nicotiana tomentosiformis were investigated. Six alkaloids were identified from al six genotypes via gas chromatograph-mass spectrometry (GC-MS). Significant differences in alkaloid content were ob-served both among different leaf positions and among cultivars. The contents of nornicotine and myosmine were positively and significantly correlated (R2=0.881), and were also separated from those of other alkaloids by clustering. Thus, the genotype plays a major role in alkaloid accumulation, indicating a high potential for manipulation of alkaloid content through traditional breeding.

  20. Genetic variation in alkaloid accumulation in leaves of Nicotiana.

    Science.gov (United States)

    Sun, Bo; Zhang, Fen; Zhou, Guo-jun; Chu, Guo-hai; Huang, Fang-fang; Wang, Qiao-mei; Jin, Li-feng; Lin, Fu-cheng; Yang, Jun

    2013-12-01

    Alkaloids are plant secondary metabolites that are widely distributed in Nicotiana species and contribute greatly to the quality of tobacco leaves. Some alkaloids, such as nornicotine and myosmine, have adverse effects on human health. To reduce the content of harmful alkaloids in tobacco leaves through conventional breeding, a genetic study of the alkaloid variation among different genotypes is required. In this study, alkaloid profiles in leaves of five Nicotiana tabacum cultivars and Nicotiana tomentosiformis were investigated. Six alkaloids were identified from all six genotypes via gas chromatograph-mass spectrometry (GC-MS). Significant differences in alkaloid content were observed both among different leaf positions and among cultivars. The contents of nornicotine and myosmine were positively and significantly correlated (R(2)=0.881), and were also separated from those of other alkaloids by clustering. Thus, the genotype plays a major role in alkaloid accumulation, indicating a high potential for manipulation of alkaloid content through traditional breeding.

  1. Pollen irradiation and possible gene transfer in Nicotiana species

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1985-01-01

    Progeny from crosses of Nicotiana langsdorffii with gamma irradiated pollen of Nicotiana alata ‘Crimson Bedder’ showed skewed segregation in the F2 favoring the maternal parent. This is probably not gene transfer in a strict sense, rather just an extreme case of reduced transmission of irradiated...... chromosomes, leading to massive overrepresentation of maternal genes. Gene transfer or mutational loss may explain some anomalous F1 plants. Segregation in the F2 progeny showed the presence of several genes from the irradiated pollen. Crosses of Nicotiana sylvestris, N. plumbaginifolia N. paniculata......, and Petunia parodii with irradiated pollen from N. alata and Petunia hybrida showed no evidence of gene transfer, nor did experiments with irradiated mentor pollen. This indicates that gene transfer with irradiated pollen between non-crossing species or between species giving sterile hybrids is probably...

  2. Predation of Apiomerus pilipes (Fabricius (Hemiptera, Reduviidae, Harpactorinae, Apiomerini over Meliponinae bees (Hymenoptera, Apidae, in the State of Amazonas, Brazil

    Directory of Open Access Journals (Sweden)

    Alexandre Coletto da Silva

    2004-12-01

    Full Text Available The present work shows the occurrence of an intense predatory activity on adults working Meliponinae bees (Hymenoptera, Apidae, by Apiomerus pilipes (Fabricius, 1787 (Hemiptera, Reduviidae, Harpactorinae, Apiomerini at a meliponary in the Instituto Nacional de Pesquisas da Amazônia (INPA, Manaus, Amazonas State, Brazil.O presente trabalho registra a ocorrência de intensa atividade predatória de Apiomerus pilipes (Fabricius, 1787 (Hemiptera, Reduviidae, Harpactorini, Apiomerini sobre operárias adultas de meliponíneos (Hymenoptera, Apidae, no meliponário experimental do Instituto Nacional de Pesquisas da Amazônia (INPA, Manaus, Estado do Amazonas, Brasil. O meliponário se encontra num fragmento de vegetação secundária no próprio INPA.

  3. Silicon delays tobacco Ringspot virus systemic symptoms in Nicotiana tabacum

    Science.gov (United States)

    Soluble silicon (Si) provides protection to plants against a variety of abiotic and biotic stress. However, the role of Si in viral infections has been elusive. To investigate the role of Si in viral infections, hydroponic studies were conducted in Nicotiana tabacum with two pathogens: Tobacco rings...

  4. Antinuclear human autoantibodies as markers in Nicotiana tabacum pollen tubes

    Directory of Open Access Journals (Sweden)

    C. Poggialini

    2014-02-01

    Full Text Available In the present paper we report on the use of antinuclear human autoantibodies as specific markers in Nicotiana tabacum pollen tubes. The antibodies have been tested by fluorescence techniques using a confocal laser scanning microscope. All the antibodies showed specifc labelling pattern and the results, although preliminary in nature, could open new perspectives of research.

  5. First observation of alternative food usage (extrafloral nectar by the assassin bug Atopozelus opsimus (Hemiptera, Reduviidae

    Directory of Open Access Journals (Sweden)

    Rhainer Guillermo-Ferreira

    2012-12-01

    Full Text Available Assassin bugs (Reduviidae are voracious insects that prey on other arthropods. Recent evidences have pointed out that these predators also feed on plant derived substances in rare opportunities. The present study describes the feeding behavior of the reduviid Atopozelus opsimus on extrafloral nectaries of Inga vera (Fabaceae in a Neotropical savanna area. It was investigated if the insects feed more frequently of extrafloral nectar or prey, and if individuals of different stages of development vary according to feeding behavior. Notably, the results suggest that the diet of all instars and adults consist mainly of extrafloral nectar (N = 1013, in detriment of captured prey ingestion (N = 18. Also, there was no variation on feeding behavior and life stage.

  6. Melt With This Kiss: Paralyzing and Liquefying Venom of The Assassin Bug Pristhesancus plagipennis (Hemiptera: Reduviidae).

    Science.gov (United States)

    Walker, Andrew A; Madio, Bruno; Jin, Jiayi; Undheim, Eivind A B; Fry, Bryan G; King, Glenn F

    2017-04-01

    Assassin bugs (Hemiptera: Heteroptera: Reduviidae) are venomous insects, most of which prey on invertebrates. Assassin bug venom has features in common with venoms from other animals, such as paralyzing and lethal activity when injected, and a molecular composition that includes disulfide-rich peptide neurotoxins. Uniquely, this venom also has strong liquefying activity that has been hypothesized to facilitate feeding through the narrow channel of the proboscis-a structure inherited from sap- and phloem-feeding phytophagous hemipterans and adapted during the evolution of Heteroptera into a fang and feeding structure. However, further understanding of the function of assassin bug venom is impeded by the lack of proteomic studies detailing its molecular composition.By using a combined transcriptomic/proteomic approach, we show that the venom proteome of the harpactorine assassin bug Pristhesancus plagipennis includes a complex suite of >100 proteins comprising disulfide-rich peptides, CUB domain proteins, cystatins, putative cytolytic toxins, triabin-like protein, odorant-binding protein, S1 proteases, catabolic enzymes, putative nutrient-binding proteins, plus eight families of proteins without homology to characterized proteins. S1 proteases, CUB domain proteins, putative cytolytic toxins, and other novel proteins in the 10-16-kDa mass range, were the most abundant venom components. Thus, in addition to putative neurotoxins, assassin bug venom includes a high proportion of enzymatic and cytolytic venom components likely to be well suited to tissue liquefaction. Our results also provide insight into the trophic switch to blood-feeding by the kissing bugs (Reduviidae: Triatominae). Although some protein families such as triabins occur in the venoms of both predaceous and blood-feeding reduviids, the composition of venoms produced by these two groups is revealed to differ markedly. These results provide insights into the venom evolution in the insect suborder

  7. Reação de híbridos somáticos de citros à infecção por Phytophthora nicotianae Reaction of citrus somatic hybrids to the infection by Phytophthora nicotianae

    Directory of Open Access Journals (Sweden)

    Meire Menezes Bassan

    2010-06-01

    Full Text Available Este trabalho avaliou a resistência à infecção de tronco e de raízes por Phytophthora nicotianae em híbridos somáticos de citros com potencial para serem utilizados como porta-enxertos. Os híbridos somáticos avaliados foram laranja 'Hamlin' (Citrus sinensis + toranja 'Indian Red' (Citrus grandis (plantas 1 e 2 e laranja 'Hamlin' (C. sinensis + toranja 'Singapura' (C. grandis. Plantas de limão 'Cravo' (Citrus limonia, laranja 'Caipira' (C. sinensis, laranja-azeda (C. aurantium e Poncirus trifoliata 'Davis A' (Poncirus trifoliata foram utilizadas como plantas-controle devido à reação conhecida à infecção pelo patógeno. Avaliações realizadas entre 30 e 60 dias após as inoculações com o patógeno incluíram o comprimento das lesões no tronco e a massa seca do sistema radicular nas plantas avaliadas. O híbrido somático laranja 'Hamlin' + toranja 'Indian Red' (planta 1 mostrou-se tolerante a P. nicotianae, indicando potencial para continuidade nas suas avaliações como porta-enxerto para citros.This study aimed to evaluate the resistance to trunk and root infection by Phytophthora nicotianae in citrus somatic hybrids with potential to be utilized as rootstocks. The somatic hybrids evaluated were 'Hamlin' sweet orange (Citrus sinensis + 'Indian Red' pummelo (Citrus grandis (plants 1 and 2, and 'Hamlin' sweet orange (C. sinensis + 'Singapura' pummelo (C. grandis. Plants of 'Rangpur' lime (Citrus limonia, 'Caipira' sweet orange (C. sinensis, sour orange (C. aurantium, and Poncirus trifoliata 'Davis A' (Poncirus trifoliata were used as control due to their known reaction to the pathogen. Evaluations performed between 30 and 60 days after pathogen inoculation included the length of trunk lesions, and root dry mass of the root system of evaluated plants. The somatic hybrid 'Hamlin' sweet orange + 'Indian Red' pummelo (plant 1 was tolerant to P. nicotianae, indicating potential to be further evaluated as a rootstock.

  8. Morfometría geométrica del borde corial y del collar de huevos de cinco especies del género Rhodnius Stal (Heteroptera, Reduviidae, Triatominae

    Directory of Open Access Journals (Sweden)

    Ximena Páez-Colasante

    2008-12-01

    Full Text Available Resumen. La taxonomía y sistemática de la subfamilia Triatominae (Heteroptera, Reduviidae se basan principalmente en caracteres biométricos y morfológicos de las formas adultas y, en algunos casos, de las ninfas. La Morfometría Geométrica es capaz de descomponer la forma en sus componentes tamaño isométrico y conformación. En el presente trabajo se utilizó laMorfometría Geométrica como herramienta de análisis de la configuración geométrica que resulta de la unión de los extremos del borde corial y del collar (configuración borde corial-collar de los huevos eclosionados de cinco especies del género Rhodnius Stal. Los huevos se observaron por Microscopía Electrónica de Barrido y las imágenes digitalizadas fueron analizadas morfogeométricamente. Estudiando el tamaño geométrico de la configuración borde corial-collar, se halló que no existen diferencias estadísticamente significativas entre las cinco especies deTriatominos. En cuanto a la conformación, se determinaron diferencias entre las especies al ser sometidas a un análisis discriminante. Se encontró además que la variación del tamaño contribuye a la variación en la conformación, es decir, efecto alométrico. Los resultados obtenidos indican que la conformación de la configuración geométrica “borde corial-collar” de los huevos de Rhodnius podría ser utilizada como herramienta taxonómica para este grupo. Morfometria geométrica da borda corial e colarinho de ovos de cinco espécies do Gênero Rhodnius (Heteroptera Reduviidae, Triatominae.Resumo. A taxonomia e sistemática da subfamília Triatominae (Heteroptera Reduviidae esta baseada principalmente em caracteres biométricos e morfológicos das formas adultas, e em alguns casos, das ninfas. A morfometria geométrica é capaz de descompor a forma em seus componentes, tamanho isométrico e conformação. No presente trabalho utilizou-se a morfometria geométrica como ferramenta de análise da configura

  9. Response of the reduviid bug, <em>Rhynocoris marginatusem> (Heteroptera: Reduviidae) to six different species of cotton pests

    DEFF Research Database (Denmark)

    Sahayaraj, Kitherian; Muthu Kumar, Subramanian; Enkegaard, Annie

    2016-01-01

    In Indian agro-ecosystems Rhynocoris marginatus (F.) is one of the most abundant predatory arthropods and feeds on a wide range of insect pests. We investigated the responses of R. marginatus to six species of cotton pests: Spodoptera litura (F.), Sylepta derogata (F.), Pericallia ricini (F.), My...

  10. Efecto de la alternación de fuentes sanguíneas sobre la fecundidad y la fertilidad de Rhodnius prolixus Stal (Heteroptera: Reduviidae

    Directory of Open Access Journals (Sweden)

    Elis José Aldana

    2009-04-01

    blood ingested and the fecundity were differents according to alternation of blood sources offered in each meal. A linear relation was found between the fecundity and the time of oviposition in every alternation of meal sources. On the other hand there was a slope increase in the second feeding of every experiment. The fertility was above 95% in each experiment. Efeito da alternação de fontes sanguíneas sobre fecundidade e fertilidade de Rhodnius prolixus Stal (Heteroptera ReduviidaeNeste trabalho evidencia-se que o sucesso reprodutivo de Rhodnius prolixus Stal esta relacionado à fonte alimentícia, então as necessidades biológicas do inseto são satisfeitas em modos diferentes segundo o tipo de alternância entre ditas fontes de alimentação; tais diferenças são reveladas mediante mudanças em fertilidade e fecundidade do inseto. Realizaram-se nove testes, em cada um foram subministradas duas alimentações, sendo que em alguns foi alternada a fonte de alimentação e nos controles foram subministradas duas alimentações com a mesma fonte. As fontes alimentícias foram pomba, galinha e humano, esta ultima mediante um aparelho de alimentação artificial; estas fontes foram escolhidas no suposto que são fontes alimentícias encontradas nos ecótopos silvestres, peridomicilio e domicilio respectivamente. Em todos os tipos de alternância foi encontrada uma relação lineal entre a fecundidade e o tempo de oviposição, que a relação entre quantidade de sangue ingerida e a fecundidade variam, que a taxa de fecundidade aumenta na segunda alimentação respeito à primeira e que a fertilidade foi superior ao 95%.

  11. Exocrine secretions of wheel bugs (Heteroptera: Reduviidae: Arilus spp.): clarification and chemistry.

    Science.gov (United States)

    Aldrich, Jeffrey R; Chauhan, Kamlesh R; Zhang, Aijun; Zarbin, Paulo H G

    2013-01-01

    Wheel bugs (Heteroptera: Reduviidae: Harpactorinae: Arilus) are general predators, the females of which have reddish-orange subrectal glands (SGs) that are eversible like the osmeteria in some caterpillars. The rancid odor of Arilus and other reduviids actually comes from Brindley's glands, which in the North (A. cristatus) and South (A. carinatus) American wheel bugs studied emit similar blends of 2-methylpropanoic, butanoic, 3-methylbutanoic, and 2-methylbutanoic acids. The Arilus SG secretions studied here are absolutely species-specific. The volatile SG components of A. carinatus include (E)-2-octenal, (E)-2-nonenal, (E)-2-decenal, (E,E)-2,4-nonadienal, (E)-2-undecenal, hexanoic acid, 4-oxo-nonanal, (E,E)-2,4-decadienal, (E,Z)-2,4- or (Z,E)-2,4-decadienal, and 4-oxo-(E)-2-nonenal; whereas in A. cristatus the SG secretion contains beta-pinene, limonene, terpinolene, terpinen-4-ol, thymol methyl ether, alpha-terpineol, bornyl acetate, methyl eugenol, beta-caryophyllene, caryophyllene oxide, and farnesol. Arilus spp. SG secretions may be sex pheromones, but verification of this hypothesis requires further testing.

  12. Heterochromatin base pair composition and diversification in holocentric chromosomes of kissing bugs (Hemiptera, Reduviidae)

    Science.gov (United States)

    Bardella, Vanessa Bellini; Pita, Sebastián; Vanzela, André Luis Laforga; Galvão, Cleber; Panzera, Francisco

    2016-01-01

    The subfamily Triatominae (Hemiptera, Reduviidae) includes 150 species of blood-sucking insects, vectors of Chagas disease or American trypanosomiasis. Karyotypic information reveals a striking stability in the number of autosomes. However, this group shows substantial variability in genome size, the amount and distribution of C-heterochromatin, and the chromosome positions of 45S rDNA clusters. Here, we analysed the karyotypes of 41 species from six different genera with C-fluorescence banding in order to evaluate the base-pair richness of heterochromatic regions. Our results show a high heterogeneity in the fluorescent staining of the heterochromatin in both autosomes and sex chromosomes, never reported before within an insect subfamily with holocentric chromosomes. This technique allows a clear discrimination of the heterochromatic regions classified as similar by C-banding, constituting a new chromosome marker with taxonomic and evolutionary significance. The diverse fluorescent patterns are likely due to the amplification of different repeated sequences, reflecting an unusual dynamic rearrangement in the genomes of this subfamily. Further, we discuss the evolution of these repeated sequences in both autosomes and sex chromosomes in species of Triatominae. PMID:27759763

  13. Assessment of the insecticidal potential of Eucalyptus urograndis essential oil against Rhodnius neglectus Lent (Hemiptera: Reduviidae).

    Science.gov (United States)

    Gomes, S P; Favero, S

    2013-08-01

    The resistance of triatomines to pyrethroids has been reported in several Latin American countries, including Brazil, indicating the need for the development of new approaches for the control of vectors of the Chagas disease. In here, we evaluated the insecticidal potential of the essential oil of Eucalyptus urograndis (Myrtaceae) against unsexed third and fourth instars of Rhodnius neglectus Lent (Hemiptera: Reduviidae) in topical application, fumigation, surface contact, and repellency . The insecticidal activity of the essential oil tested was detected by topical application (LD50 = 0.1731 μL/insect and LD99 = 0.2948 μL/insect for 24 h), fumigation (LC50 = 0.021 mL/mL air and LC99 = 0.1525 mL/mL air for 24 h) and surface contact (LC50 = 0.7073 μL/cm(2) and LC99 = 4.59 μL/cm(2) for 24 h). Mortality observed after 48-72-h exposure was very high and did not allow for any adjustment of the mortality curve. In the repellency assay, an effect was observed on 80% of tested nymphs. However, no repellency was observed after 24 h of exposure. Eucalyptus urograndis essential oil has a high insecticidal and repellent potential for R. neglectus nymphs, whether serving as a molecular model for new substances or as an alternative for the control of these insects.

  14. First record of Triatoma maculata (Erichson, 1848 (Hemiptera: Reduviidae: Triatomini in Riohacha, La Guajira – Colombia

    Directory of Open Access Journals (Sweden)

    Edith Natalia Gómez-Melendro

    2014-11-01

    Full Text Available Introduction. Knowledge of vector insect species, their habitat and geographical distribution is crucial for determining the risk of transmission of the etiologic agents that cause disease in humans, which allows defining strategies for prevention, surveillance and control in line with the characteristics of each area. Objective. To determine the presence and public health importance of vectors of Chagas disease in the indigenous settlements of Marbacella and El Horno of the Wayúu ethnic group in the municipality of Riohacha, La Guajira, Colombia. Materials and methods. From active search, installation and inspection of biosensors and occasional catches, Hemiptera: Reduviidae: Triatomini were collected intra and in the peridomicile housing of the indigenous settlements of El Horno and Marbacella of the the Wayúu ethnic group. Indices of intra and peridomestic infestation, colonization, density, dispersion and natural infection with Trypanosoma cruzi Chagas, 1909 were calculated. Results. 79.6% (n = 90 of the specimens were collected around the homes and 20.3% (n = 23 inside the homes, all corresponding to Triatoma maculata (Erichson, 1848. The natural infection indices with T. cruzi accounted for 43.5% for Marbacella and 36% for El Horno. Conclusion. This is the first reported capture of individuals of T. maculata, considered a secondary vector of Chagas disease in Colombia, naturally infected with T. cruzi in the municipality of Riohacha expanding the geographical distribution of the species in the department of La Guajira.

  15. Heterochromatin base pair composition and diversification in holocentric chromosomes of kissing bugs (Hemiptera, Reduviidae

    Directory of Open Access Journals (Sweden)

    Vanessa Bellini Bardella

    Full Text Available The subfamily Triatominae (Hemiptera, Reduviidae includes 150 species of blood-sucking insects, vectors of Chagas disease or American trypanosomiasis. Karyotypic information reveals a striking stability in the number of autosomes. However, this group shows substantial variability in genome size, the amount and distribution of C-heterochromatin, and the chromosome positions of 45S rDNA clusters. Here, we analysed the karyotypes of 41 species from six different genera with C-fluorescence banding in order to evaluate the base-pair richness of heterochromatic regions. Our results show a high heterogeneity in the fluorescent staining of the heterochromatin in both autosomes and sex chromosomes, never reported before within an insect subfamily with holocentric chromosomes. This technique allows a clear discrimination of the heterochromatic regions classified as similar by C-banding, constituting a new chromosome marker with taxonomic and evolutionary significance. The diverse fluorescent patterns are likely due to the amplification of different repeated sequences, reflecting an unusual dynamic rearrangement in the genomes of this subfamily. Further, we discuss the evolution of these repeated sequences in both autosomes and sex chromosomes in species of Triatominae.

  16. Genetic variability of Triatoma flavida and Triatoma bruneri (Hemiptera: Reduviidae by RAPD-PCR technique

    Directory of Open Access Journals (Sweden)

    Jorge Fraga

    2011-02-01

    Full Text Available The Triatominae (Hemiptera:Reduviidae contains the principal and potential Chagas disease vectors present in Mexico, Central America and South America. Triatoma flavida and T. bruneri are Cuban species. These species are closely related according to morphology and were considered synonyms until 1981, when they were separated on the grounds of external characters of the body and the morphology of male genitalia. The present study seeks to analyze genetic polymorphism of T. flavida and T. bruneri populations using RAPD techniques, and to assess the genetic relationship between these species. Ten random primers were used to evaluate the genetic variability among species using RAPD-PCR. The genetic flow among them was calculated. The dendrogram based on calculated Jaccard distances showed two clearly distinguishable clusters which coincided with the studied species. Within each species, moderate genetic differentiation (Fst 0.05-0.15 and migration rates (N > 1 were found among populations, that reveal gene flow and genetic homogeneity. Between species, the Fst value showed a high genetic differentiation and the migration rate was insufficient to maintain genetic homogeneity, and confirmed the absence of gene flow between them. Our results confirm the genetic variability among T. flavida and T. bruneri species.

  17. Genetic variability of Triatoma flavida and Triatoma bruneri (Hemiptera: Reduviidae) by RAPD-PCR technique.

    Science.gov (United States)

    Fraga, Jorge; Rodriguez, Jinnay; Fuentes, Omar; Hernández, Yenin; Castex, Mayda; Gonzalez, Raul; Fernández-Calienes, Aymé

    2011-01-01

    The Triatominae (Hemiptera:Reduviidae) contains the principal and potential Chagas disease vectors present in Mexico, Central America and South America. Triatoma flavida and T. bruneri are Cuban species. These species are closely related according to morphology and were considered synonyms until 1981, when they were separated on the grounds of external characters of the body and the morphology of male genitalia. The present study seeks to analyze genetic polymorphism of T. flavida and T. bruneri populations using RAPD techniques, and to assess the genetic relationship between these species. Ten random primers were used to evaluate the genetic variability among species using RAPD-PCR. The genetic flow among them was calculated. The dendrogram based on calculated Jaccard distances showed two clearly distinguishable clusters which coincided with the studied species. Within each species, moderate genetic differentiation (Fst 0.05-0.15) and migration rates (N > 1) were found among populations, that reveal gene flow and genetic homogeneity. Between species, the Fst value showed a high genetic differentiation and the migration rate was insufficient to maintain genetic homogeneity, and confirmed the absence of gene flow between them. Our results confirm the genetic variability among T. flavida and T. bruneri species.

  18. Resin gathering in neotropical resin bugs (Insecta: Hemiptera: Reduviidae): functional and comparative morphology.

    Science.gov (United States)

    Forero, Dimitri; Choe, Dong-Hwan; Weirauch, Christiane

    2011-02-01

    Apiomerini (Reduviidae: Harpactorinae) collect plant resins with their forelegs and use these sticky substances for prey capture or maternal care. These behaviors have not been described in detail and morphological structures involved in resin gathering, transfer, and storage remain virtually undocumented. We here describe these behaviors in Apiomerus flaviventris and document the involved structures. To place them in a comparative context, we describe and document leg and abdominal structures in 14 additional species of Apiomerini that represent all but one of the 12 recent genera in the tribe. Based on these morphological data in combination with the behavioral observations on A. flaviventris, we infer behavioral and functional hypotheses for the remaining genera within the tribe Apiomerini. Setal abdominal patches for resin storage are associated with maternal care so far only documented for species of Apiomerus. Based on the occurrence of these patches in several other genera, we propose that maternal care is widespread within the tribe. Ventral abdominal glands are widespread within female Apiomerini. We propose that their products may prevent hardening of stored resins thus providing long-term supply for egg coating. Judging from the diverse setal types and arrangements on the front legs, we predict six different behavioral patterns of resin gathering within the tribe.

  19. Genetic analysis of Phytophthora nicotianae populations from different hosts using microsatellite markers

    Science.gov (United States)

    Two hundred thirty-one isolates of P. nicotianae representing 14 populations from different host genera, including agricultural crops (Citrus, Nicotiana, and Lycopersicon), potted ornamental species in nurseries (Lavandula, Convolvulus, Myrtus, Correa and Ruta) and other plant genera of lesser econo...

  20. Variations in alkaloidal constituents of plant tissue cultures. [Nicotiana tabacum, Nicotiana rustica, Datura stramonium and Hyoscyamus niger

    Energy Technology Data Exchange (ETDEWEB)

    Essa, A.K.

    1987-01-01

    The accumulation of tobacco and Solanaceae-tropane alkaloids in calluses and suspensions of Nicotiana tabacum, Nicotiana rustica, Datura stramonium and Hyoscyamus niger was the main concern of this work. Nicotine, anabasine and anatabine had regularly been found in tobacco callus tissues grown for several passages. For the first time, N. tabacum suspensions were shown able to accumulate anabasine, anatabine, anatalline, myosmine and nicotelline whereas N. rustica calluses, N-methyl-, N,N-dimethyl- and N-methyl-N-nitroso anilines. The aim of these experiments was an attempt to affect the yields and types of alkaloids produced. The interrelationship between nicotine and protein contents in N. tabacum and N. rustica calluses was investigated. The possible role of urea and sodium propionate as precursors of nicotine in tobacco suspensions was checked by feeding the latter with carbon-14 radioactive substrates. The scope and quantum of the principle alkaloidal components present in the source N. tabacum plants and D. stramonium and H. niger seeds were investigated to stand as references against in vitro production. Identification of the products found was made by using gas chromatography-mass spectrometry (GC-MS) and/or MS.

  1. La aparición de Thyrinteina arnobia (Stoll (Lepidoptera: Geometridae de Eucalyptus y su depredación por Zelus armillatus (Lepeletier y Serville (Reduviidae: Harpactorinae

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    Marliton Barreto

    2014-04-01

    Abstract. The most important lepidopterans for eucalyptus crop are called defoliators. In Brazil, some species occur from the Amazônia to Rio Grande do Sul State and Thyrinteina arnobia arnobia (Stoll (Lepidoptera: Geometridae is cited as the most important lepidopteran defoliator in Brazil and some predatory species of the family Reduviidae are associated with various forest crops. Thus, this study recorded the ocorrence of Zelus armillatus (Lepeletier & Serville preying T. arnobia in eucaliptus crop in Sorriso city, Mato Grosso state, Brazil.

  2. Chromosomal variation, macroevolution and possible parapatric speciation in Mepraia spinolai (Porter (Hemiptera: Reduviidae

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    Frias Daniel

    1998-01-01

    Full Text Available Mepraia spinolai is an endemic species in Chile that lives in wild and domestic habitats. It is the only species of the Reduviidae family that shows alate polymorphism; females are always wingless, but males can be found with and without wings. The M. spinolai karyotype consists of 10 pairs of autosomes and a complex sex determination system. Males from the northernmost regions I and II (latitude 18°-26° South are always winged (braquipterous and are X1X2Y, with a large Y chromosome. From region III to the metropolitan region (latitude 26°-33° South, males may be either winged or wingless but appear to be polymorphic for a small neo-Y chromosome, which may have originated by fracture of the large holocentric Y chromosome found in populations from farther north. Experimental crosses suggest that the genes for wings are linked in the Y chromosome and also that there are two cytologically indistinguishable types of neo-Y chromosomes. One form (Y1 bears a gene or genes for wings while the other (Y2 lacks such genes. Males that are X1X2Y1, X1X2Y1Y1 and X1X2Y1Y2 are winged, while the absence of Y1 (X1X2Y2 and X1X2Y2Y2 results in a wingless male. These chromosomes and morphological changes are correlated with a shift of the southern population into more arid habitats of the interior in the metropolitan region and region III.

  3. Impacts of 2 species of predatory Reduviidae on bagworms in oil palm plantations.

    Science.gov (United States)

    Jamian, Syari; Norhisham, Ahmad; Ghazali, Amal; Zakaria, Azlina; Azhar, Badrul

    2017-04-01

    Integrated pest management (IPM) is widely practiced in commercial oil palm agriculture. This management system is intended to minimize the number of attacks by pest insects such as bagworms on crops, as well as curb economic loss with less dependency on chemical pesticides. One practice in IPM is the use of biological control agents such as predatory insects. In this study, we assessed the response of predatory natural enemies to pest outbreak and water stress, and document the habitat associations of potential pest predators. The abundances of 2 predatory insect species, namely Sycanus dichotomus and Cosmolestes picticeps (Hemiptera: Reduviidae), were compared bagworm outbreak sites and nonoutbreak sites within oil palm plantations. We also examined habitat characteristics that influence the abundances of both predatory species. We found that the abundance of C. picticeps was significantly higher in bagworm outbreak sites than in nonoutbreak sites. There were no significant differences in the abundance of S. dichotomus among outbreak and non-outbreak sites. Both species responded negatively to water stress in oil palm plantations. Concerning the relationship between predatory insect abundance and in situ habitat quality characteristics, our models explained 46.36% of variation for C. picticeps and 23.17% of variation for S. dichotomus. Both species of predatory insects thrived from the planting of multiple beneficial plants in oil palm plantations. The results suggest that C. picticeps can be used as a biological agent to control bagworm populations in oil palm plantations, but S. dichotomus has no or little potential for such ecosystem service. © 2015 Institute of Zoology, Chinese Academy of Sciences.

  4. PHANTASTICA regulates development of the adaxial mesophyll in Nicotiana leaves.

    Science.gov (United States)

    McHale, Neil A; Koning, Ross E

    2004-05-01

    Initiation and growth of leaf blades is oriented by an adaxial/abaxial axis aligned with the original axis of polarity in the leaf primordium. To investigate mechanisms regulating this process, we cloned the Nicotiana tabacum ortholog of PHANTASTICA (NTPHAN) and generated a series of antisense transgenics in N. sylvestris. We show that NSPHAN is expressed throughout emerging blade primordia in the wild type and becomes localized to the middle mesophyll in the expanding lamina. Antisense NSPHAN leaves show ectopic expression of NTH20, a class I KNOX gene. Juvenile transgenic leaves have normal adaxial/abaxial polarity and generate leaf blades in the normal position, but the adaxial mesophyll shows disorganized patterns of cell division, delayed maturation of palisade, and ectopic reinitiation of blade primordia along the midrib. Reversal of the phenotype with exogenous gibberellic acid suggests that NSPHAN, acting via KNOX repression, maintains determinacy in the expanding lamina and sustains the patterns of cell proliferation critical to palisade development.

  5. Silicon delays Tobacco ringspot virus systemic symptoms in Nicotiana tabacum.

    Science.gov (United States)

    Zellner, Wendy; Frantz, Jonathan; Leisner, Scott

    2011-10-15

    Soluble silicon (Si) provides protection to plants against a variety of abiotic and biotic stress. However, the effects of Si on viral infections are largely unknown. To investigate the role of Si in viral infections, hydroponic studies were conducted in Nicotiana tabacum with two pathogens: Tobacco ringspot virus (TRSV) and Tobacco mosaic virus (TMV). Plants grown in elevated Si showed a delay in TRSV systemic symptom formation and a reduction in symptomatic leaf area, compared to the non-supplemented controls. TRSV-infected plants showed significantly higher levels of foliar Si compared to mock-inoculated plants. However, the Si effect appeared to be virus-specific, since the element did not alter TMV symptoms nor did infection by this virus alter foliar Si levels. Hence, increased foliar Si levels appear to correlate with Si-modulated protection against viral infection. This is all the more intriguing since N. tabacum is classified as a low Si accumulator.

  6. Miridae, Reduviidae and Lygaeoidea (Hemiptera collected in Colonia Carlos Pellegrini (Iberá watershed, Corrientes, Argentina Reduviidae, Miridae y Lygaeoidea (Hemiptera recolectados en Colonia Carlos Pellegrini (Esteros de Iberá, Corrientes, Argentina

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    María C. Melo

    2004-07-01

    Full Text Available With the intention of contributing to the knowledge of the biodiversity in the protected areas of Argentina, terrestrial Heteroptera were collected in the Natural Reserve of the Iberá. The sampling site was the locality of Colonia Carlos Pellegrini (28° 32'S 57° 09'W and the following collection methods were used: sweeping, beating, fogging and light-trapping. A list is presented, with 90 species belonging to the superfamilies Reduvioidea (Reduviidae, Mirioidea (Miridae, and Lygaeoidea (Berytidae, Colobathristidae, Cymidae, Lygaeidae, Blissidae, Geocoridae, Pachygrontidae and Rhyparochromidae, 13 of them are new records for the Argentine Republic (Debilia fusciventris, Sindala granuligera, Pnirontis (Centromelus tabida, Cymoninus notabilis, Ochrimnus (Phaeochrimnus limbatipennis, Oncopeltus luctuosus, Ischnodemus gayi, Oedancala nana, Cistalia signoretii, Paracholula picta, Cholula lympha, Cholula maculatus, Pseudopachybrachius vinctus and 36 new records for the province of Corrientes.Con el objeto de contribuir al conocimiento de la biodiversidad en las áreas protegidas de la Argentina, se realizaron recoleccciones de Heteroptera terrestres en la Reserva Natural del Iberá. El sitio de muestreo fue la localidad de Colonia Carlos Pellegrini (28° 32'S 57°09'W y se utilizaron los siguientes métodos de recoleccion: rastreo, golpeteo, fogging y trampa de luz. Se presenta una lista con 90 especies pertenecientes a las superfamilias Reduvioidea (Reduviidae, Mirioidea (Miridae y Lygaeoidea (Berytidae, Colobathristidae, Cymidae, Lygaeidae, Blissidae, Geocoridae, Pachygrontidae y Rhyparochromidae, de las cuales 13 representan primeras citas para la República Argentina (Debilia fusciventris, Sindala granuligera, Pnirontis (Centromelus tabida, Cymoninus notabilis, Ochrimnus (Phaeochrimnus limbatipennis , Oncopeltus luctuosus , Ischnodemus gayi , Oedancala nana , Cistalia signoretii, Paracholula picta, Cholula lympha, Cholula maculatus

  7. Bioactivity of the latex from Parahancornia amapa (Apocynaceae on the development of Rhodnius nasutus (Hemiptera, Reduviidae, Triatominae under laboratory conditions

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    Marcio B. P. Lopes

    2014-12-01

    Full Text Available Bioactivity of the latex from Parahancornia amapa (Apocynaceae on the development of Rhodnius nasutus (Hemiptera, Reduviidae, Triatominae under laboratory conditions. The aim of this study was to verify the effects of the methanolic fraction of the latex from Parahancornia amapa (Apocynaceae (PALAM on individuals of the species Rhodnius nasutus Stål (Hemiptera, Triatominae. Many of the insects treated with the substance presented deformities and these may interfere in the feeding and possibly hinder the reproductive capacity. They also presented significant mortality during the molt when compared to the control group, noting a gradual increase in mortality. The treated insects also presented delayed nymphal development (5th instar and higher adult longevity.

  8. Phytophthora nicotianae diseases worldwide: new knowledge of a long-recognised pathogen

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    Franck PANABIERES

    2016-05-01

    Full Text Available Phytophthora nicotianae was first isolated from tobacco at the end of the 19th century. This organism is now considered as one of the most devastating oomycete plant pathogens, with a recognized host range of more than 255 species over five continents and a wide diversity of climates. The economic losses caused by P. nicotianae are difficult to estimate, because of the diversity of its hosts and ecological niches. For these reasons, this pathogen represents a continuous challenge to plant disease management programmes, which frequently rely solely on the use of chemicals. Phytophthora nicotianae is better adapted than its competitors to abiotic stresses, especially to climate warming. As a result, its importance is increasing. This review illustrates, with some examples, how P. nicotianae currently impacts plant economies worldwide, and how it may constitute more severe threats to agriculture and natural ecosystems in the context of global climate change.

  9. ANTIBACTERIAL ACTIVITY, PHYTOCHEMICAL SCREENING AND ANTIOXIDANT ACTIVITY OF STEM OF NICOTIANA TABACUM

    National Research Council Canada - National Science Library

    Y Sharma; D Dua; A Nagar; N S Srivastava

    2016-01-01

    .... The purpose of this present study is to investigate the antibacterial activity, phytochemical screening and the antioxidant activity of aqueous and alcoholic extracts of the stem of Nicotiana tabacum...

  10. Highly Oxygenated Flavonoids from the Leaves of Nicotiana plumbaginifolia (Solanaceae

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    Md. Shafiullah Shajib

    2017-11-01

    Full Text Available Nicotiana plumbaginifolia Viv. is an annual herb of the family Solanaceae, which grows abundantly in the weedy lands of Bangladesh . This plant possesses analgesic, antibacterial, anti-anxiety and hepatoprotective properties, and produces various phenolic compounds including flavonoids. The present study afforded determination of total phenolic and flavonoid contents, and for the first time, the isolation and characterization of highly oxygenated flavonoids, e.g., 3,3' ,5,6,7,8-hexamethoxy- 4',5'-methylenedioxyflavone (1, 3,3' ,4' ,5',5,6,7,8-octamethoxyflavone (2, exoticin, 6,7,4',5'-dimethylenedioxy-3,5,3'-trimethoxyflavone (3 and ( 3,3' ,4',5,5',8-hexamethoxy-6,7-methylenedioxyflavone (4 from the leaves of N. plumbaginifolia . All these flavonoids are rather rare natural products, and only found in a few genera, e.g.,Polygonum and Murraya. The structures of the isolated flavonoids were elucidated by comprehensive spectroscopic analyses, e.g., UV, 1H, 13C NMR, DEPT, HSQC, HMBC and MS.

  11. Description of the male genitalia of Belminus rugulosus Stål and Belminus corredori Galvão & Angulo, and comments on the holotype of Parabelminus yurupucu Lent & Wygodzinsky (Hemiptera: Heteroptera: Reduviidae: Triatominae: Bolboderini).

    Science.gov (United States)

    Gil-Santana, Hélcio R; Galvão, Cleber

    2013-12-13

    The male genitalia of Belminus rugulosus Stål and Belminus corredori Galvão & Angulo are described and illustrated for the first time. Comments on the holotype of Parabelminus yurupucu Lent & Wygodzinsky are presented (Hemiptera: Heteroptera: Reduviidae: Triatominae: Bolboderini). A discussion on previous data in the literature on male genitalia of species of Belminus Stål is provided.

  12. Current status and prospects for the study of Nicotiana genomics, genetics, and nicotine biosynthesis genes.

    Science.gov (United States)

    Wang, Xuewen; Bennetzen, Jeffrey L

    2015-02-01

    Nicotiana, a member of the Solanaceae family, is one of the most important research model plants, and of high agricultural and economic value worldwide. To better understand the substantial and rapid research progress with Nicotiana in recent years, its genomics, genetics, and nicotine gene studies are summarized, with useful web links. Several important genetic maps, including a high-density map of N. tabacum consisting of ~2,000 markers published in 2012, provide tools for genetics research. Four whole genome sequences are from allotetraploid species, including N. benthamiana in 2012, and three N. tabacum cultivars (TN90, K326, and BX) in 2014. Three whole genome sequences are from diploids, including progenitors N. sylvestris and N. tomentosiformis in 2013 and N. otophora in 2014. These and additional studies provide numerous insights into genome evolution after polyploidization, including changes in gene composition and transcriptome expression in N. tabacum. The major genes involved in the nicotine biosynthetic pathway have been identified and the genetic basis of the differences in nicotine levels among Nicotiana species has been revealed. In addition, other progress on chloroplast, mitochondrial, and NCBI-registered projects on Nicotiana are discussed. The challenges and prospects for genomic, genetic and application research are addressed. Hence, this review provides important resources and guidance for current and future research and application in Nicotiana.

  13. Efficacy of Chaetomium Species as Biological Control Agents against Phytophthora nicotianae Root Rot in Citrus.

    Science.gov (United States)

    Hung, Phung Manh; Wattanachai, Pongnak; Kasem, Soytong; Poeaim, Supattra

    2015-09-01

    Thailand is one of the largest citrus producers in Southeast Asia. Pathogenic infection by Phytophthora, however, has become one of major impediments to production. This study identified a pathogenic oomycete isolated from rotted roots of pomelo (Citrus maxima) in Thailand as Phytophthora nicotianae by the internal transcribed spacer ribosomal DNA sequence analysis. Then, we examined the in vitro and in vivo effects of Chaetomium globosum, Chaetomium lucknowense, Chaetomium cupreum and their crude extracts as biological control agents in controlling this P. nicotianae strain. Represent as antagonists in biculture test, the tested Chaetomium species inhibited mycelial growth by 50~56% and parasitized the hyphae, resulting in degradation of P. nicotianae mycelia after 30 days. The crude extracts of these Chaetomium species exhibited antifungal activities against mycelial growth of P. nicotianae, with effective doses of 2.6~101.4 µg/mL. Under greenhouse conditions, application of spores and methanol extracts of these Chaetomium species to pomelo seedlings inoculated with P. nicotianae reduced root rot by 66~71% and increased plant weight by 72~85% compared to that in the control. The method of application of antagonistic spores to control the disease was simple and economical, and it may thus be applicable for large-scale, highly effective biological control of this pathogen.

  14. A Distinct Endogenous Pararetrovirus Family in Nicotiana tomentosiformis, a Diploid Progenitor of Polyploid Tobacco1[w

    Science.gov (United States)

    Gregor, Wolfgang; Mette, M. Florian; Staginnus, Christina; Matzke, Marjori A.; Matzke, Antonius J.M.

    2004-01-01

    A distinct endogenous pararetrovirus (EPRV) family corresponding to a previously unknown virus has been identified in the genome of Nicotiana tomentosiformis, a diploid ancestor of allotetraploid tobacco (Nicotiana tabacum). The putative virus giving rise to N. tomentosiformis EPRVs (NtoEPRVs) is most similar to tobacco vein clearing virus, an episomal form of a normally silent EPRV family in Nicotiana glutinosa; it is also related to a putative virus giving rise to the NsEPRV family in Nicotiana sylvestris (the second diploid progenitor of tobacco) and in the N. sylvestris fraction of the tobacco genome. The copy number of NtoEPRVs is significantly higher in N. tomentosiformis than in tobacco. This suggests that after the polyploidization event, many copies were lost from the polyploid genome or were accumulated specifically in the diploid genome. By contrast, the copy number of NsEPRVs has remained constant in N. sylvestris and tobacco, indicating that changes have occurred preferentially in the NtoEPRV family during evolution of the three Nicotiana species. NtoEPRVs are often flanked by Gypsy retrotransposon-containing plant DNA. Although the mechanisms of NtoEPRV integration, accumulation, and/or elimination are unknown, these processes are possibly linked to retrotransposon activity. PMID:14988473

  15. Molecular genetics of alkaloid biosynthesis in Nicotiana tabacum.

    Science.gov (United States)

    Dewey, Ralph E; Xie, Jiahua

    2013-10-01

    Alkaloids represent an extensive group of nitrogen-containing secondary metabolites that are widely distributed throughout the plant kingdom. The pyridine alkaloids of tobacco (Nicotiana tabacum L.) have been the subject of particularly intensive investigation, driven largely due to the widespread use of tobacco products by society and the role that nicotine (16) (see Fig. 1) plays as the primary compound responsible for making the consumption of these products both pleasurable and addictive. In a typical commercial tobacco plant, nicotine (16) comprises about 90% of the total alkaloid pool, with the alkaloids nornicotine (17) (a demethylated derivative of nicotine), anatabine (15) and anabasine (5) making up most of the remainder. Advances in molecular biology have led to the characterization of the majority of the genes encoding the enzymes directly responsible the biosynthesis of nicotine (16) and nornicotine (17), while notable gaps remain within the anatabine (15) and anabasine (5) biosynthetic pathways. Several of the genes involved in the transcriptional regulation and transport of nicotine (16) have also been elucidated. Investigations of the molecular genetics of tobacco alkaloids have not only provided plant biologists with insights into the mechanisms underlying the synthesis and accumulation of this important class of plant alkaloids, they have also yielded tools and strategies for modifying the tobacco alkaloid composition in a manner that can result in changing the levels of nicotine (16) within the leaf, or reducing the levels of a potent carcinogenic tobacco-specific nitrosamine (TSNA). This review summarizes recent advances in our understanding of the molecular genetics of alkaloid biosynthesis in tobacco, and discusses the potential for applying information accrued from these studies toward efforts designed to help mitigate some of the negative health consequences associated with the use of tobacco products.

  16. CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum.

    Science.gov (United States)

    Gao, Junping; Wang, Genhong; Ma, Sanyuan; Xie, Xiaodong; Wu, Xiangwei; Zhang, Xingtan; Wu, Yuqian; Zhao, Ping; Xia, Qingyou

    2015-01-01

    Genome editing is one of the most powerful tools for revealing gene function and improving crop plants. Recently, RNA-guided genome editing using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system has been used as a powerful and efficient tool for genome editing in various organisms. Here, we report genome editing in tobacco (Nicotiana tabacum) mediated by the CRISPR/Cas9 system. Two genes, NtPDS and NtPDR6, were used for targeted mutagenesis. First, we examined the transient genome editing activity of this system in tobacco protoplasts, insertion and deletion (indel) mutations were observed with frequencies of 16.2-20.3% after transfecting guide RNA (gRNA) and the nuclease Cas9 in tobacco protoplasts. The two genes were also mutated using multiplexing gRNA at a time. Additionally, targeted deletions and inversions of a 1.8-kb fragment between two target sites in the NtPDS locus were demonstrated, while indel mutations were also detected at both the sites. Second, we obtained transgenic tobacco plants with NtPDS and NtPDR6 mutations induced by Cas9/gRNA. The mutation percentage was 81.8% for NtPDS gRNA4 and 87.5% for NtPDR6 gRNA2. Obvious phenotypes were observed, etiolated leaves for the psd mutant and more branches for the pdr6 mutant, indicating that highly efficient biallelic mutations occurred in both transgenic lines. No significant off-target mutations were obtained. Our results show that the CRISPR/Cas9 system is a useful tool for targeted mutagenesis of the tobacco genome.

  17. Transient Expression of Tetrameric Recombinant Human Butyrylcholinesterase in Nicotiana benthamiana

    Science.gov (United States)

    Alkanaimsh, Salem; Karuppanan, Kalimuthu; Guerrero, Andrés; Tu, Aye M.; Hashimoto, Bryce; Hwang, Min Sook; Phu, My L.; Arzola, Lucas; Lebrilla, Carlito B.; Dandekar, Abhaya M.; Falk, Bryce W.; Nandi, Somen; Rodriguez, Raymond L.; McDonald, Karen A.

    2016-01-01

    To optimize the expression, extraction and purification of plant-derived tetrameric recombinant human butyrylcholinesterase (prBChE), we describe the development and use of plant viral amplicon-based gene expression system; Tobacco Mosaic Virus (TMV) RNA-based overexpression vector (TRBO) to express enzymatically active FLAG-tagged plant made recombinant butyrylcholinesterase (rBChE) in Nicotiana benthamiana leaves using transient agroinfiltration. Two gene expression cassettes were designed to express the recombinant protein in either the ER or to the apoplastic compartment. Leaf homogenization was used to isolate ER-retained recombinant butyrylcholinesterase (prBChE-ER) while apoplast-targeted rBChE was isolated by either leaf homogenization (prBChE) or vacuum-extraction of apoplastic wash fluid (prBChE-AWF). rBChE from apoplast wash fluid had a higher specific activity but lower enzyme yield than leaf homogenate. To optimize the isolation and purification of total recombinant protein from leaf homogenates, an acidic extraction buffer was used. The acidic extraction buffer yielded >95% enzymatically active tetrameric rBChE as verified by Coomassie stained and native gel electrophoresis. Furthermore, when compared to human butyrylcholinesterase, the prBChE was found to be similar in terms of tetramerization and enzyme kinetics. The N-linked glycan profile of purified prBChE-ER was found to be mostly high mannose structures while the N-linked glycans on prBChE-AWF were primarily complex. The glycan profile of the prBChE leaf homogenates showed a mixture of high mannose, complex and paucimannose type N-glycans. These findings demonstrate the ability of plants to produce rBChE that is enzymatically active and whose oligomeric state is comparable to mammalian butyrylcholinesterase. The process of plant made rBChE tetramerization and strategies for improving its pharmacokinetics properties are also discussed. PMID:27379103

  18. Pollination triggers female gametophyte development in immature Nicotiana tabacum flowers

    Science.gov (United States)

    Brito, Michael S.; Bertolino, Lígia T.; Cossalter, Viviane; Quiapim, Andréa C.; DePaoli, Henrique C.; Goldman, Gustavo H.; Teixeira, Simone P.; Goldman, Maria H. S.

    2015-01-01

    In Nicotiana tabacum, female gametophytes are not fully developed at anthesis, but flower buds pollinated 12 h before anthesis produce mature embryo sacs. We investigated several pollination-associated parameters in N. tabacum flower buds to determine the developmental timing of important events in preparation for successful fertilization. First, we performed hand pollinations in flowers from stages 4 to 11 to study at which developmental stage pollination would produce fruits. A Peroxtesmo test was performed to correlate peroxidase activity on the stigma surface, indicative of stigma receptivity, with fruit set. Pollen tube growth and female gametophyte development were microscopically analyzed in pistils of different developmental stages. Fruits were obtained only after pollinations of flower buds at late stage 7 and older; fruit weight and seed germination capacity increased as the developmental stage of the pollinated flower approached anthesis. Despite positive peroxidase activity and pollen tube growth, pistils at stages 5 and 6 were unable to produce fruits. At late stage 7, female gametophytes were undergoing first mitotic division. After 24 h, female gametophytes of unpollinated pistils were still in the end of the first division, whereas those of pollinated pistils showed egg cells. RT-qPCR assay showed that the expression of the NtEC1 gene, a marker of egg cell development, is considerably higher in pollinated late stage 7 ovaries compared with unpollinated ovaries. To test whether ethylene is the signal eliciting female gametophyte maturation, the expression of ACC synthase was examined in unpollinated and pollinated stage 6 and late stage 7 stigmas/styles. Pollination induced NtACS expression in stage 6 pistils, which are unable to produce fruits. Our results show that pollination is a stimulus capable of triggering female gametophyte development in immature tobacco flowers and suggests the existence of a yet undefined signal sensed by the pistil. PMID

  19. Nicotiana ovule extracts in duce nuclear reconstitution of demembranated Xenopus sperm in cell-free system

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    s Nicotiana tabaccum ovule extracts induced nuclear reconstitution of demembranated Xenopus leavis sperm in a ceil-free system. Demembranated Xenopus sperm began to swell after 15 rmin of incubation with Nicotiana ovulde extracts. Accompanying the process of incubation,Xenopus sperm decondensed and their shapes changed gradually from long and ellipse to round. The completely decondensed chromatin was surrounded with membrane structure, which was a mixture envelope of a double membrane and a single membrane. Nucleosome assembly was verified by means of micrococcal nuclease digestion to reconstituted nuclei and DNA electrophoresis. Nicotiana ovule extracts supplied one more experimental model and system.The new system could promote powerfully the research on mechanisms of cell division and cell cycle regulation.

  20. FIRST REPORT OF Phytophthora nicotianae CAUSING ROOT ROT OF SOURSOP IN NORTHEASTERN BRAZIL

    Directory of Open Access Journals (Sweden)

    JAQUELINE FIGUEREDO DE OLIVEIRA COSTA

    Full Text Available ABSTRACT In 2013, soursop trees showing symptoms of root rot were observed in a field in Maceió, state of Alagoas, Brazil. It was isolated Phytophthora sp. which pathogenicity was confirmed in the host seedlings. Morphological and physiological characteristics in carrot-agar modified medium were consistent with Phytophthora nicotianae description. The PCR sequences products obtained with ITS1/ITS4 primers were compared to sequences of ribosomal DNA of Phytophthora species from the GenBank database observing high identity with other P. nicotianae isolates. A phylogenetic tree was performed to compare the isolate with other sequences of P. nicotianae, which clustering has been verified with 99% of bootstrap, confirming the morphophysiological studies. This is the first report of this pathogen on annonaceous plants in the Northeastern Brazil.

  1. Uptake of NO, NO 2 and O 3 by sunflower ( Helianthus annuus L.) and tobacco plants ( Nicotiana tabacum L.): dependence on stomatal conductivity

    Science.gov (United States)

    Neubert, A.; Kley, D.; Wildt, J.; Segschneider, H. J.; Förstel, H.

    The uptake of NO, NO 2 and O 3 by sunflowers ( Helianthus annuus L. var. giganteus) and tobacco plants ( Nicotiana tabacum L. var. Bel W3), using concentrations representative for moderately polluted air, has been determined by gas exchange experiments. Conductivities for these trace gases were measured at different light fluxes ranging from 820 μEm -2s -1 to darkness. The conductivities to water vapor and the trace gases are highly correlated. It is concluded that the uptake of NO, NO 2 and O 3 by sunflowers and tobacco plants is linearly dependent on stomatal opening. While the uptake of NO is limited by the mesophyll resistance, the uptake of NO 2 is only by diffusion through the stomata. Loss processes by deposition to the leaf surfaces are more pronounced for O 3 than for NO and NO 2.

  2. Nicotiana tabacum as model for ozone - plant surface reactions

    Science.gov (United States)

    Jud, Werner; Fischer, Lukas; Wohlfahrt, Georg; Tissier, Alain; Canaval, Eva; Hansel, Armin

    2015-04-01

    Elevated tropospheric ozone concentrations are considered a toxic threat to plants, responsible for global crop losses with associated economic costs of several billion dollars per year. The ensuing injuries have been related to the uptake of ozone through the stomatal pores and oxidative effects damaging the internal leaf tissue. A striking question of current research is the environment and plant specific partitioning of ozone loss between gas phase, stomatal or plant surface sink terms. Here we show results from ozone fumigation experiments using various Nicotiana Tabacum varieties, whose surfaces are covered with different amounts of unsaturated diterpenoids exuded by their glandular trichomes. Exposure to elevated ozone levels (50 to 150 ppbv) for 5 to 15 hours in an exceptionally clean cuvette system did neither result in a reduction of photosynthesis nor caused any visible leaf damage. Both these ozone induced stress effects have been observed previously in ozone fumigation experiments with the ozone sensitive tobacco line Bel-W3. In our case ozone fumigation was accompanied by a continuous release of oxygenated volatile organic compounds, which could be clearly associated to their condensed phase precursors for the first time. Gas phase reactions of ozone were avoided by choosing a high enough gas exchange rate of the plant cuvette system. In the case of the Ambalema variety, that is known to exude only the diterpenoid cis-abienol, ozone fumigation experiments yield the volatiles formaldehyde and methyl vinyl ketone (MVK). The latter could be unequivocally separated from isomeric methacrolein (MACR) by the aid of a Selective Reagent Ion Time-of-Flight Mass Spectrometer (SRI-ToF-MS), which was switched every six minutes from H3O+ to NO+ primary ion mode and vice versa. Consistent with the picture of an ozone protection mechanism caused by reactive diterpenoids at the leaf surface are the results from dark-light experiments. The ozone loss obtained from the

  3. Transcriptome profiling of male gametophyte development in Nicotiana tabacum

    Directory of Open Access Journals (Sweden)

    Pavel Bokvaj

    2015-03-01

    Full Text Available Pollen, an extremely reduced bicellular or tricellular male reproductive structure of flowering plants, serves as a model for numerous studies covering wide range of developmental and physiological processes. The pollen development represents a fragile and vital phase of plant ontogenesis and pollen was among the first singular plant tissues thoroughly characterized at the transcriptomic level (Honys and Twell [5]. Arabidopsis pollen developmental transcriptome has been published over a decade ago (Honys and Twell, 2004 and transcriptomes of developing pollen of other species have followed (Rice, Deveshwar et al. [2]; Triticeae, Tran et al. [11]; upland cotton, Ma et al. [8]. However, the transcriptomic data describing the development of tobacco pollen, a bicellular model for cell biology studies, have been missing. Here we provide the transcriptomic data covering three stages (Tupý et al., 1983 of wild type tobacco (Nicotiana tabacum, cv. Samsun pollen development: uninucleate microspores (UNM, stage 1, early bicellular pollen (eBCP, stage 3 and late bicellular pollen (lBCP, stage 5 as a supplement to the mature pollen (MP, 4 h-pollen tube (PT4, 24 h-pollen tubes (PT24, leaf (LF and root (RT transcriptomic data presented in our previous studies (Hafidh et al., 2012a; Hafidh et al., 2012b. We characterized these transcriptomes to refine the knowledge base of male gametophyte-enriched genes as well as genes expressed preferentially at the individual stages of pollen development. Alongside updating the list of tissue-specific genes, we have investigated differentially expressed genes with respect to early expressed genes. Pollen tube growth and competition of pollen tubes in female pistil can be viewed as a race of the fittest. Accordingly, there is an apparent evolutionary trend among higher plants to store significant material reserves and nutrients during pollen maturation. This supply ensures that after pollen germination, the pollen tube

  4. An assassin among predators: the relationship between plant-ants, their host Myrmecophytes and the Reduviidae Zelus annulosus.

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    Messika Revel

    Full Text Available Tropical plants frequently live in association with ants that protect their foliage from defoliators. Among them, myrmecophytes have evolved mutualisms with a limited number of plant-ants that they shelter and feed, and, in return, benefit from some protection. Hirtella physophora (Chrysobalanaceae, for example, houses Allomerus decemarticulatus (Myrmicinae that build gallery-shaped traps to catch large prey. In French Guiana, we frequently observed the assassin bug Zelus annulosus (Reduviidae, Harpactorinae on the leaves of H. physophora. Here, we studied the distribution of Zelus annulosus among understory plants in the Guianese rainforest and found it only on pubescent plants, including H. Physophora, whether or not it was sheltering an A. decemarticulatus colony, but only rarely on other myrmecophytes. The relationship between Z. annulosus and its host plants is, then, also mutualistic, as the plant trichomes act as an enemy-free space protecting the nymphs from large predatory ants, while the nymphs protect their host-plants from herbivorous insects. Through their relationship with A. decemarticulatus colonies, Z. annulosus individuals are protected from army ants, while furnishing nothing in return. In those cases where H. physophora sheltered both an A. decemarticulatus colony and Z. annulosus nymphs, certain plant individuals repeatedly sheltered nymphs, indicating that female bugs may select not only pubescent plants but also particular H. physophora treelets having characteristics more favourable to the development of their progeny.

  5. An assassin among predators: the relationship between plant-ants, their host Myrmecophytes and the Reduviidae Zelus annulosus.

    Science.gov (United States)

    Revel, Messika; Dejean, Alain; Céréghino, Régis; Roux, Olivier

    2010-10-01

    Tropical plants frequently live in association with ants that protect their foliage from defoliators. Among them, myrmecophytes have evolved mutualisms with a limited number of plant-ants that they shelter and feed, and, in return, benefit from some protection. Hirtella physophora (Chrysobalanaceae), for example, houses Allomerus decemarticulatus (Myrmicinae) that build gallery-shaped traps to catch large prey. In French Guiana, we frequently observed the assassin bug Zelus annulosus (Reduviidae, Harpactorinae) on the leaves of H. physophora. Here, we studied the distribution of Zelus annulosus among understory plants in the Guianese rainforest and found it only on pubescent plants, including H. Physophora, whether or not it was sheltering an A. decemarticulatus colony, but only rarely on other myrmecophytes. The relationship between Z. annulosus and its host plants is, then, also mutualistic, as the plant trichomes act as an enemy-free space protecting the nymphs from large predatory ants, while the nymphs protect their host-plants from herbivorous insects. Through their relationship with A. decemarticulatus colonies, Z. annulosus individuals are protected from army ants, while furnishing nothing in return. In those cases where H. physophora sheltered both an A. decemarticulatus colony and Z. annulosus nymphs, certain plant individuals repeatedly sheltered nymphs, indicating that female bugs may select not only pubescent plants but also particular H. physophora treelets having characteristics more favourable to the development of their progeny.

  6. Insects found in birds' nests from Argentina. Pseudoseisura lophotes Reichenbach, 1853 and Anumbius annumbi (Vieillot, 1817) (Aves: Furnariidae), hosts of Triatoma platensis Neiva, 1913 (Hemiptera: Reduviidae: Triatominae).

    Science.gov (United States)

    Paola, Turienzo

    2014-02-24

    The insect fauna of the nests of Pseudoseisura lophotes (Reichenbach, 1853) (Aves: Furnariidae) from Argentina was investigated. A total of 110 species (68 identified to species, 22 identified to genus, 20 identified to family) in 40 families of 10 orders of insects was found in these nests. Triatoma platensis Neiva, 1913 (Hemiptera: Reduviidae) was found again in nests of P. lophotes, corroborating after 73 years the first observations made by Mazza in 1936. The occurrence of the insects in nests of P. lophotes is compared with the previously known insect fauna in nests of A. annumbi, Furnarius rufus (Furnariidae), and Myiopsitta monachus (Psittacidae). The insect fauna in additional nests of Anumbius annumbi from the same and/or different localities is given, and used in comparisons. The first occurrence of Cuterebridae (Diptera) in birds' nests, their pupae as the overwintering stage, and the second simultaneous infestation by two species of Philornis (Diptera: Muscidae) on the same nestlings are presented. Other simultaneous infestations of different hematophagous arthropods (Hemiptera: Cimidae; Reduviidae: Triatominae, and Acari: Argasidae) are remarked and discussed.

  7. Identification and validation of polymorphic microsatellite loci for the analysis of Phytophthora nicotianae populations

    Science.gov (United States)

    A large number of SSR loci were screened in the genomic assemblies of 14 different isolates of Phytophthora nicotianae and primers were developed for amplification of 17 markers distributed among different contigs. These loci were highly polymorphic and amplified from genetically distant isolates of...

  8. Product Variability of the ‘Cineole Cassette'Monoterpene Synthases of Related Nicotiana Species

    Institute of Scientific and Technical Information of China (English)

    Anke F(a)hnrich; Katrin Krause; Birgit Piechulla

    2011-01-01

    Nicotiana species of the section Alatae characteristically emit the floral scent compounds of the ‘cineole cassere' comprising 1,8-cineole,limonene,myrcene,α-pinene,β-pinene,sabinene,and α-terpineol.We successfully isolated genes of Nicotiana alata and Nicotiana langsdorfii that encoded enzymes,which produced the characteristic monoterpenes of this ‘cineole cassette' with α-terpineol being most abundant in the volatile spectra.The amino acid sequences of both terpineol synthases were 99% identical.The enzymes cluster in a monophyletic branch together with the closely related cineole synthase of Nicotiana suaveolens and monoterpene synthase 1 of Solanum lycopersicum.The cyclization reactions (α-terpineol to 1,8-cineole) of the terpineol synthases of N.alata and N.langsdorfii were less efficient compared to the ‘cineole cassette′ monoterpene synthases of Arabidopsis thaliana,N.suaveolens,Salvia fruticosa,Salvia officinalis,and Citrus unshiu.The terpineol synthases of N.alata and N.langsdorfii were localized in pistils and in the adaxial and abaxial epidermis of the petals.The enzyme activities reached their maxima at the second day after anthesis when flowers were fully opened and the enzyme activity in N.alata was highest at the transition from day to night (diurnal rhythm).

  9. Phenotypical and molecular characterization of the Tomato mottle Taino virus-Nicotiana megalosiphon interaction

    NARCIS (Netherlands)

    Collazo, C.; Ramos, P.L.; Chacón, O.; Borroto, C.J.; López, Y.; Pujol, M.; Thomma, B.P.H.J.; Hein, I.; Borrás-Hidalgo, O.

    2005-01-01

    Tomato mottle Taino virus (ToMoTV) infection causes significant yield losses in plants of various Solanaceous species. In this study, the interaction between Nicotiana megalosiphon and ToMoTV was characterized on a phenotypical and molecular basis. In order to isolate genes that are differentially e

  10. Genetic characterization of Phytophthora nicotianae by the analysis of polymorphic regions of the mitochondrial DNA.

    Science.gov (United States)

    A new method based on the analysis of mitochondrial intergenic regions characterized by intraspecific variation in DNA sequences was developed and applied to the study of the plant pathogen Phytophthora nicotianae. Two regions flanked by genes trny and rns and trnw and cox2 were identified by compa...

  11. Anaerobic soil disinfestation reduces survival and infectivity of Phytophthora nicotianae chlamydospores in pepper

    Science.gov (United States)

    Phytophthora nicotianae is the principal causal agent of root and crown rot disease of pepper plants in Extremadura (western Spain), a spring-summer crop in this region. Preplant soil treatment by anaerobic soil disinfestation (ASD) may effectively control plant pathogens in many crop production sys...

  12. GENE-REGULATION IN INTERTYPIC HETEROKARYONS OF SOLANUM-TUBEROSUM AND NICOTIANA-TABACUM TISSUE PROTOPLASTS

    NARCIS (Netherlands)

    VANKESTEREN, WJP; BIJMOLT, EW; TEMPELAAR, MJ

    1994-01-01

    Activities of the beta-glucuronidase (GUS) reporter enzyme were evaluated in transgenic plants, protoplasts, and intertypic heterokaryons of Solanum tuberosum and Nicotiana tabacum. With GUS under control of the promoter of the cauliflower-mosaicvirus 35S RNA gene (CaMV), activities of the enzyme we

  13. Effects of air pollutants on Nicotiana cultivars and species used for virus studies

    Energy Technology Data Exchange (ETDEWEB)

    Grosso, J.J.; Menser, H.A.; Hodges, G.H.; McKinney, H.H.

    1971-08-01

    Three tobacco cultivars (Nicotiana tabacum) and two Nicotiana species, used for virus identification studies, were tested for susceptibility to air pollutants. The air pollutants used were (i) ozone; (ii) sulfur dioxide; and (iii) a mixture of both gases. Nicotiana rustica var. brasilia was the most susceptible cultivar to injury from individual gases, a gas mixture, and ambient polluted air. Nicotiana rustica var. brasilia displayed from 25 to 70% leaf injury when exposed to increasing ozone doses (10 to 30 pphm). Ozone injury to N. glutinosa fumigated at the same doses ranged from 10 to 60%. Samsun, Samsun (NN), and Xanthi cultivars were more resistant to ozone than N. rustica var. brasilia or N. glutinosa. Sulfur dioxide (ca. 45 pphm) added to an ozone (ca. 3.0 pphm) atmosphere induced a synergistic response evidenced as injury to all cultivars and species. Mixed gas injury was more acute on N. glutinosa and N. rustica var. brasilia than on Samsun, Samsun (NN), and Xanthi. Samsun and Samsun (NN) were more susceptible to sulfur dioxide (125 pphm) than N. glutinosa and Xanthi. Fumigations produced two injury syndromes: (i) light or dark punctate flecks, on upper leaf surfaces; or (ii) bifacial tissue collapse.

  14. In vitro grown pollen tubes of Nicotiana alata actively synthesise a fucosylated xyloglucan.

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    Edwin R Lampugnani

    Full Text Available Nicotiana alata pollen tubes are a widely used model for studies of polarized tip growth and cell wall synthesis in plants. To better understand these processes, RNA-Seq and de novo assembly methods were used to produce a transcriptome of N. alata pollen grains. Notable in the reconstructed transcriptome were sequences encoding proteins that are involved in the synthesis and remodelling of xyloglucan, a cell wall polysaccharide previously not thought to be deposited in Nicotiana pollen tube walls. Expression of several xyloglucan-related genes in actively growing pollen tubes was confirmed and xyloglucan epitopes were detected in the wall with carbohydrate-specific antibodies: the major xyloglucan oligosaccharides found in N. alata pollen grains and tubes were fucosylated, an unusual structure for the Solanaceae, the family to which Nicotiana belongs. Finally, carbohydrate linkages consistent with xyloglucan were identified chemically in the walls of N. alata pollen grains and pollen tubes grown in culture. The presence of a fucosylated xyloglucan in Nicotiana pollen tube walls was thus confirmed. The consequences of this discovery to models of pollen tube growth dynamics and more generally to polarised tip-growing cells in plants are discussed.

  15. Infestation of Palm Trees by Triatomines (Hemiptera: Reduviidae in the State of Bahia, Brazil

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    Rodrigo Gurgel-Gonçalves

    2012-12-01

    Resumo. As palmeiras desempenham papéis importantes como habitats de reprodução e alimentação para triatomíneos silvestres, vetores da doença de Chagas. A ocorrência de triatomíneos em palmeiras peridomiciliares pode aumentar o risco de invasão desses insetos em domicílios e deve ser considerada para desenvolver estratégias de prevenção. Com objetivo de investigar a infestação de palmeiras por triatomíneos no Estado da Bahia e determinar a infecção natural desses insetos por Trypanosoma spp., foram amostradas 183 palmeiras em 12 municípios entre 2006 e 2011 utilizando captura manual e/ou armadilhas adesivas iscadas com camundongos. Os triatomíneos foram detectados em 79 palmeiras (43% das espécies Copernicia prunifera (Mart. Becc., Mauritia flexuosa L. e Attalea spp. (Attalea burretiana Bondar ou Attalea salvadorensis Glassman. Em outras espécies de palmeiras (Syagrus coronata (Mill H.E. Moore, Attalea funifera Mart ex. Spreng e Elaeis guineensis Jacq não foram detectados triatomíneos. Rhodnius neglectus Lent, Triatoma sordida (Stål, e Triatoma pseudomaculata Corrêa & Espínola ocorreram em C. prunifera ao longo do rio São Francisco. No extremo oeste da Bahia, R. neglectus e Psammolestes tertius Lent & Jurberg foram detectados em M. flexuosa, enquanto Triatoma tibiamaculata (Pinto ocorreu em Attalea sp. em áreas urbanas de Salvador. No total, 180 triatomíneos foram capturados, principalmente R. neglectus. A maior taxa de infecção natural (61% foi observada em T. tibiamaculata. Os resultados indicam que pelo menos três espécies de palmeiras são habitats favoráveis para triatomíneos no estado da Bahia e ocorrem no ambiente peridomiciliar, o que pode aumentar a probabilidade de invasão de triatomíneos nas casas.

  16. Screening micro-organisms for cadmium absorption from aqueous solution and cadmium absorption properties of Arthrobacter nicotianae.

    Science.gov (United States)

    Tsuruta, Takehiko; Umenai, Daishi; Hatano, Tomonobu; Hirajima, Tsuyoshi; Sasaki, Keiko

    2014-01-01

    To obtain basic information on how microbial cells absorb cadmium from aqueous solution, we examined cadmium absorption in various micro-organisms. Of 51 micro-organism strains tested, we found that some Gram-positive bacteria, such as, Arthrobacter nicotianae and Bacillus subtilis, and some actinomycetes, such as, Streptomyces flavoviridis and S. levoris were highly capable of absorbing cadmium from an aqueous solution. A. nicotianae absorbed the largest amount of cadmium, over 800 μmol cadmium per gram of dry wt. cells. However, cadmium absorption by A. nicotianae was affected by the solution pH, cadmium concentration, and cell density. The absorption of cadmium was very rapid. Some factors that affected cadmium absorption by A. nicotianae cells were also discussed.

  17. A Redox-Mediated Modulation of Stem Bolting in Transgenic Nicotiana sylvestris Differentially Expressing the External Mitochondrial NADPH Dehydrogenase

    National Research Council Canada - National Science Library

    Yun-Jun Liu; Adriano Nunes-Nesi; Sabá V. Wallström; Ida Lager; Agnieszka M. Michalecka; Fredrik E. B. Norberg; Susanne Widell; Kenneth M. Fredlund; Alisdair R. Fernie; Allan G. Rasmusson

    2009-01-01

    .... Nicotiana sylvestris plants overexpressing potato {Solarium tuberosum) NDB1 displayed early bolting, whereas sense suppression of the same gene led to delayed bolting, with consequential changes in flowering time...

  18. Detection of a Potential New Bartonella Species “Candidatus Bartonella rondoniensis” in Human Biting Kissing Bugs (Reduviidae; Triatominae)

    Science.gov (United States)

    Laroche, Maureen; Berenger, Jean-Michel; Mediannikov, Oleg; Raoult, Didier; Parola, Philippe

    2017-01-01

    Background Among the Reduviidae family, triatomines are giant blood-sucking bugs. They are well known in Central and South America where they transmit Trypanosoma cruzi to mammals, including humans, through their feces. This parasitic protozoan is the causative agent of Chagas disease, a major public health issue in endemic areas. Because of the medical and economic impact of Chagas disease, the presence of other arthropod-borne pathogens in triatomines was rarely investigated. Methodology/Principal findings In this study, seven triatomines species involved in the transmission of T. cruzi were molecularly screened for the presence of known pathogens generally associated with arthropods, such as Rickettsia, Bartonella, Anaplasmataceae, Borrelia species and Coxiella burnetii. Of all included triatomine species, only Eratyrus mucronatus specimens tested positive for Bartonella species for 56% of tested samples. A new genotype of Bartonella spp. was detected in 13/23 Eratyrus mucronatus specimens, an important vector of T. cruzi to humans. This bacterium was further characterized by sequencing fragments of the ftsZ, gltA and rpoB genes. Depending on the targeted gene, this agent shares 84% to 91% of identity with B. bacilliformis, the agent of Carrion’s disease, a deadly sandfly-borne infectious disease endemic in South America. It is also closely related to animal pathogens such as B. bovis and B. chomelii. Conclusions As E. mucronatus is an invasive species that occasionally feeds on humans, the presence of potentially pathogenic Bartonella-infected bugs could present another risk for human health, along with the T. cruzi issue. PMID:28095503

  19. Detection of a Potential New Bartonella Species "Candidatus Bartonella rondoniensis" in Human Biting Kissing Bugs (Reduviidae; Triatominae.

    Directory of Open Access Journals (Sweden)

    Maureen Laroche

    2017-01-01

    Full Text Available Among the Reduviidae family, triatomines are giant blood-sucking bugs. They are well known in Central and South America where they transmit Trypanosoma cruzi to mammals, including humans, through their feces. This parasitic protozoan is the causative agent of Chagas disease, a major public health issue in endemic areas. Because of the medical and economic impact of Chagas disease, the presence of other arthropod-borne pathogens in triatomines was rarely investigated.In this study, seven triatomines species involved in the transmission of T. cruzi were molecularly screened for the presence of known pathogens generally associated with arthropods, such as Rickettsia, Bartonella, Anaplasmataceae, Borrelia species and Coxiella burnetii. Of all included triatomine species, only Eratyrus mucronatus specimens tested positive for Bartonella species for 56% of tested samples. A new genotype of Bartonella spp. was detected in 13/23 Eratyrus mucronatus specimens, an important vector of T. cruzi to humans. This bacterium was further characterized by sequencing fragments of the ftsZ, gltA and rpoB genes. Depending on the targeted gene, this agent shares 84% to 91% of identity with B. bacilliformis, the agent of Carrion's disease, a deadly sandfly-borne infectious disease endemic in South America. It is also closely related to animal pathogens such as B. bovis and B. chomelii.As E. mucronatus is an invasive species that occasionally feeds on humans, the presence of potentially pathogenic Bartonella-infected bugs could present another risk for human health, along with the T. cruzi issue.

  20. Importance of species of Triatominae (Heteroptera: Reduviidae) in risk of transmission of Trypanosoma cruzi in western Mexico.

    Science.gov (United States)

    Martínez-Ibarra, J A; Grant-Guillén, Y; Morales-Corona, Z Y; Haro-Rodríguez, S; Ventura-Rodríguez, L V; Nogueda-Torres, B; Bustos-Saldaña, R

    2008-05-01

    The epidemiological risk of infection by Trypanosoma cruzi Chagas in human populations of western Mexico is still under study. Although most vectors in this region and their vector capability are already known, new studies estimating the risk and the importance of individual Triatominae species (Hemiptera: Reduviidae) for T. cruzi transmission are necessary. For 1 yr, every month, > 400 human dwellings and their surroundings in eight communities of two western Mexico states were searched for triatomines. More than 1,000 specimens representing four species were collected and checked for T. cruzi infection. Based on the usual entomological indices, only the inhabitants of Gavilán El Progreso-La Villita are at serious risk of vectorial infection by T. cruzi. A population of Meccus longipennis (Usinger) was found living in peridomestic rock pile boundary walls after an insecticide spraying. It was confirmed the major role of peridomestic habitats as shelter areas for triatomines, particularly in rock pile boundary walls and chicken roosts. Triatominae presence also was verified in certain sylvatic habitats, including primarily heaps of stones. The important role of M. longipennis in the potential transmission of T. cruzi in the region and the secondary role of M. picturatus (Usinger) and Triatoma barberi Usinger also were confirmed. Null colonization of houses by T. barberi, which was collected primarily in peridomestic habitats, differs from its common intradomiciliary collection in other studies. Meccus pallidipennis (Stål) most probably does not exist in Nayarit. Meccus mazzottii (Usinger) and Meccus phyllosomus (Burmeister) are no longer found in Nayarit and Jalisco. Additional studies are necessary to determine the current epidemiological situation in other areas of western Mexico.

  1. Descrição de ovos e ninfas de Triatoma klugi (Hemiptera, Reduviidae, Triatominae Description of eggs and nymphs of Triatoma klugi (Hemiptera, Reduviidae, Triatominae

    Directory of Open Access Journals (Sweden)

    José Jurberg

    Full Text Available Os autores descreveram as características morfológicas de Triatoma klugi Carcavallo, Jurberg, Lent & Galvão, 2001 pertencente ao grupo de espécies que compõem o "complexo T. oliveirai". Até o presente tem sido difícil separar essas espécies com base nas características ninfais, o que justifica o desenvolvimento deste trabalho. Os espécimes foram coletados em frestas de rochas no morro Malavok na localidade de Linha Brasil, município de Nova Petrópolis, no Estado do Rio Grande do Sul. O local de captura dos triatomíneos situa-se entre 700 e 800 m de altitude (29º18'38''S, 51º04'57''W. Juntamente com T. oliveirai (Neiva, Pinto & Lent, 1939, espécie morfologicamente mais próxima, são as únicas do complexo que não foram encontradas, até o momento, no Estado do Mato Grosso, Mato Grosso do Sul e Goiás, Brasil. Em condições experimentais, já foi testada e comprovada a sua susceptibilidade à infecção pelo Trypanosoma cruzi (Chagas, 1909 e Trypanosoma rangeli (Tejera, 1920. A análise comparativa das ornamentações do exocório dos ovos e de três estádios ninfais de T. klugi por microscopia óptica e microscopia eletrônica de varredura mostrou algumas particularidades morfológicas, com destaque para: a face ventral da cabeça, o sulco estridulatório e os últimos segmentos abdominais (IX, X - futura genitália e XI - tubo anal. Esses dados contribuem para a ampliação dos parâmetros diferenciais visando à diagnose de T. klugi durante o seu desenvolvimento ninfal.The authors had described the morphologic characteristics of Triatoma klugi Carcavallo, Jurberg, Lent & Galvão, 2001 to the group of species belonging to "T. oliveirai complex". Until the moment it has been difficult to separate on the nymphs characteristics basis these species, what it justifies the development of this work. The specimens had been collected in openings of rocks in the Malavok mount in the locality of Linha Brazil, city of Nova Petr

  2. Nicotiana Occidentalis Chloroplast Ultrastructure imaged with Transmission Electron Microscopes Working at Different Accelerating Voltages

    OpenAIRE

    SVIDENSKÁ, Silvie

    2010-01-01

    The main goal of this thesis is to study and compare electron microscopy images of Nicotiana Occidentalis chloroplasts, obtained from two types of transmission electron microscopes,which work with different accelerating voltage of 80kV and 5kV. The two instruments, TEM JEOL 1010 and low voltage electron microscope LVEM5 are employed for experiments. In the first theoretical part, principle of electron microscopy and chloroplast morphology is described. In experimental part, electron microscop...

  3. The haploid embryoids development in anther culture of transformed tobacco (Nicotiana tabacum L.)

    OpenAIRE

    Jaroslava Dubová

    2014-01-01

    Induction of pollen embryogenesis was used for the test of cytokinin influence on the development of pollen grain. Gene for maize cytokinin-specific β-glucosidase were introduced into tobacco genome together with the methotrexate resistance marker (Brzobohatý et al. 1994). Anther culture of T1 generation of transformed Nicotiana tabacum L. plants revealed that high activity of β-glucosidase caused a reduced regeneration of pollen embryoids. The stages of pollen embryoid development were compa...

  4. Preliminary Observations on Zelus obscuridorsis (Stål (Hemiptera: Reduviidae as Predator of the Corn Leafhopper (Hemiptera: Cicadellidae in Argentina

    Directory of Open Access Journals (Sweden)

    Eduardo G. Virla

    2015-06-01

    Full Text Available The corn leafhopper Dalbulus maidis (Hemiptera: Cicadellidae, is an important corn pest in most of tropical and subtropical America. This leafhopper has a rich natural enemy complex of which parasitoids and pathogens are the most studied; knowledge on its predators is limited. We noted the presence of the native assassin bug Zelus obscuridorsis (Hemiptera: Reduviidae predating diverse motile insects, including the corn leafhopper, on corn plants cultivated in household vegetable gardens in San Miguel de Tucumán (Argentina; in order to verify its predatory actions, we exposed lab-bred individuals of D. maidis to adults of Z. obscuridorsis. The predators were starved for 24 h before trials in which the corn leafhopper in different developmental stages were exposed. Zelus obscuridorsis is highly skilled in catching specimens in motion, but it was not able to prey on eggs. The predator was capable to catch and prey on nymphs and adults.

  5. Preliminary Observations on Zelus obscuridorsis (Stål) (Hemiptera: Reduviidae) as Predator of the Corn Leafhopper (Hemiptera: Cicadellidae) in Argentina.

    Science.gov (United States)

    Virla, Eduardo G; Melo, Cecilia M; Speranza, Stefano

    2015-06-03

    The corn leafhopper Dalbulus maidis (Hemiptera: Cicadellidae), is an important corn pest in most of tropical and subtropical America. This leafhopper has a rich natural enemy complex of which parasitoids and pathogens are the most studied; knowledge on its predators is limited. We noted the presence of the native assassin bug Zelus obscuridorsis (Hemiptera: Reduviidae) predating diverse motile insects, including the corn leafhopper, on corn plants cultivated in household vegetable gardens in San Miguel de Tucumán (Argentina); in order to verify its predatory actions, we exposed lab-bred individuals of D. maidis to adults of Z. obscuridorsis. The predators were starved for 24 h before trials in which the corn leafhopper in different developmental stages were exposed. Zelus obscuridorsis is highly skilled in catching specimens in motion, but it was not able to prey on eggs. The predator was capable to catch and prey on nymphs and adults.

  6. Sincronización de Células de Tabaco (Nicotiana tabacum NT-1 Synchronization of tobacco cells (Nicotiana tabacum NT-1

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    León F Ruiz

    2010-01-01

    Full Text Available Se ha evaluado la capacidad sincronizante de afidicolina e hidroxiurea en cultivos de células de tabaco (Nicotiana tabacum NT-1. Los cultivos sincronizados son poderosas herramientas en estudios moleculares y bioquímicos relacionados al ciclo celular y comúnmente se utilizan químicos para bloquear el ciclo celular. La línea celular de tabaco (Nicotiana tabacum NT-1 proviene de la línea celular TBY-2, caracterizándose NT-1 por su menor velocidad de crecimiento y tamaño celular heterogéneo. Los resultados mostraron una sincronía del 30 % de NT-1 con afidicolina 5 mg/ml, similar a la obtenida con TBY-2. Se probaron diferentes concentraciones de hidroxiurea, obteniéndose niveles óptimos de sincronización de hasta 31 % con 0.75 mM. Se muestra también que es posible sincronizar cultivos NT-1 en porcentajes similares a cultivos TBY-2 con afidicolina e hidroxiurea. Para este propósito, se recomienda la hidroxiurea por ser químicamente más estable y económica.The synchronizing capacity of aphidicolin and hydroxyurea on NT-1 cultures of tobacco cells (Nicotiana tabacum NT-1. Synchronized cell cultures are a powerful tool in biochemistry and molecular studies related to cell cycle and commonly chemicals are used to block the biochemical cycle. The tobacco cell line NT-1 comes from cell line TBY-2, the former being characterized with a slower growth rate and heterogeneous cell size. Results show 30 % synchrony with 5 mg/ml aphidicolin, similar to that obtained with TBY-2. Different concentrations of hydroxyurea were tested, obtaining an optimal synchrony of 31 % at 0.75 mM of concentration. It is also shown that it is possible to synchronize NT-1 cultures in similar percentages to those obtained with TBY-2, with either aphidicolin or hydroxyurea. For this purpose, hydroxyurea is recommended because of its lower cost and chemical stability.

  7. Ciclo biológico de Triatoma rubrovaria (Blanchard (Reduviidae, Triatominae en laboratorio

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    Nilda V. de Argüello

    1988-01-01

    Full Text Available Fez-se uma avaliação estatística do ciclo de vida de Triatoma rubrovaria, com base em experimentos de laboratório. Foram computados a eclosão do ovo, o tempo de desenvolvimento da ninfa e sua mortalidade, e a longevidade dos adultos, machos e fêmeas; os resultados foram comparados com os da literatura existente.A statistical evaluation of the life cycle of Triatoma rubrovaria is based on cohort experiments conducted under controlled laboratory conditions. Egg hatching, nymphal development time and mortality, adult male and femals longevity were computed, and the results compared with information in the existing literature.

  8. Nicotiana small RNA sequences support a host genome origin of cucumber mosaic virus satellite RNA.

    Science.gov (United States)

    Zahid, Kiran; Zhao, Jian-Hua; Smith, Neil A; Schumann, Ulrike; Fang, Yuan-Yuan; Dennis, Elizabeth S; Zhang, Ren; Guo, Hui-Shan; Wang, Ming-Bo

    2015-01-01

    Satellite RNAs (satRNAs) are small noncoding subviral RNA pathogens in plants that depend on helper viruses for replication and spread. Despite many decades of research, the origin of satRNAs remains unknown. In this study we show that a β-glucuronidase (GUS) transgene fused with a Cucumber mosaic virus (CMV) Y satellite RNA (Y-Sat) sequence (35S-GUS:Sat) was transcriptionally repressed in N. tabacum in comparison to a 35S-GUS transgene that did not contain the Y-Sat sequence. This repression was not due to DNA methylation at the 35S promoter, but was associated with specific DNA methylation at the Y-Sat sequence. Both northern blot hybridization and small RNA deep sequencing detected 24-nt siRNAs in wild-type Nicotiana plants with sequence homology to Y-Sat, suggesting that the N. tabacum genome contains Y-Sat-like sequences that give rise to 24-nt sRNAs capable of guiding RNA-directed DNA methylation (RdDM) to the Y-Sat sequence in the 35S-GUS:Sat transgene. Consistent with this, Southern blot hybridization detected multiple DNA bands in Nicotiana plants that had sequence homology to Y-Sat, suggesting that Y-Sat-like sequences exist in the Nicotiana genome as repetitive DNA, a DNA feature associated with 24-nt sRNAs. Our results point to a host genome origin for CMV satRNAs, and suggest novel approach of using small RNA sequences for finding the origin of other satRNAs.

  9. Nicotiana small RNA sequences support a host genome origin of cucumber mosaic virus satellite RNA.

    Directory of Open Access Journals (Sweden)

    Kiran Zahid

    2015-01-01

    Full Text Available Satellite RNAs (satRNAs are small noncoding subviral RNA pathogens in plants that depend on helper viruses for replication and spread. Despite many decades of research, the origin of satRNAs remains unknown. In this study we show that a β-glucuronidase (GUS transgene fused with a Cucumber mosaic virus (CMV Y satellite RNA (Y-Sat sequence (35S-GUS:Sat was transcriptionally repressed in N. tabacum in comparison to a 35S-GUS transgene that did not contain the Y-Sat sequence. This repression was not due to DNA methylation at the 35S promoter, but was associated with specific DNA methylation at the Y-Sat sequence. Both northern blot hybridization and small RNA deep sequencing detected 24-nt siRNAs in wild-type Nicotiana plants with sequence homology to Y-Sat, suggesting that the N. tabacum genome contains Y-Sat-like sequences that give rise to 24-nt sRNAs capable of guiding RNA-directed DNA methylation (RdDM to the Y-Sat sequence in the 35S-GUS:Sat transgene. Consistent with this, Southern blot hybridization detected multiple DNA bands in Nicotiana plants that had sequence homology to Y-Sat, suggesting that Y-Sat-like sequences exist in the Nicotiana genome as repetitive DNA, a DNA feature associated with 24-nt sRNAs. Our results point to a host genome origin for CMV satRNAs, and suggest novel approach of using small RNA sequences for finding the origin of other satRNAs.

  10. Efficiency for Gene Silencing Induction in Nicotiana Species by a Viral Satellite DNA Vector

    Institute of Scientific and Technical Information of China (English)

    You-Ping Xu; Lu-Ping Zheng; Qiu-Fang Xu; Chang-Chun Wang; Xue-Ping Zhou; Zu-Jian Wu; Xin-Zhong Cai

    2007-01-01

    Virus-induced gene silencing (VIGS) is a useful technique for rapid plant gene function analysis.We recently reported a new VIGS vector modified from Tomato yellow leaf curl China virus (TYLCCNV) DNAβ (DNAm β).In this study we compared In detail DNAmβ-induced gene silencing in four Nicotiana species including N.benthamiana, N.glutinosa, N.tabacum and N.paniculata.We found that DNAmβ-induced gene silencing in the four species was distinct in developing dynamics, tissue specificity, efficiency, and constancy in the plant life span.It was most efficient in N.benthamiana, where development of VIGS was most rapid, without tissue specificity and nearly 100% efficient.DNAmβ-induced gene silencing in N.Glutinosa was also efficient despite being slightly less than in N.benthamiana.It initially occurred in veins, later was scattered to mesophyll, finally led to complete silencing in whole leaves.In both species, VIGS constantly expressed until the plants died.However, DNAmβ-mediated VIGS in the other two Nicotiana species, N.tabacum and N.paniculata, was significantly less efficient.It was strictly limited within the veins of the silenced leaves, and constantly occurred only over 3-4 weeks.The upper leaves that emerged later stopped showing the silencing phenotype, DNAm β-induced gene silencing in N.benthamiana and N.glutinosa was not significantly influenced by the growth stage when the plants were agro-inoculated,and was not sensitive to high growth temperature up to 32℃, Our results indicate that this system has great potential as a versatile VIGS system for routine functional analysis of genes in some Nicotiana species.

  11. A Novel Isoenzyme of CuZn-superoxide Dismutase from Nicotiana tobacum

    Institute of Scientific and Technical Information of China (English)

    Liang Quan SHENG; Shao Min LIU; Hou Rong XIAO; Bing Le XIA; Qing Liang LIU

    2004-01-01

    An isoenzyme of CuZn-superoxide dismutase, denoted as CuZnSODⅢ, has been separated and purified from Nicotiana Tobacum (tobacco) leaves to apparent homogeneity. Its molecular mass is 22976.6Da. It is composed of one subunit, which is consisted of 187 amine acid residues and contains 1 copper and 0.5 zinc atom. The activation energy of the thermal denaturation process has been obtained as about 143.5kJmol-1. Meanwhile, some properties of spectra were investigated.

  12. Genetic Diversity and Phylogeny of Antagonistic Bacteria against Phytophthora nicotianae Isolated from Tobacco Rhizosphere

    OpenAIRE

    Jin, Fengli; Ding, Yanqin; Ding, Wei; Reddy, M. S.; Fernando, W. G. Dilantha; Du,Binghai

    2011-01-01

    The genetic diversity of antagonistic bacteria from the tobacco rhizosphere was examined by BOXAIR-PCR, 16S-RFLP, 16S rRNA sequence homology and phylogenetic analysis methods. These studies revealed that 4.01% of the 6652 tested had some inhibitory activity against Phytophthora nicotianae. BOXAIR-PCR analysis revealed 35 distinct amplimers aligning at a 91% similarity level, reflecting a high degree of genotypic diversity among the antagonistic bacteria. A total of 25 16S-RFLP patterns were i...

  13. The haploid embryoids development in anther culture of transformed tobacco (Nicotiana tabacum L.

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    Jaroslava Dubová

    2014-01-01

    Full Text Available Induction of pollen embryogenesis was used for the test of cytokinin influence on the development of pollen grain. Gene for maize cytokinin-specific β-glucosidase were introduced into tobacco genome together with the methotrexate resistance marker (Brzobohatý et al. 1994. Anther culture of T1 generation of transformed Nicotiana tabacum L. plants revealed that high activity of β-glucosidase caused a reduced regeneration of pollen embryoids. The stages of pollen embryoid development were comparable with those of zygotic embryos. Optimal nutritive medium contained charcoal and chelated iron. The absence of iron in the initiation medium caused a distinct decrease of pollen embryoid regeneration.

  14. Sensitivity of Allium and Nicotiana in cellular and acellular comet assays to assess differential genotoxicity of direct and indirect acting mutagens.

    Science.gov (United States)

    Bandyopadhyay, Atrayee; Mukherjee, Anita

    2011-05-01

    We have evaluated the extent of DNA damage induced by direct and indirect mutagens by cellular and acellular comet assays in two plant systems, Nicotiana tabacum (wild type tobacco) and Allium cepa (common onion). The objectives of this study were: (1) to generate dose-response curves for DNA migration values from root and shoot nuclei of A. cepa and N. tabacum treated with the direct acting mutagens, ethyl methanesulphonate (EMS), hydrogen peroxide (H(2)O(2)) and the indirect acting mutagen, cadmium chloride (CdCl(2)), (2) to assess the differential response between isolated nuclei and nuclei of root and shoot and of both plants and (3) to examine the differences of sensitivity between direct and indirect acting mutagens by cellular and acellular comet assays. Similar sensitivities were evident in both plant systems to direct and indirect acting mutagens. The combination of cellular and acellular comet assays provided valuable insight to the mode of action of the genotoxicants used. The data obtained demonstrated the estimable capacity of the two plant systems to evaluate genotoxicity under different stress conditions and suggests Allium is a more desirable test system for rapid monitoring of genotoxicity.

  15. Pollination following grafting introduces efficiently Ocimum basilicum L. genes into Nicotiana tabacum L.

    Directory of Open Access Journals (Sweden)

    K. Q. Wei

    2013-09-01

    Full Text Available Tobacco is an important cash crop in the world. However, the genetic basis is comparatively narrow among the modern Nicotiana tabacum cultivars, limiting its potential for quality improvement. To introduce genes conferring desirable chemical constituents from medicinal plants, a distant hybridization test was conducted between N. tabacum and Ocimum basilicum L. Seedlings of wild type Nicotiana sylvestris and N. tabacum cultivar 78-04 respectively acted as rootstock and scion. During the flowering season, hand pollination between 78-04 as pistillate parent and O. basilicum as pollen parent was carried out under 22-25°C temperature and 70-80% of relative humidity in the greenhouse. Seed sets of 55% were obtained in 78-04, and about 400 seeds per capsule were produced. But both non-grafted and self-grafted 78-04 plants rarely resulted in fruits by hand pollination and those obtained were without seed. Similar results were obtained in different material combination. The interfamilial F1 hybrids acquired showed distinct variation with various morphological characteristics, and their hybrid nature was confirmed by isozyme and random amplified polymorphic DNA (RAPD analyses. This result indicated that pollination following grafting can facilitate gene exchange and recombination at the interfamilial level and efficiently overcome barriers of sexual incompatibility between N. tabacum and O. basilicum. Our research not only extends the genetic basis of tobacco but also will provide valuable germplasm for improvement of varieties.

  16. Low abundant spacer 5S rRNA transcripts are frequently polyadenylated in Nicotiana.

    Science.gov (United States)

    Fulnecek, Jaroslav; Kovarik, Ales

    2007-11-01

    In plants, 5S rRNA genes (5S rDNA) encoding 120-nt structural RNA molecules of ribosomes are organized in tandem arrays comprising thousands of units. Failure to correctly terminate transcription would generate longer inaccurately processed transcripts interfering with ribosome biogenesis. Hence multiple termination signals occur immediately after the 5S rRNA coding sequence. To obtain information about the efficiency of termination of 5S rDNA transcription in plants we analyzed 5S rRNA pools in three Nicotiana species, N. sylvestris, N. tomentosiformis and N. tabacum. In addition to highly abundant 120-nt 5S rRNA transcripts, we also detected RNA species composed of a genic region and variable lengths of intergenic sequences. These genic-intergenic RNA molecules occur at a frequency severalfold lower than the mature 120-nt transcripts, and are posttranscriptionally modified by polyadenylation at their 3' end in contrast to 120-nt transcripts. An absence of 5S small RNAs (smRNA) argue against a dominant role for the smRNA biosynthesis pathway in the degradation of aberrant 5S rRNA in Nicotiana. This work is the first description of polyadenylated 5S rRNA species in higher eukaryotes originating from a read-through transcription into the intergenic spacer. We propose that polyadenylation may function in a "quality control" pathway ensuring that only correctly processed molecules enter the ribosome biogenesis.

  17. Overexpression of Citrus junos mitochondrial citrate synthase gene in Nicotiana benthamiana confers aluminum tolerance.

    Science.gov (United States)

    Deng, Wei; Luo, Keming; Li, Zhengguo; Yang, Yingwu; Hu, Nan; Wu, Yu

    2009-07-01

    Aluminum (Al) toxicity is one of the major factors that limit plant growth in acid soils. Al-induced release of organic acids into rhizosphere from the root apex has been identified as a major Al-tolerance mechanism in many plant species. In this study, Al tolerance of Yuzu (Citrus Junos Sieb. ex Tanaka) was tested on the basis of root elongation and the results demonstrated that Yuzu was Al tolerant compared with other plant species. Exposure to Al triggered the exudation of citrate from the Yuzu root. Thus, the mechanism of Al tolerance in Yuzu involved an Al-inducible increase in citrate release. Aluminum also elicited an increase of citrate content and increased the expression level of mitochondrial citrate synthase (CjCS) gene and enzyme activity in Yuzu. The CjCS gene was cloned from Yuzu and overexpressed in Nicotiana benthamiana using Agrobacterium tumefaciens-mediated methods. Increased expression level of the CjCS gene and enhanced enzyme activity were observed in transgenic plants compared with the wild-type plants. Root growth experiments showed that transgenic plants have enhanced levels of Al tolerance. The transgenic Nicotiana plants showed increased levels of citrate in roots compared to wild-type plants. The exudation of citrate from roots of the transgenic plants significantly increased when exposed to Al. The results with transgenic plants suggest that overexpression of mitochondrial CS can be a useful tool to achieve Al tolerance.

  18. A taxonomic monograph of the assassin bug genus Zelus Fabricius (Hemiptera: Reduviidae): 71 species based on 10,000 specimens.

    Science.gov (United States)

    Zhang, Guanyang; Hart, Elwood R; Weirauch, Christiane

    2016-01-01

    The New World assassin bug genus Zelus Fabricius, 1803 (Insecta: Hemiptera: Heteroptera: Reduviidae: Harpactorinae: Harpactorini) is revised based on more than 10,000 specimens. Seventy-one species are recognized and twenty-four described as new: Zelus aithaleos sp. n., Zelus amblycephalus sp. n., Zelus antiguensis sp. n., Zelus auralanus sp. n., Zelus bahiaensis sp. n., Zelus banksi sp. n., Zelus casii sp. n., Zelus championi sp. n., Zelus cordazulus sp. n., Zelus fuliginatus sp. n., Zelus gilboventris sp. n., Zelus gracilipes sp. n., Zelus grandoculus sp. n., Zelus kartaboides sp. n., Zelus lewisi sp. n., Zelus panamensis sp. n., Zelus paracephalus sp. n., Zelus rosulentus sp. n., Zelus russulumus sp. n., Zelus spatulosus sp. n., Zelus truxali sp. n., Zelus umbraculoides sp. n., Zelus umbraculus sp. n., and Zelus xouthos sp. n. Five species, Zelus araneiformis Haviland, 1931, Zelus gradarius Bergroth, 1905, Zelus modestus (Stål, 1862), Zelus subfasciatus Stål, 1860 and Zelus vittaticeps Stål, 1866, are removed from Zelus and placed incertae sedis within Harpactorini. Nine new synonyms are recognized (senior synonym in parentheses): Zelus atripes Champion, 1898 syn. nov. (=Zelus conjungens [Stål, 1860]), Zelus dispar Fabricius, 1803 syn. nov. (=Zelus pedestris Fabricius, 1803), Zelus formosus Haviland, 1931 syn. nov. (=Zelus laticornis Herrich-Schaeffer, 1853), Zelus obscuridorsis (Stål, 1860) syn. nov. (=Zelus pedestris), Zelus pallidinervus Haviland, 1931 syn. nov. (=Zelus kartabensis Haviland, 1931), Zelus personatus Berg, 1879 syn. nov. (=Zelus versicolor Herrich-Schaeffer, 1848), Zelus trimaculatus Champion, 1898 syn. nov. (=Zelus means Fabricius, 1803), Zelus trimaculicollis (Stål, 1855) syn. nov. (=Zelus means), and Zelus tristis Haviland, 1931 syn. nov. (=Zelus laticornis). Zelus conjungens (Stål, 1860) stat. rev. Is resurrected from junior synonymy with zealous armillatus (Lepeletier & Seville, 1825). Zelus ambulans Stål, 1862 stat. rev. and Zelus

  19. A taxonomic monograph of the assassin bug genus Zelus Fabricius (Hemiptera: Reduviidae): 71 species based on 10,000 specimens

    Science.gov (United States)

    Hart, Elwood R; Weirauch, Christiane

    2016-01-01

    Abstract The New World assassin bug genus Zelus Fabricius, 1803 (Insecta: Hemiptera: Heteroptera: Reduviidae: Harpactorinae: Harpactorini) is revised based on more than 10,000 specimens. Seventy-one species are recognized and twenty-four described as new: Zelus aithaleos sp. n., Zelus amblycephalus sp. n., Zelus antiguensis sp. n., Zelus auralanus sp. n., Zelus bahiaensis sp. n., Zelus banksi sp. n., Zelus casii sp. n., Zelus championi sp. n., Zelus cordazulus sp. n., Zelus fuliginatus sp. n., Zelus gilboventris sp. n., Zelus gracilipes sp. n., Zelus grandoculus sp. n., Zelus kartaboides sp. n., Zelus lewisi sp. n., Zelus panamensis sp. n., Zelus paracephalus sp. n., Zelus rosulentus sp. n., Zelus russulumus sp. n., Zelus spatulosus sp. n., Zelus truxali sp. n., Zelus umbraculoides sp. n., Zelus umbraculus sp. n., and Zelus xouthos sp. n. Five species, Zelus araneiformis Haviland, 1931, Zelus gradarius Bergroth, 1905, Zelus modestus (Stål, 1862), Zelus subfasciatus Stål, 1860 and Zelus vittaticeps Stål, 1866, are removed from Zelus and placed incertae sedis within Harpactorini. Nine new synonyms are recognized (senior synonym in parentheses): Zelus atripes Champion, 1898 syn. nov. (=Zelus conjungens [Stål, 1860]), Zelus dispar Fabricius, 1803 syn. nov. (=Zelus pedestris Fabricius, 1803), Zelus formosus Haviland, 1931 syn. nov. (=Zelus laticornis Herrich-Schaeffer, 1853), Zelus obscuridorsis (Stål, 1860) syn. nov. (=Zelus pedestris), Zelus pallidinervus Haviland, 1931 syn. nov. (=Zelus kartabensis Haviland, 1931), Zelus personatus Berg, 1879 syn. nov. (=Zelus versicolor Herrich-Schaeffer, 1848), Zelus trimaculatus Champion, 1898 syn. nov. (=Zelus means Fabricius, 1803), Zelus trimaculicollis (Stål, 1855) syn. nov. (=Zelus means), and Zelus tristis Haviland, 1931 syn. nov. (=Zelus laticornis). Zelus conjungens (Stål, 1860) stat. rev. Is resurrected from junior synonymy with zealous armillatus (Lepeletier & Seville, 1825). Zelus ambulans Stål, 1862 stat. rev

  20. Embryology of Triatoma infestans (KLUG, (Hemiptera, Reduviidae, a Chagas' disease vector Embriología del Triatoma infestans, vector de la enfermedad de Chagas

    Directory of Open Access Journals (Sweden)

    Laura E. Fichera

    1992-06-01

    Full Text Available This study reports the embryogenesis of T. infestans (Hemiptera, Reduviidae. Morphological parameters of growth sequences from oviposition until hatching (12-14 d 28ºC were established. Five periods, as percent of time of development (TD, were characterized from oviposition until hatching. The most important morphological features were: 1 formation of blastoderm within 7% of TD; 2 germ band and gastrulation within 30% of TD; 3 nerve cord, limb budding, thoracic and abdominal segmentation and formation of body cavity within 50% of TD; 4 nervous system and blastokinesis end, and development of embryonic cuticle within 65% of TD; 5 differentiation of the mouth parts, fat body, and Malphigian tubules during final stage and completion of embryo at day 12 to day 14 around hatching. These signals were chosen as appropriate morphological parameters which should enable the evaluation of embryologic modifications due to the action/s of different insecticidesEn este estudio se caracterizó el desarrollo embrionario del T. infestans (Hemiptera Reduviidae. Se establecieron parámetros morfológicos secuenciales de crecimiento desde la oviposición hasta la eclosión, (12-14 d 28ºC. Cinco períodos de crecimiento fueron determinados y expresados como fracciones porcentuales del tiempo total del desarrollo (TD hasta la eclosión. Los eventos morfológicos mas importantes fueron: 1 La formación del blastodermo hasta el 7% del TD; 2 La aparición de la banda germinativa y su gastrulación transcurridos un 30% del TD; 3 El comienzo de la formación del cordón nervioso, de las extremidades, la segmentación torácica y abdominal y la formación del mixocele cuando la embriogénesis alcanzó el 50% del TD; 4 La terminación de la blastoquinesis, el desarrollo completo del sistema nervioso y la aparición de la cutícula embrionaria hasta el 65% del TD; 5 En la etapa final de la embriogenesis se observó la diferenciación del aparato mandibular, el cuerpo graso

  1. Effects of juvenile hormone analogue on ecdysis prevention induced by precocene in Rhodnius prolixus (Hemiptera: Reduviidae

    Directory of Open Access Journals (Sweden)

    Patricia de Azambuja

    1984-12-01

    Full Text Available Precocene II, added to the meal of fourth-instar larvae of Rhodnius prolixus (25 mug/ml of blood, induced an in crease in the duration of the molting cycle. This effect was related to the decrease of both the nuclear area of the prothoracic gland cells and the mitotic activity in epidermal cellS. juvenile hormone analogue applied topically (60 mug/insect together with Precocene II treatment avoided atrophy of the prothoracic glands and induced a higher number of epidermal mitosis accelerating the time of subsequent ecdysis. A possible relationship between juvenile hormone and production of ecdysone is discussed.Adicionado ao sangue alimentar na dose de 25 mug/ml o precoceno II causou um aumento no período de intermuda em ninfas de 4o. estadio de Rhodnius prolixus. Este atraso da muda foi relacionado com a diminuição da área dos núcleos das celulas das glandulas protoracicas e com a queda da atividade mitotica das células da epiderme do inseto. Um análogo de hormônio juvenil aplicado topicamente (60 mug/inseto junto com o tratamento oral com precoceno II preveniu a atrofia das glândulas protorácicas e induziu um aumento no número de mitoses nas células da epiderme, diminuindo o período de intermuda nestes insetos.A possivel relação entre a ação do hormônio juvenil e a producao de ecdisona pelas glândulas protorácicas e discutida.

  2. [Morphological and histological study of eggs of six species of the Triatoma genus (Hemiptera: Reduviidae)].

    Science.gov (United States)

    Obara, Marcos T; da Rosa, João A; da Silva, Nilza N; Ceretti, Walter; Urbinatti, Paulo R; Barata, José M S; Jurberg, José; Galvão, Cleber

    2007-01-01

    The objective of this study was to analyze the morphological and histological characteristics of eggs from six triatomine species in order to obtain a generic and specific characterization of this group of Trypanosoma cruzi vectors and better understand their phylogenetic and taxonomic aspects. The eggs of the respective species came from the collection of the Laboratório de Triatomíneos e Culicídeos da Faculdade de Saúde Pública/USP, from the Insetário do Serviço Especial de Saúde de Araraquara - SESA and from the Triatomine insectary of the Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos (FIOCRUZ), in Rio de Janeiro. Morphological studies were done with optical microscopy and scanning electron microscopy. Histological analyses used only optical microscopy. All of the species analyzed showed a predominance of hexagonal cells. The exchorion of Triatoma breyeri Del Ponte, T. costalimai Verano Galvão, and T. tibiamaculata (Pinto) consists of unornamented cells, overlapping or slightly separated, with a smooth, padded appearance. Meanwhile, in T. matogrossensis Leite Barbosa, T. sherlocki Papa, Juberg, Carcavallo, Cerqueira Barata, and T. williami Galvão, Souza Lima, the exchorion cells are ornamented on their entire surface with perforations and fissures. Egg histology showed undulations, pores, and orifices. The study helps to expand the generic and specific knowledge of the Triatominae subfamily. The characteristics identified in the exchorion of the eggshells can help separate close species. These new parameters will back the elaboration of future dichotomous keys, help to determine each vector species role and expand knowledge of the various species.

  3. Expression of a cucumber class III chitinase and Nicotiana plumbaginifolia class I glucanase genes in transgenic potato plants

    NARCIS (Netherlands)

    Moravcikova, J.; Matusikova, I.; Libantova, J.; Bauer, M.; Mlynarova, L.

    2004-01-01

    The genes encoding for a cucumber class III chitinase and Nicotiana plumbaginifolia class I glucanase were co-introduced into Slovak potato (Solanum tuberosum L.) breeding line 116/86 using Agrobacterium tumefaciens. For both transgenes the number of integrated copies and level of RNA expression wer

  4. Anti-sense expression of putrescine N-methyltransferase confirms defensive role of nicotine in Nicotiana sylvestris against Manduca sexta

    NARCIS (Netherlands)

    Voelckel, C.; Krugel, T.; Gase, K.; Heidrich, N.; Van Dam, N.M.; Winz, R.; Baldwin, I.T.

    2001-01-01

    Several lines of evidence support the defensive function of nicotine production in the Nicotiana genus against a range of herbivores, but the evidence is largely correlative. To suppress nicotine production in planta and to test its defensive function, we expressed DNA of putrescine N-methyl

  5. Digestive duet: Midgut digestive proteinases of Manduca sexta ingesting Nicotiana attenuata with manipulated trypsin proteinase inhibitor expression

    NARCIS (Netherlands)

    Zavala, J.A.; Giri, A.P.; Jongsma, M.A.; Baldwin, I.T.

    2008-01-01

    The defensive effect of endogenous trypsin proteinase inhibitors (NaTPIs) on the herbivore Manduca sexta was demonstrated by genetically altering NaTPI production in M. sexta's host plant, Nicotiana attenuata. To understand how this defense works, we studied the effects of NaTPI on M. sexta gut prot

  6. Anti-sense expression of putrescine N-methyltransferase confirms defensive role of nicotine in Nicotiana sylvestris against Manduca sexta

    NARCIS (Netherlands)

    Voelckel, C.; Krugel, T.; Gase, K.; Heidrich, N.; Van Dam, N.M.; Winz, R.; Baldwin, I.T.

    2001-01-01

    Several lines of evidence support the defensive function of nicotine production in the Nicotiana genus against a range of herbivores, but the evidence is largely correlative. To suppress nicotine production in planta and to test its defensive function, we expressed DNA of putrescine N-methyl transfe

  7. Changes in phenolic compound, TAL, PAL activity of Nicotiana rustica triggered by ethanolamine pretreatment under in vitro salt stress condition

    National Research Council Canada - National Science Library

    Ali Akbar Ehsanpour; Mohmmad Amin Toghyani; Somayeh Rajaeian

    2015-01-01

    .... For this purpose, 4-week-old Nicotiana rustica plants which had been kept in MS medium were pretreated with ethanolamine and 2 days later they were transferred to MS medium supplemented with 200 mM NaCl for 3 weeks...

  8. Expression of a cucumber class III chitinase and Nicotiana plumbaginifolia class I glucanase genes in transgenic potato plants

    NARCIS (Netherlands)

    Moravcikova, J.; Matusikova, I.; Libantova, J.; Bauer, M.; Mlynarova, L.

    2004-01-01

    The genes encoding for a cucumber class III chitinase and Nicotiana plumbaginifolia class I glucanase were co-introduced into Slovak potato (Solanum tuberosum L.) breeding line 116/86 using Agrobacterium tumefaciens. For both transgenes the number of integrated copies and level of RNA expression

  9. Spontaneous and induced loss of chromosomes in slow-growing somatic hybrid calli of Solanum tuberosum and Nicotiana plumbaginifolia

    NARCIS (Netherlands)

    Tempelaar, MJ; Drenth - Diephuis, L.J.; SAAT, TAWM; Jacobsen, E.

    1991-01-01

    Rate and extent of spontaneous and induced chromosome loss have been determined at the callus level of somatic hybrids of mutants of Solanum tuberosum and Nicotiana plumbaginifolia. AEC (amino ethyl cystein) resistance in potato and Nitrate-Reductase deficiency in N. plumbaginifolia have been used a

  10. [A hydroponic cultivation system for rapid high-yield transient protein expression in Nicotiana plants under laboratory conditions].

    Science.gov (United States)

    Mo, Qianzhen; Mai, Rongjia; Yang, Zhixiao; Chen, Minfang; Yang, Tiezhao; Lai, Huafang; Yang, Peiliang; Chen, Qiang; Zhou, Xiaohong

    2012-06-01

    To develop a hydroponic Nicotiana cultivation system for rapid and high-yield transient expression of recombinant proteins under laboratory conditions. To establish the hydroponic cultivation system, several parameters were examined to define the optimal conditions for the expression of recombinant proteins in plants. We used the green fluorescent protein (GFP) and the geminiviral plant transient expression vector as the model protein/expression vector. We examined the impact of Nicotiana species, the density and time of Agrobacterium infiltration, and the post-infiltration growth period on the accumulation of GFP. The expression levels of GFP in Nicotiana leaves were then examined by Western blotting and ELISA. Our data indicated that a hydroponic Nicotiana cultivation system with a light intensity of 9000 LX/layer, a light cycle of 16 h day/8 h night, a temperature regime of 28 degrees celsius; day/21 degrees celsius; night, and a relative humidity of 80% could support the optimal plant growth and protein expression. After agroinfiltration with pBYGFPDsRed.R/LBA4404, high levels of GFP expression were observed in both N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants cultured with this hydroponic cultivation system. An optimal GFP expression was achieved in both Nicotiana species leaves 4 days after infiltration by Agrobacterium with an OD(600) of 0.8. At a given time point, the average biomass of N. tobaccum (cv. Yuyan No.5) was significantly higher than that of N. benthamiana. The leaves from 6-week-old N. benthamiana plants and 5-week-old N. tobaccum (cv. Yuyan No.5) plants could be the optimal material for agroinfiltration. We have established a hydroponic cultivation system that allows robust growth of N. benthamiana and N. tobaccum (cv. Yuyan No.5) plants and the optimal GFP expression in the artificial climate box.

  11. In vitro cytotoxicity of Nicotiana gossei leaves, used in the Australian Aboriginal smokeless tobacco known as pituri or mingkulpa.

    Science.gov (United States)

    Moghbel, Nahid; Ryu, BoMi; Cabot, Peter J; Steadman, Kathryn J

    2016-07-08

    The Aboriginal population of Central Australia use endemic Nicotiana species to make a smokeless tobacco product known usually as pituri or mingkulpa. Nicotiana leaves are masticated with wood ash into a 'quid' that is chewed/sucked for absorption of nicotine. In addition to nicotine, smokeless tobacco products contain a spectrum of biologically active compounds that may contribute to effects on health. The objective of this study was to quantify nicotine, and related alkaloids and tobacco specific nitrosamines (TSNAs), in Nicotiana leaves used in pituri, and compare in vitro toxicity of pure nicotine with Nicotiana leaf extract at the same concentration of nicotine. An aqueous extract of dry leaves of Nicotiana gossei and a reference smokeless tobacco (CORESTA CRP2) were quantified for major pyridine alkaloids and TSNAs using HPLC-UV and LC-MS/MS. A range of extract concentrations and corresponding concentrations of nicotine standard were tested using an MTS assay to measure human lung epithelium cell (A549) survival. Cells treated for 24h with the maximum concentration of 1.5mg/ml of nicotine resulted in 77% viability. In contrast, extracts from N. gossei leaves and CRP2 containing a similar concentration of nicotine (1.3mg/ml) resulted in remarkably lower viability of 1.5 and 6%, respectively. Comparison of cytotoxicity of pure nicotine with that of the extracts revealed that nicotine was not the source of their cytotoxicity. Other biologically active compounds such as the known carcinogens NNK and NNN, derived from nicotine and nornicotine and found to be present in the smokeless tobacco extracts, may be responsible.

  12. Biology, behaviour and functional response of Cydnocoris gilvus Brum. (Heteroptera: Reduviidae: Harpactorinae a predator of Tea Mosquito Bug (Helopeltis antonii Sign. on cashew in India

    Directory of Open Access Journals (Sweden)

    K.K. Srikumar

    2014-06-01

    Full Text Available Helopeltis spp. (Hemiptera: Miridae are major sucking pests of cashew (Anacardium occidentale L. in India. Cydnocoris gilvus Brum. (Heteroptera: Reduviidae: Harpactorinae is recorded as a potential predator of Helopeltis spp. Biology, mating behaviour and functional response of C. gilvus were studied by rearing in the laboratory (temperature 26-28 0C; relative humidity 89-94 % with wax moth, Galleria mellonella, larvae. Based on laboratory rearing, the fecundity was 56.33 eggs in 8.67 batches per female. The average stadial period was 37.3 days, with a maximum of 11 days for V instar and a minimum of 4.5 days for III instars. C. gilvus took 45.5 days to complete a generation. The innate capacity of natural increase was 0.07 with a gross reproduction of 67.8 females per female. The adult exhibited a pin and jab mode of predation in a sequence of actions. The sequential action of mating comprised arousal (1.32 min, approach (12.30 min, riding over (140.48 min and copulation (85.40 min. The predator responded to increasing prey density by killing more prey than at lower prey densities

  13. Comparative study of enzymes in testes and ovaries from adult Dipetalogaster maximus (Uhler and Triatoma infestans (Klug (Hemiptera: Reduviidae. Correlation with fine structural organization

    Directory of Open Access Journals (Sweden)

    PY Scaraffia

    2009-12-01

    Full Text Available Activities of hexokinase (HK, glucose-6-phosphate dehydrogenase (G6PDH, fructose-6-phosphate kinase (F6PK, glutamate dehydrogenase (GlutDH, aspartate aminotransferase (AAT, malate dehydrogenase (MDH and glycerol-3-phosphate dehydrogenase (GPDH were determined in tissue extracts of testes and ovaries of adult Dipetalogaster maximus (Uhler and Triatoma infestans (Klug (Hemiptera: Reduviidae, insect vectors of Chagas disease. The fine structure organization of the same organs were studied by electron microscopy. Results allow the following inferences: in testes from both species, most of the glucose would be utilized through the glycolytic pathway. Amino acid catabolism for energy purposes appears to be unimportant. The number of mitochondria and the development of the rough endoplasmic reticulum in cells of the spermatogenic line indicate the occurrence of active oxidative metabolism and protein synthesis; in ovaries, levels of G6PDH indicate the existence of an active pentose pathway which would supply the NADPH required for fat and ecdysteroid synthesis. Amino acid catabolism appears to be relatively more important in ovary than in testis. Fat and glycogen are stored in follicular cells of D. maximus; oocytes of both species contain numerous fat droplets. Abundant mitocondria are present in follicular cells and oocytes. A well developed rough endoplasmic reticulum and free ribosomes are also conspicuous in these cells. The malate/aspartate H-transfer system seemed to be relatively more important than the glycerophosphate shuttle in ovaries as well in testes.

  14. Genetic Relationships and Spatial Genetic Structure Among Populations of Rhodnius prolixus (Hemiptera: Reduviidae) in Colombia and Venezuela Based on Mitochondrial Cytochrome-b Sequences.

    Science.gov (United States)

    Luna-Marín, K P; Angulo-Silva, V M; Hernández-Torres, J; Ruiz-García, M

    2016-11-26

    One hundred twenty Rhodnius prolixus (Stal) (Hemiptera: Reduviidae) specimens from 6 Colombian Departments and 1 Venezuelan State had 594-bp of the mitochondrial cytochrome-b gene sequenced to improve the understanding of evolutionary processes that shape the main vector of Chagas disease. The levels of genetic diversity for this species were low-medium with reference to other bugs. The genetic heterogeneity among the populations was very limited which means there has been extensive gene flow and/or very recent split processes. The overall sample as well as some individual populations showed evidence of recent population expansions (with the exception of Arauca, which yielded evidence of a bottleneck for a mismatch distribution). This expansion (11,000 or 2000-25,000 year ago depending of two procedures employed) coincides with the ending of the last intense glacial conditions during the Pleistocene and the beginning of the Holocene that had a warmer and wetter climate. Some of our autocorrelation analyses (AIDA and Genetic Landscape Interpolation Analysis) indicated local patches of high genetic similarity but no globally significant spatial structure. We did show an original haplotype distributed throughout the entirety of the geographical area studied.

  15. Levantamento entomofaunístico de artrópodes em algodoeiro de fibra naturalmente colorida em Ipanguaçu-RN

    Directory of Open Access Journals (Sweden)

    Bárbara Karine de Albuquerque Silva

    2016-08-01

    Full Text Available Objetivou-se com esta pesquisa identificar a diversidade de artrópodes associados à cultura do algodão Gossypium hirsutum L. com pluma colorida, sendo realizados levantamentos entomofaunístico em Ipanguaçu-RN em áreas de produção. A área experimental foi composta por 15 variedades de algodão com pluma colorida: CNPA 2009-6; CNPA 2009-11; CNPA 2009-13; CNPA 2009-16; CNPA 2009-27; CNPA 2009-42; CNPA 2009-47; CNPA 2009-48; CNPA 2009-50; CNPA 2009-59; CNPA 2009-60; CNPA 2009-62; BRS RUBI; BRS AROEIRA; BRS TOPÁZIO. O levantamento foi realizado tendo como base três métodos de coleta ativa em pontos aleatórios da área experimental. As coletas consistiram da retirada manual de folhas e maçãs diretamente da planta, além da captura de insetos em pleno voo, com auxílio de rede entomológica. Foram encontrados um total 1884 insetos adultos e 66 larvas, dispostos em 8 ordens e 22 famílias. A família Aphididae: Hemiptera foi a mais numerosa entre o material coletado, com 1720 adultos dispersos nos três métodos de coleta aplicados. Além desta, outras famílias de pragas da cultura foram encontradas, como Curculionidae: Coleoptera. Também foram coletados organismos benéficos como os polinizadores Aphidae e Megachilidae, pertencentes a ordem Hymenoptera, predadores (Coccinellidae: Coleoptera; Vespidae: Hymenoptera; Reduviidae: Hemiptera; Chrysopidae: Neuroptera e Labiidae: Dermaptera e parasitoides, como os microhimenópteros. Foram encontrados três tipos de larvas, sendo classificadas como curculioniforme as mais numerosas, apresentando um total de 57 espécimes coletados.Entomofaunistic survey of artropods in naturally colored cotton fiber in Ipanguaçu-RNAbstract: The objective of this research was to identify the diversity of arthropods associated with cotton crop Gossypium hirsutum L. with colorful plume, it was conducted entomofaunístico survey in Ipanguaçu-RN in production areas. The experimental area was composed of 15 cotton

  16. Optimization of engineered production of the glucoraphanin precursor dihomomethionine in Nicotiana benthamiana

    DEFF Research Database (Denmark)

    Crocoll, Christoph; Mirza, Nadia Muhammad Akram; Reichelt, Michael;

    2016-01-01

    Glucosinolates are natural products characteristic of the Brassicales order, which include vegetables such as cabbages and the model plant Arabidopsis thaliana. Glucoraphanin is the major glucosinolate in broccoli and associated with the health-promoting effects of broccoli consumption. Toward our...... goal of creating a rich source of glucoraphanin for dietary supplements, we have previously reported the feasibility of engineering glucoraphanin in Nicotiana benthamiana through transient expression of glucoraphanin biosynthetic genes from A. thaliana (Mikkelsen et al., 2010). As side-products, we...... to a ninefold increase compared to the highest production of this intermediate, as previously reported (Mikkelsen et al., 2010). The increased DHM production without increasing leucine-derived side-product levels provides new metabolic engineering strategies for improved glucoraphanin production...

  17. ‘COROJO 2006’: NUEVA VARIEDAD CUBANA DE TABACO NEGRO (Nicotiana tabacum L.

    Directory of Open Access Journals (Sweden)

    Vivaldo García Morejón

    2013-01-01

    Full Text Available En la Estación Experimental del Tabaco, de San Juan y Martínez, Pinar del Río durante la campaña tabacalera 2001-2002, se realizó un cruzamiento sexual con el objetivo de obtener una variedad de tabaco negro con mayor rendimiento que la comercial ‘Criollo 98’, resistente al moho azul ( Peronospora hyocyami de Bary, a la pata prieta ( Phytophthora parasitica Dast. var . Nicotianae Breda de Haan, al virus del mosaico del tabaco (VMT y a la necrosis ambiental. Después de cinco generaciones de autofecundación y selección por el método genealógico se obtuvieron siete variedades resistentes. El cruce originó la nueva variedad `Corojo 2006 ́ que superó a la `Criollo 98 ́.

  18. Molecular characterization and expression pattern of tobacco (Nicotiana tabacum ascorbate oxidase gene

    Directory of Open Access Journals (Sweden)

    Zhihua Li

    2016-11-01

    Full Text Available The complete coding sequence of one tobacco (Nicotiana tabacum gene, ascorbate oxidase (AO, was isolated by the application of reverse transcription-polymerase chain reaction. The tobacco AO gene consists of a 1722-bp open reading frame and encodes a protein of 573 amino acids. Sequence comparison analysis revealed that the tobacco AO protein shares high homology with the AO proteins of Lycopersicon esculentum (89%, Populus trichocarpa (75%, soybean (74%, castor bean (73% and peach (73%. The prediction of transmembrane helices showed that tobacco AO might be a transmembrane protein. The expression profile was studied and the results indicated that the tobacco AO gene was diversely expressed in different tobacco tissues, including leaves, stem, roots and flowers. Our experiment laid the grounds for further research on this tobacco gene.

  19. Partial Characterization of a Vicilin-Like Glycoprotein from Seeds of Flowering Tobacco (Nicotiana sylvestris

    Directory of Open Access Journals (Sweden)

    Jared Q. Gerlach

    2009-01-01

    Full Text Available A vicilin-like glycoprotein from the seeds of Nicotiana sylvestris, flowering tobacco, has been identified using nanoLC/ESI-MS/MS. Sequences from a fragment of protein demonstrated homology with vicilins from other members of the Solanaceae family, notably potato (Solanum demissum. Reducing and nonreducing SDS-PAGE analyses of the identified protein indicated that fragments resulting from in situ proteolytic processing are joined by intrachain disulphide bonds. Staining with Con A lectin was specifically inhibited by mannose suggested the presence of -linked glycosylation which was confirmed by carbohydrate compositional analysis of PVDF-bound protein subunits. HPAEC-PAD analysis of the monosaccharides released from the glycoprotein by acid hydrolysis revealed glucosamine and mannose. -acetylglucosamine termination of attached oligosaccharides was further verified by inhibitable WGA lectin staining. Immunostaining of PVDF-bound N. sylvestris proteins with antibodies against G. max total protein demonstrated cross-staining at masses corresponding to fragments from the proteolytically processed protein subunits.

  20. Protein-Protein Interaction Assays with Effector-GFP Fusions in Nicotiana benthamiana.

    Science.gov (United States)

    Petre, Benjamin; Win, Joe; Menke, Frank L H; Kamoun, Sophien

    2017-01-01

    Plant parasites secrete proteins known as effectors into host tissues to manipulate host cell structures and functions. One of the major goals in effector biology is to determine the host cell compartments and the protein complexes in which effectors accumulate. Here, we describe a five-step pipeline that we routinely use in our lab to achieve this goal, which consists of (1) Golden Gate assembly of pathogen effector-green fluorescent protein (GFP) fusions into binary vectors, (2) Agrobacterium-mediated heterologous protein expression in Nicotiana benthamiana leaf cells, (3) laser-scanning confocal microscopy assay, (4) anti-GFP coimmunoprecipitation-liquid chromatography-tandem mass spectrometry (coIP/MS) assay, and (5) anti-GFP western blotting. This pipeline is suitable for rapid, cost-effective, and medium-throughput screening of pathogen effectors in planta.

  1. Production of Recombinant Cholera Toxin B Subunit in Nicotiana benthamiana Using GENEWARE® Tobacco Mosaic Virus Vector.

    Science.gov (United States)

    Moore, Lauren; Hamorsky, Krystal; Matoba, Nobuyuki

    2016-01-01

    Here, we describe a method to produce a recombinant cholera toxin B subunit in Nicotiana benthamiana plants (CTBp) using the GENEWARE(®) tobacco mosaic virus vector system. Infectious transcripts of the vector RNA are generated in vitro and inoculated on N. benthamiana seedlings. After 11 days, CTBp is extracted in a simple tris buffer at room temperature. No protease inhibitor is required. The leaf homogenate is treated with mild heat and a pH shift to selectively precipitate host-derived proteins. CTBp is purified to >95 % homogeneity by two-step chromatography using immobilized metal affinity and ceramic hydroxyapatite resins. This procedure yields on average 400 mg of low-endotoxin CTBp from 1 kg of fresh leaf material.

  2. Wax esters of different compositions produced via engineering of leaf chloroplast metabolism in Nicotiana benthamiana.

    Science.gov (United States)

    Aslan, Selcuk; Sun, Chuanxin; Leonova, Svetlana; Dutta, Paresh; Dörmann, Peter; Domergue, Frédéric; Stymne, Sten; Hofvander, Per

    2014-09-01

    In a future bio-based economy, renewable sources for lipid compounds at attractive cost are needed for applications where today petrochemical derivatives are dominating. Wax esters and fatty alcohols provide diverse industrial uses, such as in lubricant and surfactant production. In this study, chloroplast metabolism was engineered to divert intermediates from de novo fatty acid biosynthesis to wax ester synthesis. To accomplish this, chloroplast targeted fatty acyl reductases (FAR) and wax ester synthases (WS) were transiently expressed in Nicotiana benthamiana leaves. Wax esters of different qualities and quantities were produced providing insights to the properties and interaction of the individual enzymes used. In particular, a phytyl ester synthase was found to be a premium candidate for medium chain wax ester synthesis. Catalytic activities of FAR and WS were also expressed as a fusion protein and determined functionally equivalent to the expression of individual enzymes for wax ester synthesis in chloroplasts.

  3. Polypeptide synthesis induced in Nicotiana clevelandii protoplasts by infection with raspberry ringspot nepovirus.

    Science.gov (United States)

    Acosta, O; Mayo, M A

    1993-01-01

    Infection of Nicotiana clevelandii protoplasts by raspberry ringspot nepovirus resulted in the accumulation of about 24 polypeptides that differed in M(r) and pI from polypeptides accumulating in mock-inoculated protoplasts. Similar polypeptides accumulated in protoplasts infected with the S and E strains of RRV but different infection-specific polypeptides were detected in protoplasts infected with tobacco ringspot nepovirus. The M(r) of RRV-specific polypeptides ranged from 210,000 to 18,000 and most are presumed to be derived from others by proteolytic cleavage. No evidence was found for marked changes in polypeptide abundance with time after inoculation or for any virus-specific polypeptide becoming disproportionately abundant in the medium during culture.

  4. Assessment of {sup 210}Pb concentration in Nicotiana tabacum L., burley variety, cultivated in Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Rocha, Rique J.F.X.; Silva, Carolina F.; Frujuele, Jonatan V.; Bovolini, Raquel R.; Damatto, Sandra R., E-mail: rjrocha@ipen.br, E-mail: cfsilva@ipen.br, E-mail: jonatanfrujuele@hotmail.com, E-mail: ra_bovolini@yahoo.com.br, E-mail: damatto@ipen.br [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil). Lab. de Radiometria Ambiental

    2013-07-01

    Tobacco products are extensively used throughout the world and the most consumed are cigarettes, cigars and narghile. The damaging effects that these products cause to human health are discussed worldwide and many researches are performed with the aim of relating the use of these products with various diseases. Brazil is the largest exporter and second largest producer of tobacco worldwide, according to the crop year 2009/2010 production. The tobacco plant (Nicotiana tabacum L.) is used to manufacture all derivatives and the chemical composition of the resulting tobacco varies with the type of tobacco leaves, how they are grown, the region where they are cultivated, the characteristics of preparation and the temperature variations resulting from the tobacco incomplete combustion. There is lack of information about the chemical and radiological characterization of the tobacco plant both in international and Brazilian literature. Thus a project was established with the objectives of characterizing chemically and radiologically the three varieties most cultivated in Brazil of Nicotiana tobacum L., Virginia, Burley and Common; this paper presents the preliminary results of {sup 210}Pb concentrations for the Burley variety. Plants from this variety were cultivated in pots with organic substrate and fertilizer and in a small farm in natural conditions. The entire plant was analyzed, the organic substrates, the fertilizers and the soil. The results obtained presented higher values for {sup 210}Pb in leaves when compared with the other parts of the plant. Comparing the three study areas the highest results of {sup 210}Pb concentration were obtained in the plants cultivated in the urban area probably due to its atmospheric deposition. (author)

  5. Expression, Purification, and Biophysical Characterization of a Secreted Anthrax Decoy Fusion Protein in Nicotiana benthamiana

    Science.gov (United States)

    Karuppanan, Kalimuthu; Duhra-Gill, Sifti; Kailemia, Muchena J.; Phu, My L.; Lebrilla, Carlito B.; Dandekar, Abhaya M.; Rodriguez, Raymond L.; Nandi, Somen; McDonald, Karen A.

    2017-01-01

    Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in Nicotiana benthamiana plants using a transient expression system. Using the Cauliflower Mosaic Virus (CaMV) 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation. Production kinetics were observed up to eight days post-infiltration, and maximum production of 826 mg/kg fresh leaf weight was observed on day six. Protein A affinity chromatography purification of the rCMG2-Fc-Apo protein from whole leaf extract and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis and mass spectrometry analysis confirmed the molecular integrity of the secreted protein. The N-glycosylation pattern of purified rCMG2-Fc-Apo protein was analysed; the major portion of N-glycans consists of complex type structures in both protein samples. The most abundant (>50%) N-glycan structure was GlcNAc2(Xyl)Man3(Fuc)GlcNAc2 in rCMG2-Fc-Apo recovered from whole leaf extract and apoplast wash fluid. High mannose N-glycan structures were not detected in the apoplast wash fluid preparation, which confirmed the protein secretion. Altogether, these findings demonstrate that high-level production of rCMG2-Fc-Apo can be achieved by transient production in Nicotiana benthamiana plants with apoplast targeting. PMID:28054967

  6. Expression, Purification, and Biophysical Characterization of a Secreted Anthrax Decoy Fusion Protein in Nicotiana benthamiana.

    Science.gov (United States)

    Karuppanan, Kalimuthu; Duhra-Gill, Sifti; Kailemia, Muchena J; Phu, My L; Lebrilla, Carlito B; Dandekar, Abhaya M; Rodriguez, Raymond L; Nandi, Somen; McDonald, Karen A

    2017-01-04

    Anthrax toxin receptor-mediated drug development for blocking anthrax toxin action has received much attention in recent decades. In this study, we produced a secreted anthrax decoy fusion protein comprised of a portion of the human capillary morphogenesis gene-2 (CMG2) protein fused via a linker to the fragment crystallizable (Fc) domain of human immunoglobulin G1 in Nicotiana benthamiana plants using a transient expression system. Using the Cauliflower Mosaic Virus (CaMV) 35S promoter and co-expression with the p19 gene silencing suppressor, we were able to achieve a high level of recombinant CMG2-Fc-Apo (rCMG2-Fc-Apo) protein accumulation. Production kinetics were observed up to eight days post-infiltration, and maximum production of 826 mg/kg fresh leaf weight was observed on day six. Protein A affinity chromatography purification of the rCMG2-Fc-Apo protein from whole leaf extract and apoplast wash fluid showed the homodimeric form under non-reducing gel electrophoresis and mass spectrometry analysis confirmed the molecular integrity of the secreted protein. The N-glycosylation pattern of purified rCMG2-Fc-Apo protein was analysed; the major portion of N-glycans consists of complex type structures in both protein samples. The most abundant (>50%) N-glycan structure was GlcNAc₂(Xyl)Man₃(Fuc)GlcNAc₂ in rCMG2-Fc-Apo recovered from whole leaf extract and apoplast wash fluid. High mannose N-glycan structures were not detected in the apoplast wash fluid preparation, which confirmed the protein secretion. Altogether, these findings demonstrate that high-level production of rCMG2-Fc-Apo can be achieved by transient production in Nicotiana benthamiana plants with apoplast targeting.

  7. Membrane transporters in self resistance of Cercospora nicotianae to the photoactivated toxin cercosporin.

    Science.gov (United States)

    Beseli, Aydin; Amnuaykanjanasin, Alongkorn; Herrero, Sonia; Thomas, Elizabeth; Daub, Margaret E

    2015-11-01

    The goal of this work is to characterize membrane transporter genes in Cercospora fungi required for autoresistance to the photoactivated, active-oxygen-generating toxin cercosporin they produce for infection of host plants. Previous studies implicated a role for diverse membrane transporters in cercosporin resistance. In this study, transporters identified in a subtractive cDNA library between a Cercospora nicotianae wild type and a cercosporin-sensitive mutant were characterized, including two ABC transporters (CnATR2, CnATR3), an MFS transporter (CnMFS2), a uracil transporter, and a zinc transport protein. Phylogenetic analysis showed that only CnATR3 clustered with transporters previously characterized to be involved in cercosporin resistance. Quantitative RT-PCR analysis of gene expression under conditions of cercosporin toxicity, however, showed that only CnATR2 was upregulated, thus this gene was selected for further characterization. Transformation and expression of CnATR2 in the cercosporin-sensitive fungus Neurospora crassa significantly increased cercosporin resistance. Targeted gene disruption of CnATR2 in the wild type C. nicotianae, however, did not decrease resistance. Expression analysis of other transporters in the cnatr2 mutant under conditions of cercosporin toxicity showed significant upregulation of the cercosporin facilitator protein gene (CFP), encoding an MFS transporter previously characterized as playing an important role in cercosporin autoresistance in Cercospora species. We conclude that cercosporin autoresistance in Cercospora is mediated by multiple genes, and that the fungus compensates for mutations by up-regulation of other resistance genes. CnATR2 may be a useful gene, alone or in addition to other known resistance genes, for engineering Cercospora resistance in crop plants.

  8. Plant oxidosqualene metabolism: cycloartenol synthase-dependent sterol biosynthesis in Nicotiana benthamiana.

    Science.gov (United States)

    Gas-Pascual, Elisabet; Berna, Anne; Bach, Thomas J; Schaller, Hubert

    2014-01-01

    The plant sterol pathway exhibits a major biosynthetic difference as compared with that of metazoans. The committed sterol precursor is the pentacyclic cycloartenol (9β,19-cyclolanost-24-en-3β-ol) and not lanosterol (lanosta-8,24-dien-3β-ol), as it was shown in the late sixties. However, plant genome mining over the last years revealed the general presence of lanosterol synthases encoding sequences (LAS1) in the oxidosqualene cyclase repertoire, in addition to cycloartenol synthases (CAS1) and to non-steroidal triterpene synthases that contribute to the metabolic diversity of C30H50O compounds on earth. Furthermore, plant LAS1 proteins have been unambiguously identified by peptidic signatures and by their capacity to complement the yeast lanosterol synthase deficiency. A dual pathway for the synthesis of sterols through lanosterol and cycloartenol was reported in the model Arabidopsis thaliana, though the contribution of a lanosterol pathway to the production of 24-alkyl-Δ(5)-sterols was quite marginal (Ohyama et al. (2009) PNAS 106, 725). To investigate further the physiological relevance of CAS1 and LAS1 genes in plants, we have silenced their expression in Nicotiana benthamiana. We used virus induced gene silencing (VIGS) based on gene specific sequences from a Nicotiana tabacum CAS1 or derived from the solgenomics initiative (http://solgenomics.net/) to challenge the respective roles of CAS1 and LAS1. In this report, we show a CAS1-specific functional sterol pathway in engineered yeast, and a strict dependence on CAS1 of tobacco sterol biosynthesis.

  9. Plant Oxidosqualene Metabolism: Cycloartenol Synthase–Dependent Sterol Biosynthesis in Nicotiana benthamiana

    Science.gov (United States)

    Gas-Pascual, Elisabet; Berna, Anne; Bach, Thomas J.; Schaller, Hubert

    2014-01-01

    The plant sterol pathway exhibits a major biosynthetic difference as compared with that of metazoans. The committed sterol precursor is the pentacyclic cycloartenol (9β,19-cyclolanost-24-en-3β-ol) and not lanosterol (lanosta-8,24-dien-3β-ol), as it was shown in the late sixties. However, plant genome mining over the last years revealed the general presence of lanosterol synthases encoding sequences (LAS1) in the oxidosqualene cyclase repertoire, in addition to cycloartenol synthases (CAS1) and to non-steroidal triterpene synthases that contribute to the metabolic diversity of C30H50O compounds on earth. Furthermore, plant LAS1 proteins have been unambiguously identified by peptidic signatures and by their capacity to complement the yeast lanosterol synthase deficiency. A dual pathway for the synthesis of sterols through lanosterol and cycloartenol was reported in the model Arabidopsis thaliana, though the contribution of a lanosterol pathway to the production of 24-alkyl-Δ5-sterols was quite marginal (Ohyama et al. (2009) PNAS 106, 725). To investigate further the physiological relevance of CAS1 and LAS1 genes in plants, we have silenced their expression in Nicotiana benthamiana. We used virus induced gene silencing (VIGS) based on gene specific sequences from a Nicotiana tabacum CAS1 or derived from the solgenomics initiative (http://solgenomics.net/) to challenge the respective roles of CAS1 and LAS1. In this report, we show a CAS1-specific functional sterol pathway in engineered yeast, and a strict dependence on CAS1 of tobacco sterol biosynthesis. PMID:25343375

  10. High-level diterpene production by transient expression in Nicotiana benthamiana

    Science.gov (United States)

    2013-01-01

    Background Characterization of plant terpene synthases is typically done by production of recombinant enzymes in Escherichia coli. This is often difficult due to solubility and codon usage issues. Furthermore, plant terpene synthases which are targeted to the plastids, such as diterpene synthases, have to be shortened in a more or less empirical approach to improve expression. We report here an optimized Agrobacterium-mediated transient expression assay in Nicotiana benthamiana for plant diterpene synthase expression and product analysis. Results Agrobacterium-mediated transient expression of plant diterpene synthases in N. benthamiana led to the accumulation of diterpenes within 3 days of infiltration and with a maximum at 5 days. Over 50% of the products were exported onto the leaf surface, thus considerably facilitating the analysis by reducing the complexity of the extracts. The robustness of the method was tested by expressing three different plant enzymes, cembratrien-ol synthase from Nicotiana sylvestris, casbene synthase from Ricinus communis and levopimaradiene synthase from Gingko biloba. Furthermore, co-expression of a 1-deoxy-D-xylulose-5-phosphate synthase from tomato and a geranylgeranyl diphosphate synthase from tobacco led to a 3.5-fold increase in the amount of cembratrien-ol produced, with maximum yields reaching 2500 ng/cm2. Conclusion With this optimized method for diterpene synthase expression and product analysis, a single infiltrated leaf of N. benthamiana would be sufficient to produce quantities required for the structure elucidation of unknown diterpenes. The method will also be of general use for gene function discovery, pathway reconstitution and metabolic engineering of diterpenoid biosynthesis in plants. PMID:24330621

  11. A Redox-Mediated Modulation of Stem Bolting in Transgenic Nicotiana sylvestris Differentially Expressing the External Mitochondrial NADPH Dehydrogenase1[W][OA

    National Research Council Canada - National Science Library

    Yun-Jun Liu; Adriano Nunes-Nesi; Sabá V Wallström; Ida Lager; Agnieszka M Michalecka; Fredrik E B Norberg; Susanne Widell; Kenneth M Fredlund; Alisdair R Fernie; Allan G Rasmusson

    2009-01-01

    .... Nicotiana sylvestris plants overexpressing potato (Solanum tuberosum) NDB1 displayed early bolting, whereas sense suppression of the same gene led to delayed bolting, with consequential changes in flowering time...

  12. The influence of the manifestation resistance to pests and diseases varieties of nicotiana rustica on the yield performance of vegetative and generative mass

    National Research Council Canada - National Science Library

    Кароліна Артурівна Шейдик

    2015-01-01

    It is shown the phytosanitary state of Nicotiana rustica in Transcarpathian region and it is disclosed the development of harmful organisms, including requiring a detailed study of tomato spotted wilt...

  13. The influence of the manifestation resistance to pests and diseases varieties of nicotiana rustica on the yield performance of vegetative and generative mass

    Directory of Open Access Journals (Sweden)

    Кароліна Артурівна Шейдик

    2015-04-01

    Full Text Available It is shown the phytosanitary state of Nicotiana rustica in Transcarpathian region and it is disclosed the development of harmful organisms, including requiring a detailed study of tomato spotted wilt virus and tobacco stem in view of selection process on group stability. The result of research presented in this article is to assess the Nicotiana rustica varieties for resistance and seed productivity and identification of resistant varieties for implementation in production due to lack of varieties of domestic origin.

  14. 烟草黑胫病菌对农田草本植物的寄主范围%The host range of Phytophthora nicotianae var. nicotianae in field herbs

    Institute of Scientific and Technical Information of China (English)

    马国胜; 高智谋

    2011-01-01

    烟草黑胫病是烟草生产中的重要病害,对烟草的产量和质量均影响极大[1-2]。烟草黑胫病菌Phytophthora nicotianae var.nicotianae(Breda deHaan)Tucker是引起烟草黑胫病的唯一病原菌,以往的研究均认为,

  15. Simultaneous detection and quantification of Phytophthora nicotianae and P. cactorum, and distribution analyses in strawberry greenhouses by duplex real-time PCR.

    Science.gov (United States)

    Li, Mingzhu; Inada, Minoru; Watanabe, Hideki; Suga, Haruhisa; Kageyama, Koji

    2013-01-01

    Phytophthora nicotianae and P. cactorum cause Phytophthora rot of strawberry. A duplex real-time PCR technique for simultaneous detection and quantification of the two pathogens was developed. Species-specific primers for P. nicotianae and P. cactorum were designed based on the internal transcribed spacer regions (ITS) of rDNA and the ras-related protein gene Ypt1, respectively. TaqMan probes were labeled with FAM for P. nicotianae and HEX for P. cactorum. Specificities were demonstrated using 52 isolates, including various soil-borne pathogens. Sensitivities for P. nicotianae and P. cactorum DNAs were 10 fg and 1 pg, respectively. The technique was applied to naturally infested soil and root samples; the two pathogens were detected and the target DNA concentrations were quantified. Significant correlations of DNA quantities in roots and the surrounding soils were found. The minimum soil DNA concentration predicting the development of disease symptoms was estimated as 20 pg (g soil)(-1). In three strawberry greenhouses examined, the target DNA concentrations ranged from 1 to 1,655 pg (g soil)(-1) for P. nicotianae and from 13 to 233 pg (g soil)(-1) for P. cactorum. The method proved fast and reliable, and provides a useful tool to monitor P. nicotianae and P. cactorum in plants or soils.

  16. Sôbre o órgão abdominal glandular de Arilus carinatus (Forster, 1771: (Heteroptera, Reduviidae

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    Rudolf Barth

    1961-04-01

    Full Text Available Descreve-se um órgão glandular, encontrado na fêmea de Arilus carinatus, de um tipo ainda desocnhecido em insetos. Localiza-se, em forma de um saco membranoso, nos dois lados da linha ventral, entre os 8º e 9º segmentos abdominais. O órgão é expulso por um aumento da pressão interna da cavidade abdominal e volta ao estado de repouso, no interior do corpo, por meio de contração muscular. A vesícula retal volumosa forma, em direção distal, um amplo divertículo, do qual partem dois "tubos retais" que penetram nas vesículas membranosas, tendo na superfície destas uma abertura em forma de fenda. A hipoderme do divertículo, bem como a de uma região da vesícula retal e da parte basal dos tubos retais é glandular. A secreção possui um cheiro intenso e ardido que lembra o gás de acetileno. Trata-se, provàvelmente, de uma glândula repugnatória. Não se sabe nada sôbra a ocorrência do aparelho no macho e em outras espécies de Reduviídeos.

  17. Enzymatic, expression and structural divergences among carboxyl O-methyltransferases after gene duplication and speciation in Nicotiana.

    Science.gov (United States)

    Hippauf, Frank; Michalsky, Elke; Huang, Ruiqi; Preissner, Robert; Barkman, Todd J; Piechulla, Birgit

    2010-02-01

    Methyl salicylate and methyl benzoate have important roles in a variety of processes including pollinator attraction and plant defence. These compounds are synthesized by salicylic acid, benzoic acid and benzoic acid/salicylic acid carboxyl methyltransferases (SAMT, BAMT and BSMT) which are members of the SABATH gene family. Both SAMT and BSMT were isolated from Nicotiana suaveolens, Nicotiana alata, and Nicotiana sylvestris allowing us to discern levels of enzyme divergence resulting from gene duplication in addition to species divergence. Phylogenetic analyses showed that Nicotiana SAMTs and BSMTs evolved in separate clades and the latter can be differentiated into the BSMT1 and the newly established BSMT2 branch. Although SAMT and BSMT orthologs showed minimal change coincident with species divergences, substantial evolutionary change of enzyme activity and expression patterns occurred following gene duplication. After duplication, the BSMT enzymes evolved higher preference for benzoic acid (BA) than salicylic acid (SA) whereas SAMTs maintained ancestral enzymatic preference for SA over BA. Expression patterns are largely complementary in that BSMT transcripts primarily accumulate in flowers, leaves and stems whereas SAMT is expressed mostly in roots. A novel enzyme, nicotinic acid carboxyl methyltransferase (NAMT), which displays a high degree of activity with nicotinic acid was discovered to have evolved in N. gossei from an ancestral BSMT. Furthermore a SAM-dependent synthesis of methyl anthranilate via BSMT2 is reported and contrasts with alternative biosynthetic routes previously proposed. While BSMT in flowers is clearly involved in methyl benzoate synthesis to attract pollinators, its function in other organs and tissues remains obscure.

  18. Quantitative Analysis of the Fate of Exogenous DNA in Nicotiana Protoplasts 1

    Science.gov (United States)

    Uchimiya, Hirofumi; Murashige, Toshio

    1977-01-01

    After a 5-hour incubation of protoplasts of Nicotiana tabacum L. `Xanthi' with 3H-DNA (7.26 μg/ml) from N. tabacum L. `Xanthi nc' 3.5% of the initial radioactivity was found in acid-insoluble substances of the protoplasts. The addition of DEAE-dextran and poly-l-lysine to the incubation medium nearly doubled radioactivity adsorption. The absorption was inhibited by 2,4-dinitrophenol, KCN, and low temperature (0 C); this inhibition could not be reversed by exogenous ATP. About 500 tobacco plants established from protoplasts of a normally tobacco-mosaic virus-susceptible cultivar that had been allowed to absorb DNA prepared from a resistant cultivar did not show transfer of the virus-resistant gene. A detailed analysis was performed of the disposition of exogenous DNA in plant protoplasts, by employing Escherichia coli3H-DNA and Nicotiana glutinosa protoplasts. In 5 to 20 hours, about 10% of the 3H-DNA entered the protoplasts. Competition experiments between the 3H-DNA and unlabeled DNA or thymidine showed that the entry occurred as undegraded 3H-DNA. Examination of intraprotoplast fractions revealed that 60 to 80% of the absorbed radioactivity resided in the “soluble” fraction of the cytoplasm and 20% in the nuclear fraction. The mitochondrion fraction also contained measurable radioactivity. Sizing on sucrose density gradients showed that the bulk of the absorbed E. coli DNA had been depolymerized. Of the incorporated radioactivity, 15% was accountable as DNA, exogenous as well as resynthesized, and 15% as RNA, protein, and other cell constituents. DNA/DNA hybridization test indicated that 17.6% of the re-extractable 3H-DNA retained homology with the E. coli DNA; this was equivalent to 2.6% of the absorbed radioactivity. Resynthesized receptor protoplast DNA was represented by a fraction at least 1.7% of the total absorbed radioactivity. The amount of bacterial DNA remaining in protoplasts suggests that each protoplast retained 2.3 × 10−15g donor DNA, or

  19. Use of Several Natural Products from Selected Nicotiana Species to Prevent Black Shank Disease in Tobacco

    Directory of Open Access Journals (Sweden)

    Kroumova Antoaneta B.

    2016-07-01

    Full Text Available Black shank is a major annual disease threat to all types of tobacco worldwide. It is caused by the fungus Phytophthora parasitica var. nicotianae (PPN. The major tobacco growing areas in US - Kentucky, Tennessee and North Carolina can experience devastating losses, reaching in some fields up to 100%. Thus far, the main approaches to control this disease have been creation of resistant varieties, fungicide treatments, and crop rotation. Some fungicides are reported to have negative effects on the environment. The goal of this work was to test the antifungal activity of several natural products that are synthesized by certain Nicotiana species, and secreted to the leaf surface. We hypothesized that phylloplanin, cis-abienol, labdenediol and sclareol can suppress PPN-race 0- and PPN-race 1-caused disease in Burley tobaccos KY 14 and MS KY 14 × L8LC in the greenhouse. We developed methods for leaf surface extraction, spore preparation and soil drench application of the natural compounds tested. Experiments were performed on 5–8 week-old greenhouse grown seedlings. cis-Abienol showed high inhibitory properties toward the disease. Race 0 infection was completely subdued in KY 14 while race 1 infection was reduced by 70–80%, and delayed by 6–10 days in KY 14 and MS KY14 × L8LC. Sclareol was very effective in inhibiting race 0-caused disease in both tobacco cultivars. In MS KY 14 × L8LC race 1 infection was inhibited while in KY 14 it was reduced by 85% and delayed by 6 days. Labdenediol reduced the disease by half in eight week-old KY 14 plants. Tobacco phylloplanin reduced plant infection by both races by 50–60% and delayed the disease by 6–10 days. Phylloplanin was least suppressive in both tobacco cultivars. We consider sclareol to be the best candidate for future studies due to its antifungal properties and availability. cis-Abienol, despite its good antifungal activity, is not feasible for large-scale use due to the production and

  20. Deciphering the complex leaf transcriptome of the allotetraploid species Nicotiana tabacum: a phylogenomic perspective

    Directory of Open Access Journals (Sweden)

    Bombarely Aureliano

    2012-08-01

    Full Text Available Abstract Background Polyploidization is an important mechanism in plant evolution. By analyzing the leaf transcriptomes taken from the allotetraploid Nicotiana tabacum (tobacco and parental genome donors, N. sylvesteris (S-Genome and N. tomentosiformis (T-Genome, a phylogenomic approach was taken to map the fate of homeologous gene pairs in this plant. Results A comparison between the genes present in the leaf transcriptomes of N. tabacum and modern day representatives of its progenitor species demonstrated that only 33% of assembled transcripts could be distinguished based on their sequences. A large majority of the genes (83.6% of the non parent distinguishable and 87.2% of the phylogenetic topology analyzed clusters expressed above background level (more than 5 reads showed similar overall expression levels. Homeologous sequences could be identified for 968 gene clusters, and 90% (6% of all genes of the set maintained expression of only one of the tobacco homeologs. When both homeologs were expressed, only 15% (0.5% of the total showed evidence of differential expression, providing limited evidence of subfunctionalization. Comparing the rate of synonymous nucleotide substitution (Ks and non-synonymous nucleotide substitution (Kn provided limited evidence for positive selection during the evolution of tobacco since the polyploidization event took place. Conclusions Polyploidization is a powerful mechanism for plant speciation that can occur during one generation; however millions of generations may be necessary for duplicate genes to acquire a new function. Analysis of the tobacco leaf transcriptome reveals that polyploidization, even in a young tetraploid such as tobacco, can lead to complex changes in gene expression. Gene loss and gene silencing, or subfunctionalization may explain why both homeologs are not expressed by the associated genes. With Whole Genome Duplication (WGD events, polyploid genomes usually maintain a high percentage of

  1. Wild Nicotiana Species as a Source of Cytoplasmic Male Sterility in Nicotianatabacum

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    Nikova V

    2014-12-01

    Full Text Available The results of our experiments executed to obtain tobacco male sterile lines through interspecific hybridization are summarized. Ten wild species from the genus Nicotiana: N. excelsior (exc, N. amplexicaulis (amp, N. rustica (rus, Nicotianaglauca (gla, N. velutina (vel, N. benthamiana (ben, N. maritima (mar, N. paniculata (pan, N. longiflora (lon and N. africana (afr were used as cytoplasmic donors and N. tabacum, cv. HarmanliiskaBasma (HB as a donor of the nucleus. Genetic effects of cytoplasmic-nuclear interaction of the studied species are discussed. Our results suggested that cytoplasmic male sterility (CMS was expressed when the cytoplasms of the above mentioned wild Nicotiana species were combined with the nucleus of N. tabacum. The 10 sources of CMS obtained in tobacco were characterized by altered flower phenotypes. Flowers are classified into types according the stamen, pistil and corolla modification. All these CMS sources were backcrossed to Oriental tobaccos, cvs. Tekne, Nevrokop B-12, Kroumovgrad 90 and Djebel 576, to develop corresponding CMS lines. The investigated cytoplasms produced compete male sterility in all those cultivars. The CMS lines preserved flower types, specific for every “sterile” cytoplasm. The extent of male organ modifications varied from apparently normal (but pollenless stamens in CMS (pan, (afr, some plants of (vel (mar through different degrees of malformations (shriveled anther on shortened filaments (lon, pinnate-like anthers on filaments of normal length (amp, petal - (ben, pistil- or stigma-like structures (rus, (gla to lack of male reproductive organs in (exc and in some plants of (vel, (mar, (rus and (gla. Most of the above mentioned cytoplasms had normal female gametophyte and good seed productivity. Alterations of the pistils were observed in CMS (rus, (exc and (ben causing reduction of the seed set. Electrophoresis of seed proteins of the tobacco cultivars and their CMS lines also suggested that

  2. Ecological aspects of a Cosmopolites sordidus (Germar (Coleoptera: Curculionidae population in São Miguel do Iguaçu, PR./ Aspectos ecológicos da população de Cosmopolites sordidus, (Germar (Coleoptera: Curculionidae em São Miguel do Iguaçu, PR

    Directory of Open Access Journals (Sweden)

    Cristiane Rohde

    2006-06-01

    Full Text Available The population dynamics of Cosmopolites sordidus (Germar was studied in banana crops (cultivar Nanicão at São Miguel do Iguaçu, PR, using roof-tile-like baits, between June/2003 and May/2004. Adults infected by entomopathogenic fungi were recorded, and predators were also found on the baits. A comparative bioassay was carried out between the Beauveria bassiana isolate obtained from the experiment area and from other regions in Brazil against C. sordidus adults. The population peaks of C. sordidus adults occurred in June and July, a period with mild temperatures and low precipitation. The population of C. sordidus larvae, however, was concentrated in December. The insect predators found in the area were insects of the following orders: Coleoptera (Carabidae, Dermaptera (Forficulidae, Hemiptera (Reduviidae, Hymenoptera (Formicidae, and spiders of the families Ctenidae, Clubionidae, and Lycosidae, with a population peak two months after the pest population peak. A low number of C. sordidus adults infected by B. bassiana was found, with an annual mean of 0.44%. In the laboratory, the B. bassiana isolates were pathogenic to the insects.Estudou-se a dinâmica populacional de Cosmopolites sordidus (Germar em cultivo de banana(cultivar Nanicão, em São Miguel do Iguaçu, PR, através de iscas do tipo telha, entre junho/2003 a maio/2004. Atribuiu-se conceito ao estado de conservação da isca, visando observar sua durabilidade e atratividade. Registraram-se adultos infectados por fungos entomopatogênicos, sendo também encontrados predadores nas iscas. Realizou-se um bioensaio com isolados de Beauveria bassiana da área experimental e de Centros de Pesquisa do Brasil, para avaliar a patogenicidade em C. sordidus. Verificou-se que os picos populacionais de adultos de C. sordidus ocorreram em junho e julho, período de temperaturas amenas e baixas precipitações, enquanto que as larvas foram mais abundantes em dezembro. Os insetos predadores

  3. Toxicological, Enzymatic, and Molecular Assessment of the Insecticide Susceptibility Profile of Triatoma infestans (Hemiptera: Reduviidae, Triatominae) Populations From Rural Communities of Santa Cruz, Bolivia.

    Science.gov (United States)

    Santo-Orihuela, Pablo L; Vassena, Claudia V; Carvajal, Guillermo; Clark, Eva; Menacho, Silvio; Bozo, Ricardo; Gilman, Robert H; Bern, Caryn; Marcet, Paula L

    2017-01-01

    A wide range of insecticide resistance profiles has been reported across Bolivian domestic and sylvatic populations of Triatoma infestans (Klug, 1834) (Hemiptera, Reduviidae), including some with levels proven to be a threat for vector control. In this work, the insecticide profile of domestic T. infestans was studied with standardized toxicological bioassays, in an area that has not undergone consistent vector control. F1 first-instar nymphs hatched in laboratory from bugs captured in three communities from the Santa Cruz Department were evaluated with different insecticides. Moreover, the enzymatic activity of esterases and cytochrome P450 monooxygenases was measured in individual insects to evaluate the possible mechanism of metabolic resistance to pyrethroids. In addition, the DNA sequence of sodium channel gene (kdr) was screened for two point mutations associated with pyrethroid resistance previously reported in T. infestans.All populations showed reduced susceptibility to deltamethrin and α-cypermethrin, albeit the RR50 values varied significantly among them. Increased P450 monooxygenases and permethrate esterases suggest the contribution, as detoxifying mechanisms, to the observed resistance to deltamethrin in all studied populations. No individuals presented either mutation associated to resistance in the kdr gene. The level of susceptibility to α-cypermethrin, the insecticide used by the local vector control program, falls within an acceptable range to continue its use in these populations. However, the observed RR50 values evidence the possibility of selection for resistance to pyrethroids, especially to deltamethrin. Consequently, the use of pyrethroid insecticides should be closely monitored in these communities, which should be kept under entomological surveillance and sustained interventions. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email

  4. Satellite RNA-mediated Reduction of Cucumber Mosaic Virus Genomic RNAs Accumulation in Nicotiana tabacum

    Institute of Scientific and Technical Information of China (English)

    Qiansheng LIAO; Liping ZHU; Zhiyou DU; Rong ZENG; Junli FENG; Jishuang CHEN

    2007-01-01

    Satellite RNAs (satRNAs) are molecular parasites that interfere with the pathogenesis of the helper viruses.In this study,the relative accumulation of cucumber mosaic virus (CMV)-Fny genomic RNAs with or without satRNAs were quantitatively analyzed by real-time RT-PCR.The results showed that satRs apparently attenuated the symptoms of CMV-Fny on Nicotiana tabacum by depressing the accumulation of CMV-Fny genomic RNAs,tested as open reading frames.The accumulation of CMV-Fny la,2a,2b,3a,and CP genes was much higher than that of CMV-Fny with satRs added(CMV-Fsat),at different inoculation times.CMV-Fny△2b,in which the complete 2b gene and 41 amino acids at the C-terminal of the 2a gene were deleted,caused only a slight mosaic effect on N.tabacum seedlings,similar to that of CMVFsat,but the addition of satRs to CMV-Fny△2b showed further decrease in the accumulation of CMVFny△2b genomic RNAs.Our results indicated that the attenuation of CMV,by adding satRs or deleting the 2b gene,was due to the low accumulation of CMV genomic RNAs,and that satRNA-mediated reduction of CMV genomic RNAs accumulation in N.tabacum was possibly related to the 2b gene.

  5. Functional characterization of a potassium transporter geneNrHAK1 in Nicotiana rustica

    Institute of Scientific and Technical Information of China (English)

    Zhao-kui GUO; Qian YANG; Xiu-qing WAN; Pei-qiang YAN

    2008-01-01

    The purpose of this study is to investigate the function of a novel potassium transporter gene(NrHAK1) isolated from Nicotiana rustica roots using yeast complement and real-time PCR technique. The complementary DNA (cDNA) of NrHAK1, 2 488 bp long, contains an open reading frame (ORF) of 2334 bp encoding a protein of 777 amino acids (87.6 kDa) with 12 predicted transmembrane domains. The NrHAK1 protein shows a high sequence similarity to those of high-affinity potassium transporters in Mesembryanthemum, Phytolacca acinosa, Arabidopsis thaliana, and so on. We found that the NrHAK1 gene could complement the yeast-mutant defect in K+ uptake. Among several tissues surveyed, the expression level of NrHAK1 was most abundant in the root tip and was up-regulated when exposed to potassium starvation. Moreover, the transcript accumulation was significantly reduced by adding 5 mmol/L NH4 to the solution. These results suggest that NrHAK1 plays an important role in potassium absorption in N. Rustica.

  6. Optimization of engineered production of the glucoraphanin precursor dihomo-methionine in Nicotiana benthamiana

    Directory of Open Access Journals (Sweden)

    Christoph eCrocoll

    2016-02-01

    Full Text Available Glucosinolates are natural products characteristic of the Brassicales order which include vegetables such as cabbages and the model plant Arabidopsis thaliana. Glucoraphanin is the major glucosinolate in broccoli and associated with the health-promoting effects of broccoli consumption. Towards our goal of creating a rich source of glucoraphanin for dietary supplements, we have previously reported the feasibility of engineering glucoraphanin in Nicotiana benthaminana through transient expression of glucoraphanin biosynthetic genes from Arabidopsis thaliana (Mikkelsen et al., 2010. As side-products, we obtained 5-8 fold higher levels of chain-elongated leucine-derived glucosinolates, not found in the native plant. Here, we investigated two different strategies to improve engineering of the methionine chain elongation part of the glucoraphanin pathway in N. benthamiana: 1 co-expression of the large subunit (LSU1 of the heterodimeric isopropylmalate isomerase, and 2 co-expression of BAT5 transporter for efficient transfer of intermediates across the chloroplast membrane. We succeeded in raising dihomo-methionine (DHM levels to a maximum of 432 nmol*g-1 fresh weight which is equivalent to a 9-fold increase compared to the highest production of this intermediate previously reported (Mikkelsen et al., 2010. The increased DHM production without increasing leucine-derived side-product levels provides new metabolic engineering strategies for improved glucoraphanin production in a heterologous host.

  7. Transcriptome analysis of Nicotiana tabacum infected by Cucumber mosaic virus during systemic symptom development.

    Directory of Open Access Journals (Sweden)

    Jie Lu

    Full Text Available Virus infection of plants may induce a variety of disease symptoms. However, little is known about the molecular mechanism of systemic symptom development in infected plants. Here we performed the first next-generation sequencing study to identify gene expression changes associated with disease development in tobacco plants (Nicotiana tabacum cv. Xanthi nc induced by infection with the M strain of Cucumber mosaic virus (M-CMV. Analysis of the tobacco transcriptome by RNA-Seq identified 95,916 unigenes, 34,408 of which were new transcripts by database searches. Deep sequencing was subsequently used to compare the digital gene expression (DGE profiles of the healthy plants with the infected plants at six sequential disease development stages, including vein clearing, mosaic, severe chlorosis, partial and complete recovery, and secondary mosaic. Thousands of differentially expressed genes were identified, and KEGG pathway analysis of these genes suggested that many biological processes, such as photosynthesis, pigment metabolism and plant-pathogen interaction, were involved in systemic symptom development. Our systematic analysis provides comprehensive transcriptomic information regarding systemic symptom development in virus-infected plants. This information will help further our understanding of the detailed mechanisms of plant responses to viral infection.

  8. Citrus leaf blotch virus invades meristematic regions in Nicotiana benthamiana and citrus.

    Science.gov (United States)

    Agüero, Jesús; Vives, María Carmen; Velázquez, Karelia; Ruiz-Ruiz, Susana; Juárez, Jose; Navarro, Luis; Moreno, Pedro; Guerri, José

    2013-08-01

    To invade systemically host plants, viruses need to replicate in the infected cells, spread to neighbouring cells through plasmodesmata and move to distal parts of the plant via sieve tubes to start new infection foci. To monitor the infection of Nicotiana benthamiana plants by Citrus leaf blotch virus (CLBV), leaves were agroinoculated with an infectious cDNA clone of the CLBV genomic RNA expressing green fluorescent protein (GFP) under the transcriptional control of a duplicate promoter of the coat protein subgenomic RNA. Fluorescent spots first appeared in agroinfiltrated leaves 11-12 days after infiltration, indicating CLBV replication. Then, after entering the phloem vascular system, CLBV was unloaded in the upper parts of the plant and invaded all tissues, including flower organs and meristems. GFP fluorescence was not visible in citrus plants infected with CLBV-GFP. Therefore, to detect CLBV in meristematic regions, Mexican lime (Citrus aurantifolia) plants were graft inoculated with CLBV, with Citrus tristeza virus (CTV), a virus readily eliminated by shoot-tip grafting in vitro, or with both simultaneously. Although CLBV was detected by hybridization and real-time reverse transcription-polymerase chain reaction (RT-PCR) in 0.2-mm shoot tips in all CLBV-inoculated plants, CTV was not detected. These results explain the difficulty in eliminating CLBV by shoot-tip grafting in vitro. © 2013 BSPP AND JOHN WILEY & SONS LTD.

  9. Cymbidium mosaic virus coat protein gene in antisense confers resistance to transgenic Nicotiana occidentalis.

    Science.gov (United States)

    Lim, S H; Ko, M K; Lee, S J; La, Y J; Kim, B D

    1999-12-31

    The nucleotide sequence of the 3'-terminal region of the Korean isolate of cymbidium mosaic virus (CyMV-Ca) from a naturally infected cattleya was determined. The sequence contains an open reading frame (ORF) coding for the viral coat protein (CP) at the 3'-end and three other ORFs (triple gene block or movement protein) of CyMV. The CP gene encodes a polypeptide chain of 220 amino acids with a molecular mass of 23,760 Da. The deduced CP sequence showed a strong homology with those of two CyMVs reported. A construct of the CyMV-Ca CP gene in the antisense orientation in the plant expression vector pMBP1 was transferred via Agrobacterium tumefaciens-mediated transformation into Nicotiana occidentalis which is a propagation host of CyMV. The T1 progeny of the transgenic plants were inoculated with CyMV and found to be highly resistant to CyMV infection.

  10. NMR-based structural validation of therapeutic antibody produced in Nicotiana benthamiana.

    Science.gov (United States)

    Yagi, Hirokazu; Fukuzawa, Noriho; Tasaka, Yasushi; Matsuo, Kouki; Zhang, Ying; Yamaguchi, Takumi; Kondo, Sachiko; Nakazawa, Shiori; Hashii, Noritaka; Kawasaki, Nana; Matsumura, Takeshi; Kato, Koichi

    2015-06-01

    We successfully developed a method for metabolic isotope labeling of recombinant proteins produced in transgenic tobacco. This enabled assessment of structural integrity of plant-derived therapeutic antibodies by NMR analysis. A variety of expression vehicles have been developed for the production of promising biologics, including plants, fungi, bacteria, insects, and mammals. Glycoprotein biologics often experience altered folding and post-translational modifications that are typified by variant glycosylation patterns. These differences can dramatically affect their efficacy, as exemplified by therapeutic antibodies. However, it is generally difficult to validate the structural integrity of biologics produced using different expression vehicles. To address this issue, we have developed and applied a stable-isotope-assisted nuclear magnetic resonance (NMR) spectroscopy method for the conformational characterization of recombinant antibodies produced in plants. Nicotiana benthamiana used as a vehicle for the production of recombinant immunoglobulin G (IgG) was grown in a (15)N-enriched plant growth medium. The Fc fragment derived from the (15)N-labeled antibody thus prepared was subjected to heteronuclear two-dimensional (2D) NMR measurements. This approach enabled assessment of the structural integrity of the plant-derived therapeutic antibodies by comparing their NMR spectral properties with those of an authentic IgG-Fc derived from mammalian cells.

  11. Identification of photoperiod-regulated gene in soybean and functional analysis in Nicotiana benthamiana

    Indian Academy of Sciences (India)

    Sha Ai-Hua; Chen Yin-Hua; Shan Zhi-Hui; Zhang Xiao-Juan; Wu Xue-Jun; Qiu De-Zheng; Zhou Xin-An

    2014-04-01

    Soybean (Glycine max) is a short-day crop and the photoperiod is a crucial factor regulating its flowering time. To investigate the molecular mechanism controlling the flowering time by photoperiod in soybean, cDNA-amplified fragment length polymorphism (cDNA-AFLP) was used to identify photoperiod-regulated genes in leaves of soybean growing under short-day length, neutral photoperiod and long-day length. A total of 36 transcript-derived fragments (TDFs) were identified to be regulated by photoperiod. Among them, 26 TDFs were homologues of genes with known function. These genes are involved in secondary metabolism, cellular metabolism, cell wall components metabolism, ion transport and hormone signalling. Silencing of the homologue genes in Nicotiana benthamiana for 14 TDFs was conducted by virus-induced gene silencing. The flowering time was delayed by silencing of the genes encoding rhodanese and 40S ribosomal protein S4 (RPS4). The results indicated that rhodanese and RPS4 probably play important roles in regulating flowering time.

  12. Detection of Nicotiana DNA in Tobacco Products Using a Novel Multiplex Real-Time PCR Assay.

    Science.gov (United States)

    Korchinski, Katie L; Land, Adrian D; Craft, David L; Brzezinski, Jennifer L

    2016-07-01

    Establishing that a product contains tobacco is a requirement for the U.S. Food and Drug Administration's regulation and/or prosecution of tobacco products. Therefore, a multiplex real-time PCR method was designed to determine if Nicotiana (tobacco) DNA is present in tobacco products. The PCR method simultaneously amplifies a 73 bp fragment of the cytochrome P450 monoxygenase CYP82E4 gene and 66 bp fragment in the nia-1 gene for nitrate reductase, which are detected using dual-labeled TaqMan probes. The assay is capable of detecting approximately 7.8 pg purified tobacco DNA, with a similar sensitivity for either gene target while incorporating an internal positive control (IPC). DNA was extracted from prepared tobacco products-including chewing tobacco, pipe tobacco, and snuff-or from the cut fill (no wrapper) of cigarettes and cigars. Of the 13 products analyzed, 12 were positive for both tobacco-specific markers and the IPC. DNA was also extracted from the fill of five varieties of herbal cigarettes, which were negative for both tobacco-specific gene targets and positive for the IPC. Our method expands on current assays by introducing a multiplex reaction, targeting two sequences in two different genes of interest, incorporating an IPC into the reaction, and lowering the LOD and LOQ while increasing the efficiency of the PCR.

  13. Plant coexistence can enhance phytoextraction of cadmium by tobacco (Nicotiana tabacum L.) in contaminated soil

    Institute of Scientific and Technical Information of China (English)

    Ling Liu; Yuefang Li; Jianjun Tang; Liangliang Hu; Xin Chen

    2011-01-01

    A mesocosm experiment was conducted to investigate whether plant coexistence affects cadmium (Cd) uptake by plant in contaminated soil. Tobacco (Nicotiana tabacum L. var. K326) and Japanese clover (Kummerowia striata (Thunb.) Schindl.) were used.Cadmium was applied as 3CdSO4.8H2O in solution at three levels (0, 1, and 3 mg/kg soil) to simulate an unpolluted soil and soils that were slightly and moderately polluted with Cd. Tobacco (crop), Japanese clover (non-crop), and their combination were grown under each Cd treatment. Compared to monoculture and under all Cd treatments, co-planting with Japanese clover did not affect tobacco biomass but significantly increased Cd concentration in all tobacco tissues and enhanced Cd accumulation in tobacco shoots and roots.Compared to monoculture, co-planting reduced soil pH and increased Cd bioavailability. For tobacco, co-planting with Japanese clover increased the Cd bioconcentration factor (BCF) in Cd contaminated soil. Japanese clover also accumulated substantial quantities of Cd in shoots and roots. Thus, total Cd uptake by the plants was much greater with co-planting than with monoculture. The results suggested that phytoextraction can be effectively increased through tobacco co-planting with Japanese clover in mildly Cd-contaminated soil.

  14. Techno-economic analysis of horseradish peroxidase production using a transient expression system in Nicotiana benthamiana.

    Science.gov (United States)

    Walwyn, David Richard; Huddy, Suzanne M; Rybicki, Edward P

    2015-01-01

    Despite the advantages of plant-based transient expression systems relative to microbial or mammalian cell systems, the commercial production of recombinant proteins using plants has not yet been achieved to any significant extent. One of the challenges has been the lack of published data on the costs of manufacture for products other than biopharmaceuticals. In this study, we report on the techno-economic analysis of the production of a standard commercial enzyme, namely, horseradish peroxidase (HRP), using a transient expression system in Nicotiana benthamiana. Based on the proven plant yield of 240 mg HRP/kg biomass, a biomass productivity of 15-kg biomass/m(2)/year and a process yield of 54 % (mg HRP product/mg HRP in biomass), it is apparent that HRP can be manufactured economically via transient expression in plants in a large-scale facility (>5 kg HRP/year). At this level, the process is competitive versus the existing technology (extraction of the enzyme from horseradish), and the product is of comparable or improved activity, containing only the preferred isoenzyme C. Production scale, protein yield and biomass productivity are found to be the most important determinants of overall viability.

  15. Salivary proteins of spider mites suppress defenses in Nicotiana benthamiana and promote mite reproduction.

    Science.gov (United States)

    Villarroel, Carlos A; Jonckheere, Wim; Alba, Juan M; Glas, Joris J; Dermauw, Wannes; Haring, Michel A; Van Leeuwen, Thomas; Schuurink, Robert C; Kant, Merijn R

    2016-04-01

    Spider mites (Tetranychidae sp.) are widely occurring arthropod pests on cultivated plants. Feeding by the two-spotted spider mite T. urticae, a generalist herbivore, induces a defense response in plants that mainly depends on the phytohormones jasmonic acid and salicylic acid (SA). On tomato (Solanum lycopersicum), however, certain genotypes of T. urticae and the specialist species T. evansi were found to suppress these defenses. This phenomenon occurs downstream of phytohormone accumulation via an unknown mechanism. We investigated if spider mites possess effector-like proteins in their saliva that can account for this defense suppression. First we performed an in silico prediction of the T. urticae and the T. evansi secretomes, and subsequently generated a short list of candidate effectors based on additional selection criteria such as life stage-specific expression and salivary gland expression via whole mount in situ hybridization. We picked the top five most promising protein families and then expressed representatives in Nicotiana benthamiana using Agrobacterium tumefaciens transient expression assays to assess their effect on plant defenses. Four proteins from two families suppressed defenses downstream of the phytohormone SA. Furthermore, T. urticae performance on N. benthamiana improved in response to transient expression of three of these proteins and this improvement was similar to that of mites feeding on the tomato SA accumulation mutant nahG. Our results suggest that both generalist and specialist plant-eating mite species are sensitive to SA defenses but secrete proteins via their saliva to reduce the negative effects of these defenses.

  16. Mitochondrial alternative oxidase maintains respiration and preserves photosynthetic capacity during moderate drought in Nicotiana tabacum.

    Science.gov (United States)

    Dahal, Keshav; Wang, Jia; Martyn, Greg D; Rahimy, Farkhunda; Vanlerberghe, Greg C

    2014-11-01

    The mitochondrial electron transport chain includes an alternative oxidase (AOX) that is hypothesized to aid photosynthetic metabolism, perhaps by acting as an additional electron sink for photogenerated reductant or by dampening the generation of reactive oxygen species. Gas exchange, chlorophyll fluorescence, photosystem I (PSI) absorbance, and biochemical and protein analyses were used to compare respiration and photosynthesis of Nicotiana tabacum 'Petit Havana SR1' wild-type plants with that of transgenic AOX knockdown (RNA interference) and overexpression lines, under both well-watered and moderate drought-stressed conditions. During drought, AOX knockdown lines displayed a lower rate of respiration in the light than the wild type, as confirmed by two independent methods. Furthermore, CO2 and light response curves indicated a nonstomatal limitation of photosynthesis in the knockdowns during drought, relative to the wild type. Also relative to the wild type, the knockdowns under drought maintained PSI and PSII in a more reduced redox state, showed greater regulated nonphotochemical energy quenching by PSII, and displayed a higher relative rate of cyclic electron transport around PSI. The origin of these differences may lie in the chloroplast ATP synthase amount, which declined dramatically in the knockdowns in response to drought. None of these effects were seen in plants overexpressing AOX. The results show that AOX is necessary to maintain mitochondrial respiration during moderate drought. In its absence, respiration rate slows and the lack of this electron sink feeds back on the photosynthetic apparatus, resulting in a loss of chloroplast ATP synthase that then limits photosynthetic capacity.

  17. Reduced Susceptibility to Xanthomonas citri in Transgenic Citrus Expressing the FLS2 Receptor From Nicotiana benthamiana.

    Science.gov (United States)

    Hao, Guixia; Pitino, Marco; Duan, Yongping; Stover, Ed

    2016-02-01

    Overexpression of plant pattern-recognition receptors by genetic engineering provides a novel approach to enhance plant immunity and broad-spectrum disease resistance. Citrus canker disease associated with Xanthomonas citri is one of the most important diseases damaging citrus production worldwide. In this study, we cloned the FLS2 gene from Nicotiana benthamiana cDNA and inserted it into the binary vector pBinPlus/ARS to transform Hamlin sweet orange and Carrizo citrange. Transgene presence was confirmed by polymerase chain reaction (PCR) and gene expression of NbFLS2 was compared by reverse transcription quantitative PCR. Reactive oxygen species (ROS) production in response to flg22Xcc was detected in transgenic Hamlin but not in nontransformed controls. Low or no ROS production was detected from nontransformed Hamlin seedlings challenged with flg22Xcc. Transgenic plants highly expressing NbFLS2 were selected and were evaluated for resistance to canker incited by X. citri 3213. Our results showed that the integration and expression of the NbFLS2 gene in citrus can increase canker resistance and defense-associated gene expression when challenged with X. citri. These results suggest that canker-susceptible Citrus genotypes lack strong basal defense induced by X. citri flagellin and the resistance of these genotypes can be enhanced by transgenic expression of the flagellin receptor from a resistant species.

  18. The Subcellular Localization and Functional Analysis of Fibrillarin2, a Nucleolar Protein in Nicotiana benthamiana

    Directory of Open Access Journals (Sweden)

    Luping Zheng

    2016-01-01

    Full Text Available Nucleolar proteins play important roles in plant cytology, growth, and development. Fibrillarin2 is a nucleolar protein of Nicotiana benthamiana (N. benthamiana. Its cDNA was amplified by RT-PCR and inserted into expression vector pEarley101 labeled with yellow fluorescent protein (YFP. The fusion protein was localized in the nucleolus and Cajal body of leaf epidermal cells of N. benthamiana. The N. benthamiana fibrillarin2 (NbFib2 protein has three functional domains (i.e., glycine and arginine rich domain, RNA-binding domain, and α-helical domain and a nuclear localization signal (NLS in C-terminal. The protein 3D structure analysis predicted that NbFib2 is an α/β protein. In addition, the virus induced gene silencing (VIGS approach was used to determine the function of NbFib2. Our results showed that symptoms including growth retardation, organ deformation, chlorosis, and necrosis appeared in NbFib2-silenced N. benthamiana.

  19. Short Exogenous Peptides Regulate Expression of CLE, KNOX1, and GRF Family Genes in Nicotiana tabacum.

    Science.gov (United States)

    Fedoreyeva, L I; Dilovarova, T A; Ashapkin, V V; Martirosyan, Yu Ts; Khavinson, V Kh; Kharchenko, P N; Vanyushin, B F

    2017-04-01

    Exogenous short biologically active peptides epitalon (Ala-Glu-Asp-Gly), bronchogen (Ala-Glu-Asp-Leu), and vilon (Lys-Glu) at concentrations 10(-7)-10(-9) M significantly influence growth, development, and differentiation of tobacco (Nicotiana tabacum) callus cultures. Epitalon and bronchogen, in particular, both increase growth of calluses and stimulate formation and growth of leaves in plant regenerants. Because the regulatory activity of the short peptides appears at low peptide concentrations, their action to some extent is like that of the activity of phytohormones, and it seems to have signaling character and epigenetic nature. The investigated peptides modulate in tobacco cells the expression of genes including genes responsible for tissue formation and cell differentiation. These peptides differently modulate expression of CLE family genes coding for known endogenous regulatory peptides, the KNOX1 genes (transcription factor genes) and GRF (growth regulatory factor) genes coding for respective DNA-binding proteins such as topoisomerases, nucleases, and others. Thus, at the level of transcription, plants have a system of short peptide regulation of formation of long-known peptide regulators of growth and development. The peptides studied here may be related to a new generation of plant growth regulators. They can be used in the experimental botany, plant molecular biology, biotechnology, and practical agronomy.

  20. The effect of intermittent dosing of Nicotiana glauca on teratogenesis in goats.

    Science.gov (United States)

    Welch, K D; Panter, K E; Lee, S T; Gardner, D R

    2015-01-01

    Sustained inhibition of fetal movement in livestock species, induced by several poisonous plants, can result in numerous skeletal-contracture malformations. Lupines are responsible for a condition in cattle referred to as "crooked calf syndrome" that occurs when pregnant cattle graze teratogenic lupines. Similar malformations are also seen in animals poisoned by Conium maculatum (coniine) and Nicotiana glauca (anabasine). A proposed management strategy to limit these types of birth defects includes utilizing an intermittent grazing schedule to allow short durations of grazing lupine-infested areas interrupted by movement to a lupine-free pasture. The objective of this study was to use a goat model to determine if an intermittent schedule of five continuous days on treatment followed by two days off treatment would be sufficient to decrease, or prevent, the incidence of anabasine-induced malformations. The data from this study suggest that, for N. glauca in goats, the intermittent grazing program of five days exposure with two days of non-exposure is insufficient to prevent significant skeletal malformations from occurring. However, this study did demonstrate an inverse relationship between the amount of serum anabasine in the dam and the extent of fetal movement.

  1. Arabinogalactan-Proteins of the Female Sexual Tissue of Nicotiana alata

    Science.gov (United States)

    Gell, Andrew C.; Bacic, Antony; Clarke, Adrienne E.

    1986-01-01

    Arabinogalactan-proteins (AGPs), isolated from the pistils of Nicotiana alata, an ornamental tobacco, are developmentally regulated. Both the total amount and concentration of AGP in the stigma increase during flower development, reaching 10 micrograms AGP/stigma at maturity. In contrast, AGP concentration in the style remains constant throughout the maturation period reaching 12 micrograms AGP/style at maturity. The classes of AGP present in the stigma and style during flower development, separated according to their charge by crossed-electrophoresis, are different and change during development. Pollination of flowers of N. alata with compatible or incompatible pollen results in a significant and reproducible increase in the amount of AGPs in the stigma, but not the style, compared with control unpollinated pistils. Pollination with ethanol vapor inactivated pollen also results in an increase in the amount of AGP in the stigma, but this is less than half that observed following pollination with viable pollen. There are no significant differences in the classes of AGP, based on crossed-electrophoresis, present in the pistil following pollination. Images Fig. 1 Fig. 2 PMID:16665162

  2. Lysinibacillus tabacifolii sp. nov., a novel endophytic bacterium isolated from Nicotiana tabacum leaves.

    Science.gov (United States)

    Duan, Yan-Qing; He, Song-Tao; Li, Qing-Qing; Wang, Ming-Feng; Wang, Wen-Yuan; Zhe, Wei; Cao, Yong-Hong; Mo, Ming-He; Zhai, Yu-Long; Li, Wen-Jun

    2013-06-01

    A Gram-positive, catalase- and oxidase-positive, strictly aerobic, endospore-forming rod bacterium, designated K3514(T), was isolated from the leaves of Nicotiana tabacum. The strain was able to grow at temperatures of 8-40°C, pH 5.0-10.0 and NaCl concentrations of 0-7%. The predominant quinones (>30%) of this strain were MK-7(H2) and MK-7. Phylogenetic analysis of 16S rRNA gene sequence showed that strain K3514(T) was affiliated to the genus Lysinibacillus, with its closest relatives being Lysinibacillus mangiferihumi (98.3% sequence similarity), Lysinibacillus sphaericus (97.9% sequence similarity), Lysinibacillus fusiformis (97.4% sequence similarity), and Lysinibacillus xylanilyticus (97.3% sequence similarity). However, low levels of DNA-DNA relatedness values suggested that the isolate was distinct from the other closest Lysinibacillus species. Additionally, based on analysis of morphological, physiological, and biochemical characteristics, the isolate could be differentiated from the closest known relatives. Therefore, based on polyphasic taxonomic data, the novel isolate likely represents a novel species, for which the name Lysinibacillus tabacifolii sp. nov. and the type strain K3514(T) (=KCTC 33042(T) =CCTCC AB 2012050(T)) are proposed.

  3. Light and clomazone effects on tobacco (Nicotiana tabacum) callus and leaf discs.

    Science.gov (United States)

    Camper, N D; McDonald, S K; Burrows, P M

    2003-11-01

    The effects of clomazone on the growth of tobacco (Nicotiana tabacum L. 'NC2326') callus and leaf discs were studied under four light regimes. Callus cultures and leaf discs were grown on Murashige and Skoog medium supplemented with IAA and kinetin. Light regimes were: dark grown callus kept in the dark and also transferred to the light; light grown callus kept in the light and also transferred to the dark. Two-month-old callus (cultured for 2 months from initiation) grew more rapidly than twelve-month-old callus (cultured for 12 months from initiation) under all conditions tested. Callus transferred from light to dark, or from dark to light, increased in fresh weight slower than did the callus maintained totally in light or dark. Clomazone (2-[(2-chlorophenyl)methyl]-4,4-dimethyl-3-isoxazolidinone) at 140 mg l(-1) or more was lethal to both callus and leaf discs whereas 10 mg l(-1) was stimulatory to growth. Callus tissue responded to clomazone differently depending on the light regime under which it was grown. While clomazone may be affecting the isoprenoid pathway in the callus and leaf disks resulting in growth inhibition, it is possible that other target sites are also being affected and contribute to the reduced growth.

  4. Potato virus X movement in Nicotiana benthamiana: new details revealed by chimeric coat protein variants.

    Science.gov (United States)

    Betti, Camilla; Lico, Chiara; Maffi, Dario; D'Angeli, Simone; Altamura, Maria Maddalena; Benvenuto, Eugenio; Faoro, Franco; Baschieri, Selene

    2012-02-01

    Potato virus X coat protein is necessary for both cell-to-cell and phloem transfer, but it has not been clarified definitively whether it is needed in both movement phases solely as a component of the assembled particles or also of differently structured ribonucleoprotein complexes. To clarify this issue, we studied the infection progression of a mutant carrying an N-terminal deletion of the coat protein, which was used to construct chimeric virus particles displaying peptides selectively affecting phloem transfer or cell-to-cell movement. Nicotiana benthamiana plants inoculated with expression vectors encoding the wild-type, mutant and chimeric viral genomes were examined by microscopy techniques. These experiments showed that coat protein-peptide fusions promoting cell-to-cell transfer only were not competent for virion assembly, whereas long-distance movement was possible only for coat proteins compatible with virus particle formation. Moreover, the ability of the assembled PVX to enter and persist into developing xylem elements was revealed here for the first time.

  5. Agroinfiltration reduces ABA levels and suppresses Pseudomonas syringae-elicited salicylic acid production in Nicotiana tabacum.

    Directory of Open Access Journals (Sweden)

    Arantza Rico

    Full Text Available BACKGROUND: Agrobacterium tumefaciens strain GV3101 (pMP90 is widely used in transient gene expression assays, including assays to study pathogen effectors and plant disease resistance mechanisms. However, inoculation of A. tumefaciens GV3101 into Nicotiana tabacum (tobacco leaves prior to infiltration with pathogenic and non-host strains of Pseudomonas syringae results in suppression of macroscopic symptoms when compared with leaves pre-treated with a buffer control. METHODOLOGY/FINDINGS: To gain further insight into the mechanistic basis of symptom suppression by A. tumefaciens we examined the effect of pre-treatment with A. tumefaciens on the growth of P. syringae, the production of the plant signalling molecules salicylic acid (SA and abscisic acid (ABA, and the presence of callose deposits. Pre-treatment with A. tumefaciens reduced ABA levels, P. syringae multiplication and P. syringae-elicited SA and ABA production, but promoted increased callose deposition. However, pre-treatment with A. tumefaciens did not suppress necrosis or SA production in leaves inoculated with the elicitor HrpZ. CONCLUSIONS/SIGNIFICANCE: Collectively, these results show that inoculation of N. tabacum leaves with A. tumefaciens alters plant hormone levels and plant defence responses to P. syringae, and demonstrate that researchers should consider the impact of A. tumefaciens on plant signal transduction when using A. tumefaciens-mediated transient expression assays to investigate ABA-regulated processes or pathogenicity and plant defence mechanisms.

  6. Identification of donor lines containing favorable alleles for quantitative traits in Burley tobacco (Nicotiana tabacum L.

    Directory of Open Access Journals (Sweden)

    Dražić Slobodan

    2010-01-01

    Full Text Available Well-adapted local lines might be used as donors to improve existing commercial hybrids. The objective of this study was to identify the best burley tobacco (Nicotiana tabacum L. line among a group of four lines (SA 130, Bols 334, Barlej T and TN 90 from germplasm collection, for improving an elite single hybrid Bols 335 x TN 86. Values of three quantitative traits (number of leaves, total leaf area and leaf yield were measured in two-year field experiments. Evaluation of donor lines as sources of new favorable alleles not present in the elite hybrid parent lines was conducted. According to Dudley (1984, 1987 the most significant class for improving quantitative traits is locus class G. Donor line TN 90 had positive values of the parameter μG for number of leaves and for total leaf area. Studied donor lines could not be use directly as the source of new alleles for the improvement of leaf yield in elite hybrid. The improvement of leaf yield in tobacco hybrid Bols 335 x TN 86 would be conducted indirectly via back crossing (TN 86 x TN 90 x TN 86. Significance of GxE interaction for studied traits indicates the same breeding method would be effective in broad range of environmental conditions.

  7. Jasmonate mediates salt-induced nicotine biosynthesis in tobacco (Nicotiana tabacum L.

    Directory of Open Access Journals (Sweden)

    Xiaodong Chen

    2016-04-01

    Full Text Available Jasmonate (JA, as an important signal, plays a key role in multiple processes of plant growth, development and stress response. Nicotine and related pyridine alkaloids in tobacco (Nicotiana tabacum L. are essential secondary metabolites. Whether environmental factors control nicotine biosynthesis and the underlying mechanism remains previously unreported. Here, we applied physiological and biochemical approaches to investigate how salt stress affects nicotine biosynthesis in tobacco. We found that salt stress induced the biosynthesis of JA, which subsequently triggered the activation of JA-responsive gene expression and, ultimately, nicotine synthesis. Bioinformatics analysis revealed the existence of many NtMYC2a-recognized G-box motifs in the promoter regions of NtLOX, NtAOS, NtAOC and NtOPR genes. Applying exogenous JA increased nicotine content, while suppressing JA biosynthesis reduced nicotine biosynthesis. Salt treatment could not efficiently induce nicotine biosynthesis in transgenic anti-COI1 tobacco plants. These results demonstrate that JA acts as the essential signal which triggers nicotine biosynthesis in tobacco after salt stress.

  8. Differential patterns of dehydroabietic acid biotransformation by Nicotiana tabacum and Catharanthus roseus cells.

    Science.gov (United States)

    Häkkinen, Suvi T; Lackman, Petri; Nygrén, Heli; Oksman-Caldentey, Kirsi-Marja; Maaheimo, Hannu; Rischer, Heiko

    2012-01-20

    The aim of this study was to use whole cell catalysts as tools for modification of selected resin acids in order to obtain value-added functional derivatives. The enzymatic bioconversion capacities of two plant species were tested towards dehydroabietic acid. Dehydroabietic acid (DHA) is an abundant resin acid in conifers, representing a natural wood protectant. It is also one of the constituents found in by-products of the kraft chemical pulping industry. DHA was fed to tobacco (Nicotiana tabacum) and Madagascar periwinkle (Catharanthus roseus) plant cell and tissue cultures and bioconversion product formation was monitored using NMR analysis. Both plant species took up DHA from culture medium, and various types of typical detoxification processes occurred in both cultures. In addition, diverse responses to DHA treatment were observed, including differences in uptake kinetics, chemical modification of added substrate and changes in overall metabolism of the cells. Interestingly, Catharanthus roseus, a host species for pharmaceutically valuable terpenoid indole alkaloids, exhibited a very different bioconversion pattern for exogenously applied DHA than tobacco, which does not possess a terpenoid indole pathway. In tobacco, DHA is readily glycosylated in the carbonyl group, whereas in periwinkle it is proposed that a cytochrome P450-catalyzed enzymatic detoxification reaction takes place before the formation of glycosylated product.

  9. Effect of salicylic acid on tobacco (Nicotiana rustica plant under drought conditions

    Directory of Open Access Journals (Sweden)

    Ghader Habibi

    2015-09-01

    Full Text Available Drought stress impact photosynthesis and stomatal conductance, and may reduce the overall production capacity of plants. Since exogenous application of salicylic acid (SA can partially alleviatebe increased the negative effects tolerance of drought stress by improve the metabolism pathways and increase the net photosynthesison plant photosynthesis and metabolism, the main objective of this study was to clarify the roles of SA in enhancing 28 days tobacco (Nicotiana rustica Basmas tolerance to drought stress (50% FC. The results indicated that foliar application of SA (0.5 mM influenced negatively net CO2 assimilation rate and stomatal conductance and led to reduction of shoot and root dry masses. In contrast, the stress did not reduce significantly the maximal quantum yield of photosystem II (PSII. This that can be explained by enhancement of efficiency for dissipation of excess photon energy in the PSII antenna, determined as non-photochemical quenching, and consequently further protection of PSII from photodamage. Thus, under more drought stress, the reduction of photosynthesis of tobacco plants was due mainly to reduction of stomatal conductance. Under water-deficient conditions, plants showed an increase in chlorophyll a and amino acids concentrations in the leaves when treated with SA while this change for net photosynthesis was negligible. Our results indicated showed that the foliar application of SA had no ameliorative effect on tobacco growth under drought stress, because its effect on elevation of transpiration rate did not increase net photosynthesis under drought condition.

  10. Expression of a begomoviral DNAβ gene in transgenic Nicotiana plants induced abnormal cell division

    Institute of Scientific and Technical Information of China (English)

    CUI Xiao-feng; LI Yun-qin; HU Dong-wei; ZHOU Xue-ping

    2005-01-01

    An increasing number of monopartite begomoviruses are being identified that a satellite molecule (DNAβ) is required to induce typical symptoms in host plants. DNAβ encodes a single gene (termed βC1) encoded in the complementary-sense. We have produced transgenic Nicotiana benthamiana and N. tabacum plants expressing theβC1 gene of a DNAβ associated with Tomato yellow leaf curl China virus (TYLCCNV), under the control of the Cauliflower mosaic virus 35S promoter. Transgenic plants expressing βC1 showed severe developmental abnormalities in both species. Microscopic analysis of sections of both transgenic and non-transgenic N. tabacum leaves showed abnormal outgrowths of transgenic N. tabacum to be due to disorganized cell division (hyperplasia) of spongy and palisade parenchyma. Immuno-gold labeling of sections with a polyclonal antibody against the βC1 protein showed that the βC1 protein accumulated in the nuclei of cells. The possible biological function of the βC1 protein was discussed.

  11. In vitro antioxidant properties of flavonoids and polysaccharides extract from tobacco (Nicotiana tabacum L.) leaves.

    Science.gov (United States)

    Ru, Qiao-Mei; Wang, Li-Juan; Li, Wei-Ming; Wang, Jing-Lu; Ding, Yu-Ting

    2012-09-21

    In the present study, antioxidant properties of flavonoids and polysaccharides from tobacco (Nicotiana tabacum L.) leaves were evaluated in several in vitro systems, e.g., scavenging activities on hydroxyl, superoxide anion, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals, and reducing power. Flavonoids showed much better activity than polysaccharides in scavenging activities on free radicals. When compared to the positive control, ascorbic acid, both showed weaker antioxidant potential. However, flavonoids possessed comparable superoxide anion, DPPH and ABTS radical scavenging abilities to ascorbic acid at high concentration (600 μg/mL). Meanwhile, it was found that flavonoids had prominent effects on the reducing power, which was equivalent to ascorbic acid, and was significantly higher than polysaccharides. These results clearly indicate that flavonoids are effective in scavenging free radicals and have the potential to be powerful antioxidants. Thus, tobacco leaves could be considered as a potential source of natural antioxidants for food, pharmaceutical, cosmetics or nutraceutical industries.

  12. Evaluation of hepatoprotective effect of Pistacia lentiscus, Phillyrea latifolia and Nicotiana glauca.

    Science.gov (United States)

    Janakat, Sana; Al-Merie, Hela

    2002-11-01

    The hepatoprotective effect of the boiled and non-boiled aqueous extracts of Pistacia lentiscus, Phillyrea latifolia, and Nicotiana glauca, that are alleged to be effective in the treatment of jaundice in Jordanian folk medicine, was evaluated in vivo using carbon tetrachloride (CCl(4)) intoxicated rats as an experimental model. Plant extracts were administrated orally at a dose of 4 ml/kg body weight, containing various amounts of solid matter. Only total serum bilirubin level was reduced by treatment with non-boiled aqueous extract of N. glauca leaves, while the boiled and non-boiled aqueous extracts of the N. glauca flowers were non effective. Bilirubin level and the activity of alkaline phosphatase (ALP) were both reduced upon treatment with boiled aqueous extract of P. latifolia without reducing the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Aqueous extract of P. lentiscus (both boiled and non-boiled) showed marked antihepatotoxic activity against CCl(4) by reducing the activity of the three enzymes and the level of bilirubin. The effect of the non-boiled aqueous extract was more pronounced than that of the boiled extract.

  13. A red algal cyclophilin has an effect on development and growth in Nicotiana tabacum.

    Science.gov (United States)

    Cho, Eun Kyung; Kim, Mihyang

    2008-10-01

    In this study, an algal Cyp was introduced into plant to research the effect of the gene on growth and development. cDNA GjCyp-1 was isolated from the red alga (Griffithsia japonica), and a recombinant GjCyp-1 containing a CaMV35S promoter at the amino-terminus was constructed in Nicotiana tabacum. The altered GjCyp-1 levels in plants and the expression pattern of Cyp after hormone treatment were confirmed by RNA blotting. Transcript of GjCyp-1 was induced by plant hormones such as gibberellic acid (GA(3)), indoleacetic acid (IAA), and zeatin (ZA). Constitutive overexpression of GjCyp-1 appeared to be beneficial to seed germination. The ratio of emergence of cotyledon from seeds overexpressing GjCyp-1 was almost three times higher than that of the transgenic seeds carrying only the vector. In addition, it was found that most of the seedlings overexpressing GjCyp-1 were dwarfs with altered root systems. The ratio of leaf length and width and root length from transgenic seedlings overexpressing GjCyp-1 was almost 2 and 3.5 times lower than that of the transgenic seedlings carrying only the vector, respectively. The data in this study suggest that GjCyp-1 may affect development and growth in organisms.

  14. Ocorrência de Psammolestes coreodes Bergroth, 1911 (Hemiptera: Reduviidae: Triatominae no Estado de Mato Grosso do Sul Occurrence of Psammolestes coreodes Bergroth, 1911 (Hemiptera: Reduviidae: Triatominae, in the State of Mato Grosso do Sul

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    Paulo Silva de Almeida

    2009-10-01

    Full Text Available Neste trabalho, relata-se a ocorrência de Psammolestes coreodes Bergroth, 1911 pela primeira vez no Estado de Mato Grosso do Sul. Em 2006, um espécime foi capturado em um ambiente peridomiciliar do município de Miranda, incluindo o Estado de Mato Grosso do Sul, na distribuição geográfica dessa espécie.This paper describes the occurrence of Psammolestes coreodes Bergroth, 1911, for the first time in the State of Mato Grosso do Sul. In 2006, a specimen was caught in a peridomestic environment in the municipality of Miranda, thereby including Mato Grosso do Sul within the geographical distribution of this species.

  15. Insects found in birds' nests from Argentina: Coryphistera alaudina Burmeister, 1860 (Aves: Furnariidae), their inquiline birds and mammals, new hosts for Psammolestes coreodes Bergroth, 1911 and Triatoma platensis Neiva, 1913 (Hemiptera: Reduviidae: Triatominae).

    Science.gov (United States)

    Turienzo, Paola; Di Iorio, Osvaldo

    2014-06-03

    The insect fauna in nests of Coryphistera alaudina Burmeister, 1860 (Aves: Furnariidae) were studied in the provinces of Santiago del Estero, Chaco, Córdoba, and La Pampa in Argentina. A total of 7364 insect specimens comprising 77 taxa in a total of 29 families and 7 orders was found in their nests: 40 identified to species, 23 identified to genus, and 14 identified to family. Coryphistera alaudina and some of their vertebrate inquilines are new host records for the triatomine bugs Psammolestes coreodes Bergroth, 1911 and/or Triatoma platensis Neiva, 1913 (Hemiptera: Reduviidae). The insects in the nests of C. alaudina are separated by functional guilds, and their permanence time inside the nests are presented in a new manner and discussed.

  16. Distribuição Geográfica e Dispersão Alti-latitudinal de Alguns Gêneros e Espécies da Tribo Triatomini Jeannel, 1919 (Hemiptera, Reduviidae, Triatominae

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    Galvão Cleber

    1998-01-01

    Full Text Available Geographical Distribution and Alti-latitudinal Dispersion of Some Genera and Species of the Tribe Triatomini Jeannel, 1919 (Hemiptera, Reduviidae, Triatominae - With the currently known distribution of genera and species of the tribe Triatomini Jeannel, 1919, maps and diagrams were prepared, showing the geographical area occupied by the species and their respective dispersion in grades of latitude and altitude. Two genera are not treated: Panstrongylus Berg, 1879, already published and Triatoma Laporte, 1832, that is being published partially in several articles. Genus Eratyrus Stal, 1859, has two species, while Dipetalogaster Usinger, 1939, Mepraia Mazza, Gajardo & Joerg, 1940, Paratriatoma Barber, 1938 and recently created genus Hermanlentia Jurberg & Galvão, 1997, have only one each. The study of these maps and diagrams permits a better knowledge about some ecological requirements of Chagas' disease vectors and detects gaps in the geographical distribution, where the species were not found but probably they could be prevalent

  17. Glutathione transferase from Trichoderma virens enhances cadmium tolerance without enhancing its accumulation in transgenic Nicotiana tabacum.

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    Prachy Dixit

    Full Text Available BACKGROUND: Cadmium (Cd is a major heavy metal pollutant which is highly toxic to plants and animals. Vast agricultural areas worldwide are contaminated with Cd. Plants take up Cd and through the food chain it reaches humans and causes toxicity. It is ideal to develop plants tolerant to Cd, without enhanced accumulation in the edible parts for human consumption. Glutathione transferases (GST are a family of multifunctional enzymes known to have important roles in combating oxidative stresses induced by various heavy metals including Cd. Some GSTs are also known to function as glutathione peroxidases. Overexpression/heterologous expression of GSTs is expected to result in plants tolerant to heavy metals such as Cd. RESULTS: Here, we report cloning of a glutathione transferase gene from Trichoderma virens, a biocontrol fungus and introducing it into Nicotiana tabacum plants by Agrobacterium-mediated gene transfer. Transgenic nature of the plants was confirmed by Southern blot hybridization and expression by reverse transcription PCR. Transgene (TvGST showed single gene Mendelian inheritance. When transgenic plants expressing TvGST gene were exposed to different concentrations of Cd, they were found to be more tolerant compared to wild type plants, with transgenic plants showing lower levels of lipid peroxidation. Levels of different antioxidant enzymes such as glutathione transferase, superoxide dismutase, ascorbate peroxidase, guiacol peroxidase and catalase showed enhanced levels in transgenic plants expressing TvGST compared to control plants, when exposed to Cd. Cadmium accumulation in the plant biomass in transgenic plants were similar or lower than wild-type plants. CONCLUSION: The results of the present study suggest that transgenic tobacco plants expressing a Trichoderma virens GST are more tolerant to Cd, without enhancing its accumulation in the plant biomass. It should be possible to extend the present results to crop plants for

  18. Multiplexed, targeted gene editing in Nicotiana benthamiana for glyco-engineering and monoclonal antibody production.

    Science.gov (United States)

    Li, Jin; Stoddard, Thomas J; Demorest, Zachary L; Lavoie, Pierre-Olivier; Luo, Song; Clasen, Benjamin M; Cedrone, Frederic; Ray, Erin E; Coffman, Andrew P; Daulhac, Aurelie; Yabandith, Ann; Retterath, Adam J; Mathis, Luc; Voytas, Daniel F; D'Aoust, Marc-André; Zhang, Feng

    2016-02-01

    Biopharmaceutical glycoproteins produced in plants carry N-glycans with plant-specific residues core α(1,3)-fucose and β(1,2)-xylose, which can significantly impact the activity, stability and immunogenicity of biopharmaceuticals. In this study, we have employed sequence-specific transcription activator-like effector nucleases (TALENs) to knock out two α(1,3)-fucosyltransferase (FucT) and the two β(1,2)-xylosyltransferase (XylT) genes within Nicotiana benthamiana to generate plants with improved capacity to produce glycoproteins devoid of plant-specific residues. Among plants regenerated from N. benthamiana protoplasts transformed with TALENs targeting either the FucT or XylT genes, 50% (80 of 160) and 73% (94 of 129) had mutations in at least one FucT or XylT allele, respectively. Among plants regenerated from protoplasts transformed with both TALEN pairs, 17% (18 of 105) had mutations in all four gene targets, and 3% (3 of 105) plants had mutations in all eight alleles comprising both gene families; these mutations were transmitted to the next generation. Endogenous proteins expressed in the complete knockout line had N-glycans that lacked β(1,2)-xylose and had a significant reduction in core α(1,3)-fucose levels (40% of wild type). A similar phenotype was observed in the N-glycans of a recombinant rituximab antibody transiently expressed in the homozygous mutant plants. More importantly, the most desirable glycoform, one lacking both core α(1,3)-fucose and β(1,2)-xylose residues, increased in the antibody from 2% when produced in the wild-type line to 55% in the mutant line. These results demonstrate the power of TALENs for multiplexed gene editing. Furthermore, the mutant N. benthamiana lines provide a valuable platform for producing highly potent biopharmaceutical products.

  19. Evaluation of Nicotiana tabacum plants transformed for the expression of verocytotoxic Escherichia coli antigens.

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    Angela Lombardi

    2015-07-01

    Full Text Available Two transgenic Nicotiana tabacum plants, carrying respectively the F18 adhesive fimbriae and the B subunit of verocytotoxin genes from O138 Verocytotoxic E.coli serotype were developed as a model of edible vaccine. Tobacco plants were transformed by agroinfection according to Rossi et al. (2013 stably.  The F18 adhesive fimbriae and VT2e B-subunit were placed under control of the GLOB promoter for the seed-specific protein expression. Agrobacterium tumefaciens binary vector system is an efficient tool to transform plant cells; however, the exogenous gene integrates at semi-random into the nuclear chromosome. PCR products, using specific oligonucleotides putatively encoding the B-subunit of VT2e-B and F18 fimbriae were identified on agarose gel (1.5% - 0.9% as bands with a length of 270 and 519 base pairs, respectively. We showed that the foreign VT2e-B and F18 fimbriae genes were stably integrated into the tobacco genome. Northern blot and Western blot analyses carried out respectively on total mRNA and total soluble protein extract obtained from seeds. For each line, the obtained amount of antigens is sufficient for subsequent oral immunization trials. Three lines of tobacco seeds (F18, VT2e-B, and WT were seeded in homogeneous conditions and were harvested simultaneously. Tobacco plants were analysed also by optical and electronic microscope in different phases of growth. Germination of transgenic seeds were delayed of three/five days compared to WT in two replicated experiments, suggesting that genetic manipulation may influenced mechanisms leading to germination. In conclusion the genes coding for VT2e-B and the F18 are stably maintained in the seeds and obtained tobacco seeds represent a valid strategy to ferry antigenic proteins to the gut and a promising non-invasive method of vaccination in pig industry.

  20. [Induction of polyploid in hairy roots of Nicotiana tabacum and its plant regeneration].

    Science.gov (United States)

    Hou, Lili; Shi, Heping; Yu, Wu; Tsang, Po Keung Eric; Chow, Cheuk Fai Stephen

    2014-04-01

    By genetic transformation with Agrobacterum rhizogenes and artificial chromosome doubling techniques, we studied the induction of hairy roots and their polyploidization, and subsequent plant regeneration and nicotine determination to enhance the content of nicotine in Nicotiana tabacum. The results show that hairy roots could be induced from the basal surface of leaf explants of N. tabacum 8 days after inoculation with Agrobacterium rhizogenes ATCC15834. The percentage of the rooting leaf explants was 100% 15 days after inoculation. The hairy roots could grow rapidly and autonomously on solid or liquid phytohormones-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and paper electrophoresis of opines from N. tabacum hairy roots. The highest rate of polyploidy induction, more than 64.71%, was obtained after treatment of hairy roots with 0.1% colchicine for 36 h. The optimum medium for plant regeneration from polyploid hairy roots was MS+2.0 mg/L 6-BA +0.2 mg/L NAA. Compared with the control diploid plants, the hairy roots-regenerated plants had weak apical dominance, more axillary buds and more narrow leaves; whereas the polyploid hairy root-regenerated plants had thicker stems, shorter internodes and the colour, width and thickness of leaves were significantly higher than that of the control. Observation of the number of chromosomes in their root tip cells reveals that the obtained polyploid regenerated plants were tetraploidy, with 96 (4n = 96) chromosomes. Pot-grown experiments showed compared to the control, the flowering was delayed by 21 days in diploid hairy roots-regenerated plants and polyploid hairy root-regenerated plants. GC-MS detection shows that the content of nicotine in polyploid plants was about 6.90 and 4.57 times the control and the diploid hairy roots-regenerated plants, respectively.

  1. Allometric analysis of the induced flavonols on the leaf surface of wild tobacco (Nicotiana attenuata).

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    Roda, Amy L; Oldham, Neil J; Svatos, Ales; Baldwin, Ian T

    2003-02-01

    Trichomes excrete secondary metabolites that may alter the chemical composition of the leaf surface, reducing damage caused by herbivores, pathogens and abiotic stresses. We examined the surface exudates produced by Nicotiana attenuata Torr. Ex Wats., a plant known to contain and secrete a number of secondary metabolites that are toxic or a deterrent to herbivorous insects. Extractions specific to the leaf surface, the trichomes, and the laminar components demonstrated the localization of particular compounds. Diterpene glycosides occurred exclusively in leaf mesophyll, whereas nicotine was found in both the trichomes and mesophyll. Neither rutin nor nicotine was found on the leaf surface. Quercetin and 7 methylated derivatives were found in the glandular trichomes and appeared to be excreted onto the leaf surface. We examined the elicitation of these flavonols on the leaf surface with a surface-area allometric analysis, which measures changes in metabolites independent of the effects of leaf expansion. The flavonols responded differently to wounding, methyl jasmonate (MeJA), herbivore attack and UV-C radiation, and the response patterns corresponded to their compound-specific allometries. Finding greater amounts of quercetin on younger leaves and reduced amounts after herbivore feeding and MeJA treatment, we hypothesized that quercetin may function as an attractant, helping the insects locate a preferred feeding site. Consistent with this hypothesis, mirids (Tupiocoris notatus) were found more often on mature leaves sprayed with quercetin at a concentration typical of young leaves than on unsupplemented mature leaves. The composition of metabolites on the leaf surface of N. attenuata changes throughout leaf development and in response to herbivore attack or environmental stress, and these changes are mediated in part by responses of the glandular trichomes.

  2. Herbivore induction of jasmonic acid and chemical defences reduce photosynthesis in Nicotiana attenuata.

    Science.gov (United States)

    Nabity, Paul D; Zavala, Jorge A; DeLucia, Evan H

    2013-01-01

    Herbivory initiates a shift in plant metabolism from growth to defence that may reduce fitness in the absence of further herbivory. However, the defence-induced changes in carbon assimilation that precede this reallocation in resources remain largely undetermined. This study characterized the response of photosynthesis to herbivore induction of jasmonic acid (JA)-related defences in Nicotiana attenuata to increase understanding of these mechanisms. It was hypothesized that JA-induced defences would immediately reduce the component processes of photosynthesis upon attack and was predicted that wild-type plants would suffer greater reductions in photosynthesis than plants lacking JA-induced defences. Gas exchange, chlorophyll fluorescence, and thermal spatial patterns were measured together with the production of defence-related metabolites after attack and through recovery. Herbivore damage immediately reduced electron transport and gas exchange in wild-type plants, and gas exchange remained suppressed for several days after attack. The sustained reductions in gas exchange occurred concurrently with increased defence metabolites in wild-type plants, whereas plants lacking JA-induced defences suffered minimal suppression in photosynthesis and no increase in defence metabolite production. This suppression in photosynthesis occurred only after sustained defence signalling and defence chemical mobilization, whereas a short bout of feeding damage only transiently altered components of photosynthesis. It was identified that lipoxygenase signalling interacted with photosynthetic electron transport and that the resulting JA-related metabolites reduced photosynthesis. These data represent a metabolic cost to mounting a chemical defence against herbivory and link defence-signalling networks to the differential effects of herbivory on photosynthesis in remaining leaf tissues in a time-dependent manner.

  3. Metabolite Fingerprinting in Transgenic Nicotiana tabacum Altered by the Escherichia coli Glutamate Dehydrogenase Gene

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    R. Mungur

    2005-01-01

    Full Text Available With about 200 000 phytochemicals in existence, identifying those of biomedical significance is a mammoth task. In the postgenomic era, relating metabolite fingerprints, abundances, and profiles to genotype is also a large task. Ion analysis using Fourier transformed ion cyclotron resonance mass spectrometry (FT-ICR-MS may provide a high-throughput approach to measure genotype dependency of the inferred metabolome if reproducible techniques can be established. Ion profile inferred metabolite fingerprints are coproducts. We used FT-ICR-MS-derived ion analysis to examine gdhA (glutamate dehydrogenase (GDH; EC 1.4.1.1 transgenic Nicotiana tabacum (tobacco carrying out altered glutamate, amino acid, and carbon metabolisms, that fundamentally alter plant productivity. Cause and effect between gdhA expression, glutamate metabolism, and plant phenotypes was analyzed by 13NH4+ labeling of amino acid fractions, and by FT-ICR-MS analysis of metabolites. The gdhA transgenic plants increased 13N labeling of glutamate and glutamine significantly. FT-ICR-MS detected 2 012 ions reproducible in 2 to 4 ionization protocols. There were 283 ions in roots and 98 ions in leaves that appeared to significantly change abundance due to the measured GDH activity. About 58% percent of ions could not be used to infer a corresponding metabolite. From the 42% of ions that inferred known metabolites we found that certain amino acids, organic acids, and sugars increased and some fatty acids decreased. The transgene caused increased ammonium assimilation and detectable ion variation. Thirty-two compounds with biomedical significance were altered in abundance by GDH including 9 known carcinogens and 14 potential drugs. Therefore, the GDH transgene may lead to new uses for crops like tobacco.

  4. Androgenesis Induced in Nicotiana alata and the Effect of Gamma Irradiation

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    Ayman EL-FIKI

    2015-03-01

    Full Text Available Nicotiana alata anthers cultured on different modified media based on MS, MT and N were used to obtain haploid plants through direct and indirect ways. The haploid plants resulting on MS medium ranged from 52% - 80%, on MT medium ranged from 32% - 52% and on N medium ranged from 28% - 44%. Accordingly, the best medium used for haploid induction was MS supplemented with 0.2 mgl-l NAA + 0.5 mgl-l KIN. On the other hand, MS medium supplemented with 0.4 mgl-l NAA + 0.5 mgl-l KIN or 1.0 mgl-l BAP + 0.5 mgl-l NAA were the best mediums for callus induction and plant regeneration, respectively. Morphologically, the leaf size, stem highest and diameter, flower size and diameter, anther length and number were about 67% of the diploid plants growth. Irradiated anthers with doses of 0, 2.5, 5, 7.5, 10, 15, 20 and 25 Gy caused reducing the number of haploid plants with increasing gamma radiation dose. For the haploid plants irradiated with same doses, the mortality percentage of bud survival was increasing with increasing gamma radiation dose. The irradiated callus with doses of 0, 5, 10, 15 and 20 Gy was affected negatively on growth rate and morphology. Proline content in irradiated plantlets increased with increasing gamma radiation dose. As well, total soluble protein content was increased with gamma irradiation up to 10 Gy. However, the higher doses caused a severe decrease of total soluble proteins. The production of proline and total soluble proteins in haploid plants were 48.6% and 69.5%, respectively comparing with diploid plants.

  5. HSPRO controls early Nicotiana attenuata seedling growth during interaction with the fungus Piriformospora indica.

    Science.gov (United States)

    Schuck, Stefan; Camehl, Iris; Gilardoni, Paola A; Oelmueller, Ralf; Baldwin, Ian T; Bonaventure, Gustavo

    2012-10-01

    In a previous study aimed at identifying regulators of Nicotiana attenuata responses against chewing insects, a 26-nucleotide tag matching the HSPRO (ORTHOLOG OF SUGAR BEET Hs1(pro)(-)(1)) gene was found to be strongly induced after simulated herbivory (Gilardoni et al., 2010). Here we characterized the function of HSPRO during biotic interactions in transgenic N. attenuata plants silenced in its expression (ir-hspro). In wild-type plants, HSPRO expression was not only induced during simulated herbivory but also when leaves were inoculated with Pseudomonas syringae pv tomato DC3000 and roots with the growth-promoting fungus Piriformospora indica. Reduced HSPRO expression did not affect the regulation of direct defenses against Manduca sexta herbivory or P. syringae pv tomato DC3000 infection rates. However, reduced HSPRO expression positively influenced early seedling growth during interaction with P. indica; fungus-colonized ir-hspro seedlings increased their fresh biomass by 30% compared with the wild type. Grafting experiments demonstrated that reduced HSPRO expression in roots was sufficient to induce differential growth promotion in both roots and shoots. This effect was accompanied by changes in the expression of 417 genes in colonized roots, most of which were metabolic genes. The lack of major differences in the metabolic profiles of ir-hspro and wild-type colonized roots (as analyzed by liquid chromatography time-of-flight mass spectrometry) suggested that accelerated metabolic rates were involved. We conclude that HSPRO participates in a whole-plant change in growth physiology when seedlings interact with P. indica.

  6. Expression of Aspergillus nidulans phy Gene in Nicotiana benthamiana Produces Active Phytase with Broad Specificities

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    Tae-Kyun Oh

    2014-09-01

    Full Text Available A full-length phytase gene (phy of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR, and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5, an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F, the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.

  7. Expression of Aspergillus nidulans phy gene in Nicotiana benthamiana produces active phytase with broad specificities.

    Science.gov (United States)

    Oh, Tae-Kyun; Oh, Sung; Kim, Seongdae; Park, Jae Sung; Vinod, Nagarajan; Jang, Kyung Min; Kim, Sei Chang; Choi, Chang Won; Ko, Suk-Min; Jeong, Dong Kee; Udayakumar, Rajangam

    2014-09-03

    A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.

  8. Profiling of Altered Metabolomic States in Nicotiana tabacum Cells Induced by Priming Agents

    Science.gov (United States)

    Mhlongo, Msizi I.; Steenkamp, Paul A.; Piater, Lizelle A.; Madala, Ntakadzeni E.; Dubery, Ian A.

    2016-01-01

    Metabolomics has developed into a valuable tool for advancing our understanding of plant metabolism. Plant innate immune defenses can be activated and enhanced so that, subsequent to being pre-sensitized, plants are able to launch a stronger and faster defense response upon exposure to pathogenic microorganisms, a phenomenon known as priming. Here, three contrasting chemical activators, namely acibenzolar-S-methyl, azelaic acid and riboflavin, were used to induce a primed state in Nicotiana tabacum cells. Identified biomarkers were then compared to responses induced by three phytohormones—abscisic acid, methyljasmonate, and salicylic acid. Altered metabolomes were studied using a metabolite fingerprinting approach based on liquid chromatography and mass spectrometry. Multivariate data models indicated that these inducers cause time-dependent metabolic perturbations in the cultured cells and revealed biomarkers of which the levels are affected by these agents. A total of 34 metabolites were annotated from the mass spectral data and online databases. Venn diagrams were used to identify common biomarkers as well as those unique to a specific agent. Results implicate 20 cinnamic acid derivatives conjugated to (i) quinic acid (chlorogenic acids), (ii) tyramine, (iii) polyamines, or (iv) glucose as discriminatory biomarkers of priming in tobacco cells. Functional roles for most of these metabolites in plant defense responses could thus be proposed. Metabolites induced by the activators belong to the early phenylpropanoid pathway, which indicates that different stimuli can activate similar pathways but with different metabolite fingerprints. Possible linkages to phytohormone-dependent pathways at a metabolomic level were indicated in the case of cells treated with salicylic acid and methyljasmonate. The results contribute to a better understanding of the priming phenomenon and advance our knowledge of cinnamic acid derivatives as versatile defense metabolites. PMID

  9. Production of an active anti-CD20-hIL-2 immunocytokine in Nicotiana benthamiana.

    Science.gov (United States)

    Marusic, Carla; Novelli, Flavia; Salzano, Anna M; Scaloni, Andrea; Benvenuto, Eugenio; Pioli, Claudio; Donini, Marcello

    2016-01-01

    Anti-CD20 murine or chimeric antibodies (Abs) have been used to treat non-Hodgkin lymphomas (NHLs) and other diseases characterized by overactive or dysfunctional B cells. Anti-CD20 Abs demonstrated to be effective in inducing regression of B-cell lymphomas, although in many cases patients relapse following treatment. A promising approach to improve the outcome of mAb therapy is the use of anti-CD20 antibodies to deliver cytokines to the tumour microenvironment. In particular, IL-2-based immunocytokines have shown enhanced antitumour activity in several preclinical studies. Here, we report on the engineering of an anti-CD20-human interleukin-2 (hIL-2) immunocytokine (2B8-Fc-hIL2) based on the C2B8 mAb (Rituximab) and the resulting ectopic expression in Nicotiana benthamiana. The scFv-Fc-engineered immunocytokine is fully assembled in plants with minor degradation products as assessed by SDS-PAGE and gel filtration. Purification yields using protein-A affinity chromatography were in the range of 15-20 mg/kg of fresh leaf weight (FW). Glycopeptide analysis confirmed the presence of a highly homogeneous plant-type glycosylation. 2B8-Fc-hIL2 and the cognate 2B8-Fc antibody, devoid of hIL-2, were assayed by flow cytometry on Daudi cells revealing a CD20 binding activity comparable to that of Rituximab and were effective in eliciting antibody-dependent cell-mediated cytotoxicity of human PBMC versus Daudi cells, demonstrating their functional integrity. In 2B8-Fc-hIL2, IL-2 accessibility and biological activity were verified by flow cytometry and cell proliferation assay. To our knowledge, this is the first example of a recombinant immunocytokine based on the therapeutic Rituximab antibody scaffold, whose expression in plants may be a valuable tool for NHLs treatment.

  10. Optimisation of contained Nicotiana tabacum cultivation for the production of recombinant protein pharmaceuticals.

    Science.gov (United States)

    Colgan, Richard; Atkinson, Christopher J; Paul, Matthew; Hassan, Sally; Drake, Pascal M W; Sexton, Amy L; Santa-Cruz, Simon; James, David; Hamp, Keith; Gutteridge, Colin; Ma, Julian K-C

    2010-04-01

    Nicotiana tabacum is emerging as a crop of choice for production of recombinant protein pharmaceuticals. Although there is significant commercial expertise in tobacco farming, different cultivation practices are likely to be needed when the objective is to optimise protein expression, yield and extraction, rather than the traditional focus on biomass and alkaloid production. Moreover, pharmaceutical transgenic tobacco plants are likely to be grown initially within a controlled environment, the parameters for which have yet to be established. Here, the growth characteristics and functional recombinant protein yields for two separate transgenic tobacco plant lines were investigated. The impacts of temperature, day-length, compost nitrogen content, radiation and plant density were examined. Temperature was the only environmental variable to affect IgG concentration in the plants, with higher yields observed in plants grown at lower temperature. In contrast, temperature, supplementary radiation and plant density all affected the total soluble protein yield in the same plants. Transgenic plants expressing a second recombinant protein (cyanovirin-N) responded differently to IgG transgenic plants to elevated temperature, with an increase in cyanovirin-N concentration, although the effect of the environmental variables on total soluble protein yields was the same as the IgG plants. Planting density and radiation levels were important factors affecting variability of the two recombinant protein yields in transgenic plants. Phenotypic differences were observed between the two transgenic plant lines and non-transformed N. tabacum, but the effect of different growing conditions was consistent between the three lines. Temperature, day length, radiation intensity and planting density all had a significant impact on biomass production. Taken together, the data suggest that recombinant protein yield is not affected substantially by environmental factors other than growth

  11. Host-plant-mediated effects of Nadefensin on herbivore and pathogen resistance in Nicotiana attenuata

    Directory of Open Access Journals (Sweden)

    Baldwin Ian T

    2008-10-01

    Full Text Available Abstract Background The adage from Shakespeare, "troubles, not as single spies, but in battalions come," holds true for Nicotiana attenuata, which is commonly attacked by both pathogens (Pseudomonas spp. and herbivores (Manduca sexta in its native habitats. Defense responses targeted against the pathogens can directly or indirectly influence the responses against the herbivores. Nadefensin is an effective induced defense gene against the bacterial pathogen Pseudomonas syringae pv tomato (PST DC3000, which is also elicited by attack from M. sexta larvae, but whether this defense protein influences M. sexta's growth and whether M. sexta-induced Nadefensin directly or indirectly influences PST DC3000 resistance are unknown. Results M. sexta larvae consumed less on WT and on Nadefensin-silenced N. attenuata plants that had previously been infected with PST DC3000 than on uninfected plants. WT plants infected with PST DC3000 showed enhanced resistance to PST DC3000 and decreased leaf consumption by M. sexta larvae, but larval mass gain was unaffected. PST DC3000-infected Nadefensin-silenced plants were less resistant to subsequent PST DC3000 challenge, and on these plants, M. sexta larvae consumed less and gained less mass. WT and Nadefensin-silenced plants previously damaged by M. sexta larvae were better able to resist subsequent PST DC3000 challenges than were undamaged plants. Conclusion These results demonstrate that Na-defensin directly mediates defense against PST DC3000 and indirectly against M. sexta in N. attenuata. In plants that were previously infected with PST DC3000, the altered leaf chemistry in PST DC3000-resistant WT plants and PST DC3000-susceptible Nadefensin-silenced plants differentially reduced M. sexta's leaf consumption and mass gain. In plants that were previously damaged by M. sexta, the combined effect of the altered host plant chemistry and a broad spectrum of anti-herbivore induced metabolomic responses was more

  12. Reference Gene Selection in the Desert Plant <em>Eremosparton songoricuem>m>

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    Dao-Yuan Zhang

    2012-06-01

    Full Text Available <em>Eremosparton songoricum em>(Litv. Vass. (<em>E. songoricumem> is a rare and extremely drought-tolerant desert plant that holds promise as a model organism for the identification of genes associated with water deficit stress. Here, we cloned and evaluated the expression of eight candidate reference genes using quantitative real-time reverse transcriptase polymerase chain reactions. The expression of these candidate reference genes was analyzed in a diverse set of 20 samples including various <em>E. songoricumem> plant tissues exposed to multiple environmental stresses. GeNorm analysis indicated that expression stability varied between the reference genes in the different experimental conditions, but the two most stable reference genes were sufficient for normalization in most conditions.<em> EsEFem> and <em>Esα-TUB> were sufficient for various stress conditions, <em>EsEF> and <em>EsACT> were suitable for samples of differing germination stages, and <em>EsGAPDH>and <em>Es>UBQ em>were most stable across multiple adult tissue samples. The <em>Es18Sem> gene was unsuitable as a reference gene in our analysis. In addition, the expression level of the drought-stress related transcription factor <em>EsDREB2em>> em>verified the utility of<em> E. songoricumem> reference genes and indicated that no single gene was adequate for normalization on its own. This is the first systematic report on the selection of reference genes in <em>E. songoricumem>, and these data will facilitate future work on gene expression in this species.

  13. Coat protein-mediated resistance against an Indian isolate of the Cucumber mosaic virus subgroup IB in Nicotiana benthamiana

    Indian Academy of Sciences (India)

    A Srivastava; S K Raj

    2008-06-01

    Coat protein (CP)-mediated resistance against an Indian isolate of the Cucumber mosaic virus (CMV) subgroup IB was demonstrated in transgenic lines of Nicotiana benthamiana through Agrobacterium tumefaciens-mediated transformation. Out of the fourteen independently transformed lines developed, two lines were tested for resistance against CMV by challenge inoculations. The transgenic lines exhibiting complete resistance remained symptomless throughout life and showed reduced or no virus accumulation in their systemic leaves after virus challenge. These lines also showed virus resistance against two closely related strains of CMV. This is the first report of CP-mediated transgenic resistance against a CMV subgroup IB member isolated from India.

  14. INFLUENCE OF STATIONARY MAGNETIC FIELD ON THE EARLY STAGES OF THE DEVELOPMENT OF TOBACCO SEEDS (NICOTIANA TABACUM L.

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    ANNA ALADJADJIYAN

    2003-10-01

    Full Text Available The influence of a stationary magnetic field with induction of 0,15 Т at expositions 10 min, 20 min и 30 min, on tobacco seeds (Nicotiana tabacum L., cv. Harmanly 11 has been investigated. The germination energy GE and the germination G have been determined with the aim of establishing the effect of the magnetic field on seed vitality. It has been found out that the magnetic field stimulates the development of the germ and leads to increasing the germination energy and germination. A hypothesis about the explanation of the results obtained has been proposed, especially about the stimulating effect of the magnetic field treatment.

  15. Virus-induced gene silencing reveals signal transduction components required for the Pvr9-mediated hypersensitive response in Nicotiana benthamiana.

    Science.gov (United States)

    Tran, Phu-Tri; Choi, Hoseong; Choi, Doil; Kim, Kook-Hyung

    2016-08-01

    Resistance to pathogens mediated by plant resistance (R) proteins requires different signaling transduction components and pathways. Our previous studies revealed that a potyvirus resistance gene in pepper, Pvr9, confers a hypersensitive response (HR) to pepper mottle virus in Nicotiana benthamiana. Our results show that the Pvr9-mediated HR against pepper mottle virus infection requires HSP90, SGT1, NDR1, but not EDS1. These results suggest that the Pvr9-mediated HR is possibly related to the SA pathway but not the ET, JA, ROS or NO pathways.

  16. Flower-bud formation in explants of photoperiodic and day-neutral Nicotiana biotypes and its bearing on the regulation of flower formation

    Energy Technology Data Exchange (ETDEWEB)

    Rajeevan, M.S.; Lang, A. (Michigan State Univ., East Lansing (United States))

    1993-05-15

    The capacity to form flower buds in thin-layer explants was studied in Nicotiana of several species, cultivars, and lines of differing in their response to photoperiod. This capacity was found in all biotypes examined and could extend into sepals and corolla. It varied depending on genotype, source tissue and its developmental state, and composition of the culture medium, particularly the levels of glucose, auxin, and cytokinin. It was greatest in the two day-neutral plants examined, Samsun tobacco and Nicotiana rustica, where it extended from the inflorescence region down the vegetative stem, in a basipetally decreasing gradient; it was least in the two qualitative photoperiodic plants studied, the long-day plant Nicotiana silvestris and the short-day plant Maryland Mammoth tobacco, the quantitative long-day plant Nicotiana alata and the quantitative short-day plant Nicotiana otophora line 38-G-81, where it was limited to the pedicels (and, in some cases, the sepals). Regardless of the photoperiodic response of the source plants, the response was the same in explants cultured under long and short days. The capacity to form flow buds in explants is present in all Nicotiana biotypes studied supports the idea that it is regulated by the same mechanism(s), regardless of the plant's photoperiodic character. However, flower formation in the explants is not identical with de novo flower formation in a hitherto vegetative plant: it is rather the expression of a floral state already established in the plant, although it can vary widely in extent and spatial distribution. Culture conditions that permit flower-bud formation in an explant are conditions that maintain the floral state and encourage its expression; conditions under which no flower buds are formed reduce this state and/or prevent its expression. 14 refs., 5 figs., 3 tabs.

  17. A survey of resistance to Tomato bushy stunt virus in the genus Nicotiana reveals that the hypersensitive response is triggered by one of three different viral proteins.

    Science.gov (United States)

    Angel, Carlos A; Schoelz, James E

    2013-02-01

    In this study, we screened 22 Nicotiana spp. for resistance to the tombusviruses Tomato bushy stunt virus (TBSV), Cucumber necrosis virus, and Cymbidium ringspot virus. Eighteen species were resistant, and resistance was manifested in at least two different categories. In all, 13 species responded with a hypersensitive response (HR)-type resistance, whereas another five were resistant but either had no visible response or responded with chlorotic lesions rather than necrotic lesions. Three different TBSV proteins were found to trigger HR in Nicotiana spp. in an agroinfiltration assay. The most common avirulence (avr) determinant was the TBSV coat protein P41, a protein that had not been previously recognized as an avr determinant. A mutational analysis confirmed that the coat protein rather than the viral RNA sequence was responsible for triggering HR, and it triggered HR in six species in the Alatae section. The TBSV P22 movement protein triggered HR in two species in section Undulatae (Nicotiana glutinosa and N. edwardsonii) and one species in section Alatae (N. forgetiana). The TBSV P19 RNA silencing suppressor protein triggered HR in sections Sylvestres (N. sylvestris), Nicotiana (N. tabacum), and Alatae (N. bonariensis). In general, Nicotiana spp. were capable of recognizing only one tombusvirus avirulence determinant, with the exceptions of N. bonariensis and N. forgetiana, which were each able to recognize P41, as well as P19 and P22, respectively. Agroinfiltration failed to detect the TBSV avr determinants responsible for triggering HR in N. arentsii, N. undulata, and N. rustica. This study illustrates the breadth and variety of resistance responses to tombusviruses that exists in the Nicotiana genus.

  18. Estudos sobre a resistência ao jejum e aspectos nutricionais de Triatoma lecticularia (Stal, 1859 (Hemiptera, Reduviidae, Triatominae Studies on the resistance to fasting and nutritional aspects of Triatoma lecticularia (Stal, 1859 (Hemiptera, Reduviidae, Triatominae

    Directory of Open Access Journals (Sweden)

    José Jurberg

    1989-09-01

    Full Text Available Foi feito um estudo sobre a resistência ao jejum em todas as fases evolutivas e pesagens em diferentes situações nutricionais de Triatoma lecticularia (alimentado; não alimentado; na morte após o jejum com temperatura e umidade registradas. Observou-se que os períodos de resistência das fases ninfais apresentaram médias (dias crescentes: 1º: 45,84; 2º: 61; 3º: 88,74; 4º: 123,47; 5º 162,30. Na fase adulta as médias foram aproximadas à do 3º estádio (para os machos 88,94 e para as fêmeas 83,66. O procedimento de pesagens permitiu registrar a quantidade de sangue ingerido, a perda de peso durante o jejum e o respectivo percentual em relação ao peso inicial. Esta espécie tem assinalada sua distribuição geográfica na região Neártica, onde tem sido encontrda infectada com Trypanosoma cruzi associada a Neotoma micropus Baird e Spermophilus variagatus (Erxeleben.The resistance to fasting of Triatoma lecticularia was studied in all phases of the life cycle and the insect weighed in different nutritional situations (fed, not fed, death after starvation. The temperature and humidity level were also recorded. The nymphal phases showed increasing resistance to fasting as demonstrated by the following averages (days 1st 45.84; 2nd 61; 3rd 88.74; 4th 123.44; 5th 162.30. Upon the adult phase, the averages were similar to those of the 3rd stage for the male insects 88.94 and for females 83.66. The weighing technique allowed for the measeurement of the quantity of blood ingested, the weight loss during the fast and the percentage weight lost as related to the initial weight. The species is found in the Neartic region where it has been found infected with Trypanosoma cruzi and is associated with the terrestial rodents Neotoma micropus Baird and Spermophilus variagatus (Erxelebem.

  19. Exceptional inheritance of plastids via pollen in Nicotiana sylvestris with no detectable paternal mitochondrial DNA in the progeny.

    Science.gov (United States)

    Thyssen, Gregory; Svab, Zora; Maliga, Pal

    2012-10-01

    Plastids and mitochondria, the DNA-containing cytoplasmic organelles, are maternally inherited in the majority of angiosperm species. Even in plants with strict maternal inheritance, exceptional paternal transmission of plastids has been observed. Our objective was to detect rare leakage of plastids via pollen in Nicotiana sylvestris and to determine if pollen transmission of plastids results in co-transmission of paternal mitochondria. As father plants, we used N. sylvestris plants with transgenic, selectable plastids and wild-type mitochondria. As mother plants, we used N. sylvestris plants with Nicotiana undulata cytoplasm, including the CMS-92 mitochondria that cause cytoplasmic male sterility (CMS) by homeotic transformation of the stamens. We report here exceptional paternal plastid DNA in approximately 0.002% of N. sylvestris seedlings. However, we did not detect paternal mitochondrial DNA in any of the six plastid-transmission lines, suggesting independent transmission of the cytoplasmic organelles via pollen. When we used fertile N. sylvestris as mothers, we obtained eight fertile plastid transmission lines, which did not transmit their plastids via pollen at higher frequencies than their fathers. We discuss the implications for transgene containment and plant evolutionary histories inferred from cytoplasmic phylogenies.

  20. Spatial analysis of the incidence of Phytophthora infestans (Mont. De Bary and Phytophthora nicotianae Breda de Haan on potato

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    Leónides Castellanos González

    2016-03-01

    Full Text Available The objective of this research was to conduct a spatial analysis of the incidence of Phytophthora nicotianae Breda de Haan and Phytophthora infestans (Mont. De Bary, during five seasons of potato in Fields Company Miscellaneous Crops in Horquita, Cienfuegos (2003-2004 to 2007-2008. Information about pathogens, collected by the Plant Protection Station of Yaguaramas, was used in order to do so. An alpha numerical database was made, oriented to a group of areas under center pivot irrigation machines (Kubans and Bayamón and its quadrants. Parallel to this, mapping of center pivot machines and quadrants was generated in MapInfo GIS 8.5. Several automatic geo codifications were made in order to relate the alphanumeric database and the mapping, and thematic maps were generated in the presence or absence of pathogens. The two study agents have presented high spatial variability during the five planting seasons. Both have influenced the same geographical area with similar appearance date. P. infestans has not expressed a defined dispersion pattern and it has spread at random from the primary source of incidence, while Phytophthora nicotianae has become an endemic agent which shows a dispersion pattern towards neighboring areas or areas connected by the road network from the primary sources, which have been associated with poor land leveling and late or intermediate planting seasons.

  1. Pollination ecology of the invasive tree tobacco Nicotiana glauca: comparisons across native and non-native ranges

    Directory of Open Access Journals (Sweden)

    Jeff Ollerton

    2012-10-01

    Full Text Available Interactions with pollinators are thought to play a significant role in determining whether plant species become invasive, and ecologically generalised species are predicted to be more likely to invade than more specialised species. Using published and unpublished data we assessed the floral biology and pollination ecology of the South American native Nicotiana glauca (Solanaceae which has become a significant invasive of semi-arid parts of the world. In regions where specialised bird pollinators are available, for example hummingbirds in California and sunbirds in South Africa and Israel, N. glauca interacts with these local pollinators and sets seed by both out-crossing and selfing. In areas where there are no such birds, such as the Canary Islands and Greece, abundant viable seed is set by selfing, facilitated by the shorter stigma-anther distance compared to plants in native populations. Surprisingly, in these areas without pollinating birds, the considerable nectar resources are only rarely exploited by other flower visitors such as bees or butterflies, either legitimately or by nectar robbing. We conclude that Nicotiana glauca is a successful invasive species outside of its native range, despite its functionally specialised hummingbird pollination system, because it has evolved to become more frequently self pollinating in areas where it is introduced. Its invasion success is not predictable from what is known of its interactions with pollinators in its home range.

  2. A reversed-phase HPLC-UV method developed and validated for simultaneous quantification of six alkaloids from Nicotiana spp.

    Science.gov (United States)

    Moghbel, Nahid; Ryu, BoMi; Steadman, Kathryn J

    2015-08-01

    A reversed-phase HPLC-UV method was developed, optimized, and validated for the separation and quantitation of six target alkaloids from leaves of Nicotiana species (nicotine, nornicotine, anatabine, anabasine, myosmine, and cotinine). A bidentate reversed-phase C18 column was used as stationary phase and an alkaline ammonium formate buffer and acetonitrile as mobile phase. The alkaloids were well separated in a short run time of 13min with mobile phase pH 10.5 and a small gradient of 9-13% acetonitrile, and detected using UV at 260nm. Peak parameters were acceptable for all six closely related alkaloids. The proposed method has enough linearity with correlation coefficient >0.999 within the investigated range for all tested alkaloids. Satisfactory precision was achieved for both intra- and inter-day assay, with RSD less than 2% for all alkaloid standards. Reproducibility was also within the acceptable range of RSD alkaloids. The limit of quantification was 2.8 and 4.8μg/mL for nornicotine and nicotine respectively, and below 2μg/mL for all other alkaloids. The method was successfully applied for simultaneous analysis of alkaloids in leaves of Nicotiana benthamiana.

  3. Putative Serine Protease Effectors of Clavibacter michiganensis Induce a Hypersensitive Response in the Apoplast of Nicotiana Species.

    Science.gov (United States)

    Lu, You; Hatsugai, Noriyuki; Katagiri, Fumiaki; Ishimaru, Carol A; Glazebrook, Jane

    2015-11-01

    Clavibacter michiganensis subspp. michiganensis and sepedonicus cause diseases on solanaceous crops. The genomes of both subspecies encode members of the pat-1 family of putative serine proteases known to function in virulence on host plants and induction of hypersensitive responses (HR) on nonhosts. One gene of this family in C. michiganensis subsp. sepedonicus, chp-7, is required for triggering HR in Nicotiana tabacum. Here, further investigation revealed that mutation of the putative catalytic serine residue at position 232 to threonine abolished the HR induction activity of Chp-7, suggesting that enzymatic activity is required. Purified Chp-7 triggered an HR in N. tabacum leaves in the absence of the pathogen, indicating Chp-7 itself is the HR elicitor from C. michiganensis subsp. sepedonicus. Ectopic expression of chp-7 constructs in N. tabacum leaves revealed that Chp-7 targeted to the apoplast triggered an HR while cytoplasmic Chp-7 did not, indicating that Chp-7 induces the HR in the apoplast of N. tabacum leaves. Chp-7 also induced HR in N. sylvestris, a progenitor of N. tabacum, but not in other Nicotiana species tested. ChpG, a related protein from C. michiganensis subsp. michiganensis, also triggered HR in N. tabacum and N. sylvestris. Unlike Chp-7, ChpG triggered HR in N. clevelandii and N. glutinosa.

  4. The Cotton WRKY Gene GhWRKY41 Positively Regulates Salt and Drought Stress Tolerance in Transgenic Nicotiana benthamiana.

    Directory of Open Access Journals (Sweden)

    Xiaoqian Chu

    Full Text Available WRKY transcription factors constitute a very large family of proteins in plants and participate in modulating plant biological processes, such as growth, development and stress responses. However, the exact roles of WRKY proteins are unclear, particularly in non-model plants. In this study, Gossypium hirsutum WRKY41 (GhWRKY41 was isolated and transformed into Nicotiana benthamiana. Our results showed that overexpression of GhWRKY41 enhanced the drought and salt stress tolerance of transgenic Nicotiana benthamiana. The transgenic plants exhibited lower malondialdehyde content and higher antioxidant enzyme activity, and the expression of antioxidant genes was upregulated in transgenic plants exposed to osmotic stress. A β-glucuronidase (GUS staining assay showed that GhWRKY41 was highly expressed in the stomata when plants were exposed to osmotic stress, and plants overexpressing GhWRKY41 exhibited enhanced stomatal closure when they were exposed to osmotic stress. Taken together, our findings demonstrate that GhWRKY41 may enhance plant tolerance to stress by functioning as a positive regulator of stoma closure and by regulating reactive oxygen species (ROS scavenging and the expression of antioxidant genes.

  5. Two faces of Solanaceae telomeres: a comparison between Nicotiana and Cestrum telomeres and telomere-binding proteins.

    Science.gov (United States)

    Peska, V; Sýkorová, E; Fajkus, J

    2008-01-01

    While most Solanaceae genera (e.g.Solanum, Nicotiana) possess Arabidopsis-type telomeres of (TTTAGGG)n maintained by telomerase, the genera Cestrum, Vestia and Sessea (Cestrum group) lack these telomeres. Here we show that in the Cestrum-group the activity of telomerase has been lost. Nevertheless, proteins binding the single-stranded G-rich strand of the Arabidopsis-type and related human-type (TTAGGG)n telomeric sequences are present in nuclear extracts of both Nicotiana and Cestrum species. These proteins may have a role in telomere function or other cellular activities. In addition to characterizing DNA binding specificity and molecular weights of these proteins, we searched in both N. tabacum (tobacco) and C. parqui for the presence of POT1-like proteins, involved in telomere capping and telomerase regulation. Analysis of POT1-like proteins available on public databases and cloned by us from C. parqui, revealed the N-terminal OB folds typical for this protein family and a novel, plant-specific conserved C-terminal OB-fold domain (CTOB). We propose that CTOB is involved in protein-protein interactions.

  6. Cloning of the Lycopene β-cyclase Gene in Nicotiana tabacum and Its Overexpression Confers Salt and Drought Tolerance

    Directory of Open Access Journals (Sweden)

    Yanmei Shi

    2015-12-01

    Full Text Available Carotenoids are important pigments in plants that play crucial roles in plant growth and in plant responses to environmental stress. Lycopene β cyclase (β-LCY functions at the branch point of the carotenoid biosynthesis pathway, catalyzing the cyclization of lycopene. Here, a β-LCY gene from Nicotiana tabacum, designated as Ntβ-LCY1, was cloned and functionally characterized. Robust expression of Ntβ-LCY1 was found in leaves, and Ntβ-LCY1 expression was obviously induced by salt, drought, and exogenous abscisic acid treatments. Strong accumulation of carotenoids and expression of carotenoid biosynthesis genes resulted from Ntβ-LCY1 overexpression. Additionally, compared to wild-type plants, transgenic plants with overexpression showed enhanced tolerance to salt and drought stress with higher abscisic acid levels and lower levels of malondialdehyde and reactive oxygen species. Conversely, transgenic RNA interference plants had a clear albino phenotype in leaves, and some plants did not survive beyond the early developmental stages. The suppression of Ntβ-LCY1 expression led to lower expression levels of genes in the carotenoid biosynthesis pathway and to reduced accumulation of carotenoids, chlorophyll, and abscisic acid. These results indicate that Ntβ-LCY1 is not only a likely cyclization enzyme involved in carotenoid accumulation but also confers salt and drought stress tolerance in Nicotiana tabacum.

  7. Changes in phenolic compound, TAL, PAL activity of Nicotiana rustica triggered by ethanolamine pretreatment under in vitro salt stress condition

    Directory of Open Access Journals (Sweden)

    Ali Akbar Ehsanpour

    2015-12-01

    Full Text Available Salt stress is one of the most important factors that endanger plant growth and production all around the world. Application of compounds which increase plant resistance to salt stress obviously has significant importance in reduction of economic losses. One of these compounds is the biological alcohol called ethanolamine, and the present study aimed to understand the way by which ethanolamine exerts its protecting effect against salt stress. For this purpose, 4-week-old Nicotiana rustica plants which had been kept in MS medium were pretreated with ethanolamine and 2 days later they were transferred to MS medium supplemented with 200 mM NaCl for 3 weeks. The our results revealed that Nicotiana rustica plants pretreated with exogenous ethanolamine showed elevated level of TAL activity and anthocyanin content whereas PAL activity was decreased, and reduced level of MDA, total flavonoid and phenol content in plants under salt stress condition. According to these results, it can be suggested that ethanolamine exerts its protective effect by anthocyanin content increase.

  8. Reversal by Calcium Ions of the Growth Inhibition of Debaryomyces nicotianae Caused by Antifungal Polyene Antibiotics1

    Science.gov (United States)

    Berdicevsky, Israela; Grossowicz, Nathan

    1972-01-01

    Only Debaryomyces nicotianae strain 77, of seven different yeast strains tested, was found to be resistant to heptamycin and other antifungal heptaenes when grown in a rich medium. This strain, however, like the other six, was completely susceptible to these antibiotics in a minimal medium. Addition of yeast extract to the minimal medium abolished the heptamycin effect; calcium ions fully duplicated the effect of yeast extract; Mg2+ and Mn2+ were also effective but less so than Ca2+. Ca2+ also counteracted the activity of the heptaenes ascosin and trichomycin. Complete reversal of the polyene inhibition by Ca2+ was obtained if the cation was added simultaneously with the antibiotic; addition of Ca2+ 2 hr after the polyene was without effect. Addition of Ca2+ in the absence of the polyene caused a slight, if any, growth stimulation of D. nicotianae 77. Cholesterol also counteracted polyene activity; this was due to the formation of a complex with the antibiotic which prevented the polyene from reaching the site of action—the cytoplasmic membrane. No evidence for complex formation between heptamycin and calcium was found. The importance of Ca2+ in membrane structure, as evidenced from heptaene studies, is discussed. PMID:4598328

  9. Phytoaccumulation of lead by sunflower (Helianthus annuus), tobacco (Nicotiana tabacum), and vetiver (Vetiveria zizanioides).

    Science.gov (United States)

    Boonyapookana, Benjaporn; Parkpian, Preeda; Techapinyawat, Sombun; DeLaune, R D; Jugsujinda, Aroon

    2005-01-01

    The ability of three plant species: Helianthus annuus, Nicotiana tabacum, and Vetiveria zizanioides for phytoaccumulation of Pb was studied. Plants were grown in hydroponic solution containing Pb(NO3)2 at concentration of 0.25 and 2.5 mM Pb in the presence or absence of chelating agents (EDTA or DTPA). Lead (Pb) transport and localization within the tissues of the plant species was determined using scanning electron microscope equipped with energy dispersive X-ray spectrometers (SEM-EDS). The addition of chelators increased Pb uptake as compared to plants grown in solution containing Pb alone. Lead taken up by the plant species were concentrated in both leaf and stem at the region of vascular bundles with greater amounts in the leaf portion. Lead granules were also found in the H. annuus root tissue from the epidermis layer to the central axis. After four weeks of growth a 23-fold increase in shoot Pb content for H. annuus and N. tabacum and 17-fold increase in shoot Pb for V. zizanioides resulted from plants grown in the 2.5 mM Pb-EDTA treatment. The higher Pb treatment (2.5 mM Pb containing EDTA) resulted in higher concentrations of Pb in plant tissue at the fourth week of exposure as compared to Pb treatment containing DTPA. Overall, Pb accumulation potential of H. annuus was greater than that of N. tabacum and V. zizanioides as indicated by the bioconcentration factor (171, 70, and 88, respectively). The highest measured Pb concentrations were found in H. annuus roots, stems, and leaves (2668, 843, and 3611 microg/g DW, respectively) grown in the 2.5 mM Pb-EDTA treatment. The addition of chelators caused some reduction in plant growth and biomass. Results showed that the three plant species tested have potential for use in phytoaccumulation of Pb since the Pb was concentrated in leaf and stem as compared to control plants. H. annuus however best meet the prerequisites for a hyperaccumulator plant and would have the potential for use in the restoration of

  10. Ionomic profiling of Nicotiana langsdorffii wild-type and mutant genotypes exposed to abiotic stresses.

    Science.gov (United States)

    Ardini, Francisco; Soggia, Francesco; Abelmoschi, Maria Luisa; Magi, Emanuele; Grotti, Marco

    2013-01-01

    To provide a new insight into the response of plants to abiotic stresses, the ionomic profiles of Nicotiana langsdorffii specimens have been determined before and after exposure to toxic metals (chromium) or drought conditions. The plants were genetically transformed with the rat glucocorticoid receptor (GR) or the gene for Agrobacterium rhizogenes rolC, because these modifications are known to produce an imbalance in phytohormone equilibria and a significant change in the defence response of the plant. Elemental profiles were obtained by developing and applying analytical procedures based on inductively coupled plasma atomic emission and mass spectrometry (ICP-AES/MS). In particular, the removal of isobaric interferences affecting the determination of Cr and V by ICP-MS was accomplished by use of a dynamic reaction cell, after optimization of the relevant conditions. The combined use of ICP atomic emission and mass spectrometry enabled the determination of 29 major and trace elements (Ba, Bi, Ca, Cd, Co, Cr, Cu, Eu, Fe, Ga, K, Li, Mg, Mn, Mo, Na, P, Pb, Pt, Rb, S, Sb, Sn, Sr, Te, V, W, Y, and Zn) in different parts of the plants (roots, stems, and leaves), with high accuracy and precision. Multivariate data processing and study of element distribution patterns provided new information about the ionomic response of the target organism to chemical treatment or water stress. Genetic modification mainly affected the distribution of Bi, Cr, Mo, Na, and S, indicating that these elements were involved in biochemical processes controlled by the GR or rolC genes. Chemical stress strongly affected accumulation of several elements (Ba, Ca, Fe, Ga, K, Li, Mn, Mo, Na, P, Pb, Rb, S, Sn, Te, V, and Zn) in different ways; for Ca, Fe, K, Mn, Na, and P the effect was quite similar to that observed in other studies after treatment with other transition elements, for example Cu and Cd. The effect of water deficit was less evident, mainly consisting in a decrease of Ba, Cr, Na, and Sr

  11. Production of the main celiac disease autoantigen by transient expression in Nicotiana benthamiana

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    Vanesa Soledad Marin Viegas

    2015-12-01

    Full Text Available Celiac Disease (CD is a gluten sensitive enteropathy that remains widely undiagnosed and implementation of massive screening tests is needed to reduce the long term complications associated to untreated CD. The main CD autoantigen, human tissue transglutaminase (TG2, is a challenge for the different expression systems available since its cross-linking activity affects cellular processes. Plant-based transient expression systems can be an alternative for the production of this protein. In this work, a transient expression system for the production of human TG2 in Nicotiana benthamiana leaves was optimized and reactivity of plant-produced TG2 in CD screening test was evaluated. First, a subcellular targeting strategy was tested. Cytosolic, secretory, endoplasmic reticulum (C-terminal SEKDEL fusion and vacuolar (C-terminal KISIA fusion TG2 versions were transiently expressed in leaves and recombinant protein yields were measured. ER-TG2 and vac-TG2 levels were 9 to 16 fold higher than their cytosolic and secretory counterparts. As second strategy, TG2 variants were co-expressed with a hydrophobic elastin-like polymer (ELP construct encoding for 36 repeats of the pentapeptide VPGXG in which the guest residue X were V and F in ratio 8:1. Protein bodies (PB were induced by the ELP, with a consequent 2 fold-increase in accumulation of both ER-TG2 and vac-TG2. Subsequently, ER-TG2 and vac-TG2 were produced and purified using immobilized metal ion affinity chromatography. Plant purified ER-TG2 and vac-TG2 were recognized by three anti-TG2 monoclonal antibodies that bind different epitopes proving that plant-produced antigen has immunochemical characteristics similar to those of human TG2. Lastly, an ELISA was performed with sera of CD patients and healthy controls. Both vac-TG2 and ER-TG2 were positively recognized by IgA of CD patients while they were not recognized by serum from non-celiac controls. These results confirmed the usefulness of plant

  12. EFECTO DE LA MADUREZ EN LA CRIOCONSERVACIÓN DE SEMILLAS DE Nicotiana tabacum L.

    Directory of Open Access Journals (Sweden)

    Juan L. Pérez-Rodríguez

    2016-01-01

    Full Text Available La presente investigación tuvo como objetivo determinar el efecto de la madurez en la crioconservación de semillas de Nicotiana tabacum L. cv Sancti Spíritus 96 (SS-96. El estudio se realizó en la Estación experimental del tabaco de Cabaiguán, Sancti Spíritus, perteneciente al Instituto de Investigaciones del Tabaco, entre los años 2012 y 2014. Las semillas de SS-96 se colectaron a los 14, 21, 28, 35, 42, 49 y 56 días después de la antesis (DDA y se les determinó masa fresca y masa seca de 1000 semillas, porcentaje de humedad (base masa fresca y potencia germinativa (PG, además de evaluar su tolerancia a la desecación. Al mismo tiempo, semillas de cada día de colecta se conservaron a 5 ºC o en nitrógeno líquido y, transcurridos 30 días, les fue evaluada su PG y se sometieron a las pruebas de envejecimiento acelerado y pérdida de electrólitos. Durante el desarrollo de la investigación se determinó que la madurez fisiológica de las semillas de SS-96 se alcanza a los 29,6 DDA, con un contenido de agua de 30,6 % y masa seca de 1000 semillas de 81,2 mg. Se estableció 35 DDA como el momento idóneo para la colecta de semillas en este cultivar. Una colecta antes de esta fecha provoca una disminución en la tolerancia a la desecación producto de la inmadurez de las semillas; una recolección posterior conlleva a la pérdida de vigor debido a la mayor exposición de las semillas a las condiciones ambientales.

  13. Synthesis, Crystal Structure and Luminescent Property of Cd (II Complex with <em>N-Benzenesulphonyl-L>-leucine

    Directory of Open Access Journals (Sweden)

    Xishi Tai

    2012-09-01

    Full Text Available A new trinuclear Cd (II complex [Cd3(L6(2,2-bipyridine3] [L =<em> Nem>-phenylsulfonyl-L>-leucinato] has been synthesized and characterized by elemental analysis, IR and X-ray single crystal diffraction analysis. The results show that the complex belongs to the orthorhombic, space group<em> Pem>212121 with<em> aem> = 16.877(3 Å, <em>b> em>= 22.875(5 Å, <em>c em>= 29.495(6 Å, <em>α> em>= <emem>= <emem>= 90°, <em>V> em>= 11387(4 Å3, <em>Z> em>= 4, <em>Dc>= 1.416 μg·m−3, <emem>= 0.737 mm−1, <em>F> em>(000 = 4992, and final <em>R>1 = 0.0390, <em>ωR>2 = 0.0989. The complex comprises two seven-coordinated Cd (II atoms, with a N2O5 distorted pengonal bipyramidal coordination environment and a six-coordinated Cd (II atom, with a N2O4 distorted octahedral coordination environment. The molecules form one dimensional chain structure by the interaction of bridged carboxylato groups, hydrogen bonds and p-p interaction of 2,2-bipyridine. The luminescent properties of the Cd (II complex and <em>N-Benzenesulphonyl-L>-leucine in solid and in CH3OH solution also have been investigated.

  14. [Triatoma vandae sp.n. of the oliveirai complex from the State of Mato Grosso, Brazil (Hemiptera: Reduviidae: Triatominae)].

    Science.gov (United States)

    Carcavallo, Rodolfo U; Jurberg, José; Rocha, Dayse da Silva; Galvao, Cleber; Noireau, François; Lent, Herman

    2002-07-01

    There are several specific complexes belonging to the genus Triatoma Laporte, 1832, which are generally associated to specific geographic areas. Recent publications have linked the oliveirai complex to ecosystems of Mato Grosso, which are also present in other Brazilian states and even in other bordering countries as eastern Paraguay. The study of the abundant material collected during the last years allowed the description of several new species of the oliveirai complex: T. jurbergi Carcavallo, Galvão Lent, 1998; T. baratai Carcavallo Jurberg, 2000 and T. klugi Carcavallo, Jurberg, Lent Galvão, 2001. Another new species belonging to the same complex is described here as T. vandae sp.n. It originates from the state of Mato Grosso, and has been reared in the insectary of the Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos, Departamento de Entomologia, Instituto Oswaldo Cruz-Fiocruz, Rio de Janeiro.

  15. Differences in the Detoxification Metabolism between Two clonal Lineages of the Aphid Myzus persicae (Sulzer (Hemiptera: Aphididae Reared on Tobacco (Nicotiana tabacum L. Diferencias en el Metabolismo de Detoxificación entre dos Linajes Clonales del Áfido Myzus persicae (Sulzer (Hemiptera: Aphididae creados sobre tabaco (Nicotiana tabacum L.

    Directory of Open Access Journals (Sweden)

    Marco A Cabrera-Brandt

    2010-12-01

    Full Text Available Myzus persicae (Sulzer is a highly polyphagous aphid species, with a subspecies (M. persicae nicotianae well adapted to tobacco (Nicotiana tabacum L.. We evaluated the effect of this host plant on the aphid performance and detoxification enzymes, in order to test the participation of xenobiotic metabolism on the ability of this aphid to overcome the tobacco chemical defences. Two genotypes, one corresponding to the only M. persicae nicotianae genotype reported in Chile on tobacco, and one genotype belonging to M. persicae sensu stricto were reared on tobacco and pepper (Capsicum annuum L., respectively. M. persicae nicotianae showed a significantly higher intrinsic rate of increase (r m on pepper than on tobacco, and M. persicae s.s. performed similarly, but with no reproduction at all on tobacco. In order to evaluate the effect of tobacco on detoxification enzymes, esterases, glutathione S-transferases (GST and cytochrome P-450 monooxygenases (MO were determined in both selected aphid genotypes after 12, 24, 36, 48 and 72 h of rearing on tobacco and pepper. M. persicae nicotianae exhibited the higher total esterase activities when reared on tobacco than on pepper after 48 h of rearing, while the activities of GST and MO did not show any significant difference between host-plants and duration of treatment. For M. persicae s.s., no significant differences were observed among host-plants for the studied enzymes. These results suggest a participation of the esterases, on the ability of this M. persicae nicotianae to overcome the tobacco defences.Myzus persicae (Sulzer es un áfido polífago que incluye a Myzus persicae nicotianae, una subespecie altamente adaptada sobre tabaco (Nicotiana tabacum L.. Evaluamos el efecto del tabaco sobre el desempeño biológico y sobre determinadas enzimas de detoxificación en áfidos, para estudiar su participación en la capacidad de M. persicae nicotianae de superar las defensas químicas del tabaco. Dos

  16. [Preferential localization of cadmium on the iterative DNA sequences from cultured tissues of the crown gall of tobacco (Nicotiana tabacum, var. Wisconsis 38)].

    Science.gov (United States)

    Sissoëff, I; Grisvard, J; Guillé, E; Laterjet, R

    1975-05-26

    Fractionation of total crown-gall tissue culture DNA from Nicotiana tabacum by Ag+-Cs2SO4 density gradient is described. Cadmium ions determination is performed in each fraction by anodic stripping voltammetry. The cadmium content of the DNA in the lightest density fractions is 100 to 1000 times higher than in the other fractions.

  17. NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high-level pathogen resistance under greenhouse and field conditions

    NARCIS (Netherlands)

    Portieles, R.; Ayra, C.; Gonzalez, E.; Gallo, A.; Rodriguez, R.; Chacón, O.; López, Y.; Rodriguez, M.; Castillo, J.; Pujol, M.; Enriquez, G.; Borroto, C.; Trujillo, L.; Thomma, B.P.H.J.; Borrás-Hidalgo, O.

    2010-01-01

    Plant defensins are small cysteine-rich peptides that inhibit the growth of a broad range of microbes. In this article, we describe NmDef02, a novel cDNA encoding a putative defensin isolated from Nicotiana megalosiphon upon inoculation with the tobacco blue mould pathogen Peronospora hyoscyami

  18. Back-transmission of a virus associated with apple stem pitting and pear vein yellows from Nicotiana occidentalis to apple and pear indicators

    NARCIS (Netherlands)

    Leone, G.; Lindner, J.L.; Jongedijk, G.; Meer, van der F.

    1995-01-01

    The successful back-transmission of the mechanically transmissible virus associated with apple stem pitting and pear vein yellows, from Nicotiana occidentalis to apple seedlings "Golden Delicious" under greenhouse conditions is reported. This result enabled a field experiment where isolates of apple

  19. Symptoms on apple and pear indicators after back-transmission from Nicotiana occidentalis confirm the identity of apple stem pitting virus with pear vein yellows virus

    NARCIS (Netherlands)

    Leone, G.; Lindner, J.L.; Meer, van der F.A.; Schoen, C.D.; Jongedijk, G.

    1998-01-01

    Isolates of apple stem pitting virus (ASPV) from diseased apple trees were maintained in Nicotiana occidentalis then back-transmitted mechanically from the herbaceous host to apple seedlings and indexed by double budding on apple and pear indicators for the following syndromes: apple stem pitting,

  20. Symptoms on apple and pear indicators after back-transmission from Nicotiana occidentalis confirm the identity of apple stem pitting virus with pear vein yellows virus

    NARCIS (Netherlands)

    Leone, G.; Lindner, J.L.; Meer, van der F.A.; Schoen, C.D.; Jongedijk, G.

    1998-01-01

    Isolates of apple stem pitting virus (ASPV) from diseased apple trees were maintained in Nicotiana occidentalis then back-transmitted mechanically from the herbaceous host to apple seedlings and indexed by double budding on apple and pear indicators for the following syndromes: apple stem pitting, p

  1. Recognition of an Avr3a homologue plays a major role in mediating nonhost resistance to Phytophthora capsici in Nicotiana species.

    Science.gov (United States)

    Vega-Arreguín, Julio C; Jalloh, Abubakar; Bos, Jorunn I; Moffett, Peter

    2014-08-01

    Nonhost resistance is a commonly occurring phenomenon wherein all accessions or cultivars of a plant species are resistant to all strains of a pathogen species and is likely the manifestation of multiple molecular mechanisms. Phytophthora capsici is a soil-borne oomycete that causes Phytophthora blight disease in many solanaceous and cucurbitaceous plants worldwide. Interest in P. capsici has increased considerably with the sequencing of its genome and its increasing occurrence in multiple crops. However, molecular interactions between P. capsici and both its hosts and its nonhosts are poorly defined. We show here that tobacco (Nicotiana tabacum) acts like a nonhost for P. capsici and responds to P. capsici infection with a hypersensitive response (HR). Furthermore, we have found that a P. capsici Avr3a-like gene (PcAvr3a1) encoding a putative RXLR effector protein produces a HR upon transient expression in tobacco and several other Nicotiana species. This HR response correlated with resistance in 19 of 23 Nicotiana species and accessions tested, and knock-down of PcAvr3a1 expression by host-induced gene silencing allowed infection of resistant tobacco. Our results suggest that many Nicotiana species have the capacity to recognize PcAvr3a1 via the products of endogenous disease resistance (R) genes and that this R gene-mediated response is a major component of nonhost resistance to P. capsici.

  2. Differences in intensity and specificity of hypersensitive response induction in Nicotiana spp. by INF1, INF2A, and INF2B of Phytophthora infestans

    NARCIS (Netherlands)

    Huitema, E.; Vleeshouwers, V.G.A.A.; Cakir, C.; Kamoun, S.; Govers, F.

    2005-01-01

    Elicitins form a family of structurally related proteins that induce the hypersensitive response (HR) in plants, particularly Nicotiana spp. The elicitin family is composed of several classes. Most species of the plant-pathogenic oomycete genus Phytophthora produce the well-characterized 10-kDa

  3. Ectopic expression of class 1 KNOX genes induce and adventitious shoot regeneration and alter growth and development of tobacco (Nicotiana tabacum L) and European plum (Prunus domestica L)

    Science.gov (United States)

    Transgenic plants of tobacco (Nicotiana tabacum L) and plum (Prunus domestica L) were produced by transforming with apple class 1 KNOX genes (MdKN1 and MdKN2) or corn KN1 gene. Transgenic tobacco plants were regenerated in vitro from transformed leaf discs cultured in a tissue medium lacking cytoki...

  4. NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high-level pathogen resistance under greenhouse and field conditions

    NARCIS (Netherlands)

    Portieles, R.; Ayra, C.; Gonzalez, E.; Gallo, A.; Rodriguez, R.; Chacón, O.; López, Y.; Rodriguez, M.; Castillo, J.; Pujol, M.; Enriquez, G.; Borroto, C.; Trujillo, L.; Thomma, B.P.H.J.; Borrás-Hidalgo, O.

    2010-01-01

    Plant defensins are small cysteine-rich peptides that inhibit the growth of a broad range of microbes. In this article, we describe NmDef02, a novel cDNA encoding a putative defensin isolated from Nicotiana megalosiphon upon inoculation with the tobacco blue mould pathogen Peronospora hyoscyami f.sp

  5. Back-transmission of a virus associated with apple stem pitting and pear vein yellows from Nicotiana occidentalis to apple and pear indicators

    NARCIS (Netherlands)

    Leone, G.; Lindner, J.L.; Jongedijk, G.; Meer, van der F.

    1995-01-01

    The successful back-transmission of the mechanically transmissible virus associated with apple stem pitting and pear vein yellows, from Nicotiana occidentalis to apple seedlings "Golden Delicious" under greenhouse conditions is reported. This result enabled a field experiment where isolates of apple

  6. ANÁLISIS COMPARATIVO DE INDICADORES QUÍMICOS DE LA HOJA Y DITERPENOS DE EXUDADOS FOLIARES DE Nicotiana tabacum L

    National Research Council Canada - National Science Library

    Yanelis Capdesuñer; Maribel Rivas; Janet Quiñones-Galvez; Madelin Gallo; Erinelvis Rodríguez; Juan L. Pérez; Ermis Yanes-Paz; Martha Hernández

    2016-01-01

    .... El objetivo de esta investigación es determinar la composición química de la hoja y de los extractos etanólicos obtenidos a partir de exudados foliares de diez accesiones no comerciales de Nicotiana tabacum L...

  7. <em>α>-Glucosidase Inhibitory Constituents from <em>Acanthopanax senticosusem> Harm Leaves

    Directory of Open Access Journals (Sweden)

    Hai-Xue Kuang

    2012-05-01

    Full Text Available A new triterpene glycoside, 3-<em>O-[(α>-L-rhamnopyranosyl(1→2]-[<em>β>-D-glucuronopyranosyl-6-<em>O>-methyl ester]-olean-12-ene-28-olic acid (1 and a new indole alkaloid, 5-methoxy-2-oxoindolin-3-acetic acid methyl ester (5 were isolated from the leaves of <em>Acanthopanax senticosusem> Harms along with six known compounds. The structures of the new compounds were determined by means of 2D-NMR experiments and chemical methods. All the isolated compounds were evaluated for their glycosidase inhibition activities and compound 6 showed significant <em>α>-glucosidase inhibition activity.

  8. PENGENDALIAN PENYAKIT PATIK (Cercospora nicotianae PADA TEMBAKAU NA OOGST SECARA IN-VIVO DENGAN EKSTRAK DAUN GULMA KIPAHIT (Tithonia diversifolia

    Directory of Open Access Journals (Sweden)

    Aditya Reza Apriyadi

    2013-11-01

    Full Text Available [ENGLISH] Frog eyes diseases or leaf spot caused by Cercospora nicotinae that can reduce the quality of tobacco leaves, especially when used as a cigar deckblad. Affected leaves was easyly torn and the syhmptoms developed rapidly when processed in the storage. Therefore the alternative control of plant is by mexico sun flower leaf extracts. Mexico sunflower belonging to the broadleaf weeds. It suggested contains flavonoids, tannins, terpenoids, and saponins. The results showed mexico sunflower leaf extract concentration 50 g / L was effective to control this diseases, when compared mexico sunflower leaf extract concentration 25 g / L. However, there was no different between plant sprayed with mexico sun flower extract at concentration of 50 g/L and 75 g/L. Keywords: Mexico Sunflower leaf extract; Cercospora nicotianae; Tobacco. [INDONESIAN] Penyakit patik atau bercak daun Cercospora yang disebabkan oleh jamur Cercospora nicotinae dapat mengurangi mutu daun tembakau, terutama apabila digunakan sebagai daun pembalut cerutu. Biasanya penyakit ini dikendalikan dengan pestisida kimia tetapi karena ada batasan residu kimia maksimum sebesar 2,0 ppm, maka dicari alternatif pengendaliannya. Kipahit merupakan gulma berdaun lebar yang mempunyai potensi sebagai anti jamur patogen tanaman, karena mengandung senyawa flavonoid, tannin, terpenoid, dan saponin. Dengan penyemprotan empat kali dalam interval 15 hari, diketahui ekatrak daun kipahit dengan konsentrasi 50 g/l dan 75 g/L dapat menurunkan tingkat keparahan penyakit 1% sejak 60 hst sampai 70 hst. Akan tetapi, nilai insiden penyakit diketahui 100% pada semua perlakuan ekstrak daun kipahit. Ekstrak daun kipahit dengan konsentrasi 50 g/L sudah efektif mengendalikan penyakit patik jika dibandingkan dengan ekstrak daun kipahit dengan konsentrasi 25 g/L. Namun ekstrak daun kipahit dengan konsentrasi 75 g/L tidak berbeda efektifitasnya dengan ekstrak daun kipahit dengan konsentrasi 50 g/L. Kata Kunci: Ekstrak

  9. Immobilization of Nicotiana tabacum plant cell suspensions within calcium alginate gel beads for the production of enhanced amounts of scopolin.

    Science.gov (United States)

    Gilleta; Roisin; Fliniaux; Jacquin-Dubreuil; Barbotin; Nava-Saucedo

    2000-02-01

    Scopolin-producing cells of Nicotiana tabacum were immobilized within Ca-alginate gel beads. Free cell suspensions accumulated scopolin within cytoplasmic compartments and cell disruption was necessary to recover scopolin. On the contrary, immobilized plant cells excreted considerable amounts of scopolin. Scopolin diffused throughout the gel matrix and reached the culture media. A large fraction of produced scopolin could then be recovered from the culture medium without disrupting cells. Immobilized N. tabacum cells produced more scopolin than free cell suspensions did (3.8 mg/g fresh weight biomass [into the culture media] versus 0.2 mg/g fresh weight biomass [intracellular]). Variation of the immobilization conditions revealed a marked influence on the behavior of N. tabacum plant cells: production of scopolin and enhanced excretion, cell growth, and morphological aspect of plant cell colonies. This excretion phenomenon could be used advantageously at an industrial production level.

  10. Identification of a mitochondrial external NADPH dehydrogenase by overexpression in transgenic ¤Nicotiana sylvestris¤

    DEFF Research Database (Denmark)

    Michalecka, A.M.; Agius, S.C.; Møller, I.M.;

    2004-01-01

    (P)H dehydrogenases, was introduced into Nicotiana sylvestris. Transgenic lines with high transcript and protein levels for St-NDB1 had up to threefold increased activity of external NADPH dehydrogenase in isolated mitochondria as compared to the wild type (WT). In two lines, the external NADPH dehydrogenase activity...... for NADPH and dependent on calcium for activity. Transgenic lines overexpressing St-ndb1 had specifically increased protein levels for alternative oxidase and uncoupling protein, as compared to the WT and one co-suppressing line. This indicates cross-talk in the expressional control, or metabolic conditions...... influencing it, for the different categories of energy-dissipating proteins that bypass oxidative phosphorylation. The potential effects of external NADPH oxidation on other cellular processes are discussed....

  11. Cloning and functional analysis of chloroplast division gene NtFtsZ2-1 in Nicotiana tabacum

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    FtsZ protein plays an important role in the division of chloroplasts. With the finding and functional analysis of higher plant FtsZ proteins, people have deepened the understanding in the molecular mechanism of chloroplast division. Multiple ftsZ genes are diversified into two families in higher plants, ftsZ1 and ftsZ2. On the basis of the research on ftsZ1 family, we analyzed the function of NtFtsZ2-1 gene in Nicotiana tabacum. Microscopic analysis of the sense and antisense NtFtsZ2-1 transgenic tobacco plants revealed that the chloroplasts were abnormal in size and also in number when compared with wild-type tobacco chloroplasts. Our investigations confirmed that the NtFtsZ2-1 gene is involved in plant chloroplast division.

  12. Macrocyclic Trichothecenes from Myrothecium roridum Strain M10 with Motility Inhibitory and Zoosporicidal Activities against Phytophthora nicotianae.

    Science.gov (United States)

    Mondol, Muhammad Abdul Mojid; Surovy, Musrat Zahan; Islam, M Tofazzal; Schüffler, Anja; Laatsch, Hartmut

    2015-10-14

    The cytotoxicity of the extract obtained from Myrothecium roridum M10 and a characteristic (1)H signal at δH ∼8 led to the assumption that verrucarin/roridin-type compounds were present. Upscaling on rice medium led to the isolation of four new metabolites: verrucarins Y (1) and Z (6) (macrocyclic trichothecenes), bilain D (12) (a diketopiperazine derivative), and hamavellone C (14) (an unusual cyclopropyl diketone). In addition, nine known trichothecenes [verrucarin A (3), 16-hydroxyverrucarin A (5), verrucarin B (7), 16-hydroxyverrucarin B (8), verrucarin J (2), verrucarin X (4), roridin A (9), roridin L-2 (10), and trichoverritone (11)] and a bicyclic lactone [myrotheciumone A (15)] were identified. Their structures and configurations were determined by spectroscopic methods, published data, Mosher's method, and considering biosyntheses. Some trichothecenes showed motility inhibition followed by lysis of the zoospores against devastating Phytophthora nicotianae within 5 min. Compounds 2, 3, 7, and 9 also exhibited potent activities against Candida albicans and Mucor miehei.

  13. Cloning, expression analysis and recombinant expression of a gene encoding a polygalacturonase-inhibiting protein from tobacco, Nicotiana tabacum

    Directory of Open Access Journals (Sweden)

    Chengsheng Zhang

    2016-05-01

    Full Text Available Polygalacturonase inhibiting proteins (PGIPs are major defensive proteins produced by plant cell walls that play a crucial role in pathogen resistance by reducing polygalacturonase (PG activity. In the present study, a novel PGIP gene was isolated from tobacco (Nicotiana tabacum, hereafter referred as NtPGIP. A full-length NtPGIP cDNA of 1,412 bp with a 186 bp 5′-untranslated region (UTR, and 209 bp 3′-UTR was cloned from tobacco, NtPGIP is predicted to encode a protein of 338 amino acids. The NtPGIP sequence from genomic DNA showed no introns and sequence alignments of NtPGIP’s deduced amino acid sequence showed high homology with known PGIPs from other plant species. Moreover, the putative NtPGIP protein was closely clustered with several Solanaceae PGIPs. Further, the expression profile of NtPGIP was examined in tobacco leaves following stimulation with the oomycete Phytophthora nicotianae and other stressors, including salicylic acid (SA, abscisic acid (ABA, salt, and cold treatment. The results showed that all of the treatments up-regulated the expression of NtPGIP at different times. To understand the biochemical activity of NtPGIP gene, a full-length NtPGIP cDNA sequence was subcloned into a pET28a vector and transformed into E. coli BL21 (DE3. Recombinant proteins were successfully induced by 1.0 nmol/L IPTG and the purified proteins effectively inhibited Phytophthora capsici PG activity. The results of this study suggest that NtPGIP may be a new candidate gene with properties that could be exploited in plant breeding.

  14. Cloning and characterization of a 9-lipoxygenase gene induced by pathogen attack from Nicotiana benthamiana for biotechnological application

    Directory of Open Access Journals (Sweden)

    Schwab Wilfried

    2011-03-01

    Full Text Available Abstract Background Plant lipoxygenases (LOXs have been proposed to form biologically active compounds both during normal developmental stages such as germination or growth as well as during responses to environmental stress such as wounding or pathogen attack. In our previous study, we found that enzyme activity of endogenous 9-LOX in Nicotiana benthamiana was highly induced by agroinfiltration using a tobacco mosaic virus (TMV based vector system. Results A LOX gene which is expressed after treatment of the viral vectors was isolated from Nicotiana benthamiana. As the encoded LOX has a high amino acid identity to other 9-LOX proteins, the gene was named as Nb-9-LOX. It was heterologously expressed in yeast cells and its enzymatic activity was characterized. The yeast cells expressed large quantities of stable 9-LOX (0.9 U ml-1 cell cultures which can oxygenate linoleic acid resulting in high yields (18 μmol ml-1 cell cultures of hydroperoxy fatty acid. The product specificity of Nb-9-LOX was examined by incubation of linoleic acid and Nb-9-LOX in combination with a 13-hydroperoxide lyase from watermelon (Cl-13-HPL or a 9/13-hydroperoxide lyase from melon (Cm-9/13-HPL and by LC-MS analysis. The result showed that Nb-9-LOX possesses both 9- and 13-LOX specificity, with high predominance for the 9-LOX function. The combination of recombinant Nb-9-LOX and recombinant Cm-9/13-HPL produced large amounts of C9-aldehydes (3.3 μmol mg-1 crude protein. The yield of C9-aldehydes from linoleic acid was 64%. Conclusion The yeast expressed Nb-9-LOX can be used to produce C9-aldehydes on a large scale in combination with a HPL gene with 9-HPL function, or to effectively produce 9-hydroxy-10(E,12(Z-octadecadienoic acid in a biocatalytic process in combination with cysteine as a mild reducing agent.

  15. Study of photosynthesis process in the presence of low concentrations of clomazone herbicide in tobacco (Nicotiana tabacum

    Directory of Open Access Journals (Sweden)

    Darwish, Majd

    2013-02-01

    Full Text Available The effect of chemical residues of clomazone on photosynthetic processes has been studiedby using several low concentrations of the herbicide (0, 1, 0.1, 0.01, 0.001, 0.0001 and 0.00001 µM and seedlings of two varieties of tobacco (Nicotiana tabacum L. cv. Virginie vk51, Nicotiana tabacum, L. cv. Xanthi. The content of photosynthetic pigments, the parameters of the chlorophyll-a fluorescence and the JIP-test were performed on an adult leaf (AL and a young leaf (YL, that gave a complementary designto know the action's mode of clomazone on the plant physiological processes. Clomazone reducedthe total chlorophyll (a+b, carotenoids pigments (reduction in size antenna pigments judged by an increase in the chlorophyll a/b ratio in young leaves more than adults leaves. The maximal photochemical efficiency (Fv/Fm of photosystem II (PSII decreased significantly in youngleavescompared to adult leaves and in (Virginie variety than (Xanthi variety. Among the parameters calculated of the JIP-test most affected by the treatment, PIabs, 1-VJ, ABS/RC, DI0/RC, TR0/RC, ET0/RC, ET0/ABS, which indicated acomparable effects of clomazone(1μM, 0.1µM, 0.01µM between the two types of leaves and the varieties used. More, the results showed that the concentration ( 1μM was the most effective amongthe other low concentrations used and the (Virginie variety ismore sensitive than the (Xanthivariety. We conclude that clomazone has probably two combined functions (physiological, toxic judged by the different behavior of both types of leaves in the presence of the herbicide.

  16. Genetic Diversity in Haploid Nicotiana alata Induced by Gamma Irradiation, Salt Tolerance and Detection of These Differences by RAPD

    Directory of Open Access Journals (Sweden)

    Ayman EL-FIKI

    2016-03-01

    Full Text Available Haploid plants of Nicotiana alata were cultured in vitro on MS medium with IAA + KIN. The resulting plantlets were irradiated using gamma radiation doses of 10, 15, 20 and 25 Gy. Single node pieces were cut and transferred onto fresh MS medium. Gamma radiation doses caused the death of 9% and up to 28% of explants. NaCl concentrations caused the death of 8% up to 36% of explants, while the combined effect between gamma radiation doses and salinity had an impact suffused on the percentage of survival. The combined effect of gamma radiation doses 20 Gy and 25 Gy on NaCl concentrations of 100, 150 and 200 mM were deadly. Even more, the combined effect of gamma radiation doses and salinity had a severe negative impact on both the proline content and total soluble protein. Random amplified polymorphic DNA (RAPD analysis was used to determine the degree of genetic variation in treated haploid Nicotiana alata plants. Total genomic DNAs from different haploid plantlets treated were amplified using five arbitrary primers. Two hundred and seventy bands were detected from plantlets irradiated with doses of 15, 20 and 25 Gy, with polymorphic band number 226 (83.7%. The total number of bands resulted from plant grew on 150 mM and 200 mM NaCl were 260 bands with polymorphic bands 185 (85.6%. However, the total number of bands produced from combined effects between gamma rays and salinity (20 Gy X 50 mM NaCl, 20 Gy X 100 mM NaCl and 25 Gy X 50 mM NaCl were 270, with polymorphic band number 231 (85.5%. High similarity between treatments was revealed. Treatments relationships were estimated through cluster analysis (UPGMA based on RAPD data.

  17. Expressing foreign genes in the pistil: a comparison of S-RNase constructs in different Nicotiana backgrounds.

    Science.gov (United States)

    Murfett, J; McClure, B A

    1998-06-01

    Transgenic plant experiments have great potential for extending our understanding of the role of specific genes in controlling pollination. Often, the intent of such experiments is to over-express a gene and test for effects on pollination. We have examined the efficiency of six different S-RNase constructs in Nicotiana species and hybrids. Each construct contained the coding region, intron, and downstream sequences from the Nicotiana alata S(A2)-RNase gene. Among the six expression constructs, two utilized the cauliflower mosaic virus (CaMV) 35S promoter with duplicated enhancer, and four utilized promoters from genes expressed primarily in pistils. The latter included promoters from the tomato Chi2;1 and 9612 genes, a promoter from the N. alata S(A2)-RNase gene, and a promoter from the Brassica SLG-13 gene. Some or all of the constructs were tested in N. tabacum, N. plumbaginifolia, N. plumbaginifolia x SI N. alata S(C10)S(c10) hybrids, N. langsdorffii, and N. langsdorffii x SC N. alata hybrids. Stylar specific RNase activities and S(A2)-RNase transcript levels were determined in transformed plants. Constructs including the tomato Chi2;1 gene promoter or the Brassica SLG-13 promoter provided the highest levels of S(A2)-RNase expression. Transgene expression patterns were tightly regulated, the highest level of expression was observed in post-anthesis styles. Expression levels of the S(A2)-RNase transgenes was dependent on the genetic background of the host. Higher levels of S(A2)-RNase expression were observed in N. plumbaginifolia x SC N. alata hybrids than in N. plumbaginifolia.

  18. Effects of down-regulating ornithine decarboxylase upon putrescine-associated metabolism and growth in Nicotiana tabacum L.

    Science.gov (United States)

    Dalton, Heidi L; Blomstedt, Cecilia K; Neale, Alan D; Gleadow, Ros; DeBoer, Kathleen D; Hamill, John D

    2016-05-01

    Transgenic plants of Nicotiana tabacum L. homozygous for an RNAi construct designed to silence ornithine decarboxylase (ODC) had significantly lower concentrations of nicotine and nornicotine, but significantly higher concentrations of anatabine, compared with vector-only controls. Silencing of ODC also led to significantly reduced concentrations of polyamines (putrescine, spermidine and spermine), tyramine and phenolamides (caffeoylputrescine and dicaffeoylspermidine) with concomitant increases in concentrations of amino acids ornithine, arginine, aspartate, glutamate and glutamine. Root transcript levels of S-adenosyl methionine decarboxylase, S-adenosyl methionine synthase and spermidine synthase (polyamine synthesis enzymes) were reduced compared with vector controls, whilst transcript levels of arginine decarboxylase (putrescine synthesis), putrescine methyltransferase (nicotine production) and multi-drug and toxic compound extrusion (alkaloid transport) proteins were elevated. In contrast, expression of two other key proteins required for alkaloid synthesis, quinolinic acid phosphoribosyltransferase (nicotinic acid production) and a PIP-family oxidoreductase (nicotinic acid condensation reactions), were diminished in roots of odc-RNAi plants relative to vector-only controls. Transcriptional and biochemical differences associated with polyamine and alkaloid metabolism were exacerbated in odc-RNAi plants in response to different forms of shoot damage. In general, apex removal had a greater effect than leaf wounding alone, with a combination of these injury treatments producing synergistic responses in some cases. Reduced expression of ODC appeared to have negative effects upon plant growth and vigour with some leaves of odc-RNAi lines being brittle and bleached compared with vector-only controls. Together, results of this study demonstrate that ornithine decarboxylase has important roles in facilitating both primary and secondary metabolism in Nicotiana.

  19. Estadísticos poblacionales de Triatoma sordida Stäl 1859 (Hemiptera : Reduviidae en condiciones experimentales Population statistics of Triatoma sordida Stäl 1859 (Hemiptera: Reduviidae in experimental conditions

    Directory of Open Access Journals (Sweden)

    Elena B. Oscherov

    1996-10-01

    Full Text Available Se realizó el seguimiento longitudinal de cuatro cohortes de 100 huevos cada una. Los insectos fueron criados a 28°C ± 3°C y 63 % ± 10% de humedad relativa, y alimentados cada siete días sobre gallina (Gallus domesticus, durante 40 min. Las cohortes fueron controladas una vez por semana. Se observó un valor constante de supervivencia en las cuatro cohortes, sin picos importantes. La expectativa de vida de los machos, en promedio, fue de 37,4 semanas y la de las hembras 36,8. El tiempo generacional demandó 61,7 semanas. La tasa reproductiva neta indica que esta población se incrementará 143,3 veces con cada generación. La tasa intrínseca de crecimiento natural fue de 0,082 por semana. El máximo valor reproductivo se verificó, en promedio, a la tercer semana del ingreso al estado adulto. La distribución estable de edades de T. sordida sería de 32% huevos, 26% ninfas de primer estadio, 19% ninfas de segundo estadio, 13% ninfas de tercer estadio, 6% ninfas de cuarto estadio, 3% ninfas de quinto estadio y 1% de adultos. Es la primera cita para Argentina sobre la tabla de vida de T. sordida. El análisis de estos parámetros permite considerar que esta especie se comporta como "K estratega", en condiciones experimentales.Foram estudados diferentes parâmetros populacionais de Triatoma sordida em condições de laboratório. Realizou-se um seguimento longitudinal de quatro coortes de 100 ovos cada uma; os insetos foram criados a 28°C ± 3°C e 63% ± 10% de umidade relativa, e alimentados cada sete dias sobre galinha (Gallus domesticus durante 40'. As coortes foram controladas uma vez por semana. A expectativa de vida média dos machos foi de 37,4 semanas e a das fêmeas de 36,8.O tempo médio de desenvolvimento foi de 61,7 semanas. A taxa reprodutiva líquida indica que esta população aumentará 143,2 vezes durante cada geração. A taxa intrínseca do crecimento natural foi de 0,082 por semana. A média do valor máximo reprodutivo

  20. Three New Myrsinol Diterpenes from <em>Euphorbia proliferaem> and Their Neuroprotective Activities

    Directory of Open Access Journals (Sweden)

    Yuanqiang Guo

    2012-08-01

    Full Text Available Three new myrsinol diterpenes were isolated from the roots of<em> em>>Euphorbia proliferaem>. Their structures were elucidated as 2<em>α-O>-isobutyryl-3<em>β>,5<em>α>,7<em>β>,10,15<em>β-penta-O>-acetyl-14<em>α-O>-benzoyl-10,18-dihydromyrsinol (1, 2<em>α-O>-isobutyryl-3<em>β-O>-propion-yl-5<em>α>,7<em>β>,10,15<em>β-tetra-O>-acetyl-10,18-dihydromyrsinol (2, and 2<em>α>,14<em>α-di-O>-benzoyl-3<em>β>,5<em>α>,7<em>β>,10,15<em>β-penta-O>-acetyl-10,18-dihydromyrsinol (3 on the basis of spectroscopic data analyses (IR, ESI-MS, HR-ESI-MS, and 1D and 2D NMR. Their neuroprotective activities were evaluated and compounds 1 and 2 showed neuroprotective effects against MPP+-induced neuronal cell death in SH-SY5Y cells.

  1. Morfometria comparada de Triatoma infestans, T. rubrovaria e T. platensis (Hemiptera, Reduviidae, Triatominae do Uruguai Comparative Morphometry among Triatoma infestans, T. rubrovaria and T. platensis (Hemiptera, Reduviidae, Triatominae from Uruguay

    Directory of Open Access Journals (Sweden)

    Carolina M. dos Santos

    2009-03-01

    Full Text Available No Uruguai, são conhecidas 10 espécies de triatomíneos com importância epidemiológica variável, dentre as quais destacamse Triatoma infestans (Klug, 1834 e T. rubrovaria (Blanchard, 1843, consideradas as principais espécies vetoras. Triatoma platensis Neiva, 1913, apesar de ser uma espécie silvestre, pode conviver com T. infestans em galinheiros, chegando a gerar híbridos férteis. Estas três espécies fazem parte do "complexo T. infestans", formado por cinco espécies. Devido à importância epidemiológica dessas espécies no Uruguai e com a finalidade de ampliar o conjunto dos parâmetros morfológicos diferenciais destas espécies apresentamos uma abordagem morfométrica comparativa da cabeça e do pronoto. Foram medidas quatro variáveis da cabeça e quatro do pronoto a partir de 50 machos e 50 fêmeas, de cada espécie, provenientes de criação de laboratório. O programa JMP foi utilizado para as análises de Componentes Principais (ACP e Discriminante (AD. Os resultados demonstraram que os dois primeiros componentes principais (CP1 e CP2 exibem aproximadamente 77% da variação total. Os caracteres mais importantes para a distinção das espécies foram a região anteocular (RAO e a região pós-ocular (RPO, indicando que a morfometria da cabeça se mostrou mais eficiente que o pronoto para diferenciar estas espécies. A análise discriminante possibilitou separar T. platensis e T. rubrovaria mas não foi capaz de separá-las de T. infestans.In Uruguay, we can find 10 species with different epidemiological importance, amongst them we emphasize Triatoma infestans (Klug, 1834 and T. rubrovaria (Blanchard, 1843, considered main vectors. Triatoma platensis Neiva, 1913, that although being a wild species can cohabit with T. infestans in chicken coops. These three species, belong to "T. infestans complex", wich is composed by five species. Because the epidemiological importance of these species in Uruguay and, with the aim to

  2. The C-terminal half of Phytophthora infestans RXLR effector AVR3a is sufficient to trigger R3a-mediated hypersensitivity and suppress INF1-induced cell death in Nicotiana benthamiana.

    Science.gov (United States)

    Bos, Jorunn I B; Kanneganti, Thirumala-Devi; Young, Carolyn; Cakir, Cahid; Huitema, Edgar; Win, Joe; Armstrong, Miles R; Birch, Paul R J; Kamoun, Sophien

    2006-10-01

    The RXLR cytoplasmic effector AVR3a of Phytophthora infestans confers avirulence on potato plants carrying the R3a gene. Two alleles of Avr3a encode secreted proteins that differ in only three amino acid residues, two of which are in the mature protein. Avirulent isolates carry the Avr3a allele, which encodes AVR3aKI (containing amino acids C19, K80 and I103), whereas virulent isolates express only the virulence allele avr3a, encoding AVR3aEM (S19, E80 and M103). Only the AVR3aKI protein is recognized inside the plant cytoplasm where it triggers R3a-mediated hypersensitivity. Similar to other oomycete avirulence proteins, AVR3aKI carries a signal peptide followed by a conserved motif centered on the consensus RXLR sequence that is functionally similar to a host cell-targeting signal of malaria parasites. The interaction between Avr3a and R3a can be reconstructed by their transient co-expression in Nicotiana benthamiana. We exploited the N. benthamiana experimental system to further characterize the Avr3a-R3a interaction. R3a activation by AVR3aKI is dependent on the ubiquitin ligase-associated protein SGT1 and heat-shock protein HSP90. The AVR3aKI and AVR3aEM proteins are equally stable in planta, suggesting that the difference in R3a-mediated death cannot be attributed to AVR3aEM protein instability. AVR3aKI is able to suppress cell death induced by the elicitin INF1 of P. infestans, suggesting a possible virulence function for this protein. Structure-function experiments indicated that the 75-amino acid C-terminal half of AVR3aKI, which excludes the RXLR region, is sufficient for avirulence and suppression functions, consistent with the view that the N-terminal region of AVR3aKI and other RXLR effectors is involved in secretion and targeting but is not required for effector activity. We also found that both polymorphic amino acids, K80 and I103, of mature AVR3a contribute to the effector functions.

  3. Neonatal Phosphate Nutrition Alters <em>in em>Vivo> and <em>in em>Vitro> Satellite Cell Activity in Pigs

    Directory of Open Access Journals (Sweden)

    Chad H. Stahl

    2012-05-01

    Full Text Available Satellite cell activity is necessary for postnatal skeletal muscle growth. Severe phosphate (PO4 deficiency can alter satellite cell activity, however the role of neonatal PO4 nutrition on satellite cell biology remains obscure. Twenty-one piglets (1 day of age, 1.8 ± 0.2 kg BW were pair-fed liquid diets that were either PO4 adequate (0.9% total P, supra-adequate (1.2% total P in PO4 requirement or deficient (0.7% total P in PO4 content for 12 days. Body weight was recorded daily and blood samples collected every 6 days. At day 12, pigs were orally dosed with BrdU and 12 h later, satellite cells were isolated. Satellite cells were also cultured <em>in vitroem> for 7 days to determine if PO4 nutrition alters their ability to proceed through their myogenic lineage. Dietary PO4 deficiency resulted in reduced (<em>P> < 0.05 sera PO4 and parathyroid hormone (PTH concentrations, while supra-adequate dietary PO4 improved (<em>P> < 0.05 feed conversion efficiency as compared to the PO4 adequate group. <em>In vivoem> satellite cell proliferation was reduced (<em>P> < 0.05 among the PO4 deficient pigs, and these cells had altered <em>in vitroem> expression of markers of myogenic progression. Further work to better understand early nutritional programming of satellite cells and the potential benefits of emphasizing early PO4 nutrition for future lean growth potential is warranted.

  4. Constituents from <em>Vigna em>vexillata> and Their Anti-Inflammatory Activity

    Directory of Open Access Journals (Sweden)

    Guo-Feng Chen

    2012-08-01

    Full Text Available The seeds of <em>Vigna em>genus are important food resources and there have already been many reports regarding their bioactivities. In our preliminary bioassay, the chloroform layer of methanol extracts of<em> V. vexillata em>demonstrated significant anti-inflammatory bioactivity. Therefore, the present research is aimed to purify and identify the anti-inflammatory principles of <em>V. vexillataem>. One new sterol (1 and two new isoflavones (2,3 were reported from the natural sources for the first time and their chemical structures were determined by the spectroscopic and mass spectrometric analyses. In addition, 37 known compounds were identified by comparison of their physical and spectroscopic data with those reported in the literature. Among the isolates, daidzein (23, abscisic acid (25, and quercetin (40 displayed the most significant inhibition of superoxide anion generation and elastase release.

  5. Improved phytoaccumulation of cadmium by genetically modified tobacco plants (Nicotiana tabacum L.). Physiological and biochemical response of the transformants to cadmium toxicity

    Energy Technology Data Exchange (ETDEWEB)

    Gorinova, N. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria)]. E-mail: noraig60@yahoo.co.uk; Nedkovska, M. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Todorovska, E. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Simova-Stoilova, L. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Stoyanova, Z. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Georgieva, K. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Demirevska-Kepova, K. [Institute of Plant Physiology, Bulgarian Academy of Sciences, 1113 Sofia (Bulgaria); Atanassov, A. [AgroBioInstitute, 8 Dragan Tzankov Blvd., 1164 Sofia (Bulgaria); Herzig, R. [Phytotech-Foundation PT-F, Quartiergasse 12, CH 3013 Bern (Switzerland)

    2007-01-15

    The response of tobacco plants (Nicotiana tabacum L.)-non-transformed and transformed with a metallothionein gene MThis from Silene vulgaris L. - to increase cadmium supply in the nutrient solution was compared. The transgenic plants accumulated significantly more Cd both in the roots and the leaves. Visual toxicity symptoms and disturbance in water balance were correlated with Cd tissue content. Treatment with 300 {mu}M CdCl{sub 2} resulted in inhibition of photosynthesis and mobilization of the ascorbate-glutathione cycle. Treatment with 500 {mu}M CdCl{sub 2} led to irreversible damage of photosynthesis and oxidative stress. An appearance of a new peroxidase isoform and changes in the leaf polypeptide pattern were observed at the highest Cd concentration. The level of non-protein thiols gradually increased following the Cd treatment both in transgenic and non-transformed plants. - Genetic transformation of Nicotiana tabacum L. by metallothionein gene improved phytoaccumulation of cadmium.

  6. The Antimicrobial Efficacy of <em>Elaeis guineensisem>: Characterization, <em>in Vitroem> and <em>in Vivoem> Studies

    Directory of Open Access Journals (Sweden)

    Sreenivasan Sasidharan

    2012-04-01

    Full Text Available The urgent need to treat multi-drug resistant pathogenic microorganisms in chronically infected patients has given rise to the development of new antimicrobials from natural resources. We have tested <em>Elaeis guineensis em>Jacq> em>(Arecaceae methanol extract against a variety of bacterial, fungal and yeast strains associated with infections. Our studies have demonstrated that <em>E. guineensisem> exhibits excellent antimicrobial activity <em>in vitroem> and <em>in vivoem> against the bacterial and fungal strains tested. A marked inhibitory effect of the <em>E. guineensisem> extracts was observed against <em>C. albicansem> whereby <em>E. guineensisem> extract at ½, 1, or 2 times the MIC significantly inhibited <em>C. albicansem> growth with a noticeable drop in optical density (OD of the bacterial culture. This finding confirmed the anticandidal activity of the extract on <em>C. albicansem>. Imaging using scanning (SEM and transmission (TEM electron microscopy was done to determine the major alterations in the microstructure of the extract-treated <em>C. albicansem>. The main abnormalities noted via SEM and TEM studies were the alteration in morphology of the yeast cells. <em>In vivoem> antimicrobial activity was studies in mice that had been inoculated with <em>C. albicansem> and exhibited good anticandidal activity. The authors conclude that the extract may be used as a candidate for the development of anticandidal agent.<em> em>

  7. NaStEP: a proteinase inhibitor essential to self-incompatibility and a positive regulator of HT-B stability in Nicotiana alata pollen tubes.

    Science.gov (United States)

    Jiménez-Durán, Karina; McClure, Bruce; García-Campusano, Florencia; Rodríguez-Sotres, Rogelio; Cisneros, Jesús; Busot, Grethel; Cruz-García, Felipe

    2013-01-01

    In Solanaceae, the self-incompatibility S-RNase and S-locus F-box interactions define self-pollen recognition and rejection in an S-specific manner. This interaction triggers a cascade of events involving other gene products unlinked to the S-locus that are crucial to the self-incompatibility response. To date, two essential pistil-modifier genes, 120K and High Top-Band (HT-B), have been identified in Nicotiana species. However, biochemistry and genetics indicate that additional modifier genes are required. We recently reported a Kunitz-type proteinase inhibitor, named NaStEP (for Nicotiana alata Stigma-Expressed Protein), that is highly expressed in the stigmas of self-incompatible Nicotiana species. Here, we report the proteinase inhibitor activity of NaStEP. NaStEP is taken up by both compatible and incompatible pollen tubes, but its suppression in Nicotiana spp. transgenic plants disrupts S-specific pollen rejection; therefore, NaStEP is a novel pistil-modifier gene. Furthermore, HT-B levels within the pollen tubes are reduced when NaStEP-suppressed pistils are pollinated with either compatible or incompatible pollen. In wild-type self-incompatible N. alata, in contrast, HT-B degradation occurs preferentially in compatible pollinations. Taken together, these data show that the presence of NaStEP is required for the stability of HT-B inside pollen tubes during the rejection response, but the underlying mechanism is currently unknown.

  8. Molecular cloning and functional characterization of the lycopene ε-cyclase gene via virus-induced gene silencing and its expression pattern in Nicotiana tabacum.

    Science.gov (United States)

    Shi, Yanmei; Wang, Ran; Luo, Zhaopeng; Jin, Lifeng; Liu, Pingping; Chen, Qiansi; Li, Zefeng; Li, Feng; Wei, Chunyang; Wu, Mingzhu; Wei, Pan; Xie, He; Qu, Lingbo; Lin, Fucheng; Yang, Jun

    2014-08-22

    Lycopene ε-cyclase (ε-LCY) is a key enzyme that catalyzes the synthesis of α-branch carotenoids through the cyclization of lycopene. Two cDNA molecules encoding ε-LCY (designated Ntε-LCY1 and Ntε-LCY2) were cloned from Nicotiana tabacum. Ntε-LCY1 and Ntε-LCY2 are encoded by two distinct genes with different evolutionary origins, one originating from the tobacco progenitor, Nicotiana sylvestris, and the other originating from Nicotiana tomentosiformis. The two coding regions are 97% identical at the nucleotide level and 95% identical at the amino acid level. Transcripts of Ntε-LCY were detectable in both vegetative and reproductive organs, with a relatively higher level of expression in leaves than in other tissues. Subcellular localization experiments using an Ntε-LCY1-GFP fusion protein demonstrated that mature Ntε-LCY1 protein is localized within the chloroplast in Bright Yellow 2 suspension cells. Under low-temperature and low-irradiation stress, Ntε-LCY transcript levels substantially increased relative to control plants. Tobacco rattle virus (TRV)-mediated silencing of ε-LCY in Nicotiana benthamiana resulted in an increase of β-branch carotenoids and a reduction in the levels of α-branch carotenoids. Meanwhile, transcripts of related genes in the carotenoid biosynthetic pathway observably increased, with the exception of β-OHase in the TRV-ε-lcy line. Suppression of ε-LCY expression was also found to alleviate photoinhibition of Potosystem II in virus-induced gene silencing (VIGS) plants under low-temperature and low-irradiation stress. Our results provide insight into the regulatory role of ε-LCY in plant carotenoid biosynthesis and suggest a role for ε-LCY in positively modulating low temperature stress responses.

  9. Molecular Cloning and Functional Characterization of the Lycopene ε-Cyclase Gene via Virus-Induced Gene Silencing and Its Expression Pattern in Nicotiana tabacum

    Directory of Open Access Journals (Sweden)

    Yanmei Shi

    2014-08-01

    Full Text Available Lycopene ε-cyclase (ε-LCY is a key enzyme that catalyzes the synthesis of α-branch carotenoids through the cyclization of lycopene. Two cDNA molecules encoding ε-LCY (designated Ntε-LCY1 and Ntε-LCY2 were cloned from Nicotiana tabacum. Ntε-LCY1 and Ntε-LCY2 are encoded by two distinct genes with different evolutionary origins, one originating from the tobacco progenitor, Nicotiana sylvestris, and the other originating from Nicotiana tomentosiformis. The two coding regions are 97% identical at the nucleotide level and 95% identical at the amino acid level. Transcripts of Ntε-LCY were detectable in both vegetative and reproductive organs, with a relatively higher level of expression in leaves than in other tissues. Subcellular localization experiments using an Ntε-LCY1-GFP fusion protein demonstrated that mature Ntε-LCY1 protein is localized within the chloroplast in Bright Yellow 2 suspension cells. Under low-temperature and low-irradiation stress, Ntε-LCY transcript levels substantially increased relative to control plants. Tobacco rattle virus (TRV-mediated silencing of ε-LCY in Nicotiana benthamiana resulted in an increase of β-branch carotenoids and a reduction in the levels of α-branch carotenoids. Meanwhile, transcripts of related genes in the carotenoid biosynthetic pathway observably increased, with the exception of β-OHase in the TRV-ε-lcy line. Suppression of ε-LCY expression was also found to alleviate photoinhibition of Potosystem II in virus-induced gene silencing (VIGS plants under low-temperature and low-irradiation stress. Our results provide insight into the regulatory role of ε-LCY in plant carotenoid biosynthesis and suggest a role for ε-LCY in positively modulating low temperature stress responses.

  10. N-Glycosylation of cholera toxin B subunit in Nicotiana benthamiana: impacts on host stress response, production yield and vaccine potential

    OpenAIRE

    Hamorsky, Krystal Teasley; Kouokam, J. Calvin; Jurkiewicz, Jessica M.; Nelson, Bailey; Moore, Lauren J.; Husk, Adam S.; Kajiura, Hiroyuki; Fujiyama, Kazuhito; Matoba, Nobuyuki

    2015-01-01

    Plant-based transient overexpression systems enable rapid and scalable production of subunit vaccines. Previously, we have shown that cholera toxin B subunit (CTB), an oral cholera vaccine antigen, is N-glycosylated upon expression in transgenic Nicotiana benthamiana. Here, we found that overexpression of aglycosylated CTB by agroinfiltration of a tobamoviral vector causes massive tissue necrosis and poor accumulation unless retained in the endoplasmic reticulum (ER). However, the re-introduc...

  11. C3HC4-type RING finger protein NbZFP1 is involved in growth and fruit development in Nicotiana benthamiana.

    Directory of Open Access Journals (Sweden)

    Wenxian Wu

    Full Text Available C3HC4-type RING finger proteins constitute a large family in the plant kingdom and play important roles in various physiological processes of plant life. In this study, a C3HC4-type zinc finger gene was isolated from Nicotiana benthamiana. Sequence analysis indicated that the gene encodes a 24-kDa protein with 191 amino acids containing one typical C3HC4-type zinc finger domain; this gene was named NbZFP1. Transient expression of pGDG-NbZFP1 demonstrated that NbZFP1 was localized to the chloroplast, especially in the chloroplasts of cells surrounding leaf stomata. Virus-induced gene silencing (VIGS analysis indicated that silencing of NbZFP1 hampered fruit development, although the height of the plants was normal. An overexpression construct was then designed and transferred into Nicotiana benthamiana, and PCR and Southern blot showed that the NbZFP1 gene was successfully integrated into the Nicotiana benthamiana genome. The transgenic lines showed typical compactness, with a short internode length and sturdy stems. This is the first report describing the function of a C3HC4-type RING finger protein in tobacco.

  12. [Effects of seven RNA silencing suppressors on heterologous expression of green fluorescence protein expression mediated by a plant virus-based system in Nicotiana benthamiana].

    Science.gov (United States)

    Wang, Sheng; Dong, Jie; Cao, Min; Mu, Hongzhen; Ding, Guoping; Zhang, Hong

    2012-11-01

    To test the effects of 7 virus-encoded RNA silencing suppressors (RSSs) for enhancement of a plant virus-based vector system-mediated heterologous expression of green fluorescence protein (GFP) in Nicotiana benthamiana. Seven transient expression vectors for the 7 RSSs were constructed and co-inoculated on the leaves of Nicotiana benthamiana with PVXdt-GFP vector, a novel Potato virus X-based plant expression vector, through agroinfiltration. The protein and mRNA expression levels of the reporter gene GFP in the co-inoculated Nicotiana leaves were examined by Western blotting, ELISA and RT-qPCR to assess the effect of the RSSs for GFP expression enhancement. The 7 RSSs differed in the degree and duration of enhancement of heterologous GFP expression, and the p19 protein of Tomato bushy stunt virus (TBSV) induced the highest expression of GFP. African cassava mosaic virus AC2 protein and Rice yellow mettle virus P1 protein produced no obvious enhancement GFP expression. Transient co-expression of RSSs suppresses host silencing response to allow high-level and long-term expression of heterologous genes in plant, but the optimal RSS has to be identified for each plant virus-based expression vector system.

  13. Study on the Cultural Characteristics of Phytophthora parasitica var. nicotianae%烟草黑胫病菌培养特性的研究

    Institute of Scientific and Technical Information of China (English)

    张文友; 刘铭; 尹福强; 赵云飞; 刘朝科

    2013-01-01

    研究了采自攀西地区的烟草黑胫病菌在不同培养基、不同温度及不同pH值等条件下培养特性。结果表明:烟草黑胫病菌最适合在燕麦培养基上生长,菌丝生长快。菌丝在20~36℃都能生长,28℃生长最佳。在不同pH值的培养基培养下,菌丝最适合在pH值为5.5的培养基上生长。%Phytophthora parasitica var.nicotianae was cultivated under the conditions of different cultural media ,pH value and temperature ,then observed its growth status and morphological characteristics.The results showed that oatmeal agar was most suitable for the growth of the strains of P. Nicotianae.The hyphae can grow at 20~36℃,the temperature of 28℃and the pH value of 5.5 were the best for the growth of the strains of P.Nicotianae.

  14. Efficient Heterologous Transformation of <em>Chlamydomonas> reinhardtiiem> <em>npq2em> Mutant with the Zeaxanthin Epoxidase Gene Isolated and Characterized from<em> Chlorella zofingiensisem>

    Directory of Open Access Journals (Sweden)

    Herminia Rodríguez

    2012-09-01

    Full Text Available In the violaxanthin cycle, the violaxanthin de-epoxidase and zeaxanthin epoxidase catalyze the inter-conversion between violaxanthin and zeaxanthin in both plants and green algae. The zeaxanthin epoxidase gene from the green microalga <em>Chlorella zofingiensisem> (<em>Czzep> has been isolated<em>. em>This gene encodes a polypeptide of 596 amino acids. A single copy of <em>Czzep> has been found in the <em>C. zofingiensisem> genome by Southern blot analysis. qPCR analysis has shown that transcript levels of <em>Czzep> were increased after zeaxanthin formation under high light conditions. The functionality of <em>Czzep> gene by heterologous genetic complementation in the <em>Chlamydomonas> mutant <em>npq2em>, which lacks zeaxanthin epoxidase (ZEP activity and accumulates zeaxanthin in all conditions, was analyzed. The <em>Czzep> gene was adequately inserted in the pSI105 vector and expressed in <em>npq2em>. The positive transformants were able to efficiently convert zeaxanthin into violaxanthin, as well as to restore their maximum quantum efficiency of the PSII (Fv/Fm. These results show that <em>Chlamydomonas> can be an efficient tool for heterologous expression and metabolic engineering for biotechnological applications.

  15. Dermatoses em renais cronicos em terapia dialitica

    Directory of Open Access Journals (Sweden)

    Luis Alberto Batista Peres

    2014-03-01

    Full Text Available Objetivo: As desordens cutâneas e das mucosas são comuns em pacientes em hemodiálise a longo prazo. A diálise prolonga a expectativa de vida, dando tempo para a manifestação destas anormalidades. Os objetivos deste estudo foram avaliar a prevalência de problemas dermatológicos em pacientes com doença renal crônica (DRC em hemodiálise. Métodos: Cento e quarenta e cinco pacientes com doença renal crônica em hemodiálise foram estudados. Todos os pacientes foram completamente analisados para as alterações cutâneas, de cabelos, mucosas e unhas por um único examinador e foram coletados dados de exames laboratoriais. Os dados foram armazenados em um banco de dados do Microsolft Excel e analisados por estatística descritiva. As variáveis contínuas foram comparadas pelo teste t de Student e as variáveis categóricas utilizando o teste do qui-quadrado ou o teste Exato de Fischer, conforme adequado. Resultados: O estudo incluiu 145 pacientes, com idade média de 53,6 ± 14,7 anos, predominantemente do sexo masculino (64,1% e caucasianos (90,0%. O tempo médio de diálise foi de 43,3 ± 42,3 meses. As principais doenças subjacentes foram: hipertensão arterial em 33,8%, diabetes mellitus em 29,6% e glomerulonefrite crônica em 13,1%. As principais manifestações dermatológicas observadas foram: xerose em 109 (75,2%, equimose em 87 (60,0%, prurido em 78 (53,8% e lentigo em 33 (22,8% pacientes. Conclusão: O nosso estudo mostrou a presença de mais do que uma dermatose por paciente. As alterações cutâneas são frequentes em pacientes em diálise. Mais estudos são necessários para melhor caracterização e manejo destas dermatoses.

  16. Bioassay-Guided Isolation and Identification of Cytotoxic Compounds from <em>Gymnosperma> <em>glutinosum> Leaves

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    Cristina Rodríguez-Padilla

    2012-09-01

    Full Text Available Bioassay-guided fractionation of hexane extracts of<em> Gymnosperma glutinosumem> (Asteraceae leaves, collected in North Mexico, afforded the known compounds hentriacontane (1 and (+-13<em>S>,14<em>R>,15-trihydroxy-<em>ent>-labd-7-ene (2, as well as the new <em>ent>-labdane diterpene (−-13<em>S>,14<em>R>,15-trihydroxy-7-oxo-<em>ent>-labd-8(9-ene (3. In addition, D-glycero-D-galactoheptitol (4 was isolated from the methanolic extract of this plant. Their structures were established on the basis of high-field 1D- and 2D NMR methods supported by HR-MS data. The cytotoxic activity was determined by using the <em>in vitroem> L5178Y-R lymphoma murine model. Hentriacontane (1 and the new <em>ent>-labdane 3 showed weak cytotoxicity, whereas the <em>ent>-labdane 2 showed significant (<em>p> < 0.05 and concentration dependent cytotoxicity (up to 78% against L5178Y-R cells at concentrations ranging from 7.8 to 250 µg/mL.

  17. A pleiotropic drug resistance transporter in Nicotiana tabacum is involved in defense against the herbivore Manduca sexta.

    Science.gov (United States)

    Bienert, Manuela D; Siegmund, Stephanie E G; Drozak, Anna; Trombik, Tomasz; Bultreys, Alain; Baldwin, Ian T; Boutry, Marc

    2012-12-01

    Pleiotropic drug resistance (PDR) transporters are a group of membrane proteins belonging to the ABCG sub-family of ATP binding cassette (ABC) transporters. There is clear evidence for the involvement of plant ABC transporters in resistance to fungal and bacterial pathogens, but not in the biotic stress response to insect or herbivore attack. Here, we describe a PDR transporter, ABCG5/PDR5, from Nicotiana tabacum. GFP fusion and subcellular fractionation studies revealed that ABCG5/PDR5 is localized to the plasma membrane. Staining of transgenic plants expressing the GUS reporter gene under the control of the ABCG5/PDR5 transcription promoter and immunoblotting of wild-type plants showed that, under standard growth conditions, ABCG5/PDR5 is highly expressed in roots, stems and flowers, but is only expressed at marginal levels in leaves. Interestingly, ABCG5/PDR5 expression is induced in leaves by methyl jasmonate, wounding, pathogen infiltration, or herbivory by Manduca sexta. To address the physiological role of ABCG5/PDR5, N. tabacum plants silenced for the expression of ABCG5/PDR5 were obtained. No phenotypic modification was observed under standard conditions. However, a small increase in susceptibility to the fungus Fusarium oxysporum was observed. A stronger effect was observed in relation to herbivory: silenced plants allowed better growth and faster development of M. sexta larvae than wild-type plants, indicating an involvement of this PDR transporter in resistance to M. sexta herbivory.

  18. Isolation and molecular characterization of an ethylene response factor NtERF1-1 in Nicotiana tabacum cv. Xanthi

    Indian Academy of Sciences (India)

    Jun-Shan Gao; Li Hu; Peng Xie; Yan Meng; Yong-Ping Cai; Yi Lin

    2014-12-01

    Apetala2/Ethylene Response Factors (AP2/ERF) play important roles in regulating gene expression under abiotic and biotic stress in the plant kingdom. Here, we isolated a member of the AP2/ERF transcription factors, NtERF1-1, from Nicotiana tabcum cv. Xanthi NN carrying the N gene, which is resistant to Tobacco mosaic virus (TMV). NtERF1-1 encoded a putative protein of 229 amino acids with a predicted molecular mass of 24.58 kDa. Nucleotide sequence analysis showed that NtERF1-1 contained a conserved DNA-binding domain at the N-terminal. Comparison of amino acid sequences revealed that NtERF1-1 possessed high similarities to ERFs from diverse plants. Semi-quantitative and real-time quantitative RT-PCR analyses indicated that NtERF1-1 was up-regulated following TMV infection. In addition, we speculated that NtERF1-1 might participate in the signal transduction pathway of defence response inducted by the interaction between the gene and TMV.

  19. The ABC transporter ATR1 is necessary for efflux of the toxin cercosporin in the fungus Cercospora nicotianae.

    Science.gov (United States)

    Amnuaykanjanasin, Alongkorn; Daub, Margaret E

    2009-02-01

    The Cercospora nicotianae mutant deficient for the CRG1 transcription factor has marked reductions in both resistance and biosynthesis of the toxin cercosporin. We cloned and sequenced full-length copies of two genes, ATR1 and CnCFP, previously identified from a subtractive library between the wild type (WT) and a crg1 mutant. ATR1 is an ABC transporter gene and has an open reading frame (ORF) of 4368bp with one intron. CnCFP encodes a MFS transporter with homology to Cercospora kikuchii CFP, previously implicated in cercosporin export, and has an ORF of 1975bp with three introns. Disruption of ATR1 indicated atr1-null mutants had dramatic reductions in cercosporin production (25% and 20% of WT levels) in solid and liquid cultures, respectively. The ATR1 disruptants also showed moderately higher sensitivity to cercosporin. Constitutive expression of ATR1 in the crg1 mutant restored cercosporin biosynthesis and moderately increased resistance. In contrast, CnCFP overexpression in the mutant did not restore toxin production, however, it moderately enhanced toxin resistance. The results together indicate ATR1 acts as a cercosporin efflux pump in this fungus and plays a partial role in resistance.

  20. The dihydrolipoyl acyltransferase gene BCE2 participates in basal resistance against Phytophthora infestans in potato and Nicotiana benthamiana.

    Science.gov (United States)

    Wang, Hongyang; Sun, Chunlian; Jiang, Rui; He, Qin; Yang, Yu; Tian, Zhejuan; Tian, Zhendong; Xie, Conghua

    2014-07-01

    Dihydrolipoyl acyltransferase (EC 2.3.1.12), a branched-chain α-ketoacid dehydrogenase E2 subunit (BCE2), catalyzes the transfer of the acyl group from the lipoyl moiety to coenzyme A. However, the role of BCE2 responding to biotic stress in plant is not clear. In this study, we cloned and characterized a BCE2 gene from potato, namely StBCE2, which was previously suggested to be involved in Phytophthora infestans-potato interaction. We found that the expression of StBCE2 was strongly induced by both P. infestans isolate HB09-14-2 and salicylic acid. Besides, when the homolog of StBCE2 in Nicotiana benthamiana named NbBCE2 was silenced, plants showed increased susceptibility to P. infestans and reduced accumulation of hydrogen peroxide (H2O2). Furthermore, we found that a marker gene NbrbohB involved in the production of reactive oxygen species, was also suppressed in NbBCE2-silenced plants. However, silencing of NbBCE2 had no significant effect on the hypersensitive responses trigged by INF1, R3a-AVR3a(KI) pair or Rpi-vnt1.1-AVR-vnt1.1 pair. Our results suggest that BCE2 is associated with the basal resistance to P. infestans by regulating H2O2 production.

  1. Mechanisms of Optimal Defense Patterns in Nicotiana attenuata: Flowering Attenuates Herbivory-elicited Ethylene and Jasmonate Signaling

    Institute of Scientific and Technical Information of China (English)

    Celia Diezel; Silke AIImann; lan T. Baldwin

    2011-01-01

    To defend themselves against herbivore attack,plants produce secondary metabolites,which are variously inducible and constitutively deployed,presumably to optimize their fitness benefits in light of their fitness costs.Three phytohormones,jasmonates (JA) and their active forms,the JA-isoleucine (JA-Ile) and ethylene (ET),are known to play central roles in the elicitation of induced defenses,but little is known about how this mediation changes over ontogeny.The Optimal Defense Theory (ODT) predicts changes in the costs and benefits of the different types of defenses and has been usefully extrapolated to their modes of deployment.Here we studied whether the herbivore-induced accumulation of JA,JA-lle and ET changed over ontogeny in Nicotiana attenuata,a native tobacco in which inducible defenses are particularly well studied.Herbivore-elicited ET production changed dramatically during six developmental stages,from rosette through flowering,decreasing with the elongation of the first corollas during flower development.This decrease was largely recovered within a day after flower removal by decapitation.A similar pattern was found for the herbivore-induced accumulation of JA and JA-IIe.These results are consistent with ODT predictions and suggest that the last steps in floral development control the inducibility of at least three plant hormones,optimizing defense-growth tradeoffs.

  2. Shifting Nicotiana attenuata’s diurnal rhythm does not alter its resistance to the specialist herbivore Manduca sexta

    Institute of Scientific and Technical Information of China (English)

    Jasmin Herden; Stefan Meldau; Sang-Gyu Kim; Grit Kunert; Youngsung Joo; Ian T Baldwin; Meredith C Schuman

    2016-01-01

    Arabidopsis thaliana plants are less resistant to attack by the generalist lepidopteran herbivore Trichoplusia ni when plants and herbivores are entrained to opposite, versus identical diurnal cycles and tested under constant conditions. This effect is associated with circadian fluctuations in levels of jasmonic acid, the transcription factor MYC2, and glucosino-late contents in leaves. We tested whether a similar effect could be observed in a different plant–herbivore system:the wild tobacco Nicotiana attenuata and its co-evolved specialist herbivore, Manduca sexta. We measured larval growth on plants under both constant and diurnal conditions following identical or opposite entrainment, profiled the metabolome of attacked leaf tissue, quantified specific metabolites known to reduce M. sexta growth, and monitored M. sexta feeding activity under all experimental conditions. Entrainment did not consistently affect M. sexta growth or plant defense induction. However, both were reduced under constant dark conditions, as was M. sexta feeding activity. Our data indicate that the response induced by M. sexta in N. attenuata is robust to diurnal cues and independent of plant or herbivore entrain-ment. We propose that while the patterns of constitutive or general damage-induced defense may undergo circadian fluctuation, the orchestration of specific induced responses is more complex.

  3. Rapid Transient Production of a Monoclonal Antibody Neutralizing the Porcine Epidemic Diarrhea Virus (PEDV) in Nicotiana benthamiana and Lactuca sativa.

    Science.gov (United States)

    Rattanapisit, Kaewta; Srijangwad, Anchalee; Chuanasa, Taksina; Sukrong, Suchada; Tantituvanont, Angkana; Mason, Hugh S; Nilubol, Dachrit; Phoolcharoen, Waranyoo

    2017-06-02

    Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea, vomiting, dehydration, weight loss, and high mortality rate in neonatal piglets. Porcine epidemic diarrhea (PED) has been reported in Europe, America, and Asia including Thailand. The disease causes substantial losses to the swine industry in many countries. Presently, there is no effective PEDV vaccine available. In this study, we developed a plant-produced monoclonal antibody (mAb) 2C10 as a prophylactic candidate to prevent the PEDV infection. Recently, plant expression systems have gained interest as an alternative for the production of antibodies because of many advantages, such as low production cost, lack of human and animal pathogen, large scalability, etc. The 2C10 mAb was transiently expressed in Nicotiana benthamiana and lettuce using geminiviral vector. After purification by protein A affinity chromatography, the antibody was tested for the binding and neutralizing activity against PEDV. Our result showed that the plant produced 2C10 mAb can bind to the virus and also inhibit PEDV infection in vitro. These results show excellent potential for a plant-expressed 2C10 as a PEDV prophylaxis and a diagnostic for PEDV infection. Georg Thieme Verlag KG Stuttgart · New York.

  4. Cloning the bacterial bphC gene into Nicotiana tabacum to improve the efficiency of phytoremediation of polychlorinated biphenyls

    Science.gov (United States)

    Novakova, Martina; Mackova, Martina; Antosova, Zuzana; Viktorova, Jitka; Szekeres, Miklos; Demnerova, Katerina

    2010-01-01

    The aim of this work was to construct transgenic plants with increased capabilities to degrade organic pollutants, such as polychlorinated biphenyls. The environmentally important gene of bacterial dioxygenase, the bphC gene, was chosen to clone into a plant of Nicotiana tabacum. The chosen bphC gene encodes 2,3-dihydroxybiphenyl-1,2-dioxygenase, which cleaves the aromatic ring of dihydroxybiphenyl, and we cloned it in fusion with the gene for β-glucuronidase (GUS), luciferase (LUC) or with a histidine tail. Several genetic constructs were designed and prepared and the possible expression of desired proteins in tobacco plants was studied by transient expression. We used genetic constructs successfully expressing dioxygenase's genes we used for preparation of transgenic tobacco plants by agrobacterial infection. The presence of transgenic DNA , mRNA and protein was determined in parental and the first filial generation of transgenic plants with the bphC gene. Properties of prepared transgenic plants will be further studied. PMID:21468210

  5. Relationship of microbial communities and suppressiveness of Trichoderma fortified composts for pepper seedlings infected by Phytophthora nicotianae.

    Science.gov (United States)

    Ros, Margarita; Raut, Iulia; Santisima-Trinidad, Ana Belén; Pascual, Jose Antonio

    2017-01-01

    The understanding of the dynamic of soil-borne diseases is related to the microbial composition of the rhizosphere which is the key to progress in the field of biological control. Trichoderma spp. is commonly used as a biological control agent. The use of next generation sequencing approaches and quantitative PCR are two successful approaches to assess the effect of using compost as substrate fortified with two Trichoderma strains (Trichoderma harzianum or Trichoderma asperellum) on bacterial and fungal communities in pepper rhizosphere infected with Phytophthora nicotianae. The results showed changes in the bacterial rhizosphere community not attributed to the Trichoderma strain, but to the pathogen infection, while, fungi were not affected by pathogen infection and depended on the type of substrate. The Trichoderma asperellum fortified compost was the most effective combination against the pathogen. This could indicate that the effect of fortified composts is greater than compost itself and the biocontrol effect should be attributed to the Trichoderma strains rather than the compost microbiota, although some microorganisms could help with the biocontrol effect.

  6. Silencing of host basal defense response-related gene expression increases susceptibility of Nicotiana benthamiana to Clavibacter michiganensis subsp. michiganensis.

    Science.gov (United States)

    Balaji, Vasudevan; Sessa, Guido; Smart, Christine D

    2011-03-01

    Clavibacter michiganensis subsp. michiganensis is an actinomycete, causing bacterial wilt and canker disease of tomato (Solanum lycopersicum). We used virus-induced gene silencing (VIGS) to identify genes playing a role in host basal defense response to C. michiganensis subsp. michiganensis infection using Nicotiana benthamiana as a model plant. A preliminary VIGS screen comprising 160 genes from tomato known to be involved in defense-related signaling identified a set of 14 genes whose suppression led to altered host-pathogen interactions. Expression of each of these genes and three additional targets was then suppressed in larger-scale VIGS experiments and the effect of silencing on development of wilt disease symptoms and bacterial growth during an N. benthamiana-C. michiganensis subsp. michiganensis compatible interaction was determined. Disease susceptibility and in planta bacterial population size were enhanced by silencing genes encoding N. benthamiana homologs of ubiquitin activating enzyme, snakin-2, extensin-like protein, divinyl ether synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase 2, and Pto-like kinase. The identification of genes having a role in the host basal defense-response to C. michiganensis subsp. michiganensis advances our understanding of the plant responses activated by C. michiganensis subsp. michiganensis and raises possibilities for devising novel and effective molecular strategies to control bacterial canker and wilt in tomato.

  7. Effects and Effectiveness of Two RNAi Constructs for Resistance to Pepper golden mosaic virus in Nicotiana benthamiana Plants

    Directory of Open Access Journals (Sweden)

    Diana Medina-Hernández

    2013-11-01

    Full Text Available ToChLPV and PepGMV are Begomoviruses that have adapted to a wide host range and are able to cause major diseases in agronomic crops. We analyzed the efficacy of induced resistance to PepGMV in Nicotiana benthamiana plants with two constructs: one construct with homologous sequences derived from PepGMV, and the other construct with heterologous sequences derived from ToChLPV. Plants protected with the heterologous construct showed an efficacy to decrease the severity of symptoms of 45%, while plants protected with the homologous construct showed an efficacy of 80%. Plants protected with the heterologous construct showed a reduction of incidence of 42.86%, while the reduction of incidence in plants protected with the homologous construct was 57.15%. The efficacy to decrease viral load was 95.6% in plants protected with the heterologous construct, and 99.56% in plants protected with the homologous construct. We found, in both constructs, up-regulated key components of the RNAi pathway. This demonstrates that the efficacy of the constructs was due to the activation of the gene silencing mechanism, and is reflected in the decrease of viral genome copies, as well as in recovery phenotype. We present evidence that both constructs are functional and can efficiently induce transient resistance against PepGMV infections. This observation guarantees a further exploration as a strategy to control complex Begomovirus diseases in the field.

  8. Mp10 and Mp42 from the aphid species Myzus persicae trigger plant defenses in Nicotiana benthamiana through different activities.

    Science.gov (United States)

    Rodriguez, Patricia A; Stam, Remco; Warbroek, Tim; Bos, Jorunn I B

    2014-01-01

    Aphids are phloem-feeding insects that, like other plant parasites, deliver effectors inside their host to manipulate host responses. The Myzus persicae (green peach aphid) candidate effectors Mp10 and Mp42 were previously found to reduce aphid fecundity upon intracellular transient overexpression in Nicotiana benthamiana. We performed functional analyses of these proteins to investigate whether they activate defenses through similar activities. We employed a range of functional characterization experiments based on intracellular transient overexpression in N. benthamiana to determine the subcellular localization of Mp10 and Mp42 and investigate their role in activating plant defense signaling. Mp10 and Mp42 showed distinct subcellular localization in planta, suggesting that they target different host compartments. Also, Mp10 reduced the levels of Agrobacterium-mediated overexpression of proteins. This reduction was not due to an effect on Agrobacterium viability. Transient overexpression of Mp10 but not Mp42 activated jasmonic acid and salicylic acid signaling pathways and decreased susceptibility to the hemibiotrophic plant pathogen Phytophthora capsici. We found that two candidate effectors from the broad-host-range aphid M. persicae can trigger aphid defenses through different mechanisms. Importantly, we found that some (candidate) effectors such as Mp10 interfere with Agrobacterium-based overexpression assays, an important tool to study effector activity and function.

  9. Effects of selenium on the growth and photosynthetic characteristics of flue-cured tobacco (Nicotiana tabacum L.

    Directory of Open Access Journals (Sweden)

    Chaoqiang Jiang

    2015-03-01

    Full Text Available The objective of this study was to investigate the effect of Selenium (Se supply (0, 3, 6, 12, 24 mg kg−1 on the growth, photosynthetic characteristics, Se accumulation and distribution of flue-cured tobacco (Nicotiana tabacum L.. Results showed that low-dose Se treatments (≤6 mg kg−1 stimulated plant growth but high-dose Se treatments (≥12 mg kg−1 hindered plant growth. Optimal Se dose (6 mg kg−1 stimulated plant growth by reducing MDA content and improving photosynthetic capability. However, excess Se (24 mg kg−1 increased MDA content by 28%, decreased net photosynthetic rate and carboxylation efficiency by 34% and 39%, respectively. The Se concentration in the roots, stems, and leaves of the tobacco plants significantly increased with increasing Se application. A linear correlation (R = 0.95, P leaf > stem. The Se concentration in the roots was 3.17 and 7.57 times higher than that in the leaves and stems, respectively, after treatment with 24 mg kg−1 Se. In conclusion, the present study suggested that optimal Se dose (6 mg kg−1 improved the plant growth mainly by enhancing photosynthesis, stomatal conductance, carboxylation efficiency and Rubisco content in the flue-cured tobacco leaves. However, the inhibition of excess Se on tobacco growth might be due to high accumulation of Se in roots and the damage of photosynthesis in leaves.

  10. Heterologous expression of the BABY BOOM AP2/ERF transcription factor enhances the regeneration capacity of tobacco (Nicotiana tabacum L.).

    Science.gov (United States)

    Srinivasan, Chinnathambi; Liu, Zongrang; Heidmann, Iris; Supena, Ence Darmo Jaya; Fukuoka, Hiro; Joosen, Ronny; Lambalk, Joep; Angenent, Gerco; Scorza, Ralph; Custers, Jan B M; Boutilier, Kim

    2007-01-01

    Gain-of-function studies have shown that ectopic expression of the BABY BOOM (BBM) AP2/ERF domain transcription factor is sufficient to induce spontaneous somatic embryogenesis in Arabidopsis (Arabidopsis thaliana (L.) Heynh) and Brassica napus (B. napus L.) seedlings. Here we examined the effect of ectopic BBM expression on the development and regenerative capacity of tobacco (Nicotiana tabacum L.) through heterologous expression of Arabidopsis and B. napus BBM genes. 35S::BBM tobacco lines exhibited a number of the phenotypes previously observed in 35S::BBM Arabidopsis and B. napus transgenics, including callus formation, leaf rumpling, and sterility, but they did not undergo spontaneous somatic embryogenesis. 35S::BBM plants with severe ectopic expression phenotypes could not be assessed for enhanced regeneration at the seedling stage due to complete male and female sterility of the primary transformants, therefore fertile BBM ectopic expression lines with strong misexpression phenotypes were generated by expressing a steroid-inducible, post-translationally controlled BBM fusion protein (BBM:GR) under the control of a 35S promoter. These lines exhibited spontaneous shoot and root formation, while somatic embryogenesis could be induced from in-vitro germinated seedling hypocotyls cultured on media supplemented with cytokinin. Together these results suggest that ectopic BBM expression in transgenic tobacco also activates cell proliferation pathways, but differences exist between Arabidopsis/B. napus and N. tabacum with respect to their competence to respond to the BBM signalling molecule.

  11. Role of Rice stripe virus NSvc4 in cell-to-cell movement and symptom development in Nicotiana benthamiana

    Directory of Open Access Journals (Sweden)

    Yi eXu

    2012-12-01

    Full Text Available Our previous work has demonstrated that the NSvc4 protein of Rice stripe virus (RSV functions as a cell-to-cell movement protein. However, the mechanisms whereby RSV traffics through plasmodesmata (PD are unknown. Here we provide evidence that the NSvc4 moves on the actin filament and endoplasmic reticulum (ER network, but not microtubules, to reach cell wall PD. Disruption of cytoskeleton using different inhibitors altered NSvc4 localization to PD, thus impeding RSV infection of Nicotiana benthamiana. Sequence analyses and deletion mutagenesis experiment revealed that the N-terminal 125 amino acids (AAs of the NSvc4 determine PD targeting and that a transmembrane domain spanning AAs 106 to 125 is critical for PD localization. We also found that the NSvc4 protein can localize to chloroplasts in infected cells. Analyses using deletion mutants revealed that the N-terminal 73 AAs are essential for chloroplast localization. Furthermore, expression of NSvc4 from a Potato virus X (PVX vector resulted in more severe disease symptoms than PVX alone in systemically infected N. benthamiana leaves. Expression of NSvc4 in Spodoptera frugiperda 9 (Sf-9 cells did not elicit tubule formation, but instead resulted in punctate foci at the plasma membrane. These findings shed new light on our understanding of the movement mechanisms whereby RSV infects host plants.

  12. Dicer-Like 4 Is Involved in Restricting the Systemic Movement of Zucchini yellow mosaic virus in Nicotiana benthamiana.

    Science.gov (United States)

    Cordero, Teresa; Cerdán, Lidia; Carbonell, Alberto; Katsarou, Konstantina; Kalantidis, Kriton; Daròs, José-Antonio

    2017-01-01

    Zucchini yellow mosaic virus (ZYMV) induces serious diseases in cucurbits. To create a tool to screen for resistance genes, we cloned a wild ZYMV isolate and inserted the visual marker Rosea1 to obtain recombinant clone ZYMV-Ros1. While in some plant-virus combinations Rosea1 induces accumulation of anthocyanins in infected tissues, ZYMV-Ros1 infection of cucurbits did not lead to detectable anthocyanin accumulation. However, the recombinant virus did induce dark red pigmentation in infected tissues of the model plant Nicotiana benthamiana. In this species, ZYMV-Ros1 multiplied efficiently in local inoculated tissue but only a few progeny particles established infection foci in upper leaves. We used this system to analyze the roles of Dicer-like (DCL) genes, core components of plant antiviral RNA silencing pathways, in ZYMV infection. ZYMV-Ros1 local replication was not significantly affected in single DCL knockdown lines nor in double DCL2/4 and triple DCL2/3/4 knockdown lines. ZYMV-Ros1 systemic accumulation was not affected in knockdown lines DCL1, DCL2, and DCL3. However in DCL4 and also in DCL2/4 and DCL2/3/4 knockdown lines, ZYMV-Ros1 systemic accumulation dramatically increased, which highlights the key role of DCL4 in restricting virus systemic movement. The effect of DCL4 on ZYMV systemic movement was confirmed with a wild-type version of the virus.

  13. [The role of Cd-binding proteins and phytochelatins in the formation of cadmium resistance in Nicotiana plumbaginifolia cell lines].

    Science.gov (United States)

    Fenik, S I; Solodushko, V G; Kaliniak, T B; Blium, Ia B

    2007-01-01

    Nicotiana plumbaginifolia callus lines with the equal resistance to cadmium have been produced under different selective conditions--either without inhibition of the phytochelatin synthesis (line Cd-R) or in the presence of the inhibitor butionine sulfoximine (line Cd-Ri). The level of phytochelatin synthesis in the line Cd-R five-fold exceeded the control value and in the line Cd-Ri it was twice as much as in the control. It was shown that in the control line mainly three cadmium-binding proteins are expressed of the molecular weihgts 41, 34 and 19 kD. The common feature of the both resistant lines is the expression of the cadmium-binding proteins of 40, 37 and 19 kD. The resistant lines differ with respect to the synthesis of relatively low-molecular cadmium-binding proteins. The proteins of the molecular weights 12.5, 11.5 and 9 kD are expressed in the line Cd-R, while the proteins of 13 and 10 kD are expressed in the line Cd-Ri. It was supposed that both the phytochelatins and the Cd-binding proteins contribute to the resisitance of N. plumbaginifolia callus lines to cadmium and the lack of the phytochelatins can be equilibrated by the changes in the low-molecular Cd-binding protein synthesis.

  14. Theobroxide Treatment Inhibits Wild Fire Disease Occurrence in Nicotiana benthamiana by the Overexpression of Defense-related Genes

    Directory of Open Access Journals (Sweden)

    Soon Young Ahn

    2013-03-01

    Full Text Available Theobroxide, a novel compound isolated from a fungus Lasiodiplodia theobromae, stimulates potato tuber formation and induces flowering of morning glory by initiating the jasmonic acid synthesis pathway. To elucidate the effect of theobroxide on pathogen resistance in plants, Nicotiana benthamiana plants treated with theobroxide were immediately infiltrated with Pseudomonas syringae pv. tabaci. Exogenous application of theobroxide inhibited development of lesion symptoms, and growth of the bacterial cells was significantly retarded. Semi-quantitative RT-PCRs using the primers of 18 defense-related genes were performed to investigate the molecular mechanisms of resistance. Among the genes, the theobroxide treatment increased the expression of pathogenesis-related protein 1a (PR1a, pathogenesis-related protein 1b (PR1b, glutathione S-transferase (GST, allen oxide cyclase (AOC, and lipoxyganase (LOX. All these data strongly indicate that theobroxide treatment inhibits disease development by faster induction of defense responses, which can be possible by the induction of defense-related genes including PR1a, PR1b, and GST triggered by the elevated jasmonic acid.

  15. Narboh D, a Respiratory Burst Oxidase Homolog in Nicotiana attenuata, is Required for Late Defense Responses After Herbivore Attack

    Institute of Scientific and Technical Information of China (English)

    Jinsong Wu; Lei Wang; Hendrik Wünsche; Ian T.Baldwin

    2013-01-01

    The superoxide (O2-)-generating NADPH oxidases are crucial for the defense of plants against attack from pathogens; however,it remains unknown whether they also mediate responses against chewing insect herbivores.The transcripts of the respiratory burst NADPH oxidase homolog Narboh D in Nicotiana attenuate are rapidly and transiently elicited by wounding,and are amplified when Manduca sexta oral secretions (OS) are added to the wounds.The fatty-acid-amino-acid-conjugates (FACs),demonstrably the major elicitors in M.sexta OS,are responsible for the increase in Narboh D transcripts.Silencing Narboh D significantly reduced reactive oxygen species (ROS) levels after OS elicitation,but neither OS-elicited jasmonic acid (JA) or JA-isoleucine (JA-Ile) bursts,pivotal hormones that regulates plant resistance to herbivores,nor early transcripts of herbivore defense-related genes (NaJAR4 and NaPAL1),were influenced.However,late OS-elicited increases in trypsin proteinase inhibitors (TPIs),as well as the transcript levels of defense genes such as polyphenol oxidase,TPI and Thionin were significantly reduced.In addition,Narboh D-silenced plants were more vulnerable to insect herbivores,especially the larvae of the generalist Spodoptera littoralis.We thus conclude that Narboh D-based defenses play an important role in late herbivore-elicited responses.

  16. Norovirus Narita 104 Virus-Like Particles Expressed in Nicotiana benthamiana Induce Serum and Mucosal Immune Responses

    Directory of Open Access Journals (Sweden)

    Lolita George Mathew

    2014-01-01

    Full Text Available Narita 104 virus is a human pathogen belonging to the norovirus (family Caliciviridae genogroup II. Noroviruses cause epidemic gastroenteritis worldwide. To explore the potential of developing a plant-based vaccine, a plant optimized gene encoding Narita 104 virus capsid protein (NaVCP was expressed transiently in Nicotiana benthamiana using a tobacco mosaic virus expression system. NaVCP accumulated up to approximately 0.3 mg/g fresh weight of leaf at 4 days postinfection. Initiation of hypersensitive response-like symptoms followed by tissue necrosis necessitated a brief infection time and was a significant factor limiting expression. Transmission electron microscopy of plant-derived NaVCP confirmed the presence of fully assembled virus-like particles (VLPs. In this study, an optimized method to express and partially purify NaVCP is described. Further, partially purified NaVCP was used to immunize mice by intranasal delivery and generated significant mucosal and serum antibody responses. Thus, plant-derived Narita 104 VLPs have potential for use as a candidate subunit vaccine or as a component of a multivalent subunit vaccine, along with other genotype-specific plant-derived VLPs.

  17. Norovirus Narita 104 Virus-Like Particles Expressed in Nicotiana benthamiana Induce Serum and Mucosal Immune Responses

    Science.gov (United States)

    Mathew, Lolita George; Herbst-Kralovetz, Melissa M.; Mason, Hugh S.

    2014-01-01

    Narita 104 virus is a human pathogen belonging to the norovirus (family Caliciviridae) genogroup II. Noroviruses cause epidemic gastroenteritis worldwide. To explore the potential of developing a plant-based vaccine, a plant optimized gene encoding Narita 104 virus capsid protein (NaVCP) was expressed transiently in Nicotiana benthamiana using a tobacco mosaic virus expression system. NaVCP accumulated up to approximately 0.3 mg/g fresh weight of leaf at 4 days postinfection. Initiation of hypersensitive response-like symptoms followed by tissue necrosis necessitated a brief infection time and was a significant factor limiting expression. Transmission electron microscopy of plant-derived NaVCP confirmed the presence of fully assembled virus-like particles (VLPs). In this study, an optimized method to express and partially purify NaVCP is described. Further, partially purified NaVCP was used to immunize mice by intranasal delivery and generated significant mucosal and serum antibody responses. Thus, plant-derived Narita 104 VLPs have potential for use as a candidate subunit vaccine or as a component of a multivalent subunit vaccine, along with other genotype-specific plant-derived VLPs. PMID:24949472

  18. Pretreatment with alternation of light/dark periods improves the tolerance of tobacco (Nicotiana tabacum) to clomazone herbicide.

    Science.gov (United States)

    Darwish, Majd; Lopez-Lauri, Félicie; El Maataoui, Mohamed; Urban, Laurent; Sallanon, Huguette

    2014-05-01

    This work analyses the effects of alternation of light/dark periods pretreatment (AL) in tobacco plantlets (Nicotiana tabacum L. cv.Virginie vk51) growing in solution with low concentration of the clomazone herbicide. The experimentation has been carried out by exposing the plantlets to successive and regulated periods of light (16min light/8min dark cycles, PAR 50μmolm(-2)s(-1)) for three days. The photosynthesis efficiency was determined by mean of the chlorophyll fluorescence and JIP-test. The AL pretreatment improved the clomazone tolerance; this has been observed by the increase in the leaf area of the plant, the maximal photochemical quantum efficiency of PSII (Fv/Fm), the actual PSII efficiency (ФPSII), the performance index (PIabs), the electron flux beyond Quinone A (1-VJ), and also by the diminution of the energy dissipating into heat (DI0/RC). Furthermore, AL pretreatment led to low accumulation of hydrogen peroxide (H2O2) which proves that the scavenging enzymatic system have been activated before clomazone treatment. In the plantlets pretreated with AL, with regard to the ascorbate content, some of antioxidant enzyme whose function is associated with it have continued to scavenge reactive oxygen species (ROS) induced by clomazone, such as ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and glutathione reductase (GR). So, the observed photooxidative damages induced by clomazone herbicide were noticeably reduced.

  19. Behaviour of Potassium and Trace Elements in Rhizosphere of Flue—Cured Tobacco (Nicotiana Tabacum L.)

    Institute of Scientific and Technical Information of China (English)

    HUGUO-SONG; CAOZHI-HONG; 等

    1993-01-01

    The study on the behaviour of potassium,phosphate and trace elements,Fe,Mn,Cu and Zn,in the rhizosphere of different varieties of flue-cured tobacco (Nicotiana tabacum L.)with high and low potassium application rate with rhizobag technique showed that soil available K,soil available P,and slow available K was in depletion status,whereas DTPA extractable Fe,Mn,Zn and Cu accumulated obviously in rhizosphere.The depletion and accumulation rates of mineral nutrients differed in degree with K application rate,soil type,and tobacco variety.The content of available K in both rhizosphere and bulk soil and K concentration in tobacco leaf increased significantly,and the available P in rhizosphere dropped with more K application.The DTPA-Fe content of red soil much lower in pH was higher than that of calcareous soil in bulk soil.But the DTPA-Fe content of calcareous soil was much higher than that of red soil in rhizosphere,which was considered perhaps to be mainly related to releasing of Fe phytosiderophore.Nitrate coule increase depletion of a vailable K in rhizosphere and also soil pH in comparision with ammonium.

  20. Deep sequencing analysis reveals a TMV mutant with a poly(A) tract reduces host defense responses in Nicotiana benthamiana.

    Science.gov (United States)

    Guo, Song; Wong, Sek-Man

    2017-07-15

    Tobacco mosaic virus (TMV) possesses an upstream pseudoknotted domain (UPD), which is important for replication. After substituting the UPD with an internal poly(A) tract (43 nt), a mutant TMV-43A was constructed. TMV-43A replicated slower than TMV and induced a non-lethal mosaic symptom in Nicotiana benthamiana. In this study, deep sequencing was performed to detect the differences of small RNA profiles between TMV- and TMV-43A-infected N. benthamiana. The results showed that TMV-43A produced lesser amount of virus-derived interfering RNAs (vsiRNAs) than that of TMV. However, the distributions of vsiRNAs generation hotspots between TMV and TMV-43A were similar. Expression of genes related to small RNA biogenesis in TMV-43A-infected N. benthamiana was significantly lower than that of TMV, which leads to generation of lesser vsiRNAs. The expressions of host defense response genes were up-regulated after TMV infection, as compared to TMV-43A-infected plants. Host defense response to TMV-43A infection was lower than that to TMV. The absence of UPD might contribute to the reduced host response to TMV-43A. Our study provides valuable information in the role of the UPD in eliciting host response genes after TMV infection in N. benthamiana. (187 words). Copyright © 2017 Elsevier B.V. All rights reserved.

  1. The sclerophyllous Eucalyptus camaldulensis and herbaceous Nicotiana tabacum have different mechanisms to maintain high rates of photosynthesis

    Directory of Open Access Journals (Sweden)

    Wei Huang

    2016-11-01

    Full Text Available It is believed that high levels of mesophyll conductance (gm largely contribute to the high rates of photosynthesis in herbaceous C3 plants. However, some sclerophyllous C3 plants that display low levels of gm have high rates of photosynthesis, and the underlying mechanisms responsible for high photosynthetic rates in sclerophyllous C3 plants are unclear. In the present study, we examined photosynthetic characteristics in two high-photosynthesis plants (the sclerophyllous Eucalyptus camaldulensis and the herbaceous Nicotiana tabacum using measurements of gas exchange and chlorophyll fluorescence. Under saturating light intensities, both species had similar rates of CO2 assimilation at 400 μmol mol-1 CO2 (A400. However, E. camaldulensis exhibited significantly lower gm and chloroplast CO2 concentration (Cc than N. tabacum. A quantitative analysis revealed that, in E. camaldulensis, the gm limitation was the most constraining factor for photosynthesis. By comparison, in N. tabacum, the biochemical limitation was the strongest, followed by gm and gs limitations. In conjunction with a lower Cc, E. camaldulensis up-regulated the capacities of photorespiratory pathway and alternative electron flow. Furthermore, the rate of alternative electron flow was positively correlated with the rates of photorespiration and ATP supply from other flexible mechanisms, suggesting the important roles of photorespiratory pathway and alternative electron flow in sustaining high rate of photosynthesis in E. camaldulensis. These results highlight the different mechanisms used to maintain high rates of photosynthesis in the sclerophyllous E. camaldulensis and the herbaceous N. tabacum.

  2. The Sclerophyllous Eucalyptus camaldulensis and Herbaceous Nicotiana tabacum Have Different Mechanisms to Maintain High Rates of Photosynthesis

    Science.gov (United States)

    Huang, Wei; Tong, You-Gui; Yu, Guo-Yun; Yang, Wei-Xian

    2016-01-01

    It is believed that high levels of mesophyll conductance (gm) largely contribute to the high rates of photosynthesis in herbaceous C3 plants. However, some sclerophyllous C3 plants that display low levels of gm have high rates of photosynthesis, and the underlying mechanisms responsible for high photosynthetic rates in sclerophyllous C3 plants are unclear. In the present study, we examined photosynthetic characteristics in two high-photosynthesis plants (the sclerophyllous Eucalyptus camaldulensis and the herbaceous Nicotiana tabacum) using measurements of gas exchange and chlorophyll fluorescence. Under saturating light intensities, both species had similar rates of CO2 assimilation at 400 μmol mol−1 CO2 (A400). However, E. camaldulensis exhibited significantly lower gm and chloroplast CO2 concentration (Cc) than N. tabacum. A quantitative analysis revealed that, in E. camaldulensis, the gm limitation was the most constraining factor for photosynthesis. By comparison, in N. tabacum, the biochemical limitation was the strongest, followed by gm and gs limitations. In conjunction with a lower Cc, E. camaldulensis up-regulated the capacities of photorespiratory pathway and alternative electron flow. Furthermore, the rate of alternative electron flow was positively correlated with the rates of photorespiration and ATP supply from other flexible mechanisms, suggesting the important roles of photorespiratory pathway, and alternative electron flow in sustaining high rate of photosynthesis in E. camaldulensis. These results highlight the different mechanisms used to maintain high rates of photosynthesis in the sclerophyllous E. camaldulensis and the herbaceous N. tabacum. PMID:27933083

  3. Experimental and computational studies of Nicotiana tabacum leaves extract as green corrosion inhibitor for mild steel in acidic medium

    Directory of Open Access Journals (Sweden)

    Jeetendra Bhawsar

    2015-09-01

    Full Text Available In the present work corrosion inhibition of mild steel in 2 M H2SO4 solution by Nicotiana tabacum extract was studied by weight loss method. It has been found that the extract acts as an effective corrosion inhibitor for mild steel in Sulfuric acid medium. The inhibition process is attributed to the formation of an adsorbed film of inhibitor on the metal surface which protects the metal against corrosion. The inhibition efficiency (%IE and surface coverage (θ of N. tabacum extract increased with increase in inhibitor concentration but decreased with increasing the temperature. The adsorption of extract on the mild steel surface was found to obey Langmuir’s adsorption isotherm. The free energy value (ΔGads indicated that the adsorption of inhibitor molecules was typical of physisorption. The results obtained show that N. tabacum Extract could serve as an excellent eco-friendly green corrosion inhibitor. Quantum chemical parameters such as highest occupied molecular orbital energy (EHOMO, lowest unoccupied molecular orbital energy (ELUMO, energy gap (ΔE, dipole moment (μ and Mulliken charges were calculated. Quantum chemical calculations also supported experimental data and the adsorption of inhibitor molecules onto the metal surface.

  4. Temporal and spatial resolution of activated plant defense responses in leaves of Nicotiana benthamiana infected with Dickeya dadantii

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    María Luisa ePérez-Bueno

    2016-01-01

    Full Text Available The necrotrophic bacteria Dickeya dadantii is the causal agent of soft-rot disease in a broad range of hosts. The model plant Nicotiana benthamiana, commonly used as experimental host for a very broad range of plant pathogens, is susceptible to infection by D. dadantii. The inoculation with D. dadantii at high dose seems to overcome the plant defense capacity, inducing maceration and death of the tissue, although restricted to the infiltrated area. By contrast, the output of the defense response to low dose inoculation is inhibition of maceration and limitation in the growth, or even eradication, of bacteria. Responses of tissue invaded by bacteria (neighbouring the infiltrated areas after 2-3 days post-inoculation included: i inhibition of photosynthesis in terms of photosystem II efficiency; ii activation of energy dissipation as non-photochemical quenching in photosystem II, which is related to the activation of plant defense mechanisms; and iii accumulation of secondary metabolites in cell walls of the epidermis (lignins and the apoplast of the mesophyll (phytoalexins. Infiltrated tissues showed an increase in the content of the main hormones regulating stress responses, including abscisic acid (ABA, jasmonic acid (JA and salicylic acid (SA. We propose a mechanism involving the three hormones by which N. benthamiana could activate an efficient defense response against D. dadantii.

  5. The combination of plant translational enhancers and terminator increase the expression of human glucocerebrosidase in Nicotiana benthamiana plants.

    Science.gov (United States)

    Limkul, Juthamard; Misaki, Ryo; Kato, Ko; Fujiyama, Kazuhito

    2015-11-01

    Gaucher's disease is a lysosomal storage disorder caused by mutations in the gene encoding glucocerebrosidase (GCase). It is currently treated by enzyme replacement therapy using recombinant GCase expressed in mammalian cells. Plant production systems are among the most attractive alternatives for pharmaceutical protein production due to such advantages as low-cost, high-scalability, and safety from human pathogen contamination. Because of its high biomass yield, Nicotiana benthamiana could be an economical recombinant GCase production system. In this study, a translational enhancer and suitable terminator were utilized to obtain a powerful expression system for GCase production in N. benthamiana plants. Six plasmid constructs were used. The highest activity of 44.5units/mg protein (after subtraction of endogenous glucosidase activity of the wild-type plant) was observed in transgenic plants transformed with pAt-GC-HSP combined with a 5' untranslated region of the Arabidopsis alcohol dehydrogenase gene with the Arabidopsis heat shock protein terminator. These transgenic plant lines could pave the way to a stable plant-production system for low-cost, high-yield human GCase production.

  6. em arquitetura e urbanismo

    Directory of Open Access Journals (Sweden)

    Ana Maria Sala Minucci Martins

    2006-01-01

    Full Text Available Está sendo desenvolvido projeto de pesquisa denominado Geometria Fractal e suas Aplicações em Arquitetura e Urbanismo, com o fito de estudar e desenvolver ferramentas analíticas e propositivas para serem aplicadas em arquitetura e urbanismo, com base em conceitos provenientes da geometria fractal.

  7. Antimicrobial Activity of Geranium Oil against Clinical Strains<em> em>of <em>Staphylococcus aureusem>

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    Monika Sienkiewicz

    2012-08-01

    Full Text Available The aim of this work was to investigate the antibacterial properties of geranium oil obtained from <em>Pelargonium graveolensem> Ait. (family <em>Geraniaceae>, against one standard <em>S. aureus em>strain ATCC 433000 and seventy clinical <em>S. aureusem> strains. The agar dilution method was used for assessment of bacterial growth inhibition at various concentrations of geranium oil. Susceptibility testing of the clinical strains to antibiotics was carried out using the disk-diffusion and E-test methods. The results of our experiment showed that the oil from <em>P. graveolensem> has strong activity against all of the clinical <em>S. aureusem> isolates—including multidrug resistant strains, MRSA strains and MLSB-positive strains—exhibiting MIC values of 0.25–2.50 μL/mL.

  8. Phytophthora nicotianae var. nicotianae on tomatoes

    NARCIS (Netherlands)

    Weststeijn, G.

    1973-01-01

    Around 1960 some disorders which initially were considered to be of a physiological nature were found in tomato plants grown in glasshouses in the Netherlands. One complex of symptoms was a brown rot of the lateral roots and the tap root, often followed by decomposition of the stem base ti

  9. Phytophthora nicotianae var. nicotianae on tomatoes

    NARCIS (Netherlands)

    Weststeijn, G.

    1973-01-01

    Around 1960 some disorders which initially were considered to be of a physiological nature were found in tomato plants grown in glasshouses in the Netherlands. One complex of symptoms was a brown rot of the lateral roots and the tap root, often followed by decomposition of the stem base tissue and d

  10. Phytophthora nicotianae var. nicotianae on tomatoes

    NARCIS (Netherlands)

    Weststeijn, G.

    1973-01-01

    Around 1960 some disorders which initially were considered to be of a physiological nature were found in tomato plants grown in glasshouses in the Netherlands. One complex of symptoms was a brown rot of the lateral roots and the tap root, often followed by decomposition of the stem base

  11. EM International. Volume 1

    Energy Technology Data Exchange (ETDEWEB)

    1993-07-01

    It is the intent of EM International to describe the Office of Environmental Restoration and Waste Management`s (EM`s) various roles and responsibilities within the international community. Cooperative agreements and programs, descriptions of projects and technologies, and synopses of visits to international sites are all highlighted in this semiannual journal. Focus on EM programs in this issue is on international collaboration in vitrification projects. Technology highlights covers: in situ sealing for contaminated sites; and remote sensors for toxic pollutants. Section on profiles of countries includes: Arctic contamination by the former Soviet Union, and EM activities with Germany--cooperative arrangements.

  12. Microsatellite Loci in the Gypsophyte <em>Lepidium subulatum em>(Brassicaceae, and Transferability to Other <em>Lepidieae>

    Directory of Open Access Journals (Sweden)

    José Gabriel Segarra-Moragues

    2012-09-01

    Full Text Available Polymorphic microsatellite markers were developed for the Ibero-North African, strict gypsophyte <em>Lepidium subulatumem> to unravel the effects of habitat fragmentation in levels of genetic diversity, genetic structure and gene flow among its populations. Using 454 pyrosequencing 12 microsatellite loci including di- and tri-nucleotide repeats were characterized in <em>L. subulatumem>. They amplified a total of 80 alleles (2–12 alleles per locus in a sample of 35 individuals of <em>L. subulatumem>, showing relatively high levels of genetic diversity, <em>H>O = 0.645, <em>H>E = 0.627. Cross-species transferability of all 12 loci was successful for the Iberian endemics <em>Lepidium cardaminesem>, <em>Lepidium stylatumem>, and the widespread, <em>Lepidium graminifoliumem> and one species each of two related genera, <em>Cardaria drabaem> and <em>Coronopus didymusem>. These microsatellite primers will be useful to investigate genetic diversity, population structure and to address conservation genetics in species of <em>Lepidium>.

  13. New Trifluoromethyl Triazolopyrimidines as Anti-<em>Plasmodium> <em>falciparum> Agents

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    Núbia Boechat

    2012-07-01

    Full Text Available According to the World Health Organization, half of the World’s population, approximately 3.3 billion people, is at risk for developing malaria. Nearly 700,000 deaths each year are associated with the disease. Control of the disease in humans still relies on chemotherapy. Drug resistance is a limiting factor, and the search for new drugs is important. We have designed and synthesized new 2-(trifluoromethyl[1,2,4]triazolo[1,5-<em>a>]pyrimidine derivatives based on bioisosteric replacement of functional groups on the anti-malarial compounds mefloquine and amodiaquine. This approach enabled us to investigate the impact of: (i ring bioisosteric replacement; (ii a CF3 group substituted at the 2-position of the [1,2,4]triazolo[1,5-<em>a>]pyrimidine scaffold and (iii a range of amines as substituents at the 7-position of the of heterocyclic ring; on <em>in vitroem> activity against <em>Plasmodium falciparumem>. According to docking simulations, the synthesized compounds are able to interact with <em>P. falciparumem> dihydroorotate dehydrogenase (<em>Pf>DHODH through strong hydrogen bonds. The presence of a trifluoromethyl group at the 2-position of the [1,2,4]triazolo[1,5-<em>a>]pyrimidine ring led to increased drug activity. Thirteen compounds were found to be active, with IC50 values ranging from 0.023 to 20 µM in the anti-HRP2 and hypoxanthine assays. The selectivity index (SI of the most active derivatives 5, 8, 11 and 16 was found to vary from 1,003 to 18,478.

  14. Screening and Analysis of the Potential Bioactive Components in Rabbit Plasma after Oral Administration of Hot-Water Extracts from Leaves of <em>B>ambusa em>>textilis em>McClure

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    Jia Sun

    2012-07-01

    Full Text Available <em>Bambusa textilisem> McClure is a traditional Chinese medicinal plant belonging to the Bambusoideae subfamily and used to treat chronic fever and infectious diseases. To investigate the bioactive compounds absorbed in the rabbit blood after oral administration of hot-<em>water extractem>>s from em>>the leaves of em>>B. textilisem> McClure, a validated chromatographic fingerprint method was established using LC-Q-TOF-MS. Twenty compounds in bamboo leaves and three potential bioactive compounds in rabbit plasma were detected. Of the twenty detected compounds <em>in vitroem>, fifteen of which were tentatively identified either by comparing the retention time and mass spectrometry data with that of reference compounds or by reviewing the literature. Three potential bioactive compounds, including (<em>E-p>-coumaric acid, (<em>Z-p>-coumaric acid, and apigenin-8-<em>C>-β-D-(2"-<em>O>-α-L-rhamnosyl-gluco-pyranoside, were detected in both <em>the leaves of em>>B. textilis em>McClure and rabbit plasma. Of the three compounds, apigenin-8-<em>C>-β-D-(2"-<em>O>-α-L-rhamnosylglucopyranoside was identified based on its UV, MS, and NMR spectra. This study provides helpful chemical information for further pharmacology and active mechanism research on <em>B. textilisem> McClure.

  15. UJI TOLERANSI TANAMAN TEMBAKAU (Nicotiana tabacum L. TERHADAP CEKAMAN KADMIUM (Cd, TIMBAL (Pb, DAN TEMBAGA (Cu PADA KULTUR CAIR

    Directory of Open Access Journals (Sweden)

    S Rosidah

    2014-11-01

    Full Text Available Penelitian ini menyelidiki respon fisiologis, anatomis, dan morfologis tanaman tembakau (Nicotiana tabacum L. terhadap cekaman logam berat Cu, Cd, dan Pb. Sampel yang digunakan adalah tembakau umur 3-4 minggu yang dikecambahkan secara in vitro dan kemudian dipapar logam berat selama 14 hari. Desain penelitian yang digunakan yaitu rancangan acak lengkap dengan satu faktor, yaitu konsentrasi logam Cu (0 µM, 50 µM, 100 µM, 150 µM & 200 µM, Cd (0 µM, 50 µM, 100 µM, 200 µM & 300 µM, dan Pb (0 µM, 5 µM, 20 µM, 50 µM & 100 µM. Parameter yang digunakan: pertambahan panjang akar, pertambahan jumlah akar, akumulasi logam dalam akar, lokalisasi penimbunan dalam akar, dan warna daun. Bertambahnya konsentrasi logam menghambat pertumbuhan akar dan menyebabkan deposit logam pada jaringan akar dan gejala klorosis. Hasil uji Atomic Absorbtion Spectrophotometry (AAS menunjukkan semakin besar konsentrasi semakin banyak akumulasi logam pada jaringan akar. Akan tetapi, akumulasi Cd pada konsentrasi 200 µM lebih besar dibanding pada konsentrasi 300 µM. Analisis kualitatif membuktikan bahwa cekaman Cu tidak berpengaruh signifikan terhadap warna daun, sedangkan pada cekaman Cd (100, 150 dan 200 µM dan Pb (150 µM daun mengalami klorosis. Pada konsentrasi logam yang rendah seperti 50 µM Cu, 50 µM Cd, dan 5 µM Pb tidak berbeda nyata dengan kontrol. Dengan demikian disimpulkan bahwa tembakau mampu mentoleransi cekaman logam pada konsentrasi yang rendah.  This research investigated the physiological, anatomical, and morphological responses of tobacco (Nicotiana tabacum L. on stresses of heavy metals Cu, Cd, and Pb. The samples were 3- to 4-week tobacco plants germinated in vitro and then were exposed to heavy metals for 14 days. This study used a completed random design with single factor, i.e. the concentrations of Cu (0 µM, 50 µM, 100 µM, 150 µM & 200 µM, Cd (0 µM, 50 µM, 100 µM, 200 µM & 300 µM, and Pb (0 µM, 5 µM, 20 µM, 50 µM & 100

  16. Genotoxicity of <em>Euphorbia hirtaem>: An <em>Allium cepaem> Assay

    Directory of Open Access Journals (Sweden)

    Kwan Yuet Ping

    2012-06-01

    Full Text Available The potential genotoxic effects of methanolic extracts of <em>Euphorbia hirta em>which is commonly used in traditional medicine to treat a variety of diseased conditions including asthma, coughs, diarrhea and dysentery was investigated using <em>Allium cepaem> assay. The extracts of 125, 250, 500 and 1,000 µg/mL were tested on root meristems of <em>A. cepaem>. Ethylmethanesulfonate was used as positive control and distilled water was used as negative control. The result showed that mitotic index decreased as the concentrations of <em>E. hirtaem> extract increased. A dose-dependent increase of chromosome aberrations was also observed. Abnormalities scored were stickiness, c-mitosis, bridges and vagrant chromosomes. Micronucleated cells were also observed at interphase. Result of this study confirmed that the methanol extracts of <em>E. hirta em>exerted significant genotoxic and mitodepressive effects at 1,000 µg/mL.

  17. Salicylic acid and jasmonic acid are essential for systemic resistance against tobacco mosaic virus in Nicotiana benthamiana.

    Science.gov (United States)

    Zhu, Feng; Xi, De-Hui; Yuan, Shu; Xu, Fei; Zhang, Da-Wei; Lin, Hong-Hui

    2014-06-01

    Systemic resistance is induced by pathogens and confers protection against a broad range of pathogens. Recent studies have indicated that salicylic acid (SA) derivative methyl salicylate (MeSA) serves as a long-distance phloem-mobile systemic resistance signal in tobacco, Arabidopsis, and potato. However, other experiments indicate that jasmonic acid (JA) is a critical mobile signal. Here, we present evidence suggesting both MeSA and methyl jasmonate (MeJA) are essential for systemic resistance against Tobacco mosaic virus (TMV), possibly acting as the initiating signals for systemic resistance. Foliar application of JA followed by SA triggered the strongest systemic resistance against TMV. Furthermore, we use a virus-induced gene-silencing-based genetics approach to investigate the function of JA and SA biosynthesis or signaling genes in systemic response against TMV infection. Silencing of SA or JA biosynthetic and signaling genes in Nicotiana benthamiana plants increased susceptibility to TMV. Genetic experiments also proved the irreplaceable roles of MeSA and MeJA in systemic resistance response. Systemic resistance was compromised when SA methyl transferase or JA carboxyl methyltransferase, which are required for MeSA and MeJA formation, respectively, were silenced. Moreover, high-performance liquid chromatography-mass spectrometry analysis indicated that JA and MeJA accumulated in phloem exudates of leaves at early stages and SA and MeSA accumulated at later stages, after TMV infection. Our data also indicated that JA and MeJA could regulate MeSA and SA production. Taken together, our results demonstrate that (Me)JA and (Me)SA are required for systemic resistance response against TMV.

  18. Silencing of DS2 aminoacylase-like genes confirms basal resistance to Phytophthora infestans in Nicotiana benthamiana.

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    Nakano, Masahito; Nishihara, Masahiro; Yoshioka, Hirofumi; Ohnishi, Kouhei; Hikichi, Yasufumi; Kiba, Akinori

    2014-01-01

    Nicotiana benthamiana is a potential host to several plant pathogens, and immature leaves of N. benthamiana are susceptible to Phytophthora infestans. In contrast, mature leaves of N. benthamiana are weakly susceptible and show basal resistance to P. infestans. We screened a gene-silenced mature plant showing high resistance to P. infestans, designated as DS2 (Disease suppression 2). The deduced amino acid sequence of cDNA responsible for DS2 encoded a putative aminoacylase. Growth of P. infestans decreased in DS2 plants. Trypan blue staining revealed inhibited hyphae growth of P. infestans with an increased number of dead cells under the penetration site in DS2 plants. Consistent with growth inhibition of P. infestans, defense responses such as reactive oxygen generation and expression of a salicylic acid-dependent PR-1a increased markedly in DS2 plants compared with that of control plants. DS2 phenotype was compromised in NahG plants, suggesting DS2 phenotype depends on the salicylic acid signaling pathway. Accelerated defense response was observed in DS2 plants elicited by INF1 elicitin as well as by NbMEK2(DD), which is the constitutive active form of NbMEK2, and act as a downstream regulator of INF1 perception. On the other hand, INF1- and NbMEK2(DD)-induced defense responses were prevented by DS2-overexpressing transgenic tobacco. These results suggest that DS2 negatively regulates plant defense responses against P. infestans via NbMEK2 and SA-dependent signaling pathway in N. benthamiana.

  19. Transcriptional and biochemical responses of monoacylglycerol acyltransferase-mediated oil synthesis and associated senescence-like responses in Nicotiana benthamiana.

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    Uday Kumar Divi

    2014-05-01

    Full Text Available Triacylglycerol (TAG accumulates in plant seeds as a major renewable source of carbon for food, fuel and industrial feedstock. Approaches to enhance TAG content by altering lipid pathways and genes in vegetative parts have gained significant attention for biofuel and other applications. However, consequences of these modifications are not always studied in detail. In an attempt to increase TAG levels in leaves we previously demonstrated that a novel substrate, monoacylglycerol (MAG, can be used for the biosynthesis of diacylglycerol (DAG and TAG. Transient expression of the Mus musculus monoacylglycerol acyltransferase MGAT1 and 2 in the model plant Nicotiana benthamiana increased TAG levels at 5 days post infiltration (dpi. Here we show that increased TAG and DAG levels can be achieved as early as 2 dpi. In addition, the MGAT1 infiltrated areas showed senescence-like symptoms from 3 dpi onwards. To unravel underlying molecular mechanisms, Illumina deep sequencing was carried out (a for de-novo assembling and annotation of N. benthamiana leaf transcripts and (b to characterize MGAT1-responsive transcriptome. We found that MGAT1-responsive genes are involved in several processes including TAG biosynthesis, photosynthesis, cell-wall, cutin, suberin, wax and mucilage biosynthesis, lipid and hormone metabolism. Comparative analysis with transcript profiles from other senescence studies identified characteristic gene expression changes involved in senescence induction. We confirmed that increased TAG and observed senescence-symptoms are due to the MAG depletion caused by MGAT1 activity and suggest a mechanism for MGAT1 induced TAG increase and senescence-like symptoms. The data generated will serve as a valuable resource for oil and senescence related studies and for future N. benthamiana transcriptome studies.

  20. Growth and stress response in Arabidopsis thaliana, Nicotiana benthamiana, Glycine max, Solanum tuberosum and Brassica napus cultivated under polychromatic LEDs.

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    Janda, Martin; Navrátil, Oldřich; Haisel, Daniel; Jindřichová, Barbora; Fousek, Jan; Burketová, Lenka; Čeřovská, Noemi; Moravec, Tomáš

    2015-01-01

    The use of light emitting diodes (LEDs) brings several key advantages over existing illumination technologies for indoor plant cultivation. Among these are that LEDs have predicted lifetimes from 50-100.000 hours without significant drops in efficiency and energy consumption is much lower compared to traditional fluorescent tubes. Recent advances allow LEDs to be used with customized wavelengths for plant growth. However, most of these LED growth systems use mixtures of chips emitting in several narrow wavelengths and frequently they are not compatible with existing infrastructures. This study tested the growth of five different plant species under phosphor coated LED-chips fitted into a tube with a standard G13 base that provide continuous visible light illumination with enhanced blue and red light. The LED system was characterized and compared with standard fluorescence tubes in the same cultivation room. Significant differences in heat generation between LEDs and fluorescent tubes were clearly demonstrated. Also, LED lights allowed for better control and stability of preset conditions. Physiological properties such as growth characteristics, biomass, and chlorophyll content were measured and the responses to pathogen assessed for five plant species (both the model plants Arabidopsis thaliana, Nicotiana bentamiana and crop species potato, oilseed rape and soybean) under the different illumination sources. We showed that polychromatic LEDs provide light of sufficient quality and intensity for plant growth using less than 40% of the electricity required by the standard fluorescent lighting under test. The tested type of LED installation provides a simple upgrade pathway for existing infrastructure for indoor plant growth. Interestingly, individual plant species responded differently to the LED lights so it would be reasonable to test their utility to any particular application.

  1. Elicitation of hypersensitive responses in Nicotiana glutinosa by the suppressor of RNA silencing protein P0 from poleroviruses.

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    Wang, Ken-Der; Empleo, Roman; Nguyen, Tan Tri V; Moffett, Peter; Sacco, Melanie Ann

    2015-06-01

    Plant disease resistance (R) proteins that confer resistance to viruses recognize viral gene products with diverse functions, including viral suppressors of RNA silencing (VSRs). The P0 protein from poleroviruses is a VSR that targets the ARGONAUTE1 (AGO1) protein for degradation, thereby disrupting RNA silencing and antiviral defences. Here, we report resistance against poleroviruses in Nicotiana glutinosa directed against Turnip yellows virus (TuYV) and Potato leafroll virus (PLRV). The P0 proteins from TuYV (P0(T) (u) ), PLRV (P0(PL) ) and Cucurbit aphid-borne yellows virus (P0(CA) ) were found to elicit a hypersensitive response (HR) in N. glutinosa accession TW59, whereas other accessions recognized P0(PL) only. Genetic analysis showed that recognition of P0(T) (u) by a resistance gene designated RPO1 (Resistance to POleroviruses 1) is inherited as a dominant allele. Expression of P0 from a Potato virus X (PVX) expression vector transferred recognition to the recombinant virus on plants expressing RPO1, supporting P0 as the unique Polerovirus factor eliciting resistance. The induction of HR required a functional P0 protein, as P0(T) (u) mutants with substitutions in the F-box motif that abolished VSR activity were unable to elicit HR. We surmised that the broad P0 recognition seen in TW59 and the requirement for the F-box protein motif could indicate detection of P0-induced AGO1 degradation and disruption of RNA silencing; however, other viral silencing suppressors, including the PVX P25 that also causes AGO1 degradation, failed to elicit HR in N. glutinosa. Investigation of P0 elicitation of RPO1 could provide insight into P0 activities within the cell that trigger resistance. © 2014 BSPP AND JOHN WILEY & SONS LTD.

  2. High levels of jasmonic acid antagonize the biosynthesis of gibberellins and inhibit the growth of Nicotiana attenuata stems.

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    Heinrich, Maria; Hettenhausen, Christian; Lange, Theo; Wünsche, Hendrik; Fang, Jingjing; Baldwin, Ian T; Wu, Jianqiang

    2013-02-01

    Hormones play pivotal roles in regulating plant development, growth, and stress responses, and cross-talk among different hormones fine-tunes various aspects of plant physiology. Jasmonic acid (JA) is important for plant defense against herbivores and necrotic fungi and also regulates flower development; in addition, Arabidopsis mutants over-producing JA usually have stunted stems and wound-induced jasmonates suppress Arabidopsis growth, suggesting that JA is also involved in stem elongation. Gibberellins (GAs) promote stem and leaf growth and modulate seed germination, flowering time, and the development of flowers, fruits, and seeds. However, little is known about the interaction between the JA and GA pathways. Two calcium-dependent protein kinases, CDPK4 and CDPK5, are important suppressors of JA accumulation in a wild tobacco species, Nicotiana attenuata. The stems of N. attenuata silenced in CDPK4 and CDPK5 (irCDPK4/5 plants) had dramatically increased levels of JA and exhibited stunted elongation and had very high contents of secondary metabolites. Genetic analysis indicated that the high JA levels in irCDPK4/5 stems accounted for the suppressed stem elongation and the accumulation of secondary metabolites. Supplementation of GA(3) to irCDPK4/5 plants largely restored normal stem growth to wild-type levels. Measures of GA levels indicated that over-accumulation of JA in irCDPK4/5 stems inhibited the biosynthesis of GAs. Finally, we show that JA antagonizes GA biosynthesis by strongly inhibiting the transcript accumulation of GA20ox and possibly GA13ox, the key genes in GA production, demonstrating that high JA levels antagonize GA biosynthesis in stems.

  3. Pithy protection: Nicotiana attenuata's jasmonic acid-mediated defenses are required to resist stem-boring weevil larvae.

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    Diezel, Celia; Kessler, Danny; Baldwin, Ian T

    2011-04-01

    Folivory is the best studied plant-herbivore interaction, but it is unclear whether the signaling and resistance traits important for the defense of leaves are also important for other plant parts. Larvae of the tobacco stem weevil, Trichobaris mucorea, burrow into stems of Nicotiana attenuata and feed on the pith. Transgenic N. attenuata lines silenced in signaling and foliar defense traits were evaluated in a 2-year field study for resistance against attack by naturally occurring T. mucorea larva. Plants silenced in early jasmonic acid (JA) biosynthesis (antisense [as]-lipoxygenase3 [lox3]; inverted repeat [ir]-allene oxide cyclase), JA perception (as-coronatine insensitive1), proteinase inhibitors (ir-pi), and nicotine (ir-putrescine methyl-transferase) direct defenses and lignin (ir-cad) biosynthesis were infested more frequently than wild-type plants. Plants unable to emit C(6) aldehydes (as-hpl) had lower infestation rates, while plants silenced in late steps in JA biosynthesis (ir-acyl-coenzyme A oxidase, ir-opr) and silenced in diterpene glycoside production (ir-geranylgeranyl pyrophosphate synthase) did not differ from wild type. Pith choice assays revealed that ir-putrescine methyl-transferase, ir-coronatine insensitive1, and ir-lox3 pith, which all had diminished nicotine levels, were preferred by larvae compared to wild-type pith. The lack of preference for ir-lox2 and ir-cad piths, suggest that oviposition attraction and vascular defense, rather than pith palatability accounts for the higher attack rates observed for these plants. We conclude that traits that influence a plant's apparency, stem hardness, and pith direct defenses all contribute to resistance against this herbivore whose attack can be devastating to N. attenuata's fitness.

  4. Organ- and Growing Stage-Specific Expression of Solanesol Biosynthesis Genes in Nicotiana tabacum Reveals Their Association with Solanesol Content

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    Ning Yan

    2016-11-01

    Full Text Available Solanesol is a noncyclic terpene alcohol that is composed of nine isoprene units and mainly accumulates in solanaceous plants, especially tobacco (Nicotiana tabacum L.. In the present study, RNA-seq analyses of tobacco leaves, stems, and roots were used to identify putative solanesol biosynthesis genes. Six 1-deoxy-d-xylulose 5-phosphate synthase (DXS, two 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR, two 2-C-methyl-d-erythritol 4-phosphate cytidylyltransferase (IspD, four 4-diphosphocytidyl-2-C-methyl-d-erythritol kinase (IspE, two 2-C-methyl-d-erythritol 2,4-cyclo-diphosphate synthase (IspF, four 1-hydroxy-2-methyl-2-(E-butenyl 4-diphosphate synthase (IspG, two 1-hydroxy-2-methyl-2-(E-butenyl 4-diphosphate reductase (IspH, six isopentenyl diphosphate isomerase (IPI, and two solanesyl diphosphate synthase (SPS candidate genes were identified in the solanesol biosynthetic pathway. Furthermore, the two N. tabacum SPS proteins (NtSPS1 and NtSPS2, which possessed two conserved aspartate-rich DDxxD domains, were highly homologous with SPS enzymes from other solanaceous plant species. In addition, the solanesol contents of three organs and of leaves from four growing stages of tobacco plants corresponded with the distribution of chlorophyll. Our findings provide a comprehensive evaluation of the correlation between the expression of different biosynthesis genes and the accumulation of solanesol, thus providing valuable insight into the regulation of solanesol biosynthesis in tobacco.

  5. Differential RNAi responses of Nicotiana benthamiana individuals transformed with a hairpin-inducing construct during Plum pox virus challenge.

    Science.gov (United States)

    Montes, Christian; Castro, Álvaro; Barba, Paola; Rubio, Julia; Sánchez, Evelyn; Carvajal, Denisse; Aguirre, Carlos; Tapia, Eduardo; DelÍ Orto, Paola; Decroocq, Veronique; Prieto, Humberto

    2014-10-01

    Gene silencing and large-scale small RNA analysis can be used to develop RNA interference (RNAi)-based resistance strategies for Plum pox virus (PPV), a high impact disease of Prunus spp. In this study, a pPPViRNA hairpin-inducing vector harboring two silencing motif-rich regions of the PPV coat protein (CP) gene was evaluated in transgenic Nicotiana benthamiana (NB) plants. Wild-type NB plants infected with a chimeric PPV virus (PPV::GFP) exhibited affected leaves with mosaic chlorosis congruent to GFP fluorescence at 21 day post-inoculation; transgenic lines depicted a range of phenotypes from fully resistant to susceptible. ELISA values and GFP fluorescence intensities were used to select transgenic-resistant (TG-R) and transgenic-susceptible (TG-S) lines for further characterization of small interfering RNAs (siRNAs) by large-scale small RNA sequencing. In infected TG-S and untransformed (WT) plants, the observed siRNAs were nearly exclusively 21- and 22-nt siRNAs that targeted the whole PPV::GFP genome; 24-nt siRNAs were absent in these individuals. Challenged TG-R plants accumulated a full set of 21- to 24-nt siRNAs that were primarily associated with the selected motif-rich regions, indicating that a trans-acting siRNAs process prevented viral multiplication. BLAST analysis identified 13 common siRNA clusters targeting the CP gene. 21-nt siRNA sequences were associated with the 22-nt siRNAs and the scarce 23- and 24-nt molecules in TG-S plants and with most of the observed 22-, 23-, and 24-nt siRNAs in TG-R individuals. These results validate the use of a multi-hot spot silencing vector against PPV and elucidate the molecules by which hairpin-inducing vectors initiate RNAi in vivo.

  6. Digestive duet: midgut digestive proteinases of Manduca sexta ingesting Nicotiana attenuata with manipulated trypsin proteinase inhibitor expression.

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    Jorge A Zavala

    Full Text Available BACKGROUND: The defensive effect of endogenous trypsin proteinase inhibitors (NaTPIs on the herbivore Manduca sexta was demonstrated by genetically altering NaTPI production in M. sexta's host plant, Nicotiana attenuata. To understand how this defense works, we studied the effects of NaTPI on M. sexta gut proteinase activity levels in different larval instars of caterpillars feeding freely on untransformed and transformed plants. METHODOLOGY/ PRINCIPAL FINDINGS: Second and third instars larvae that fed on NaTPI-producing (WT genotypes were lighter and had less gut proteinase activity compared to those that fed on genotypes with either little or no NaTPI activity. Unexpectedly, NaTPI activity in vitro assays not only inhibited the trypsin sensitive fraction of gut proteinase activity but also halved the NaTPI-insensitive fraction in third-instar larvae. Unable to degrade NaTPI, larvae apparently lacked the means to adapt to NaTPI in their diet. However, caterpillars recovered at least part of their gut proteinase activity when they were transferred from NaTPI-producing host plants to NaTPI-free host plants. In addition extracts of basal leaves inhibited more gut proteinase activity than did extracts of middle stem leaves with the same protein content. CONCLUSIONS/ SIGNIFICANCE: Although larvae can minimize the effects of high NaTPI levels by feeding on leaves with high protein and low NaTPI activity, the host plant's endogenous NaTPIs remain an effective defense against M. sexta, inhibiting gut proteinase and affecting larval performance.

  7. Three nicotine demethylase genes mediate nornicotine biosynthesis in Nicotiana tabacum L.: functional characterization of the CYP82E10 gene.

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    Lewis, Ramsey S; Bowen, Steven W; Keogh, Matthew R; Dewey, Ralph E

    2010-12-01

    In most tobacco (Nicotiana tabacum L.) plants, nornicotine is a relatively minor alkaloid, comprising about 2-5% of the total pyridine alkaloid pool in the mature leaf. Changes in gene expression at an unstable locus, however, can give rise to plants that produce high levels of nornicotine, specifically during leaf senescence and curing. Minimizing the nornicotine content in tobacco is highly desirable, because this compound serves as the direct precursor in the synthesis of N'-nitrosonornicotine, a potent carcinogen in laboratory animals. Nornicotine is likely produced almost entirely via the N-demethylation of nicotine, in a process called nicotine conversion that is catalyzed by the enzyme nicotine N-demethylase (NND). Previous studies have identified CYP82E4 as the specific NND gene responsible for the unstable conversion phenomenon, and CYP82E5v2 as a putative minor NND gene. Here, by discovery and characterization of CYP82E10, a tobacco NND gene, is reported. PCR amplification studies showed that CYP82E10 originated from the N. sylvestris ancestral parent of modern tobacco. Using a chemical mutagenesis strategy, knockout mutations were induced and identified in all three tobacco NND genes. By generating a series of mutant NND genotypes, the relative contribution of each NND gene toward the nornicotine content of the plant was assessed. Plants possessing knockout mutations in all three genes displayed nornicotine phenotypes that were much lower (∼0.5% of total alkaloid content) than that found in conventional tobacco cultivars. The introduction of these mutations into commercial breeding lines promises to be a viable strategy for reducing the levels of one of the best characterized animal carcinogens found in tobacco products.

  8. Cadmium and zinc activate adaptive mechanisms in Nicotiana tabacum similar to those observed in metal tolerant plants.

    Science.gov (United States)

    Vera-Estrella, Rosario; Gómez-Méndez, María F; Amezcua-Romero, Julio C; Barkla, Bronwyn J; Rosas-Santiago, Paul; Pantoja, Omar

    2017-04-28

    Tobacco germinated and grew in the presence of high concentrations of cadmium and zinc without toxic symptoms. Evidence suggests that these ions are sequestered into the vacuole by heavy metal/H (+) exchanger mechanisms. Heavy metal hyperaccumulation and hypertolerance are traits shared by a small set of plants which show specialized physiological and molecular adaptations allowing them to accumulate and sequester toxic metal ions. Nicotiana tabacum was used to test its potential as a metal-accumulator in a glass house experiment. Seed germination was not affected in the presence of increasing concentrations of zinc and cadmium. Juvenile and adult plants could concentrate CdCl2 and ZnSO4 to levels exceeding those in the hydroponic growth medium and maintained or increased their leaf dry weight when treated with 0.5- or 1-mM CdCl2 or 1-mM ZnSO4 for 5 days. Accumulation of heavy metals did not affect the chlorophyll and carotenoid levels, while variable effects were observed in cell sap osmolarity. Heavy metal-dependent H(+) transport across the vacuole membrane was monitored using quinacrine fluorescence quenching. Cadmium- or zinc-dependent fluorescence recovery revealed that increasing concentrations of heavy metals stimulated the activities of the tonoplast Cd(2+) or Zn(2+)/H(+) exchangers. Immunodetection of the V-ATPase subunits showed that the increased proton transport by zinc was not due to changes in protein amount. MTP1 and MTP4 immunodetection and semiquantitative RT-PCR of NtMTP1, NtNRAMP1, and NtZIP1 helped to identify the genes that are likely involved in sequestration of cadmium and zinc in the leaf and root tissue. Finally, we demonstrated that cadmium and zinc treatments induced an accumulation of zinc in leaf tissues. This study shows that N. tabacum possesses a hyperaccumulation response, and thus could be used for phytoremediation purposes.

  9. Genetic and molecular requirements for function of the Pto/Prf effector recognition complex in tomato and Nicotiana benthamiana.

    Science.gov (United States)

    Balmuth, Alexi; Rathjen, John P

    2007-09-01

    The Pto gene of tomato (Solanum lycopersicum) confers specific recognition of the unrelated bacterial effector proteins AvrPto and AvrPtoB. Pto resides in a constitutive molecular complex with the nucleotide binding site-leucine rich repeats protein Prf. Prf is absolutely required for specific recognition of both effectors. Here, using stable transgenic lines, we show that expression of Pto from its genomic promoter in susceptible tomatoes was sufficient to complement recognition of Pseudomonas syringae pv. tomato (Pst) bacteria expressing either avrPto or avrPtoB. Pto kinase activity was absolutely required for specific immunity. Expression of the Pto N-myristoylation mutant, pto(G2A), conferred recognition of Pst (avrPtoB), but not Pst (avrPto), although bacterial growth in these lines was intermediate between resistant and susceptible lines. Overexpression of pto(G2A) complemented recognition of avrPto. Transgenic tomato plants overexpressing wild-type Pto exhibited constitutive growth phenotypes, but these were absent in lines overexpressing pto(G2A). Therefore, Pto myristoylation is a quantitative factor for effector recognition in tomato, but is absolutely required for overexpression phenotypes. Native expression of Pto in the heterologous species Nicotiana benthamiana did not confer resistance to P. syringae pv. tabaci (Pta) expressing avrPto or avrPtoB, but recognition of both effectors was complemented by Prf co-expression. Thus, specific resistance conferred solely by Pto in N. benthamiana is an artefact of overexpression. Finally, pto(G2A) did not confer recognition of either avrPto or avrPtoB in N. benthamiana, regardless of the presence of Prf. Thus, co-expression of Prf in N. benthamiana complements many but not all aspects of normal Pto function.

  10. Genetic modification of alternative respiration in Nicotiana benthamiana affects basal and salicylic acid-induced resistance to potato virus X

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    Verchot-Lubicz Jeanmarie

    2011-02-01

    Full Text Available Abstract Background Salicylic acid (SA regulates multiple anti-viral mechanisms, including mechanism(s that may be negatively regulated by the mitochondrial enzyme, alternative oxidase (AOX, the sole component of the alternative respiratory pathway. However, studies of this mechanism can be confounded by SA-mediated induction of RNA-dependent RNA polymerase 1, a component of the antiviral RNA silencing pathway. We made transgenic Nicotiana benthamiana plants in which alternative respiratory pathway capacity was either increased by constitutive expression of AOX, or decreased by expression of a dominant-negative mutant protein (AOX-E. N. benthamiana was used because it is a natural mutant that does not express a functional RNA-dependent RNA polymerase 1. Results Antimycin A (an alternative respiratory pathway inducer and also an inducer of resistance to viruses and SA triggered resistance to tobacco mosaic virus (TMV. Resistance to TMV induced by antimycin A, but not by SA, was inhibited in Aox transgenic plants while SA-induced resistance to this virus appeared to be stronger in Aox-E transgenic plants. These effects, which were limited to directly inoculated leaves, were not affected by the presence or absence of a transgene constitutively expressing a functional RNA-dependent RNA polymerase (MtRDR1. Unexpectedly, Aox-transgenic plants infected with potato virus X (PVX showed markedly increased susceptibility to systemic disease induction and virus accumulation in inoculated and systemically infected leaves. SA-induced resistance to PVX was compromised in Aox-transgenic plants but plants expressing AOX-E exhibited enhanced SA-induced resistance to this virus. Conclusions We conclude that AOX-regulated mechanisms not only play a role in SA-induced resistance but also make an important contribution to basal resistance against certain viruses such as PVX.

  11. Comparative study on macro- and micro-elements concentration in Nicotiana tabacum and Faba siliquis plants by ICP-MS

    Science.gov (United States)

    Balazs, Zoltan; Voica, Cezara; Dehelean, Adriana; Magdas, Dana Alina; Ristoiu, Dumitru

    2015-12-01

    Plants are important components of ecosystems as they transfer elements from abiotic into biotic environments. The concentration of macro and micro-elements in tobacco leaves (Nicotiana tabacum) and bean (Faba siliquis) was analyzed using ICP-MS technique. The results obtained indicated that the mean concentration of Mg, P, K and Ca in tobacco leaves was 0.965, 0.812, 4.412 and 2.694 g.kg-1, respectively, while in bean samples were 0.899, 2.024, 6.725 and 1.387 g.kg-1, respectively. Mn concentration ranged from 156.835 mg.kg-1 to 234.593 mg.kg-1 in tobacco leaves and from 116.174 mg.kg-1 to 440.423 mg.kg-1 in bean samples. The results for Cu and Zn were between 7.262 mg.kg-1 and 105.738 mg.kg-1, 68.549 mg.kg-1 and 113.720 mg.kg-1 (tobacco leaves); and 6.830 mg.kg-1 and 46.034 mg.kg-1, 50.166 mg.kg-1 and 77.242 mg.kg-1 (bean samples), respectively. In analyzed samples, Pb, Cd and As concentrations ranged between <0.001-0.717 mg.kg-1, 0.046 mg.kg-1 -6.218 mg.kg-1, <0.001-0.381 mg.kg-1. The paper discusses the transfer of metal ions (Mn, As, Cd, Cu, Pb and Zn, respectively) from soil to these plants in terms of transfer factors (TF).

  12. Posttranslational processing of a new class of hydroxyproline-containing proteins. Prolyl hydroxylation and C-terminal cleavage of tobacco (Nicotiana tabacum) vacuolar chitinase.

    Science.gov (United States)

    Sticher, L; Hofsteenge, J; Neuhaus, J M; Boller, T; Meins, F

    1993-04-01

    The fungicidal class I chitinases (EC 3.2.1.14) are believed to be important in defending plants against microbial pathogens. The vacuolar isoforms of tobacco (Nicotiana tabacum), chitinases A and B, are the first examples of a new type of hydroxyproline-containing protein with intracellular location, enzymic activity, and a small number of hydroxyprolyl residues restricted to a single, short peptide sequence. We have investigated the posttranslational processing and intracellular transport of transgene-encoded chitinase A in callus cultures of Nicotiana tabacum L. cv Havana 425 and leaves of Nicotiana sylvestris Spegazzini and Comes. Pulse-chase experiments and cell fractionation show that chitinase A is processed in two distinct steps. In the first step, the nascent protein undergoes an increase in apparent M(r) of approximately 1500 detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Experiments with the inhibitor of prolyl hydroxylation, alpha,alpha'-dipyridyl, and pulse-chase labeling of cells expressing recombinant forms of chitinase A indicate that the anomalous increase in M(r) is due to hydroxylation of prolyl residues. This step occurs in the endomembrane system before sorting for secretion and vacuolar transport and does not appear to be required for correct targeting of chitinase A to the vacuole. The second step is a proteolytic cleavage. Sequencing of tryptic peptides of the mature proteins indicates that during processing essentially all molecules of chitinase A and B lose a C-terminal heptapeptide, which has been shown to be a vacuolar targeting signal. This appears to occur primarily in the endomembrane system late in intracellular transport. A model for the posttranslational modification of chitinase A is proposed.

  13. Nectar sugars and amino acids in day- and night-flowering Nicotiana species are more strongly shaped by pollinators’ preferences than organic acids and inorganic ions

    Science.gov (United States)

    Tiedge, Kira; Lohaus, Gertrud

    2017-01-01

    Floral nectar contains mainly sugars but also amino acids, organic acids, inorganic ions and secondary compounds to attract pollinators. The genus Nicotiana exhibits great diversity among species in floral morphology, flowering time, nectar compositions, and predominant pollinators. We studied nectar samples of 20 Nicotiana species, composed equally of day- and night-flowering plants and attracting different groups of pollinators (e.g. hummingbirds, moths or bats) to investigate whether sugars, amino acids, organic acids and inorganic ions are influenced by pollinator preferences. Glucose, fructose and sucrose were the only sugars found in the nectar of all examined species. Sugar concentration of the nectar of day-flowering species was 20% higher and amino acid concentration was 2-3-fold higher compared to the nectar of night-flowering species. The sucrose-to-hexose ratio was significantly higher in night-flowering species and the relative share of sucrose based on the total sugar correlated with the flower tube length in the nocturnal species. Flowers of different tobacco species contained varying volumes of nectar which led to about 150-fold higher amounts of total sugar per flower in bat- or sunbird-pollinated species than in bee-pollinated or autogamous species. This difference was even higher for total amino acids per flower (up to 1000-fold). As a consequence, some Nicotiana species invest large amounts of organic nitrogen for certain pollinators. Higher concentrations of inorganic ions, predominantly anions, were found in nectar of night-flowering species. Therefore, higher anion concentrations were also associated with pollinator types active at night. Malate, the main organic acid, was present in all nectar samples but the concentration was not correlated with pollinator type. In conclusion, statistical analyses revealed that pollinator types have a stronger effect on nectar composition than phylogenetic relations. In this context, nectar sugars and amino

  14. Nectar sugars and amino acids in day- and night-flowering Nicotiana species are more strongly shaped by pollinators' preferences than organic acids and inorganic ions.

    Science.gov (United States)

    Tiedge, Kira; Lohaus, Gertrud

    2017-01-01

    Floral nectar contains mainly sugars but also amino acids, organic acids, inorganic ions and secondary compounds to attract pollinators. The genus Nicotiana exhibits great diversity among species in floral morphology, flowering time, nectar compositions, and predominant pollinators. We studied nectar samples of 20 Nicotiana species, composed equally of day- and night-flowering plants and attracting different groups of pollinators (e.g. hummingbirds, moths or bats) to investigate whether sugars, amino acids, organic acids and inorganic ions are influenced by pollinator preferences. Glucose, fructose and sucrose were the only sugars found in the nectar of all examined species. Sugar concentration of the nectar of day-flowering species was 20% higher and amino acid concentration was 2-3-fold higher compared to the nectar of night-flowering species. The sucrose-to-hexose ratio was significantly higher in night-flowering species and the relative share of sucrose based on the total sugar correlated with the flower tube length in the nocturnal species. Flowers of different tobacco species contained varying volumes of nectar which led to about 150-fold higher amounts of total sugar per flower in bat- or sunbird-pollinated species than in bee-pollinated or autogamous species. This difference was even higher for total amino acids per flower (up to 1000-fold). As a consequence, some Nicotiana species invest large amounts of organic nitrogen for certain pollinators. Higher concentrations of inorganic ions, predominantly anions, were found in nectar of night-flowering species. Therefore, higher anion concentrations were also associated with pollinator types active at night. Malate, the main organic acid, was present in all nectar samples but the concentration was not correlated with pollinator type. In conclusion, statistical analyses revealed that pollinator types have a stronger effect on nectar composition than phylogenetic relations. In this context, nectar sugars and amino

  15. Non-host resistance induced by the Xanthomonas effector XopQ is widespread within the genus Nicotiana and functionally depends on EDS1

    Directory of Open Access Journals (Sweden)

    Norman Adlung

    2016-11-01

    Full Text Available Most Gram-negative plant pathogenic bacteria translocate effector proteins (T3Es directly into plant cells via a conserved type III secretion system, which is essential for pathogenicity in susceptible plants. In resistant plants, recognition of some T3Es is mediated by corresponding resistance (R genes or R proteins and induces effector triggered immunity (ETI that often results in programmed cell death reactions. The identification of R genes and understanding their evolution/distribution bears great potential for the generation of resistant crop plants. We focus on T3Es from Xanthomonas campestris pv. vesicatoria (Xcv, the causal agent of bacterial spot disease on pepper and tomato plants. Here, 86 Solanaceae lines mainly of the genus Nicotiana were screened for phenotypical reactions after Agrobacterium tumefaciens-mediated transient expression of 21 different Xcv effectors to (i identify new plant lines for T3E characterization, (ii analyze conservation/evolution of putative R genes and (iii identify promising plant lines as repertoire for R-gene isolation. The effectors provoked different reactions on closely related plant lines indicative of a high variability and evolution rate of potential R genes. In some cases, putative R genes were conserved within a plant species but not within superordinate phylogenetical units. Interestingly, the effector XopQ was recognized by several Nicotiana spp. lines, and Xcv infection assays revealed that XopQ is a host range determinant in many Nicotiana species. Non-host resistance against Xcv and XopQ recognition in N. benthamiana required EDS1, strongly suggesting the presence of a TIR domain-containing XopQ-specific R protein in these plant lines. XopQ is a conserved effector among most xanthomonads, pointing out the XopQ-recognizing RxopQ as candidate for targeted crop improvement.

  16. Cultivos celulares de nicotiana tabacum L.cv.BY-2 como sistema modelo en el estudio de la adaptación al estrés salino.

    OpenAIRE

    García de la Garma García, Jesús

    2013-01-01

    El objetivo general de esta tesis doctoral es ampliar los conocimientos sobre los mecanismos que permiten la adaptación de los cultivos celulares de Nicotiana tabacum cv. BY-2 al estrés abiótico, concretamente estrés salino. Para ello, hemos establecido como objetivos específicos los siguientes: 1. Estudiar diferentes mecanismos fisiológicos, bioquímicos y celulares de adaptación a salinidad. 2. Estudiar el papel que desempeñan las giberelinas (GAs) en el proceso de adaptación a salini...

  17. Assimilation of phytate-phosphorus by the extracellular phytase activity of tobacco (Nicotiana tabacum) is affected by the availability of soluble phytate

    OpenAIRE

    2006-01-01

    Phytate, the major organic phosphorus in soil, is not readily available to plants as a source of phosphorus (P). It is either complexed with cations or adsorbed to various soil components. The present study was carried out to investigate the extracellular phytase activities of tobacco (Nicotiana tabacum variety GeXin No.1) and its ability to assimilate external phytate-P. Whereas phytase activities in roots, shoots and growth media of P i-fed 14-day-old seedlings were only 1.3-4.9% of total a...

  18. "Nicotiana tabacum" L. cv Xanthi como sistema heterólogo para la producción de lactógeno placentario humano (hPL)

    OpenAIRE

    Urreta Gómez, Iratxe

    2012-01-01

    152 p. : il., col. El lactógeno placentario humano (hPL) es una hormona polipeptídica que forma parte de la pequeña lista de factores del crecimiento capaces de tratar la diabetes tipo 1 mediante el transplante de islotes humanos. Esta hormona tiene la capacidad de mejorar la supervivencia de los islotes humanos antes y/o después de su transplante. En este trabajo se aborda la producción de hPL en sistemas heterólogos vegetales basados en Nicotiana tabacum cv Xanthi. El cDNA del hPL ha sid...

  19. Transfer of the cytochrome P450-dependent dhurrin pathway from Sorghum bicolor into Nicotiana tabacum chloroplasts for light-driven synthesis

    DEFF Research Database (Denmark)

    Gnanasekaran, Thiyagarajan; Karcher, Daniel; Nielsen, Agnieszka Janina Zygadlo;

    2016-01-01

    . For this purpose, we stably engineered the dhurrin pathway from Sorghum bicolor into the chloroplasts of Nicotiana tabacum (tobacco). Dhurrin is a cyanogenic glucoside and its synthesis from the amino acid tyrosine is catalysed by two membrane-bound cytochrome P450 enzymes (CYP79A1 and CYP71E1) and a soluble...... compared to 6% in sorghum. The results obtained pave the way for plant P450s involved in the synthesis of economically important compounds to be engineered into the thylakoid membrane of chloroplasts, and demonstrate that their full catalytic cycle can be driven directly by photosynthesis-derived electrons....

  20. 烟草黑胫病菌产孢性状的研究%Study on the Producing Spores Properties of Phytophthora Parasitica var nicotianae

    Institute of Scientific and Technical Information of China (English)

    周志成

    2005-01-01

    研究了湖南烟区烟草黑胫病菌(Phytophthora parasitica var nicotianae)分离菌株的产孢性状.结果表明,Py96-1和Py96-2两菌株用燕麦片培养基,在25~28℃温度下,培养21~28 d,产孢菌丝生长最好,菌丝茂密,产生孢子囊多.

  1. The hrpZ Gene of Pseudomonas syringae pv. phaseolicola Enhances Resistance to Rhizomania Disease in Transgenic Nicotiana benthamiana and Sugar Beet

    OpenAIRE

    Ourania I Pavli; Kelaidi, Georgia I.; Tampakaki, Anastasia P.; Skaracis,George N.

    2011-01-01

    To explore possible sources of transgenic resistance to the rhizomania-causing Beet necrotic yellow vein virus (BNYVV), Nicotiana benthamiana plants were constructed to express the harpin of Pseudomonas syringae pv. phaseolicola (HrpZ(Psph)). The HrpZ protein was expressed as an N-terminal fusion to the PR1 signal peptide (SP/HrpZ) to direct harpin accumulation to the plant apoplast. Transgene integration was verified by mPCR in all primary transformants (T0), while immunoblot analysis confir...

  2. Alberprosenia malheiroi Serra, Atzingen & Serra, 1987 (Hemiptera, Reduviidae: redescrição e bionomia Alberprosenia malheiroi Serra, Atzingen & Serra, 1987 (Hemiptera, Rediviidae: redescription and bionomics

    Directory of Open Access Journals (Sweden)

    Rodolfo U. Carcavallo

    1995-12-01

    Full Text Available Sobre Alberprosenia malheiroi n. sp. deu-se a conhecer uma diagnose resumida em 1980, porém sem valor bibliográfico. Em 1987 publicou-se a mesma diagnose anexando uma foto e alguns comentários, porém sem realizar uma descrição formal. Descreve-se para esta espécie, os adultos, os estádios imaturos, determina-se a série sintípica e apresenta-se dados bionômicos e de criação em insetário. As diferenças mais evidentes com A. goyovargasi, a única espécie que se conhecia do gênero até então, são a coloração geral negra, o espaço interocular maior que o tamanho de um olho visto dorsalmente, os tubérculos do colar com o ápice agudo e o tamanho maior, na nova espécie, quase o dobro do da primeira espécie. Os ovos são pequenos, fixados ao substrato em grupos de 3 ou 4, elipsóides, não achatados lateralmente, com o opérculo proeminente, convexo, sem estruturas evidentes. As ninfas apresentam em todos os estádios caracteres típicos do gênero e da tribo, com região anteocular menos longa que a post-ocular e característica pilosidade do tegumento que vai se acentuando a cada estádio. A. malheiroi n. sp. foi capturado em ecótopos silvestres em palmeiras em floresta no Estado do Pará, associados com morcegos ou aves. Nenhum dos exemplares estava infectado com Trypanosoma cruzi. Esses triatomíneos foram mantidos em insetários a ± 25°C e ± 60% UR, são insetos ágeis e voam com relativa facilidade. Alimentaram-se bem em pombos e morcegos e não aceitaram alimentação em ratos, camundongos ou hamsters. O período de incubação dos ovos foi em média treze dias e o tempo de evolução do período ninfal foi em média cento e trinta e dois dias.Alberprosenia malheiroi n. sp. first became known through an informal publication of a paper submitted a Congress, in 1980. The same diagnosis with a photograph was published in 1987, but no formal description has yeet been made. The present authors describe adults and immature

  3. Emergency Medical Service (EMS) Stations

    Data.gov (United States)

    Kansas Data Access and Support Center — EMS Locations in Kansas The EMS stations dataset consists of any location where emergency medical services (EMS) personnel are stationed or based out of, or where...

  4. Isolation and Characterization of a Lycopene ε-Cyclase Gene of <em>Chlorella em>(Chromochloris> <em>zofingiensis>. Regulation of the Carotenogenic Pathway by Nitrogen and Light

    Directory of Open Access Journals (Sweden)

    Maria Angeles Vargas

    2012-09-01

    Full Text Available The isolation and characterization of the lycopene ε-cyclase gene from the green microalga <em>Chlorella em>(Chromochloris> <em>zofingiensis> (<em>Czlcy-e> was performed. This gene is involved in the formation of the carotenoids α-carotene and lutein. <em>Czlcy-e> gene encoded a polypeptide of 654 amino acids. A single copy of <em>Czlcy-e> was found in <em>C. zofingiensisem>. Functional analysis by heterologous complementation in <em>Escherichia coliem> showed the ability of this protein to convert lycopene to δ-carotene. In addition, the regulation of the carotenogenic pathway by light and nitrogen was also studied in <em>C. zofingiensisem>. High irradiance stress did not increase mRNA levels of neither lycopene β<em>->cyclase gene (<em>lcy-b> nor lycopene ε-cyclase gene<em> em>(lcy-e> as compared with low irradiance conditions, whereas the transcript levels of <em>psy>, <em>pds>, <em>chyB> and <em>bkt> genes were enhanced, nevertheless triggering the synthesis of the secondary carotenoids astaxanthin, canthaxanthin and zeaxanthin and decreasing the levels of the primary carotenoids α-carotene, lutein, violaxanthin and β-carotene. Nitrogen starvation <em>per seem> enhanced mRNA levels of all genes considered, except <em>lcy-e and pdsem>, but did not trigger the synthesis of astaxanthin, canthaxanthin nor zeaxanthin. The combined effect of both high light and nitrogen starvation stresses enhanced significantly the accumulation of these carotenoids as well as the transcript levels of <em>bkt> gene, as compared with the effect of only high irradiance stress.

  5. Estudos anatômicos e histológicos sobre a subfamilia Triatominae (Heteroptera, reduviidae: cérebro e seus nervos de Triatoma infestans

    Directory of Open Access Journals (Sweden)

    Rudolf Barth

    1976-01-01

    Full Text Available A fim de apresentar um conhecimento detalhado da estrutura e função dos diferentes sistemas nervosos dos Triatominae, começamos neste estudo com a descrição do sistema senso-motor. Como primeira parte escolhemos o cérebro e os nervos deste. A forma externa do cérebro é predeterminada pela anatomia da cápsula craniana. O cérebro localiza-se na parte posterior da cabeça, embaixo e por trás dos dois ocelos. É caracterizado por um encurtamento em sentido longitudinal, de modo que as conectivas entre sincérebro e Gnatocérebro são extremamente curtas, porém largas, além disto, a terceira comissura é incluída na massa do segmento mandibular do Gnatocérebro. As três massas ópticas são de formação típica, bem como a comissura óptica. O corpo central é grande e está em ligação direta com a maioria dos centros do Protocérebro e do Deutocérebro. O corpo pedunculado possui apenas um único glomérulo em forma de cogumelo pedunculado sem sinal de formação de um cálice como em formigas e outros insetos mais evoluídos. As extremidades do pedúnculo desfazem-se embaixo do Corpo central em um grande complexo de anexos claviformes como encontrado nos Machilidae. O Deutocérebro é caracterizado por um grande centro antenal, composto de numerosos pequenos glomérulos em situação periférica. O Tritocérebro é pequeno, mostrando na sua parte anterior duas conectivas frontais muito curtas, de modo que o gãnglio frontal se situa perto do Tritocérebro. A massa compacta do Gnatocérebro mostra sua composição por três pares de gânglios apenas em séries de cortes. Além dos Lobos ópticos, saem do cérebro 8 pares de nervos e 3 nervos ímpares: Do Protocérebro: Os nervos dos ocelos e o nervo do Corpus allatum; Do Deutocérebro: Os nervos das antenas e os da faringe; Do tritocérebro: Os nervos labrais e, através do gânglio frontal, o nervo recurrente e o nervo dos músculos da faringe; Do gnatocérebro: Os nervos das mand

  6. Induction of systemic disease resistance in Nicotiana benthamiana by the cyclodipeptides cyclo (l-Pro-l-Pro) and cyclo (d-Pro-d-Pro).

    Science.gov (United States)

    Wu, Liming; Wu, Huijun; Chen, Lina; Zhang, Hongyue; Gao, Xuewen

    2017-01-01

    Cyclodipeptides, formed from two amino acids by cyclodehydration, are produced naturally by many organisms, and are known to possess a large number of biological activities. In this study, we found that cyclo (l-Pro-l-Pro) and cyclo (d-Pro-d-Pro) (where Pro is proline) could induce defence responses and systemic resistance in Nicotiana benthamiana. Treatment with the two cyclodipeptides led to a reduction in disease severity by Phytophthora nicotianae and Tobacco mosaic virus (TMV) infections compared with controls. Both cyclopeptides triggered stomatal closure, induced reactive oxygen species production and stimulated cytosolic calcium ion and nitric oxide production in guard cells. In addition, the application of cyclodipeptides significantly up-regulated the expression of the plant defence gene PR-1a and the PR-1a protein, and increased cellular salicylic acid (SA) levels. These results suggest that the SA-dependent defence pathway is involved in cyclodipeptide-mediated pathogen resistance in N. benthamiana. We report the systemic resistance induced by cyclodipeptides, which sheds light on the potential of cyclodipeptides for the control of plant diseases.

  7. Antigen production using heterologous expression of dengue virus-2 non-structural protein 1 (NS1) in Nicotiana tabacum (Havana) for immunodiagnostic purposes.

    Science.gov (United States)

    Amaro, Marilane O F; Xisto, Mariana F; Dias, Ana Carolina F; Versiani, Alice F; Cardoso, Silvia A; Otoni, Wagner C; da Silva, Cynthia C; De Paula, Sérgio O

    2015-06-01

    Expression of dengue-2 virus NS1 protein in Nicotiana tabacum plants for development of dengue immunodiagnostic kits. Dengue is one of the most important diseases caused by arboviruses in the world. A significant increase in its geographical distribution has been noticed over the last 20 years, with continuous transmission of several serotypes and emergence of the hemorrhagic fever in areas where the disease was previously not prevalent. Although the methodological processes for dengue diagnosis are in deep development and improvement, a limitation for the realization of dengue diagnostic tests is the difficulty of large-scale production of the antigen to be used in diagnostic tests. Due to this demand, the purpose of this study was to obtain the non-structural protein 1 (NS1) from dengue-2 serotype by heterologous expression in Nicotiana tabacum (Havana). After confirmation of the NS1 protein gene integration in the plant genome, the heterologous protein was characterized using SDS-PAGE and immunoblotting. In an immunoenzymatic test, the recombinant NS1 protein presents an antigen potential for development of dengue immunodiagnostic kits.

  8. Effectiveness of Biopesticide Derived fromCassia spectabilis and Nicotiana tabacum Leaves Against the Main Insect Pests of Coffee and Its Effect On Other Arthropods

    Directory of Open Access Journals (Sweden)

    Soekadar Wiryadiputra

    2006-05-01

    Full Text Available An experiment on the effectiveness of biopesticide made of ramayana (Cassia spectabilis and tobacco leaves (Nicotiana tabacum it called as Casnic, on Hypothenemus hampeiand Planococcus citriand its effect on the arthropods population fauna in coffee plantation had been conducted in Pest Laboratory of Indonesian Coffee and Cocoa Researh Institute and Bangelan Plantation in Malang. The treatment consisted of three level concentrations of botanical pesticide(15 ml, 30 ml, and 60 ml/lof water, Beauveria bassiana(at a dose 100 g spore per ha, metidation spraying (2 ml formulation per litre of water and two control treatments (with and without soap. The field experiment was arranged in randomized complete block design (RCBD with four replications. The results showed that botanical pesticide at the concentration of 30.0 ml stock solution per litre of water and applied four times with monthly interval was effective in controlling coffee berry borer (Hypothenemus hampei and coffee mealy bug (Planococcus citri. The effectiveness was not significantly different compared to metidation and Beauveria bassianaat the dose of 0.8 l and 100 g formulation per ha per application. Application of the botanical pesticide did not show negative effect on the usefull predatory insects and other arthropods population in coffee ecosystem. Key words : Botanical pesticide, Cassia spectabilis, Nicotiana tabacum, Coffee, Hypothenemus hampei, Planococcus citri, Arthropods.

  9. Two aspartate residues at the putative p10 subunit of a type II metacaspase from Nicotiana tabacum L. may contribute to the substrate-binding pocket.

    Science.gov (United States)

    Acosta-Maspons, Alexis; Sepúlveda-García, Edgar; Sánchez-Baldoquín, Laura; Marrero-Gutiérrez, Junier; Pons, Tirso; Rocha-Sosa, Mario; González, Lien

    2014-01-01

    Metacaspases are cysteine proteases present in plants, fungi, prokaryotes, and early branching eukaryotes, although a detailed description of their cellular function remains unclear. Currently, three-dimensional (3D) structures are only available for two metacaspases: Trypanosoma brucei (MCA2) and Saccharomyces cerevisiae (Yca1). Furthermore, metacaspases diverged from animal caspases of known structure, which limits straightforward homology-based interpretation of functional data. We report for the first time the identification and initial characterization of a metacaspase of Nicotiana tabacum L., NtMC1. By combining domain search, multiple sequence alignment (MSA), and protein fold-recognition studies, we provide compelling evidences that NtMC1 is a plant metacaspase type II, and predict its 3D structure using the crystal structure of two type I metacaspases (MCA2 and Yca1) and Gsu0716 protein from Geobacter sulfurreducens as template. Analysis of the predicted 3D structure allows us to propose Asp353, at the putative p10 subunit, as a new member of the aspartic acid triad that coordinates the P1 arginine/lysine residue of the substrate. Nevertheless, site-directed mutagenesis and expression analysis in bacteria and Nicotiana benthamiana indicate the functionality of both Asp348 and Asp353. Through the co-expression of mutant and wild-type proteins by transient expression in N. benthamiana leaves we found that polypeptide processing seems to be intramolecular. Our results provide the first evidence in plant metacaspases concerning the functionality of the putative p10 subunit.

  10. Characterization of cDNA for PMT: a Partial Nicotine Biosynthesis-Related Gene Isolated from Indonesian Local Tobacco (Nicotiana tabacum cv. Sindoro1

    Directory of Open Access Journals (Sweden)

    Sesanti Basuki

    2013-12-01

    Full Text Available Nicotine is the major alkaloid compound in cultivated tobacco (Nicotiana tabacum that could potentially be converted into carcinogenic compound (nor-nicotine. The PMT gene encoding putrescine N-methyltransferase (PMT is one of the two key genes that play a prominent role in nicotine biosynthesis. The aimed of this study was to isolate and characterize the cDNA sequence originated from Indonesian local tobacco cv. Sindoro1 (Ntpmt_Sindoro1. The results showed that the Ntpmt_Sindoro1 was 1124 bp in length. This cDNA fragment encodes for 374 amino acid residues. The predicted polypeptide from the cDNA is a hidrophilic protein, and has a predicted molecular weight of 40.95 kD. The predicted amino acids sequence also showed high similarity to the PMT gene product Nicotiana sp. available in the GenBank data base. The amino acid sequences also exert conserved residues specifically exhibited only by PMT gene originated from N. tabacum. Clustering analysis revealed that Ntpmt_Sindoro1 belongs to the same clade as the PMT3 gene, a member of the N. tabacum PMT gene family. The Ntpmt_Sindoro1 cDNA sequence covering exon1-exon8 of the PMT gene fragment has been registered in the GenBank data base, under the accession number JX978277.

  11. Non-host Plant Resistance against Phytophthora capsici Is Mediated in Part by Members of the I2 R Gene Family in Nicotiana spp.

    Science.gov (United States)

    Vega-Arreguín, Julio C.; Shimada-Beltrán, Harumi; Sevillano-Serrano, Jacobo; Moffett, Peter

    2017-01-01

    The identification of host genes associated with resistance to Phytophthora capsici is crucial to developing strategies of control against this oomycete pathogen. Since there are few sources of resistance to P. capsici in crop plants, non-host plants represent a promising source of resistance genes as well as excellent models to study P. capsici – plant interactions. We have previously shown that non-host resistance to P. capsici in Nicotiana spp. is mediated by the recognition of a specific P. capsici effector protein, PcAvr3a1 in a manner that suggests the involvement of a cognate disease resistance (R) genes. Here, we have used virus-induced gene silencing (VIGS) and transgenic tobacco plants expressing dsRNA in Nicotiana spp. to identify candidate R genes that mediate non-host resistance to P. capsici. Silencing of members of the I2 multigene family in the partially resistant plant N. edwardsonii and in the resistant N. tabacum resulted in compromised resistance to P. capsici. VIGS of two other components required for R gene-mediated resistance, EDS1 and SGT1, also enhanced susceptibility to P. capsici in N. edwardsonii, as well as in the susceptible plants N. benthamiana and N. clevelandii. The silencing of I2 family members in N. tabacum also compromised the recognition of PcAvr3a1. These results indicate that in this case, non-host resistance is mediated by the same components normally associated with race-specific resistance. PMID:28261255

  12. MORFOMETRÍA COMPARADA DE SEMILLAS DE NICOTIANA (SOLANACEAE E IDENTIFICACIÓN DE SEMILLAS CARBONIZADAS PROVENIENTES DE UN SITIO ARQUEOLÓGICO EN CHILE CENTRAL

    Directory of Open Access Journals (Sweden)

    M. Teresa Planella

    2012-01-01

    Full Text Available La presencia frecuente de pipas para fumar en sitios arqueológicos del Período Alfarero Temprano deChile central y las evidencias en relación con la costumbre de fumar especies de Nicotiana halladas en sitios prehispánicos de otros lugares de las Américas, muestran la necesidad de contar con una metodología para identificar las especies de este género usadas en Chile. En este trabajo se ha realizado un estudio morfométrico en semillas de especies de Nicotiana que sirve de referencia para comparaciones con semillas de origen arqueológico. La forma y tamaño de la semilla, el patrón de ornamentación dado por las células epidérmicas y la ubicación del hilum resultaron ser caracteres relevantes para identificaciones confiables. Utilizando estos caracteres, se determinaron como N. corymbosa a las semillas recuperadas en el sitio arqueológico Las Morrenas 1, ubicado en Chile central.

  13. Effects of aluminum oxide nanoparticles on the growth, development, and microRNA expression of tobacco (Nicotiana tabacum.

    Directory of Open Access Journals (Sweden)

    Caitlin E Burklew

    Full Text Available Nanoparticles are a class of newly emerging environmental pollutions. To date, few experiments have been conducted to investigate the effect nanoparticles may have on plant growth and development. It is important to study the effects nanoparticles have on plants because they are stationary organisms that cannot move away from environmental stresses like animals can, therefore they must overcome these stresses by molecular routes such as altering gene expression. microRNAs (miRNA are a newly discovered, endogenous class of post-transcriptional gene regulators that function to alter gene expression by either targeting mRNAs for degradation or inhibiting mRNAs translating into proteins. miRNAs have been shown to mediate abiotic stress responses such as drought and salinity in plants by altering gene expression, however no study has been performed on the effect of nanoparticles on the miRNA expression profile; therefore our aim in this study was to classify if certain miRNAs play a role in plant response to Al(2O(3 nanoparticle stress. In this study, we exposed tobacco (Nicotiana tabacum plants (an important cash crop as well as a model organism to 0%, 0.1%, 0.5%, and 1% Al(2O(3 nanoparticles and found that as exposure to the nanoparticles increased, the average root length, the average biomass, and the leaf count of the seedlings significantly decreased. We also found that miR395, miR397, miR398, and miR399 showed an extreme increase in expression during exposure to 1% Al(2O(3 nanoparticles as compared to the other treatments and the control, therefore these miRNAs may play a key role in mediating plant stress responses to nanoparticle stress in the environment. The results of this study show that Al(2O(3 nanoparticles have a negative effect on the growth and development of tobacco seedlings and that miRNAs may play a role in the ability of plants to withstand stress to Al(2O(3 nanoparticles in the environment.

  14. A genetic system for Citrus Tristeza Virus using the non-natural host Nicotiana benthamiana: an update

    Directory of Open Access Journals (Sweden)

    Silvia eAmbrós

    2013-07-01

    Full Text Available In nature Citrus tristeza virus (CTV, genus Closterovirus, infects only the phloem cells of species of Citrus and related genera. Finding that the CTV T36 strain replicated in Nicotiana benthamiana (NB protoplasts and produced normal virions allowed development of the first genetic system based on protoplast transfection with RNA transcribed from a full-genome cDNA clone, a laborious and uncertain system requiring several months for each experiment. We developed a more efficient system based on agroinfiltration of NB leaves with CTV-T36-based binary plasmids, which caused systemic infection in this non-natural host within a few weeks yielding in the upper leaves enough CTV virions to readily infect citrus by slash inoculation. Stem agroinoculation of citrus and NB plants with oncogenic strains of Agrobacterium tumefaciens carrying a CTV-T36 binary vector with a GUS marker, induced GUS positive galls in both species. However, while most NB tumours were CTV positive and many plants became systemically infected, no coat protein or viral RNA was detected in citrus tumours, even though CTV cDNA was readily detected by PCR in the same galls. This finding suggests i strong silencing or CTV RNA processing in transformed cells impairing infection progress, and ii the need for using NB as an intermediate host in the genetic system. To maintain CTV-T36 in NB or assay other CTV genotypes in this host, we also tried to graft-transmit the virus from infected to healthy NB, or to mechanically inoculate NB leaves with virion extracts. While these trials were mostly unsuccessful on non-treated NB plants, agroinfiltration with silencing suppressors enabled for the first time infecting NB plants by side-grafting and by mechanical inoculation with virions, indicating that previous failure to infect NB was likely due to virus silencing in early infection steps. Using NB as a CTV host provides new possibilities to study virus-host interactions with a simple and

  15. Molecular Characterization of Two Lysophospholipid:acyl-CoA Acyltransferases Belonging to the MBOAT Family in Nicotiana benthamiana.

    Directory of Open Access Journals (Sweden)

    Donghui Zhang

    Full Text Available In the remodeling pathway for the synthesis of phosphatidylcholine (PC, acyl-CoA-dependent lysophosphatidylcholine (lysoPC acyltransferase (LPCAT catalyzes the reacylation of lysoPC. A number of genes encoding LPCATs have been cloned and characterized from several plants in recent years. Using Arabidopsis and other plant LPCAT sequences to screen the genome database of Nicotiana benthamiana, we identified two cDNAs encoding the putative tobacco LPCATs (NbLPCAT1 and NbLPCAT2. Both of them were predicted to encode a protein of 463 amino acids with high similarity to LPCATs from other plants. Protein sequence features such as the presence of at least eight putative transmembrane regions, four highly conserved signature motifs and several invariant residues indicate that NbLPCATs belong to the membrane bound O-acyltransferase family. Lysophospholipid acyltransferase activity of NbLPCATs was confirmed by testing lyso-platelet-activating factor (lysoPAF sensitivity through heterologous expression of each full-length cDNA in a yeast mutant Y02431 (lca1△ disrupted in endogenous LPCAT enzyme activity. Analysis of fatty acid profiles of phospholipids from the NbLPCAT-expressing yeast mutant Y02431 cultures supplemented with polyunsaturated fatty acids suggested more incorporation of linoleic acid (18:2n6, LA and α-linolenic acid (18:3n3, ALA into PC compared to yeast mutant harbouring empty vector. In vitro enzymatic assay demonstrated that NbLPCAT1had high lysoPC acyltransferase activity with a clear preference for α-linolenoyl-CoA (18:3, while NbLPCAT2 showed a high lysophosphatidic acid (lysoPA acyltransferase activity towards α-linolenoyl-CoA and a weak lysoPC acyltransferase activity. Tissue-specific expression analysis showed a ubiquitous expression of NbLPCAT1 and NbLPCAT2 in roots, stems, leaves, flowers and seeds, and a strong expression in developing flowers. This is the first report on the cloning and characterization of lysophospholipid

  16. Rapid modification of the insect elicitor N-linolenoyl-glutamate via a lipoxygenase-mediated mechanism on Nicotiana attenuata leaves

    Directory of Open Access Journals (Sweden)

    VanDoorn Arjen

    2010-08-01

    Full Text Available Abstract Background Some plants distinguish mechanical wounding from herbivore attack by recognizing specific constituents of larval oral secretions (OS which are introduced into plant wounds during feeding. Fatty acid-amino acid conjugates (FACs are major constituents of Manduca sexta OS and strong elicitors of herbivore-induced defense responses in Nicotiana attenuata plants. Results The metabolism of one of the major FACs in M. sexta OS, N-linolenoyl-glutamic acid (18:3-Glu, was analyzed on N. attenuata wounded leaf surfaces. Between 50 to 70% of the 18:3-Glu in the OS or of synthetic 18:3-Glu were metabolized within 30 seconds of application to leaf wounds. This heat-labile process did not result in free α-linolenic acid (18:3 and glutamate but in the biogenesis of metabolites both more and less polar than 18:3-Glu. Identification of the major modified forms of this FAC showed that they corresponded to 13-hydroxy-18:3-Glu, 13-hydroperoxy-18:3-Glu and 13-oxo-13:2-Glu. The formation of these metabolites occurred on the wounded leaf surface and it was dependent on lipoxygenase (LOX activity; plants silenced in the expression of NaLOX2 and NaLOX3 genes showed more than 50% reduced rates of 18:3-Glu conversion and accumulated smaller amounts of the oxygenated derivatives compared to wild-type plants. Similar to 18:3-Glu, 13-oxo-13:2-Glu activated the enhanced accumulation of jasmonic acid (JA in N. attenuata leaves whereas 13-hydroxy-18:3-Glu did not. Moreover, compared to 18:3-Glu elicitation, 13-oxo-13:2-Glu induced the differential emission of two monoterpene volatiles (β-pinene and an unidentified monoterpene in irlox2 plants. Conclusions The metabolism of one of the major elicitors of herbivore-specific responses in N. attenuata plants, 18:3-Glu, results in the formation of oxidized forms of this FAC by a LOX-dependent mechanism. One of these derivatives, 13-oxo-13:2-Glu, is an active elicitor of JA biosynthesis and differential

  17. MAPK-dependent JA and SA signalling in Nicotiana attenuata affects plant growth and fitness during competition with conspecifics

    Directory of Open Access Journals (Sweden)

    Meldau Stefan

    2012-11-01

    Full Text Available Abstract Background Induced defense responses to herbivores are generally believed to have evolved as cost-saving strategies that defer the fitness costs of defense metabolism until these defenses are needed. The fitness costs of jasmonate (JA-mediated defenses have been well documented. Those of the early signaling units mediating induced resistance to herbivores have yet to be examined. Early signaling components that mediate herbivore-induced defense responses in Nicotiana attenuata, have been well characterized and here we examine their growth and fitness costs during competition with conspecifics. Two mitogen-activated protein kinases (MAPKs, salicylic acid (SA-induced protein kinase (SIPK and wound-induced protein kinase (WIPK are rapidly activated after perception of herbivory and both kinases regulate herbivory-induced JA levels and JA-mediated defense metabolite accumulations. Since JA-induced defenses result in resource-based trade-offs that compromise plant productivity, we evaluated if silencing SIPK (irSIPK and WIPK (irWIPK benefits the growth and fitness of plants competiting with wild type (WT plants, as has been shown for plants silenced in JA-signaling by the reduction of Lipoxygenase 3 (LOX3 levels. Results As expected, irWIPK and LOX3-silenced plants out-performed their competing WT plants. Surprisingly, irSIPK plants, which have the largest reductions in JA signaling, did not. Phytohormone profiling of leaves revealed that irSIPK plants accumulated higher levels of SA compared to WT. To test the hypothesis that these high levels of SA, and their presumed associated fitness costs of pathogen associated defenses in irSIPK plants had nullified the JA-deficiency-mediated growth benefits in these plants, we genetically reduced SA levels in irSIPK plants. Reducing SA levels partially recovered the biomass and fitness deficits of irSIPK plants. We also evaluated whether the increased fitness of plants with reduced SA or JA levels

  18. <em>Angiostrongylus vasorumem> in red foxes (<em>Vulpes vulpesem> and badgers (<em>Meles melesem> from Central and Northern Italy

    Directory of Open Access Journals (Sweden)

    Marta Magi

    2010-06-01

    Full Text Available Abstract During 2004-2005 and 2007-2008, 189 foxes (<em>Vulpes vulpesem> and 6 badgers (<em>Meles melesem> were collected in different areas of Central Northern Italy (Piedmont, Liguria and Tuscany and examined for <em>Angiostrongylus vasorumem> infection. The prevalence of the infection was significantly different in the areas considered, with the highest values in the district of Imperia (80%, Liguria and in Montezemolo (70%, southern Piedmont; the prevalence in Tuscany was 7%. One badger collected in the area of Imperia turned out to be infected, representing the first report of the parasite in this species in Italy. Further studies are needed to evaluate the role played by fox populations as reservoirs of infection and the probability of its spreading to domestic dogs.
    Riassunto <em>Angiostrongylus vasorumem> nella volpe (<em>Vulpes vulpesem> e nel tasso (<em>Meles melesem> in Italia centro-settentrionale. Nel 2004-2005 e 2007-2008, 189 volpi (<em>Vulpes vulpesem> e 6 tassi (<em>Meles melesem> provenienti da differenti aree dell'Italia settentrionale e centrale (Piemonte, Liguria Toscana, sono stati esaminati per la ricerca di <em>Angiostrongylus vasorumem>. La prevalenza del nematode è risultata significativamente diversa nelle varie zone, con valori elevati nelle zone di Imperia (80% e di Montezemolo (70%, provincia di Cuneo; la prevalenza in Toscana è risultata del 7%. Un tasso proveniente dall'area di Imperia è risultato positivo per A. vasorum; questa è la prima segnalazione del parassita in tale specie in Italia. Ulteriori studi sono necessari per valutare il potenziale della volpe come serbatoio e la possibilità di diffusione della parassitosi ai cani domestici.

    doi:10.4404/hystrix-20.2-4442

  19. <em>An entem>-Kaurane-Type Diterpene in <em>Croton antisyphiliticusem> Mart.

    Directory of Open Access Journals (Sweden)

    Ana Maria Pereira

    2012-07-01

    Full Text Available <em>Croton antisyphiliticus em>is a medicinal plant widely used in the treatment of microbial infections, especially those affecting the genital tract. Crude extract, fractions and pure compound isolated from roots of this species were investigated to validate their antimicrobial activity against <em>Escherichia coliem> and <em>Staphylococcus aureusem>. The compound <em>ent>-kaur-16-en-18-oic acid was isolated as a major component (0.7% of crude extract, and its MIC value determined against <em>S. aureusem> (ATCC 6538 was 250 μg/mL. This is the first phytochemical work on the species monitored with antimicrobial assay.

  20. The Tomato spotted wilt virus cell-to-cell movement protein (NSM) triggers a hypersensitive response in Sw-5 containing resistant tomato lines and Nicotiana benthamiana transformed with the functional Sw-5b resistance gene copy.

    NARCIS (Netherlands)

    Hallwass, M.; Silva de Oliveira, A.; Dianese, E.C.; Lohuis, D.; Boiteux, L.S.; Inoue-Nagata, A.K.; Resende, de R.O.; Kormelink, R.J.M.

    2014-01-01

    Although the Sw-5 gene cluster has been cloned, and Sw-5b has been identified as the functional gene copy that confers resistance to Tomato spotted wilt virus (TSWV), its avirulence (Avr) determinant has not been identified to date. Nicotiana tabacum SR1 plants transformed with a copy of the Sw-5b

  1. Ectopic expression of Malus domestica class 1 knox genes altered growth and development of Nicotiana tabacum and Prunus domestica, and induced adventitious shoot regeneration from leaf explants without exogenous cytokinin

    Science.gov (United States)

    Transgenic tobacco (Nicotiana tabacum L) and plum (Prunus domestica L) plants were regenerated by transforming with apple class 1 KNOX genes (MdKNP1 and MdKNP2) or a corn KN1 (ZmKN1) gene. Transgenic tobacco plants were produced in vitro from transformed leaf discs in the absence of cytokinin in th...

  2. Expression of Selected <em>Ginkgo em>>biloba em>Heat Shock Protein Genes After Cold Treatment Could Be Induced by Other Abiotic Stress

    Directory of Open Access Journals (Sweden)

    Feng Xu

    2012-05-01

    Full Text Available Heat shock proteins (HSPs play various stress-protective roles in plants. In this study, three <em>HSP> genes were isolated from a suppression subtractive hybridization (SSH cDNA library of <em>Ginkgo bilobaem> leaves treated with cold stress. Based on the molecular weight, the three genes were designated <em>GbHSP16.8em>, <em>GbHSP17em> and <em>GbHSP70em>. The full length of the three genes were predicted to encode three polypeptide chains containing 149 amino acids (Aa, 152 Aa, and 657 Aa, and their corresponding molecular weights were predicted as follows: 16.67 kDa, 17.39 kDa, and 71.81 kDa respectively. The three genes exhibited distinctive expression patterns in different organs or development stages. <em>GbHSP16.8em> and <em>GbHSP70em> showed high expression levels in leaves and a low level in gynoecia, <em>GbHSP17em> showed a higher transcription in stamens and lower level in fruit. This result indicates that <em>GbHSP16.8em> and <em>GbHSP70 em>may play important roles in <em>Ginkgo> leaf development and photosynthesis, and <em>GbHSP17em> may play a positive role in pollen maturation. All three <em>GbHSPs> were up-regulated under cold stress, whereas extreme heat stress only caused up-regulation of <em>GbHSP70em>, UV-B treatment resulted in up-regulation of <em>GbHSP16.8em> and <em>GbHSP17em>, wounding treatment resulted in up-regulation of <em>GbHSP16.8em> and <em>GbHSP70em>, and abscisic acid (ABA treatment caused up-regulation of <em>GbHSP70em> primarily.

  3. Pesquisando em fontes visuais

    OpenAIRE

    Proença, Caio Carvalho; FFCH

    2012-01-01

    O presente ensaio pretende demonstrar reflexões sobre os usos da fotografia como fonte. Na primeira parte do ensaio procura-se demonstrar a experiência de um primeiro contato com a fonte visual, em especial a fotografia, para posteriormente entrar em contato com as diferentes formas metodológicas presente na pesquisa desta fonte. Em um segundo momento mostra-se alguns apontamentos sobre o método de pesquisa voltado à fonte visual fotográfica que auxiliam a pesquisa, contemplando a discussão s...

  4. Meteorologia em linha

    OpenAIRE

    2015-01-01

    DISPOSITIVO TÉCNICO-PEDAGÓGICO: CURSO DE METEOROLOGIA EM LINHA. Designamos por curso o sistema que resulta da relação entre um conjunto de componentes, dando unidade e identidade ao mesmo, incorporados na plataforma Versal: conjunto de 30 lições em vídeo, recursos complementares, valorização e contextualização do curso, orientação e acompanhamento desenhados e previstos, interação com os utilizadores do curso a partir do momento da sua edição aberta em linha. (https://versal.com/c/1fzwaz/m...

  5. A hipnose em triatletas

    OpenAIRE

    Szenészi, Daniela Scharamm

    2005-01-01

    Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Filosofia e Ciências Humanas. Programa de Pós-Graduação em Psicologia. Esta pesquisa objetivou investigar em atletas de triatlon a percepção das características da visualização da prova de Ironman e os seus componentes psicofisiológicos durante o transe hipnótico. Foram estudados 7 atletas do sexo masculino em 6 sessões de hipnose. Após cada sessão foi feita uma entrevista semi-estruturada e aplicado um questionário...

  6. Osteoartrites em equinos

    OpenAIRE

    Rocha, Francisco José Martins

    2008-01-01

    Dissertação de Mestrado Integrado em Medicina Veterinária A Osteoartrite (OA) é a principal causa de claudicação no cavalo de desporto e lazer, sendo uma afecção que tem grandes repercussões económicas. Este trabalho descreve algumas das características importantes da estrutura articular, bem como da sua fisiologia. Define a OA e todas as estruturas envolvidas no seu processo. Os mecanismos fisiopatológicos põem em evidência os factores de risco em causa e que determinam tod...

  7. Literatura em quadrinhos

    OpenAIRE

    Ferreira,Tânia Regina

    2015-01-01

    Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Comunicação e Expressão, Programa de Pós-Graduação em Literatura, Florianópolis, 2015. O objetivo desta tese é fazer o estudo do desfecho do romance de Lima Barreto, Triste fim de Policarpo Quaresma - (centenário de publicação em livro 1915-2015) - adaptado para as quatro versões homônimas em quadrinhos por: Edgar Vasques e Flávio Braga (Desiderata, 2010); Cesar Lobo e Luiz Antonio Aguiar (Ática, 2010); Lailson de Holanda...

  8. Melhores medicamentos em Pediatria

    OpenAIRE

    2014-01-01

    Resumo: Existem actualmente muitos medicamentos que são utilizados em crianças sem terem sido suficientemente estudados nas diferentes sub-populações pediátricas, com consequências preocupantes. O reconhecimento deste facto levou à criação de regras específicas na investigação de medicamentos pediátricos nos EUA, já em 1997. De igual forma, o Regulamento de Medicamentos para Uso Pediátrico aprovado em De zembro de 2006 pelo Parlamento Europeu, tem como objectivo a resolução deste problema no ...

  9. Characterization of <em>Erysiphe necatorem>-Responsive Genes in Chinese Wild <em>Vitis> <em>quinquangularis>

    Directory of Open Access Journals (Sweden)

    Xiping Wang

    2012-09-01

    Full Text Available Powdery mildew (PM, caused by fungus <em>Erysiphe necatorem>, is one of the most devastating diseases of grapevine. To better understand grapevine-PM interaction and provide candidate resources for grapevine breeding, a suppression subtractive hybridization (SSH cDNA library was constructed from <em>E. necatorem>-infected leaves of a resistant Chinese wild <em>Vitis quinquangularisem> clone “Shang-24”. A total of 492 high quality expressed sequence tags (ESTs were obtained and assembled into 266 unigenes. Gene ontology (GO analysis indicated that 188 unigenes could be assigned with at least one GO term in the biological process category, and 176 in the molecular function category. Sequence analysis showed that a large number of these genes were homologous to those involved in defense responses. Genes involved in metabolism, photosynthesis, transport and signal transduction were also enriched in the library. Expression analysis of 13 selected genes by qRT-PCR revealed that most were induced more quickly and intensely in the resistant material “Shang-24” than in the sensitive <em>V. pseudoreticulata em>clone “Hunan-1” by<em> E. necatorem> infection. The ESTs reported here provide new clues to understand the disease-resistance mechanism in Chinese wild grapevine species and may enable us to investigate <em>E. necatorem>-responsive genes involved in PM resistance in grapevine germplasm.

  10. Synthesis, Reactions and Evaluation of the Antimicrobial Activity of Some 4-(<em>p>-Halophenyl-4<em>H>-naphthopyran, Pyranopyrimidine and Pyranotriazolopyrimidine Derivatives

    Directory of Open Access Journals (Sweden)

    Ashraf H. F. Abd El-Wahab

    2012-07-01

    Full Text Available A series of naphthopyran derivatives 3a–f were prepared. Reaction of <em>2-em>amino-4-(<em>p>-chlorophenyl-7-methoxy-4<em>H>-naphtho[2,1-<em>b>]pyran-3-carbonitrile (3b with Ac2O afforded two products, 2-acetylamino-7-methoxy-4-(<em>p>-chlorophenyl-4<em>H>-naphtho-[2,1-<em>b>]pyran-3-carbonitrile (4 and 10,11-dihydro-3-methoxy-9-methyl-12-(<em>p>-chloro-phenyl-12<em>H>-naphtho[2,1-<em>b>]pyran[2,3-<em>d>]pyrimidine-11-one (5 and treatment of 3b with benzoyl chloride gave the pyranopyrimidin-11-one derivative 6. While treatment of 3b with formamide afforded 11-amino-3-methoxy-12-(<em>p>-chlorophenyl-12<em>H>-naphtho[2,1-<em>b>]pyrano[2,3-<em>d>]pyrimidine (7. Reaction of 3b with triethyl orthoformate gave the corresponding 2-ethoxymethyleneamino-7-methoxy-4-(<em>p>-chlorophenyl-4<em>H>-naphtho-[2,1-<em>b>]pyran-3-carbonitrile (8. Hydrazinolysis of 8 in EtOH at room temperature yielded 10-amino-10,11-dihydro-11-imino-3-methoxy-12-(<em>p>-chlorophenyl-12<em>H>-naphtho[2,1-b]pyrano-[2,3-d]pyrimidine (9, while aminolysis of 8 with methylamine or dimethylamine gave the corresponding pyranopyrimidine and <em>N,N>-dimethylaminomethylene derivatives 10 and 11. Condensation of 9 with some carboxylic acid derivatives afforded triazolopyrimidine derivatives 12–16, while reaction of 9 with benzaldehyde gave 10-benzalamino-10,11-dihydro-11-imino-3-methoxy-12-(<em>p>-chlorophenyl12<em>H>-naphtho[2,1-<em>b>]pyrano[2,3-<em>d>]pyrimidine (17. The structures of the newly synthesized compounds were confirmed by spectral data. The

  11. Spectral reflectance, chlorophyll fluorescence and virological investigations of tobacco plants (Nicotiana tabacum L.) infected with Tobacco mosaic virus (TMV)

    Science.gov (United States)

    Krezhova, Dora; Hristova, Dimitrina; Iliev, Ilko; Yanev, Tony

    Application of multispectral remote sensing techniques to plant condition monitoring has been adopted for various purposes. Remote sensing is a reliable tool for detecting signs of vege-tation stress and diseases. Spectral reflectance and chlorophyll fluorescence are functions of tissue optical properties and biological status of the plants, and illumination conditions. The mean reflectance spectrum depends on the relative composition of all the pigments in the leaf including chlorophylls, carotenoids etc. Chlorophyll fluorescence results from the primary re-actions of photosynthesis and during the last decade it finds widening application as a means for revelation of stress and diseases. The changes in chlorophyll function take place before the alteration in chlorophyll content to occur so that changes in the fluorescence signal arise before any visible signs are apparent. The aim of our investigations was to study the development and spreading out of a viral infection on the leaves of two cultivars tobacco plants (Nicotiana tabacum L.) infected with Tobacco mosaic virus (TMV). We applied two remote sensing tech-niques (spectral reflectance and chlorophyll fluorescence measurements) for evaluation of the changes in the optical properties of the plants in accordance to their physiological status. The serological analyses via the Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) were made with appropriate kits (Leowe, Germany) for quantitative assessment of the concentration of viruses in the plants. The tobacco plants were grown in green house under controlled conditions. The first cultivar Nevrocop 1146 is known as resistive to the TMV, i.e. it shows hypersensitive response. The second cultivar named Krumovgrad is normally sen-sitive to the TMV. At growth stage 4-6 expanded leaf, up to one leaf from 20 plants for each cultivar were inoculated with TMV. The leaves opposite to the infected ones formed the group of control (untreated) leaves. The

  12. International EMS Systems

    DEFF Research Database (Denmark)

    Langhelle, Audun; Lossius, Hans Morten; Silfvast, Tom;

    2004-01-01

    Emergency medicine service (EMS) systems in the five Nordic countries have more similarities than differences. One similarity is the involvement of anaesthesiologists as pre-hospital physicians and their strong participation for all critically ill and injured patients in-hospital. Discrepancies do...... exist, however, especially within the ground and air ambulance service, and the EMS systems face several challenges. Main problems and challenges emphasized by the authors are: (1) Denmark: the dispatch centres are presently not under medical control and are without a national criteria based system....... Access to on-line medical advice of a physician is not available; (2) Finland: the autonomy of the individual municipalities and their responsibility to cover for primary and specialised health care, as well as the EMS, and the lack of supporting or demanding legislation regarding the EMS; (3) Iceland...

  13. Preconceito em disfarce

    Directory of Open Access Journals (Sweden)

    João de Almeida

    2001-02-01

    Full Text Available

    0 presente estudo pretende analisar os significados básicos do texto "Preto e Branco", de F. Sabino, valendo-se de alguns modelos teóricos lingüísticos, principalmente de Greimas e de Pottier, em seqüência que vai da estrutura da narrativa e do percurso gerativo de sentido até as relações sêmicas, em especial.

  14. Biology of Triatominae (Reduviidae Hemiptera from North of Formosa County (Goiás-Brazil I. Length of life cycle of Triatoma sordida (Stal. 1859

    Directory of Open Access Journals (Sweden)

    Dalva A. Mello

    1976-12-01

    Full Text Available In the present paper the life cycle of Triatoma sordida was studied. The mean length from egg to adult was 213 days. The mean length in days from each stage was: 24.3 (± 1.30 for the first. 32.8 (± 1.45 (2nd, 36.1 (± 1.50 (3rd, 44.6 (± 1.85 (4th and 52.0 (± 1.92 (5th. The mean egg incubation períod was 23.2 (± 1.40. Overall mortality was 18.8% and egg viability was 82.5%.Foi estudado no presente trabalho o ciclo biológico do Triatoma sordida A média da duração do ciclo de ovo a adulto foi 213 dias. A média de duração em dias para cada estágio foi 24,3 (± 1,30 para o primeiro, 32,8 (± 1,45 para o segundo, 36,1 (± 1,50 para o terceiro, 44,6 (± 1,85 para o quarto e 52,0 (± 1,92 para o quinto. A média do período de incubação dos ovos foi 23,2 (± 1,40. A mortalidade em todos ps estágios foi 18,8% e a viabilidade dos ovos foi 82,5%.

  15. <em>In Vitro em>and <em>in em>Vivo> Antitumor Effect of Trachylobane-360, a Diterpene from<em> Xylopia langsdorffianaem>

    Directory of Open Access Journals (Sweden)

    João Carlos Lima Rodrigues Pita

    2012-08-01

    Full Text Available Trachylobane-360 (<em>ent>-7α-acetoxytrachyloban-18-oic acid was isolated from <em>Xylopia langsdorffianaem>. Studies have shown that it has weak cytotoxic activity against tumor and non-tumor cells. This study investigated the <em>in vitroem>> em>and <em>in vivoem> antitumor effects of trachylobane-360, as well as its cytotoxicity in mouse erythrocytes. In order to evaluate the <em>in vivoem> toxicological aspects related to trachylobane-360 administration, hematological, biochemical and histopathological analyses of the treated animals were performed. The compound exhibited a concentration-dependent effect in inducing hemolysis with HC50 of 273.6 µM, and a moderate <em>in vitroem>> em>concentration-dependent inhibitory effect on the proliferation of sarcoma 180 cells with IC50 values of 150.8 µM and 150.4 µM, evaluated by the trypan blue exclusion test and MTT reduction assay, respectively. The <em>in vivoem> inhibition rates of sarcoma 180 tumor development were 45.60, 71.99 and 80.06% at doses of 12.5 and 25 mg/kg of trachylobane-360 and 25 mg/kg of 5-FU, respectively. Biochemical parameters were not altered. Leukopenia was observed after 5-FU treatment, but this effect was not seen with trachylobane-360 treatment. The histopathological analysis of liver and kidney showed that both organs were mildly affected by trachylobane-360 treatment. Trachylobane-360 showed no immunosuppressive effect. In conclusion, these data reinforce the anticancer potential of this natural diterpene.

  16. Heterologous expression of the isopimaric acid pathway in Nicotiana benthamiana and the effect of N-terminal modifications of the involved cytochrome P450 enzyme

    DEFF Research Database (Denmark)

    Gnanasekaran, Thiyagarajan; Vavitsas, Konstantinos; Andersen-Ranberg, Johan;

    2015-01-01

    in the chloroplast and subsequently oxidized by a cytochrome P450, CYP720B4. RESULTS: We transiently expressed the isopimaric acid pathway in Nicotiana benthamiana leaves and enhanced its productivity by the expression of two rate-limiting steps in the pathway (providing the general precursor of diterpenes). This co...... enzymes. CONCLUSIONS: It is possible to localize a diterpenoid pathway from spruce fully within the chloroplast of N. benthamiana and a few modifications of the N-terminal sequences of the CYP720B4 can facilitate the expression of plant P450s in the plastids. The coupling of terpene biosynthesis closer......BACKGROUND: Plant terpenoids are known for their diversity, stereochemical complexity, and their commercial interest as pharmaceuticals, food additives, and cosmetics. Developing biotechnology approaches for the production of these compounds in heterologous hosts can increase their market...

  17. A new virus-induced gene silencing vector based on Euphorbia mosaic virus-Yucatan peninsula for NPR1 silencing in Nicotiana benthamiana and Capsicum annuum var. Anaheim.

    Science.gov (United States)

    Villanueva-Alonzo, Hernan J; Us-Camas, Rosa Y; López-Ochoa, Luisa A; Robertson, Dominique; Guerra-Peraza, Orlene; Minero-García, Yereni; Moreno-Valenzuela, Oscar A

    2013-05-01

    Virus-induced gene silencing is based on the sequence-specific degradation of RNA. Here, a gene silencing vector derived from EuMV-YP, named pEuMV-YP:ΔAV1, was used to silence ChlI and NPR1 genes in Nicotiana benthamiana. The silencing of the ChlI transcripts was efficient in the stems, petioles and leaves as reflected in tissue bleaching and reduced transcript levels. The silencing was stable, reaching the flowers and fruits, and was observed throughout the life cycle of the plants. Additionally, the silencing of the NPR1 gene was efficient in both N. benthamiana and Capsicum annuum. After silencing, the plants' viral symptoms increased to levels similar to those seen in wild-type plants. These results suggest that NPR1 plays a role in the compatible interactions of EuMV-YP N. benthamiana and EuMV-C. annum var. anaheim.

  18. Heterologous expression of the isopimaric acid pathway in Nicotiana benthamiana and the effect of N-terminal modifications of the involved cytochrome P450 enzyme

    DEFF Research Database (Denmark)

    Gnanasekaran, Thiyagarajan; Vavitsas, Konstantinos; Andersen-Ranberg, Johan

    2015-01-01

    in the chloroplast and subsequently oxidized by a cytochrome P450, CYP720B4. RESULTS: We transiently expressed the isopimaric acid pathway in Nicotiana benthamiana leaves and enhanced its productivity by the expression of two rate-limiting steps in the pathway (providing the general precursor of diterpenes). This co...... enzymes. CONCLUSIONS: It is possible to localize a diterpenoid pathway from spruce fully within the chloroplast of N. benthamiana and a few modifications of the N-terminal sequences of the CYP720B4 can facilitate the expression of plant P450s in the plastids. The coupling of terpene biosynthesis closer......BACKGROUND: Plant terpenoids are known for their diversity, stereochemical complexity, and their commercial interest as pharmaceuticals, food additives, and cosmetics. Developing biotechnology approaches for the production of these compounds in heterologous hosts can increase their market...

  19. Coat protein gene and 3′ non-coding region of tobacco mosaic virus and tomato mosaic virus are associated with viral pathogenesis in Nicotiana tabacum

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    The camellia isolate of tomato mosaic virus (ToMV-TL) can induce local necrotic lesions on the inoculated leaves in Nicotiana tabacum, whereas the broad bean isolate of tobacco mosaic virus (TMV-B) produces the mosaic symptom on systemic leaves. To examine viral determinant for differential infection phenotype in N. tabacum, the coat protein gene and the 3′ non-coding region of TMV was replaced with that of ToMV, the chimeric virus induced similar local necrotic lesions to that induced by ToMV. The results indicate that the coat protein gene and the 3′ non-coding region of TMV and ToMV influence the virus-induced pathogenesis in N. tabacum.

  20. Alamethicin permeabilizes the plasma membrane and mitochondria but not the tonoplast in tobacco (Nicotiana tabacum L. cv Bright Yellow) suspension cells

    DEFF Research Database (Denmark)

    Matic, S.; Geisler, D.A.; Møller, I.M.

    2005-01-01

    The ion channel-forming peptide AlaM (alamethicin) is known to permeabilize isolated mitochondria as well as animal cells. When intact tobacco (Nicotiana tabacum L.) Bright Yellow-2 cells were treated with AlaM, the cells became permeable for low-molecular-mass molecules as shown by induced leakage...... remained intact, as indicated by an unaffected tonoplast proton gradient. Low-flux permeabilization of plasma membranes and mitochondria at moderate AlaM concentrations was reversible and did not affect cell vigour. Higher AlaM concentrations induced cell death. After the addition of catalase that removes...... of NAD(P)(+). After the addition of cofactors and substrates, activities of cytosolic as well as mitochondrial respiratory enzymes could be directly determined inside the permeabilized cells. However, at an AlaM concentration at which the cytoplasmic enzymes were maximally accessible, the vacuole...

  1. The Agrobacterium tumefaciens Ti Plasmid Virulence Gene virE2 Reduces Sri Lankan Cassava Mosaic Virus Infection in Transgenic Nicotiana benthamiana Plants

    Directory of Open Access Journals (Sweden)

    Thulasi Raveendrannair Resmi

    2015-05-01

    Full Text Available Cassava mosaic disease is a major constraint to cassava cultivation worldwide. In India, the disease is caused by Indian cassava mosaic virus (ICMV and Sri Lankan cassava mosaic virus (SLCMV. The Agrobacterium Ti plasmid virulence gene virE2, encoding a nuclear-localized, single-stranded DNA binding protein, was introduced into Nicotiana benthamiana to develop tolerance against SLCMV. Leaf discs of transgenic N. benthamiana plants, harboring the virE2 gene, complemented a virE2 mutation in A. tumefaciens and produced tumours. Three tested virE2 transgenic plants displayed reduction in disease symptoms upon agroinoculation with SLCMV DNA A and DNA B partial dimers. A pronounced reduction in viral DNA accumulation was observed in all three virE2 transgenic plants. Thus, virE2 is an effective candidate gene to develop tolerance against the cassava mosaic disease and possibly other DNA virus diseases.

  2. Transfer of the cytochrome P450-dependent dhurrin pathway from Sorghum bicolor into Nicotiana tabacum chloroplasts for light-driven synthesis

    DEFF Research Database (Denmark)

    Gnanasekaran, Thiyagarajan; Karcher, Daniel; Nielsen, Agnieszka Janina Zygadlo;

    2016-01-01

    Plant chloroplasts are light-driven cell factories that have great potential to act as a chassis for metabolic engineering applications. Using plant chloroplasts, we demonstrate how photosynthetic reducing power can drive a metabolic pathway to synthesise a bio-active natural product....... For this purpose, we stably engineered the dhurrin pathway from Sorghum bicolor into the chloroplasts of Nicotiana tabacum (tobacco). Dhurrin is a cyanogenic glucoside and its synthesis from the amino acid tyrosine is catalysed by two membrane-bound cytochrome P450 enzymes (CYP79A1 and CYP71E1) and a soluble...... glucosyltransferase (UGT85B1), and is dependent on electron transfer from a P450 oxidoreductase. The entire pathway was introduced into the chloroplast by integrating CYP79A1, CYP71E1, and UGT85B1 into a neutral site of the N. tabacum chloroplast genome. The two P450s and the UGT85B1 were functional when expressed...

  3. Molecular and functional analysis of phosphomannomutase (PMM) from higher plants and genetic evidence for the involvement of PMM in ascorbic acid biosynthesis in Arabidopsis and Nicotiana benthamiana

    DEFF Research Database (Denmark)

    Qian, W; Yu, C; Qin, H

    2007-01-01

    Phosphomannomutase (PMM) catalyzes the interconversion of mannose-6-phosphate and mannose-1-phosphate. However, systematic molecular and functional investigations on PMM from higher plants have hitherto not been reported. In this work, PMM cDNAs were isolated from Arabidopsis, Nicotiana benthamiana...... was constitutively expressed in both vegetative and reproductive organs. Reducing the PMM expression level through virus-induced gene silencing caused a substantial decrease in ascorbic acid (AsA) content in N. benthamiana leaves. Conversely, raising the PMM expression level in N. benthamiana using viral-vector......-mediated ectopic expression led to a 20-50% increase in AsA content. Consistent with this finding, transgenic expression of an AtPMM-GFP fusion protein in Arabidopsis also increased AsA content by 25-33%. Collectively, this study improves our understanding on the molecular and functional properties of plant PMM...

  4. Effects of Initial Infestation Levels ofCallosobruchus maculatus(F.) (Coleoptera:Chrysomelidae) on Cowpea and Use ofNicotiana tabacum L. Aqueous Extract as Grain Protectant

    Institute of Scientific and Technical Information of China (English)

    Musa A K; Odunayo A; Adeyeye O E

    2015-01-01

    This study determined the effects of initial infestation of cowpea seeds (Ife brown variety) with different insect densities (0, 2, 4 and 6 pairs per 50 g seeds) ofCallosobruchus maculatus(F.) and evaluated the effects of aqueous leaf extract of Nicotiana tabacum L. onC. maculatus in the laboratory. It was observed that adult beetle population increased significantly (p<0.05) with increase in insect density. The increase in population of beetles and corresponding weight loss of the seeds in different levels of infestation showed that the cowpea variety was susceptible to beetle infestation, emergence and survival of progeny. Significantly more adults emerged on higher infestation compared to lower and no infestation. In Nigeria,Nicotiana tabacum L. is a locally available plant, with known insecticidal properties. The plant leaf extract was easily extracted with water and confirmed its effectiveness as a protective agent for stored cowpea seeds. Experiment was conducted to assess the effects of aqueous extracts of N. tabacumat 0, 0.1, 0.2 and 0.3 mL•50 g-1 cowpea seeds onC. maculatus. Data was recorded and showed varying levels of effectiveness againstC. maculatus. Result showed that seed appearance was dependent on levels of insect population, while N. tabacum aqueous extract exerted effects on survival ofC. maculatus. Aqueous leaf extract ofN. tabacum probably contained some insecticidal properties which might have significantly conferred beetle mortality and reduced beetle emergence leading to a decrease in seed weight loss.

  5. 烟草疫霉分离及生长培养基的选择%Selection of Isolating and Culturing Medium for Phytophthora nicotianae

    Institute of Scientific and Technical Information of China (English)

    苏凯; 桑维钧; 张新强; 王慧

    2013-01-01

    Different base medium adding different combination of antibiotics were applied for isolation and culture of Phytophthora nicotianae.The results showed that selective medium with Vs base medium (100mL/L V8juice+0.02 g/L CaCO3+2 g/Lagar) adding 10 mg/mL ampicillin +5 mg/mL nysfungin+5 mg/mL carbendazim had the best isolation effect as the successful isolation rate was 100%,and the hypha grew well on this medium.After separation,the hypha could be cultured in Vs base medium as the hypha was white,tight and strong with big bacteria clone and strong growth potential.%采用不同的基本培养基添加抗生素组合对烟草疫霉(Phytophthora nicotianae)进行分离培养.结果表明,以V8培养基(100 mL/L Vs汁+0.02 g/L CaCO3+2 g/L洋菜)为基本培养基附加10 mg/mL氨苄青霉素+5 mg/mL制霉素+5 mg/mL多菌灵的选择培养基分离烟草疫霉的效果较好,分离成功率达100%,菌丝的生长状态也较好.菌丝分离后在V8基本培养基中培养,菌丝紧密、浓白、粗壮,菌落大、长势旺.

  6. New microsome-associated HT-family proteins from Nicotiana respond to pollination and define an HT/NOD-24 protein family

    Institute of Scientific and Technical Information of China (English)

    Katsuhiko Kondo; Bruce McClure

    2008-01-01

    HT-family proteins have been identified in Nicotiana, Solanum,and Petunia.HT-B-type proteins are implicated in S-RNase-based self-incompatibility,but the functions of other family members are unknown.Screening for cDNA sequences with an expression pattern similar to HT-B in Nicotiana alata revealed a new group of small HT-family proteins.designated HT-M.HT-M proteins resemble HT-B in several respects:their pistil-specific expression pattern iS indistinguish-able from HT-B,they pellet with a microsome fraction,and their abundance decreases after pollination.Unlike HT-B,there iS no S-specificity to this response,and RNAi experiments show that HT-M proteins are not necessary for self-incompatibility.Identification of a third group of pistil-specific HT-family proteins helps better define the characteristics of the family and allowed identification of putative new family members.By searching the databases with only the most conserved HT-family sequence elements,the signal sequence and cysteine motifs,we identified nodulin-24-1ike proteins and several small glycine-rich proteins as putative HT-family members.Like HT-M and HT-B,nodulin-24 iS membrane associated.We propose that the conserved features in HT-family proteins are important for targeting or modification and refer to the broader family that includes both HT-and nodulin-24-like proteins as the HT/NOD-24-family.

  7. Purification, Characterization and Antioxidant Activities <em>in Vitroem>> em>and <em>in Vivoem> of the Polysaccharides from <em>Boletus edulisem> Bull

    Directory of Open Access Journals (Sweden)

    Yijun Fan

    2012-07-01

    Full Text Available A water-soluble polysaccharide (BEBP was extracted from <em>Boletus edulis em>Bull using hot water extraction followed by ethanol precipitation. The polysaccharide BEBP was further purified by chromatography on a DEAE-cellulose column, giving three major polysaccharide fractions termed BEBP-1, BEBP-2 and BEBP-3. In the next experiment, the average molecular weight (Mw, IR and monosaccharide compositional analysis of the three polysaccharide fractions were determined. The evaluation of antioxidant activities both <em>in vitroem> and <em>in vivo em>suggested that BEBP-3 had good potential antioxidant activity, and should be explored as a novel potential antioxidant.

  8. Sulla presenza di <em>Sorex antinoriiem>, <em>Neomys anomalusem> (Insectivora, Soricidae e <em>Talpa caecaem> (Insectivora, Talpidae in Umbria

    Directory of Open Access Journals (Sweden)

    A.M. Paci

    2003-10-01

    Full Text Available Lo scopo del contributo è di fornire un aggiornamento sulla presenza del Toporagno del Vallese <em>Sorex antinoriiem>, del Toporagno acquatico di Miller <em>Neomys anomalusem> e della Talpa cieca <em>Talpa caecaem> in Umbria, dove le specie risultano accertate ormai da qualche anno. A tal fine sono stati rivisitati i reperti collezionati e la bibliografia conosciuta. Toporagno del Vallese: elevato di recente a livello di specie da Brünner et al. (2002, altrimenti considerato sottospecie del Toporagno comune (<em>S. araneus antinoriiem>. È conservato uno di tre crani incompleti (mancano mandibole ed incisivi superiori al momento prudenzialmente riferiti a <em>Sorex> cfr. <em>antinorii>, provenienti dall?Appennino umbro-marchigiano settentrionale (dintorni di Scalocchio - PG, 590 m. s.l.m. e determinati sulla base della pigmentazione rossa degli ipoconi del M1 e M2; Toporagno acquatico di Miller: tre crani (Breda in Paci e Romano op. cit. e un esemplare intero (Paci, ined. sono stati trovati a pochi chilometri di distanza gli uni dall?altro tra i comuni di Assisi e Valfabbrica, in ambienti mediocollinari limitrofi al Parco Regionale del M.te Subasio (Perugia. In provincia di Terni la specie viene segnalata da Isotti (op. cit. per i dintorni di Orvieto. Talpa cieca: sono noti una femmina e un maschio raccolti nel comune di Pietralunga (PG, rispettivamente in una conifereta a <em>Pinus nigraem> (m. 630 s.l.m. e nelle vicinanze di un bosco misto collinare a prevalenza di <em>Quercus cerrisem> (m. 640 s.l.m.. Recentemente un terzo individuo è stato rinvenuto nel comune di Sigillo (PG, all?interno del Parco Regionale di M.te Cucco, sul margine di una faggeta a 1100 m s.l.m. In entrambi i casi l?areale della specie è risultato parapatrico con quello di <em>Talpa europaeaem>.

  9. Glycosylation of Vanillin and 8-Nordihydrocapsaicin by Cultured <em>Eucalyptus perrinianaem> Cells

    Directory of Open Access Journals (Sweden)

    Naoji Kubota

    2012-05-01

    Full Text Available Glycosylation of vanilloids such as vanillin and 8-nordihydrocapsaicin by cultured plant cells of <em>Eucalyptus perrinianaem> was studied. Vanillin was converted into vanillin 4-<em>O>-b-D-glucopyranoside, vanillyl alcohol, and 4-<em>O>-b-D-glucopyranosylvanillyl alcohol by <em>E. perriniana em>cells. Incubation of cultured <em>E. perrinianaem> cells with 8-nor- dihydrocapsaicin gave 8-nordihydrocapsaicin 4-<em>O>-b-D-glucopyranoside and 8-nordihydro- capsaicin 4-<em>O>-b-D-gentiobioside.

  10. A specimen of <em>Sorex> cfr. <em>samniticus> in Barn Owl's pellets from Murge plateau (Apulia, Italy / Su di un <em>Sorex> cfr. <em>samniticus> (Insectivora, Soricidae rinvenuto in borre di <em>Tyto albaem> delle Murge (Puglia, Italia

    Directory of Open Access Journals (Sweden)

    Giovanni Ferrara

    1992-07-01

    Full Text Available Abstract In a lot of Barn Owl's pellets from the Murge plateau a specimen of <em>Sorex> sp. was detected. Thank to some morphological and morphometrical features, the cranial bones can be tentatively attributed to <em>Sorex samniticusem> Altobello, 1926. The genus <em>Sorex> was not yet included in the Apulia's fauna southwards of the Gargano district; the origin and significance of the above record is briefly discussed, the actual presence of a natural population of <em>Sorex> in the Murge being not yet proved. Riassunto Viene segnalato il rinvenimento di un esemplare di <em>Sorex> cfr. <em>samniticus> da borre di <em>Tyto albaem> delle Murge. Poiché il genere non era stato ancora segnalato nella Puglia a sud del Gargano, viene discusso il significato faunistico del reperto.

  11. Study of the <em>in Vitroem> Antiplasmodial, Antileishmanial and Antitrypanosomal Activities of Medicinal Plants from Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Nawal M. Al-Musayeib

    2012-09-01

    Full Text Available The present study investigated the <em>in vitroem> antiprotozoal activity of sixteen selected medicinal plants. Plant materials were extracted with methanol and screened <em>in vitroem> against erythrocytic schizonts of <em>Plasmodium falciparumem>, intracellular amastigotes of <em>Leishmania infantum em>and <em>Trypanosoma cruzi em>and free trypomastigotes of<em> T. bruceiem>. Cytotoxic activity was determined against MRC-5 cells to assess selectivity<em>. em>The criterion for activity was an IC50 < 10 µg/mL (4. Antiplasmodial activity was found in the<em> em>extracts of<em> em>>Prosopis julifloraem> and <em>Punica granatumem>. Antileishmanial activity<em> em>against <em>L. infantumem> was demonstrated in <em>Caralluma sinaicaem> and <em>Periploca aphylla.em> Amastigotes of<em> T. cruzi em>were affected by the methanol extract of<em> em>>Albizia lebbeckem>> em>pericarp, <em>Caralluma sinaicaem>,> Periploca aphylla em>and <em>Prosopius julifloraem>. Activity against<em> T. brucei em>was obtained in<em> em>>Prosopis julifloraem>. Cytotoxicity (MRC-5 IC50 < 10 µg/mL and hence non-specific activities were observed for<em> em>>Conocarpus lancifoliusem>.>

  12. Rational management of tobacco (Nicotiana tabacum L. crop residues to obtain organic compostManejo racional de resíduos da cultura do fumo (Nicotiana tabacum L. para obtenção de composto orgânico

    Directory of Open Access Journals (Sweden)

    Antonio Carlos Souza Borges Filho

    2011-10-01

    Full Text Available Crop residues may be used to produce organic composts for agricultural use. The aim of this study was to evaluate the effect of two inoculums on the decomposition speed of tobacco residues (stems. The experiment was carried out in a completely randomized design, with three treatments and six replications. The residue used in the study was passed through a forage chopper and mixed to the inoculums in a concrete truck mixer. The treatments evaluated were: TF + EB + RB - tobacco residue, bovine manure and bovine rumen; TF + EB + MP - tobacco residue, bovine manure and Microsept-Dust and TF + EB - tobacco residue and bovine manure. The temperature and humidity of the piles were monitored weekly. The piles were initially turned up side down every seven days, and then every fourteen days till closing 60 days. Samples for chemical analysis of the composted material were collected at 30, 60 and 90 days after the beginning of the composting process. The temperature of the piles stayed around 57 to 60 ºC in the period between the 20 to 40 days after the beginning of the composting process. The mix containing cattle rumen (RB presented the lowest temperature after forty days. At the end of the composting the mix containing Microsept-Dust (MP presented the lowest humidity. The C/N relation of the composts obtained was around 10/1 and 11/1. The three treatments presented pH values close to the neutrality in the mature stage. A period of sixty days seems to be sufficient to obtain a mature and stable biocompost of tobacco waste using manure independent of the addition of inoculums.Os resíduos vegetais podem ser utilizados na produção de adubo orgânico por meio da compostagem. O objetivo deste estudo foi avaliar o efeito de dois inoculantes sobre a velocidade de decomposição do resíduo da cultura do fumo. O experimento foi conduzido em delineamento inteiramente casualizado, com três tratamentos e seis repetições, sendo o resíduo picado em picadeira

  13. Molecular heterogeneity of photosystem I. psaD, psaE, psaF, psaH, and psaL are all present in isoforms in Nicotiana spp.

    Science.gov (United States)

    Obokata, J; Mikami, K; Hayashida, N; Nakamura, M; Sugiura, M

    1993-08-01

    The protein composition of photosystem I (PSI) was examined in Nicotiana spp. by high-resolution polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and immunoblot analysis. Five PSI proteins show polymorphism in an amphidiploid species, Nicotiana tabacum, but not in its ancestral diploid species, Nicotiana sylvestris and Nicotiana tomentosiformis. These Nicotiana spp. appear to have at least 18 PSI proteins per genome that range in molecular mass from 3 to 20 kD. They include the products of nuclear genes psaD, psaE, psaF, psaG, psaH, psaK, and psaL, the product of chloroplast gene psaC, N-terminally blocked proteins of 4.5 and 3.0 kD, and an unidentified protein of 12.5 kD. The psaD, psaF, psaH, and psaL products have two isoforms each that are distinguished by different mobilities in polyacrylamide gel electrophoresis, and the psaE product has four isoforms. The two isoforms of the psaD product have distinct amino acid sequences, indicating that they are encoded by different genes within the genome. Four isoforms of the psaE products can be classified into two groups by N-terminal amino acid sequence, indicating that at least two psaE genes are present in the genome. To examine whether the polymorphic nature of PSI is peculiar to Nicotiana spp., we carried out immunoblot analysis of the psaD and psaE products in isogenic lines of tomato (Lycopersicon esculentum), Arabidopsis thaliana, red bean (Vigna angularis), and corn (Zea mays). Two electrophoretically distinct isoforms were found for the psaD products of tomato, A. thaliana, and corn, and two isoforms of psaE products were detected in tomato, A. thaliana, and red bean. These results suggest that the nuclear-encoded subunits of PSI, except for the psaG and psaK products, generally have two isoforms.

  14. EMS in Mauritius.

    Science.gov (United States)

    Ramalanjaona, Georges; Brogan, Gerald X

    2009-02-01

    Mauritius lies in the southwest Indian Ocean about 1250 miles from the African coast and 500 miles from Madagascar. Mauritius (estimated population 1,230,602) became independent from the United Kingdom in 1968 and has one of the highest GDP per capita in Africa. Within Mauritius there is a well established EMS system with a single 999 national dispatch system. Ambulances are either publicly or privately owned. Public ambulances are run by the Government (SAMU). Megacare is a private subscriber only ambulance service. The Government has recently invested in new technology such as telemedicine to further enhance the role of EMS on the island. This article describes the current state of EMS in Mauritius and depicts its development in the context of Government effort to decentralise and modernise the healthcare system.

  15. Acidose ruminal em caprinos

    OpenAIRE

    Cunha, José Diogo de Oliveira e silva Ribeiro da

    2012-01-01

    Dissertação de Mestrado Integrado em Medicina Veterinária Com este trabalho pretendeu-se fazer uma revisão acerca da acidose ruminal em caprinos, visto haver pouca bibliografia sobre o tema. Este trabalho foi baseado na revisão bibliográfica de artigos científicos e completado através da observação de casos clínicos ocorridos durante o estágio curricular. O objectivo deste estudo foi realizar uma breve revisão da anatomia e fisiologia do tracto gastrointestinal dos caprinos assim com...

  16. Ulisses em Kafka

    Directory of Open Access Journals (Sweden)

    Stéphane Mosès

    2015-02-01

    Full Text Available ResumoEm sua introdução para Exegese de uma lenda, Stéphane Mosès afirma que a obra de Kafka não necessita de novas interpretações, mas sim de uma “análise rigorosa de sua lógica subjacente”. Assim, em “Ulisses em Kafka”, investigandoa dialética formal de “O silêncio das sereias”, Mosès procura expor a estrutura narrativa que comandaria esse breve texto em que o escritor tcheco acrescenta à astucia de Ulisses a ingenuidade como elemento de salvação.Palavras-chave: Kafka; dialética; salvação.AbstractIn his introduction to Exegèse d’une légende, Stéphane Mosès affirms that Kafka’s works are not in need of new interpretations, but rather, of a “rigorous analysis of their underlying logic”. Thus, in “Ulisses chez Kafka”, by investigating the formal dialectic of “Das Schweigen der Sirenen”, Mosès seeks to exhibit the narrative structure that would supposedly guide this short text in which the Czech author adds naïveté to Ulisses’s cunning intelligence as an element of salvation.Keywords: Kafka; dialectic; salvation.Stéphane Mosès foi professor emérito da Universidade Hebraica de Jerusalém, onde ensinou Literatura Alemã e Comparada, entre os seus principais interesses estavam o pensamento de Franz Rosenzweig e de Walter Benjamin e as literaturas de Franz Kafka e de Paul Célan. Autor de diversos livros, entre eles: L'ange de l'histoire. Rosenzweig, Benjamin, Scholem, Exégèse d'une légende, lectures de Kafka, e Rêves de Freud. Six lectures.Rodrigo Ielpo é doutor em literatura francesa pela UFRJ e em História e Semiologia do texto e da imagem pela Université Paris VII, com pós-doutorado em Teoria Literária pela UNICAMP. Professor Adjunto do Departamento de Letras Neolatinas da Universidade Federal do Rio de Janeiro, possui publicações e traduções nos seguintes temas: teoria e literatura francesa contemporâneas, escrita e processos de subjetivação.rodrigoielpo@gmail.com 

  17. International EMS Systems

    DEFF Research Database (Denmark)

    Langhelle, Audun; Lossius, Hans Morten; Silfvast, Tom

    2004-01-01

    exist, however, especially within the ground and air ambulance service, and the EMS systems face several challenges. Main problems and challenges emphasized by the authors are: (1) Denmark: the dispatch centres are presently not under medical control and are without a national criteria based system......Emergency medicine service (EMS) systems in the five Nordic countries have more similarities than differences. One similarity is the involvement of anaesthesiologists as pre-hospital physicians and their strong participation for all critically ill and injured patients in-hospital. Discrepancies do...

  18. Frenectomia em foco

    OpenAIRE

    Vieira, Joana Isabel Monteiro de Paiva

    2012-01-01

    Projeto de Pós Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Medicina Dentária O freio é uma estrutura anatómica oral classificada em freio labial (médio maxilar ou mandibular, lateral maxilar ou mandibular) ou lingual que se pode tornar num problema quando a sua inserção cria impedimentos estruturais e dificuldades a vários níveis. Um freio labial anormal pode causar algumas anomalias ou problemas, tais como u...

  19. Cyberbullying em adolescentes brasileiros

    OpenAIRE

    Wendt, Guilherme Welter

    2012-01-01

    O cyberbullying é entendido como uma forma de comportamento agressivo que ocorre através dos meios eletrônicos de interação (computadores, celulares, sites de relacionamento virtual), sendo realizado de maneira intencional por uma pessoa ou grupo contra alguém em situação desigual de poder e, ainda, com dificuldade em se defender. Os estudos disponíveis até o presente momento destacam que o cyberbullying é um fator de risco para o desenvolvimento de sintomas de ansiedade, depressão, ideação s...

  20. Corantes vitais em cromovitrectomia

    OpenAIRE

    Dib, Eduardo; Rodrigues,Eduardo Büchelle; Maia, Maurício [UNIFESP; Meyer, Carsten H; Penha, Fernando Marcondes [UNIFESP; Furlani, Bruno de Albuquerque [UNIFESP; Costa, Elaine de Paula Fiod [UNIFESP; Farah, Michel Eid

    2009-01-01

    O objetivo do artigo é apresentar os dados atuais da aplicação de corantes vitais durante cirurgia vitreorretiniana, "cromovitrectomia", bem como uma revisão da literatura atual sobre o assunto no tocante às técnicas de aplicação, indicações e complicações em cromovitrectomia. Um grande número de publicações tem abordado o perfil tóxico da indocianina verde na cromovitrectomia. Dados experimentais mostram uma toxicidade dose-dependente da mesma em várias populações de células retinianas. Nova...

  1. O recasamento em Portugal

    OpenAIRE

    Lobo, Cristina; Conceição, Cristina Palma

    2003-01-01

    Identificar tendências do fenómeno do recasamento em Portugal, nas duas últimas décadas, constitui o objectivo principal deste artigo. Para essa identificação contribuiu o conhecimento da amplitude do próprio fenómeno, isto é, a percentagem de recasamentos no total dos casamentos em Portugal, e ainda um conjunto de variáveis que permitiu conhecer a caracterização social dos protagonistas do recasamento, por um lado, e delinear a diversidade dos seus trajectos conjugais após um divórcio, por o...

  2. Frenectomia em foco

    OpenAIRE

    Vieira, Joana Isabel Monteiro de Paiva

    2012-01-01

    Projeto de Pós Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Medicina Dentária O freio é uma estrutura anatómica oral classificada em freio labial (médio maxilar ou mandibular, lateral maxilar ou mandibular) ou lingual que se pode tornar num problema quando a sua inserção cria impedimentos estruturais e dificuldades a vários níveis. Um freio labial anormal pode causar algumas anomalias ou problemas, tais como u...

  3. Acidose ruminal em caprinos

    OpenAIRE

    Cunha, José Diogo de Oliveira e silva Ribeiro da

    2012-01-01

    Dissertação de Mestrado Integrado em Medicina Veterinária Com este trabalho pretendeu-se fazer uma revisão acerca da acidose ruminal em caprinos, visto haver pouca bibliografia sobre o tema. Este trabalho foi baseado na revisão bibliográfica de artigos científicos e completado através da observação de casos clínicos ocorridos durante o estágio curricular. O objectivo deste estudo foi realizar uma breve revisão da anatomia e fisiologia do tracto gastrointestinal dos caprinos assim com...

  4. Arreglo de los polígonos del exocorion de huevos eclosionados de algunas especies de los géneros Triatoma Laporte, Meccus Stal y Nesotriatoma Usinger (Heteroptera: Reduviidae.

    Directory of Open Access Journals (Sweden)

    Glauco López

    2009-12-01

    Full Text Available Resumen. En este trabajo se estudiaron estructural y cuantitativamente las celdas exocoriales de huevos eclosionados de Triatoma infestans (Klug, Triatoma lenti Sherlock & Serafim, Triatoma maculata (Erichson, Triatoma sordida (Stal, Meccus picturatus (Usinger, Meccus longipennis (Usinger, Meccus pallidipennis (Stal and Nesotriatoma flavida (Neiva. Los huevos se analizaron por regiones: caudal, media y cefálica. Se encontró que el hexágono fue el polígono más abundante en todas las regiones del huevo y en todas las especies, seguido del pentágono y del heptágono; se encontraron diferencias estadísticamente significativas en el número tanto de hexágonos y pentágonos entre regiones de una misma especie y entre la misma región de las diferentes especies, sin embargo, el número promedio de heptágonos no varió al comparar la región media entre las diferentes especies y al comparar las diferentes regiones en una misma especie. No se encontraron diferencias estadísticamente significativas en el promedio de polígonos, segmentos totales, heptágonos, hexágonos y pentágonos, al comparar las especies examinadas del género Meccus. Se propone un protocolo de identificación y análisis de teselaciones, con el cual se encontró que en 108 campos examinados de 385.000 µm2 cada uno, se identificaron teselaciones compartidas en los 108 campos observados con determinadas combinaciones de heptágonos, hexágonos y pentágonos, y se observaron teselaciones no compartidas en la totalidad de los campos, con combinaciones de polígonos de cuatro a nueve lados, y cada teselación se representó por un grafo. Se discuten las implicaciones taxonómicas, funcionales, filogenéticas y construccionales de los arreglos poligonales descritos.Arragement of exochorial polygons of hatched eggs in several species of the genera Triatoma Laporte, Meccus Stal y Nesotriatoma Usinger (Heteroptera: Reduviidae. Abstract. In this study a structural and quantitative

  5. Horizontal Transmission of the Entomopathogen Fungus <em>Metarhizium anisopliae em>in <em>Microcerotermes diversusem> Groups

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    Mohammad Saied Mossadegh

    2012-08-01

    Full Text Available An experiment was carried out in order to investigate fungal conidia transmission of <em>Metarhizium anisopliae em>(Metschnikoff> em>Sorokin from vector (donor to healthy <em>Microcerotermes diversusem> Silvestri (Iso.: Termitidae and determine the best donor/concentration ratio for transmission. After preliminary trials, concentrations of 3.1 × 104, 3.9 × 105, 3.2 × 106 and 3.5 × 108 conidia mL−1 were selected for testing. The experiment was performed at three donor : Recipient ratios of 10, 30 and 50%. The highest mortality of recipient workers was observed after 14 days at the concentration of 3.5 × 108 conidia mL−1 and donor ratio of 50%. The mortality of recipient workers was less than 20% at all concentrations at a donor ratio of 10%. Our observations indicate social behavior of <em>M. diversusem>, such as grooming, can be effective in promoting epizootic outbreaks in a colony. While the current results suggest good potential for efficacy, the use of <em>M. anisopliaeem> as a component of integrated pest management of <em>M. diversusem> still needs to be proven under field conditions.

  6. Effet comparé des poudres de Nicotiana tabacum L, Cymbopogon citratus (D.C. Stapf et de l'huile de Ricinus communis L sur la conservation des graines de Vigna unguiculata (L Walp

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    Gakuru, S.

    1995-01-01

    Full Text Available Compared Effect of Nicotiana tabacum L, Cymbopogon citratus (D.C. Stapf Powders and Castor Oil Ricinus communis L. on Conservation of Cowpea Vigna Unguiculata (L. Walp Grains. The effect of powder of tobacco Nicotiana tabacum L. and citronella grass Cymbopogon citratus (D.C. Stapf and castor oil Ricinus communis L. on conservation of cowpea Vigna unguiculata (L. Walp. grains was investigated in Kisangani, Zaire. After 5 months of conservation, infestation rates by bean weevil Acanthoscelides obtectus Say were 72.5 %, 74.5 %, 49.5 % and 5 % respectively for the check, the samples treated by 1 % of citronella grass and tobacco powder and 1 % of castor oil. The powder dose of 7.5 % did not give more interesting results.

  7. Clinical Relevance of <em>CDH1em> and <em>CDH13em> DNA-Methylation in Serum of Cervical Cancer Patients

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    Günther K. Bonn

    2012-07-01

    Full Text Available This study was designed to investigate the DNA-methylation status of <em>E>-cadherin (<em>CDH1em> and <em>H>-cadherin (<em>CDH13em> in serum samples of cervical cancer patients and control patients with no malignant diseases and to evaluate the clinical utility of these markers. DNA-methylation status of <em>CDH1em> and <em>CDH13em> was analyzed by means of MethyLight-technology in serum samples from 49 cervical cancer patients and 40 patients with diseases other than cancer. To compare this methylation analysis with another technique, we analyzed the samples with a denaturing high performance liquid chromatography (DHPLC PCR-method. The specificity and sensitivity of <em>CDH1em> DNA-methylation measured by MethyLight was 75% and 55%, and for <em>CDH13em> DNA-methylation 95% and 10%. We identified a specificity of 92.5% and a sensitivity of only 27% for the <em>CDH1em> DHPLC-PCR analysis. Multivariate analysis showed that serum <em>CDH1em> methylation-positive patients had a 7.8-fold risk for death (95% CI: 2.2–27.7; <em>p> = 0.001 and a 92.8-fold risk for relapse (95% CI: 3.9–2207.1; <em>p> = 0.005. We concluded that the serological detection of <em>CDH1em> and <em>CDH13em> DNA-hypermethylation is not an ideal diagnostic tool due to low diagnostic specificity and sensitivity. However, it was validated that <em>CDH1em> methylation analysis in serum samples may be of potential use as a prognostic marker for cervical cancer patients.

  8. A polygalacturonase-inhibiting protein from grapevine reduces the symptoms of the endopolygalacturonase BcPG2 from Botrytis cinerea in Nicotiana benthamiana leaves without any evidence for in vitro interaction

    NARCIS (Netherlands)

    Joubert, D.A.; Kars, I.; Wagemakers, L.; Bergmann, C.; Kemp, G.; Vivier, M.A.; Kan, van J.A.L.

    2007-01-01

    Six endopolygalacturonases from Botrytis cinerea (BcPG1 to BcPG6) as well as mutated forms of BcPG1 and BcPG2 were expressed transiently in leaves of Nicotiana benthamiana using agroinfiltration. Expression of BcPG1, BcPG2, BcPG4, BcPG5, and mutant BcPG1-D203A caused symptoms, whereas BcPG3, BcPG6,

  9. Host range of symptomatology of <em>Pepino mosaicem> virus strains occurring in Europe

    DEFF Research Database (Denmark)

    Blystad, Dag-Ragnar; van der Vlugt, René; Alfaro-Fernández, Ana;

    2015-01-01

    for the three strains tested at 10 different European locations with both international and local cultivars showed that eggplant is an alternative host of PepMV. Sweet pepper is not an important host of PepMV, but potato can be infected when the right isolate is matched with a specific cultivar. Nicotiana...

  10. Virus-Induced Gene Silencing Using Tobacco Rattle Virus as a Tool to Study the Interaction between Nicotiana attenuata and Rhizophagus irregularis.

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    Karin Groten

    Full Text Available Most land plants live in a symbiotic association with arbuscular mycorrhizal fungi (AMF that belong to the phylum Glomeromycota. Although a number of plant genes involved in the plant-AMF interactions have been identified by analyzing mutants, the ability to rapidly manipulate gene expression to study the potential functions of new candidate genes remains unrealized. We analyzed changes in gene expression of wild tobacco roots (Nicotiana attenuata after infection with mycorrhizal fungi (Rhizophagus irregularis by serial analysis of gene expression (SuperSAGE combined with next generation sequencing, and established a virus-induced gene-silencing protocol to study the function of candidate genes in the interaction. From 92,434 SuperSAGE Tag sequences, 32,808 (35% matched with our in-house Nicotiana attenuata transcriptome database and 3,698 (4% matched to Rhizophagus genes. In total, 11,194 Tags showed a significant change in expression (p2-fold change after infection. When comparing the functions of highly up-regulated annotated Tags in this study with those of two previous large-scale gene expression studies, 18 gene functions were found to be up-regulated in all three studies mainly playing roles related to phytohormone metabolism, catabolism and defense. To validate the function of identified candidate genes, we used the technique of virus-induced gene silencing (VIGS to silence the expression of three putative N. attenuata genes: germin-like protein, indole-3-acetic acid-amido synthetase GH3.9 and, as a proof-of-principle, calcium and calmodulin-dependent protein kinase (CCaMK. The silencing of the three plant genes in roots was successful, but only CCaMK silencing had a significant effect on the interaction with R. irregularis. Interestingly, when a highly activated inoculum was used for plant inoculation, the effect of CCaMK silencing on fungal colonization was masked, probably due to trans-complementation. This study demonstrates that

  11. Virus-Induced Gene Silencing Using Tobacco Rattle Virus as a Tool to Study the Interaction between Nicotiana attenuata and Rhizophagus irregularis.

    Science.gov (United States)

    Groten, Karin; Pahari, Nabin T; Xu, Shuqing; Miloradovic van Doorn, Maja; Baldwin, Ian T

    2015-01-01

    Most land plants live in a symbiotic association with arbuscular mycorrhizal fungi (AMF) that belong to the phylum Glomeromycota. Although a number of plant genes involved in the plant-AMF interactions have been identified by analyzing mutants, the ability to rapidly manipulate gene expression to study the potential functions of new candidate genes remains unrealized. We analyzed changes in gene expression of wild tobacco roots (Nicotiana attenuata) after infection with mycorrhizal fungi (Rhizophagus irregularis) by serial analysis of gene expression (SuperSAGE) combined with next generation sequencing, and established a virus-induced gene-silencing protocol to study the function of candidate genes in the interaction. From 92,434 SuperSAGE Tag sequences, 32,808 (35%) matched with our in-house Nicotiana attenuata transcriptome database and 3,698 (4%) matched to Rhizophagus genes. In total, 11,194 Tags showed a significant change in expression (p2-fold change) after infection. When comparing the functions of highly up-regulated annotated Tags in this study with those of two previous large-scale gene expression studies, 18 gene functions were found to be up-regulated in all three studies mainly playing roles related to phytohormone metabolism, catabolism and defense. To validate the function of identified candidate genes, we used the technique of virus-induced gene silencing (VIGS) to silence the expression of three putative N. attenuata genes: germin-like protein, indole-3-acetic acid-amido synthetase GH3.9 and, as a proof-of-principle, calcium and calmodulin-dependent protein kinase (CCaMK). The silencing of the three plant genes in roots was successful, but only CCaMK silencing had a significant effect on the interaction with R. irregularis. Interestingly, when a highly activated inoculum was used for plant inoculation, the effect of CCaMK silencing on fungal colonization was masked, probably due to trans-complementation. This study demonstrates that large

  12. Time-Dependent Damage Investigation of Rock Mass in an <em>In> em>Situ> Experimental Tunnel

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    Jing Chen

    2012-08-01

    Full Text Available In underground tunnels or caverns, time-dependent deformation or failure of rock mass, such as extending cracks, gradual rock falls, <em>etc>., are a costly irritant and a major safety concern if the time-dependent damage of surrounding rock is serious. To understand the damage evolution of rock mass in underground engineering, an <em>in situem> experimental testing was carried out in a large belowground tunnel with a scale of 28.5 m in width, 21 m in height and 352 m in length. The time-dependent damage of rock mass was detected in succession by an ultrasonic wave test after excavation. The testing results showed that the time-dependent damage of rock mass could last a long time, <em>i.e.>, nearly 30 days. Regression analysis of damage factors defined by wave velocity, resulted in the time-dependent evolutional damage equation of rock mass, which corresponded with logarithmic format. A damage viscoelastic-plastic model was developed to describe the exposed time-dependent deterioration of rock mass by field test, such as convergence of time-dependent damage, deterioration of elastic modules and logarithmic format of damage factor. Furthermore, the remedial measures for damaged surrounding rock were discussed based on the measured results and the conception of damage compensation, which provides new clues for underground engineering design.

  13. Ultrasound-Assisted Extraction of Syringin from the Bark of <em>Ilex em>rotunda> Thumb Using Response Surface Methodology

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    Hui Wang

    2012-06-01

    Full Text Available In this work, a rapid extraction method based on ultrasound-assisted extraction (UAE of syringin from the bark of <em>Ilex rotunda em>Thumb using response surface methodology (RSM is described. The syringin was analyzed and quantified by high performance liquid chromatography coupled with UV detection (HPLC-UV. The extraction solvent, extraction temperature and extraction time, the three main factors for UAE, were optimized with Box-Behnken design (BBD to obtain the highest extraction efficiency. The optimal conditions were the use of a sonication frequency of 40 kHz, 65% methanol as the solvent, an extraction time of 30 min and an extraction temperature of 40 °C. Using these optimal conditions, the experimental values agreed closely with the predicted values. The analysis of variance (ANOVA indicated a high goodness of model fit and the success of the RSM method for optimizing syringin extraction from the bark of <em>I>. em>rotunda>.

  14. Classe social: conceitos e esquemas operacionais em pesquisa em saude

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    Rita Barradas Barata

    2013-08-01

    Full Text Available Discute-se a utilização do conceito de classe em pesquisas em saúde, as diferentes abordagens sociológicas de estratificação social e de estrutura de classes, o potencial explicativo do conceito em estudos de determinação social e desigualdades em saúde, os modelos de operacionalização elaborados para uso em pesquisas sociológicas, demográficas ou de saúde e os limites e possibilidades desses modelos. Foram destacados quatro modelos de operacionalização: de Singer para estudo da distribuição de renda no Brasil, adaptado por Barros para uso em pesquisas epidemiológicas; de Bronfman & Tuirán para o censo demográfico mexicano, adaptado por Lombardi et al para pesquisas epidemiológicas; de Goldthorpe para estudos socioeconômicos ingleses, adaptado pela Sociedade Espanhola de Epidemiologia; e o modelo de Wright para pesquisa em sociologia e ciência política, também usado em inquéritos populacionais em saúde. Em conclusão, conceitualmente cada um dos modelos apresentados é coerente com a concepção teórica que os embasam, mas não há como optar por qualquer deles, descartando os demais.

  15. Efecto de extractos vegetales de <em>Polygonum hydropiperoidesem>, <em>Solanum nigrumem> y <em>Calliandra pittieriem> sobre el gusano cogollero (<em>Spodoptera frugiperdaem>

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    Lizarazo H. Karol

    2008-12-01

    Full Text Available

    El gusano cogollero <em>Spodoptera frugiperdaem> es una de las plagas que más afectan los cultivos en la región de Sumapaz (Cundinamarca, Colombia. En la actualidad se controla principalmente aplicando productos de síntesis química, sin embargo la aplicación de extractos vegetales surge como una alternativa de menor impacto sobre el ambiente. Este control se emplea debido a que las plantas contienen metabolitos secundarios que pueden inhibir el desarrollo de los insectos. Por tal motivo, la presente investigación evaluó el efecto insecticida y antialimentario de extractos vegetales de barbasco <em>Polygonum hydropiperoidesem> (Polygonaceae, carbonero <em>Calliandra pittieriem> (Mimosaceae y hierba mora <em>Solanum nigrumem> (Solanaceae sobre larvas de <em>S. frugiperdaem> biotipo maíz. Se estableció una cría masiva del insecto en el laboratorio utilizando una dieta natural con hojas de maíz. Posteriormente se obtuvieron extractos vegetales utilizando solventes de alta polaridad (agua y etanol y media polaridad (diclorometano los cuales se aplicaron sobre las larvas de segundo instar. Los resultados más destacados se presentaron con extractos de <em>P. hydropiperoidesem>, obtenidos con diclorometano en sus diferentes dosis, con los cuales se alcanzó una mortalidad de 100% 12 días después de la aplicación y un efecto antialimentario representado por un consumo de follaje de maíz inferior al 4%, efectos similares a los del testigo comercial (Clorpiriphos.

  16. Prima segnalazione di <em>Brachylaima fulvusem> Dujardin, 1843 (Digenea, Brachylaimidae in <em>Sorex> cfr. <em>antinorii> Bonaparte, 1848 (Insectivora, Soricidae in Italia

    Directory of Open Access Journals (Sweden)

    Juan Carlos Casanova

    2003-10-01

    Full Text Available Un?indagine parassitologica è stata condotta su esemplari di <em>Sorex> cfr. <em>antinorii> Bonaparte, 1840 (Insectivora, Soricidae catturati in Sila Grande (Valle Capra; 16.29.30; 39.21.40. Uno degli esemplari è stato trovato parassitato da Trematodi digenei rinvenuti nell?esofago e nello stomaco. I parassiti isolati sono stati fissati con liquido di Bouin, colorati con carminio acetico di Semichon, fissati in una serie di alcooli, chiarificati in xilolo e montati con Balsamo del Canada. I parassiti sono stati identificati come membri della Famiglia Brachylaimidae Joyeux & Foley (1930 appartenenti al genere <em>Brachylaima> Dujardin (1843. La morfologia generale ed i caratteri metrici hanno permesso di identificarli come <em>B. fulvusem> Dujardin, 1843 (lunghezza corpo: 2.39-2.46; larghezza corpo: 0.72-0.78; diametro trasversale ventosa orale: 212.48- 238.08x240.64-266.24; ventosa ventrale: 250.88-258.56x250.88-266.24; faringe 120.32-168.96; testicoli: 151.04-189.44x168.96-189.44; ovario: 122.88x171.52; uova: 26-30x2-17. Il range dei dati morfometrici coincide con quelli riscontrati da diversi autori negli esemplari di <em>B. fulvusem> in Europa centrale e occidentale (Jourdane, 1971; Lewis, 1969; Zarnowski,1960; Mas-Coma & Gallego, 1975. Dopo la descrizione originale, la sistematica di <em>B. fulvusem> Blanchard, 1847 non è stata mai confutata, sebbene varie specie dei generi <em>Harmostomum> Braun, 1899 e <em>Panopistus Sinitzinem>, 1931 siano state proposte come sinonimi di <em>B. fulvusem> da diversi autori. Zarnowski (1960 considera aperta la questione di identità di <em>H. (H. dujardiniem> Baer, 1928 e propone la sinonimia di <em>B. oesophageiem> Shaldibin, 1953 e <em>B. fulvusem>. Lewis (1969, al contrario, ha riconvalidato <em>B. oesophageiem> come specie, utilizzando criteri che, però, Jourdane (1971 ritiene, non idonei a differenziare le due specie. Mas-Coma & Gallego (1975, considerano <em

  17. PERDA DE SOLO EM ESTRADAS RURAIS EM ÁREA DE CULTIVO DE TABACO NA BACIA DO ARROIO BOA VISTA- PARANÁ

    Directory of Open Access Journals (Sweden)

    Valdemir Antoneli

    2016-06-01

    Full Text Available O objetivo desta pesquisa foi mensurar a perda de solos em estradas rurais na Bacia do Arroio Boa Vista no município de Guamiranga Paraná. A área de estudos é ocupada por pequenas propriedades agrícolas, com 85% da área utilizada para o cultivo do tabaco (Nicotiana tabacun. O cultivo desta Salanaceae é realizado de setembro a março indicando variação nas atividades agrícolas na região, com períodos de intensificação de atividade (safra e redução das atividades (entressafra. Esta variação nas atividades pode interferir nas perdas de solo nas estradas. Para avaliar as perdas de solo das estradas foram instaladas 6 parcelas de 600 m2 e, em cada uma foi construída uma caixa de contenção nas margens das estradas para deposição de sedimentos. Foi mensurada também a perda de solos dos barrancos por meio da instalação 14 parcelas de pinos de erosão com um total de 140 pinos. O monitoramento foi realizado durante um ano, sendo avaliado período de safra e entresafra. A perda média de solo foi de 27,6 kg/m2/ano. Deste total 4,9 kg/m2/ano (17,8% foi produzido pelos barrancos e 22,7 kg/m2/ano (82,2% oriundo do leito das estradas. Cerca de 68,2% da perda de solo foram registradas nos períodos de intensificação das atividades agrícolas (setembro a março, enquanto que 32,8% das perdas foram registradas nos períodos de entre safra.

  18. <em>Bacillus anthracisem> Factors for Phagosomal Escape

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    Irene Zornetta

    2012-07-01

    Full Text Available The mechanism of phagosome escape by intracellular pathogens is an important step in the infectious cycle. During the establishment of anthrax, <em>Bacillus anthracisem> undergoes a transient intracellular phase in which spores are engulfed by local phagocytes. Spores germinate inside phagosomes and grow to vegetative bacilli, which emerge from their resident intracellular compartments, replicate and eventually exit from the plasma membrane. During germination, <em>B. anthracisem> secretes multiple factors that can help its resistance to the phagocytes. Here the possible role of <em>B. anthracisem> toxins, phospholipases, antioxidant enzymes and capsules in the phagosomal escape and survival, is analyzed and compared with that of factors of other microbial pathogens involved in the same type of process.

  19. Distribution and change patterns of free IAA, ABP 1 and PM H⁺-ATPase during ovary and ovule development of Nicotiana tabacum L.

    Science.gov (United States)

    Chen, Dan; Deng, Yingtian; Zhao, Jie

    2012-01-15

    Auxin plays key roles in flower induction, embryogenesis, seed formation and seedling development, but little is known about whether auxin regulates the development of ovaries and ovules before pollination. In the present report, we measured the content of free indole-3-acetic (IAA) in ovaries of Nicotiana tabacum L., and localized free IAA, auxin binding protein 1 (ABP1) and plasma membrane (PM) H⁺-ATPase in the ovaries and ovules. The level of free IAA in the developmental ovaries increased gradually from the stages of ovular primordium to the functional megaspore, but slightly decreased when the embryo sacs formed. Immunoenzyme labeling clearly showed that both IAA and ABP1 were distributed in the ovules, the edge of the placenta, vascular tissues and the ovary wall, while PM H⁺-ATPase was mainly localized in the ovules. By using immunogold labeling, the subcellular distributions of IAA, ABP1 and PM H⁺-ATPase in the ovules were also shown. The results suggest that IAA, ABP1 and PM H⁺-ATPase may play roles in the ovary and ovule initiation, formation and differentiation.

  20. Silencing ribulose-1,5-bisphosphate carboxylase/oxygenase expression does not disrupt nitrogen allocation to defense after simulated herbivory in Nicotiana attenuata.

    Science.gov (United States)

    Stanton, Mariana A; Ullmann-Zeunert, Lynn; Wielsch, Natalie; Bartram, Stefan; Svatoš, Aleš; Baldwin, Ian T; Groten, Karin

    2013-01-01

    Ribulose-1,5-bisphosphate carboxylase/ oxygenase (RuBisCO) is the most abundant protein on the planet and in addition to its central role in photosynthesis it is thought to function as a nitrogen (N)-storage protein and a potential source of N for defense biosynthesis in plants. In a recent study in the wild tobacco Nicotiana attenuata, we showed that the decrease in absolute N invested in soluble proteins and RuBisCO elicited by simulated herbivory was much larger than the N-requirements of nicotine and phenolamide biosynthesis; (15)N flux studies revealed that N for defensive phenolamide synthesis originates from recently assimilated N rather than from RuBisCO turnover. Here we show that a transgenic line of N. attenuata silenced in the expression of RuBisCO (asRUB) invests similar or even larger amounts of N into phenolamide biosynthesis compared with wild type plants, consistent with our previous conclusion that recently assimilated N is channeled into phenolamide synthesis after elicitation. We suggest that the decrease in leaf proteins after simulated herbivory is a tolerance mechanism, rather than a consequence of N-demand for defense biosynthesis.

  1. Recombinant jacalin-like plant lectins are produced at high levels in Nicotiana benthamiana and retain agglutination activity and sugar specificity.

    Science.gov (United States)

    Fernandez-del-Carmen, Asun; Juárez, Paloma; Presa, Silvia; Granell, Antonio; Orzáez, Diego

    2013-02-20

    The plant kingdom is an underexplored source of valuable proteins which, like plant lectins, display unique interacting specificities. Furthermore, plant protein diversity remains under-exploited due to the low availability and heterogeneity of native sources. All these hurdles could be overcome with recombinant production. A narrow phylogenetic gap between the native source and the recombinant platform is likely to facilitate proper protein processing and stability; therefore, the plant cell chassis should be specially suited for the recombinant production of many plant native proteins. This is illustrated herein with the recombinant production of two representatives of the plant jacalin-related lectin (JRLs) protein family in Nicotiana benthamiana using state-of-the-art magnICON technology. Mannose-specific Banlec JRL was produced at very high levels in leaves, reaching 1.0mg of purified protein per gram of fresh weight and showing strong agglutination activity. Galactose-specific jacalin JRL, with its complicated processing requirements, was also successfully produced in N. benthamiana at levels of 0.25 mg of purified protein per gram of fresh weight. Recombinant Jacalin (rJacalin) proved efficient in the purification of human IgA1, and was able to discriminate between plant-made and native IgA1 due to their differential glycosylation status. Together, these results show that the plant cell factory should be considered a primary option in the recombinant production of valuable plant proteins.

  2. Influence of retinoblastoma-related gene silencing on the initiation of DNA replication by African cassava mosaic virus Rep in cells of mature leaves in Nicotiana benthamiana plants

    Directory of Open Access Journals (Sweden)

    Bruce Gareth

    2011-12-01

    Full Text Available Abstract Background Geminiviruses mainly infect terminally differentiated tissues and cells in plants. They need to reprogramme host cellular machinery for DNA replication. This process is thought to be mediated by inactivation of cell-cycle repressor proteins and by induction of host DNA synthesis protein expression through actions of the geminviral replication initiator protein (Rep. Findings Exploiting a Nicotiana benthamiana pOri2 line, which is transformed with a transgene consisting of a direct repeat of the African cassava mosaic virus (ACMV-replication origin (Ori flanking a non-viral DNA region, and virus-induced RNA silencing (VIGS, the impact of host gene expression on replication of the ACMV-derived replicon was investigated. The ACMV Rep trans-replicated the viral episomal replicon in leaves of young but not older pOri2 plants. Upon VIGS-mediated down-regulation of N. benthamiana NbRBR1, the retinoblastoma-related protein gene coding for a negative cell-cycle suppressor, recovered the ability of ACMV Rep for trans DNA replication, whereas the silencing of NbPCNA coding for the sliding clamp of DNA polymerase had no effect. Conclusions These results suggest that the cellular machinery for DNA replication in differentiated tissues of older leaves cannot be reprogrammed by Rep alone but may need other uncharacterised viral and plant factors.

  3. The transmitting tissue of Nicotiana tabacum is not essential to pollen tube growth, and its ablation can reverse prezygotic interspecific barriers.

    Science.gov (United States)

    Smith, Alan G; Eberle, Carrie A; Moss, Nicole G; Anderson, Neil O; Clasen, Benjamin M; Hegeman, Adrian D

    2013-12-01

    The Nicotiana tabacum transmitting tissue is a highly specialized file of metabolically active cells that is the pathway for pollen tubes from the stigma to the ovules where fertilization occurs. It is thought to be essential to pollen tube growth because of the nutrients and guidance it provides to the pollen tubes. It also regulates gametophytic self-incompatibility in the style. To test the function of the transmitting tissue in pollen tube growth and to determine its role in regulating prezygotic interspecific incompatibility, genetic ablation was used to eliminate the mature transmitting tissue, producing a hollow style. Despite the absence of the mature transmitting tissue and greatly reduced transmitting-tissue-specific gene expression, self-pollen tubes had growth to the end of the style. Pollen tubes grew at a slower rate in the transmitting-tissue-ablated line during the first 24 h post-pollination. However, pollen tubes grew to a similar length 40 h post-pollination with and without a transmitting tissue. Ablation of the N. tabacum transmitting tissue significantly altered interspecific pollen tube growth. These results implicate the N. tabacum transmitting tissue in facilitating or inhibiting interspecific pollen tube growth in a species-dependent manner and in controlling prezygotic reproductive barriers.

  4. Evaluation of tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots for the production of geraniol, the first committed step in terpenoid indole alkaloid pathway.

    Science.gov (United States)

    Ritala, Anneli; Dong, Lemeng; Imseng, Nicole; Seppänen-Laakso, Tuulikki; Vasilev, Nikolay; van der Krol, Sander; Rischer, Heiko; Maaheimo, Hannu; Virkki, Arho; Brändli, Johanna; Schillberg, Stefan; Eibl, Regine; Bouwmeester, Harro; Oksman-Caldentey, Kirsi-Marja

    2014-04-20

    The terpenoid indole alkaloids are one of the major classes of plant-derived natural products and are well known for their many applications in the pharmaceutical, fragrance and cosmetics industries. Hairy root cultures are useful for the production of plant secondary metabolites because of their genetic and biochemical stability and their rapid growth in hormone-free media. Tobacco (Nicotiana tabacum L. cv. Petit Havana SR1) hairy roots, which do not produce geraniol naturally, were engineered to express a plastid-targeted geraniol synthase gene originally isolated from Valeriana officinalis L. (VoGES). A SPME-GC-MS screening tool was developed for the rapid evaluation of production clones. The GC-MS analysis revealed that the free geraniol content in 20 hairy root clones expressing VoGES was an average of 13.7 μg/g dry weight (DW) and a maximum of 31.3 μg/g DW. More detailed metabolic analysis revealed that geraniol derivatives were present in six major glycoside forms, namely the hexose and/or pentose conjugates of geraniol and hydroxygeraniol, resulting in total geraniol levels of up to 204.3 μg/g DW following deglycosylation. A benchtop-scale process was developed in a 20-L wave-mixed bioreactor eventually yielding hundreds of grams of biomass and milligram quantities of geraniol per cultivation bag. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Salinity-induced accumulation of endogenous H2S and NO is associated with modulation of the antioxidant and redox defense systems in Nicotiana tabacum L. cv. Havana.

    Science.gov (United States)

    da Silva, Cristiane Jovelina; Batista Fontes, Elizabeth Pacheco; Modolo, Luzia Valentina

    2017-03-01

    Salinity is one of the abiotic factors that most affect crop growth and production. This study focused on the effect of high salinity on the endogenous levels of the signaling molecules hydrogen sulfite (H2S) and nitric oxide (NO) in Nicotiana tabacum leaves and the extent of these for the biochemically-driven plant tolerance to such abiotic stress. The NaCl treatment for 10days led to an expressive augment of H2S and NO levels. This increase was correlated with the raise of l-Cys and l-Arg and the induction of l-cysteine desulfhydrase, cyanoalanine synthase, cysteine synthase, nitrate reductase and arginase, enzymes known to be involved in the biosynthesis of H2S or NO. The enzymatic antioxidant system (superoxide dismutase and catalase activity) was boosted and the non-enzymatic antioxidant glutathione was intensively oxidized in leaves upon stress allowing plants to cope with oxidative stress. Lower stomatal conductance was observed in stressed plants in comparison with control ones. Moreover, the high activity of antioxidant enzymes and high rate of glutathione oxidation following salt stress were considerably decreased upon NO or H2S scavenging. Thus, increment in NO and H2S levels and their interplay, along with metabolic and physiological changes, contributed to tobacco survival to extreme salinity conditions. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  6. Accumulation of human EGF in nectar of transformed plants of Nicotiana langsdorffii x N. sanderae and transfer to honey by bees.

    Science.gov (United States)

    Helsper, J P F G; Ruyter-Spira, C P; Kwakman, P H S; Bleeker, W K; Keizer, L C P; Bade, J B; Te Velde, A A; Zaat, S A J; Verbeek, M; Creemers-Molenaar, J

    2011-09-01

    Honey has been used successfully in wound healing for thousands of years. The peptide hormone human epidermal growth factor (hEGF) is also known to have a beneficial effect in various wound healing processes via mechanisms that differ from those for honey. In this study, we show that hEGF can be incorporated into honey via nectar. Plants of Nicotiana langsdorffii x N. sanderae were transformed with the gene for hEGF, equipped with a nectary-targeted promoter and a signal sequence for secretion to nectar. These plants accumulated hEGF in the nectar. The maximum hEGF concentration recorded with ELISA in these plants is 2.5 ng·ml⁻¹. There is a significant linear relationship (Pplants did not allow production of honey from their nectar, we used feeding solutions, spiked with synthetic hEGF, to study transfer of this peptide into honey through bee activity. Transfer of hEGF from a feeding solution to honey by bees occurred with retention of the hEGF concentration and the capacity to induce hEGF-receptor phosphorylation. These observations indicate that plants can function as a production platform for honey containing biologically active peptides, which may enhance wound healing and other biological processes.

  7. Expression of a Recombinant Anti-HIV and Anti-Tumor Protein, MAP30, in Nicotiana tobacum Hairy Roots: A pH-Stable and Thermophilic Antimicrobial Protein.

    Science.gov (United States)

    Moghadam, Ali; Niazi, Ali; Afsharifar, Alireza; Taghavi, Seyed Mohsen

    2016-01-01

    In contrast to conventional antibiotics, which microorganisms can readily evade, it is nearly impossible for a microbial strain that is sensitive to antimicrobial proteins to convert to a resistant strain. Therefore, antimicrobial proteins and peptides that are promising alternative candidates for the control of bacterial infections are under investigation. The MAP30 protein of Momordica charantia is a valuable type I ribosome-inactivating protein (RIP) with anti-HIV and anti-tumor activities. Whereas the antimicrobial activity of some type I RIPs has been confirmed, less attention has been paid to the antimicrobial activity of MAP30 produced in a stable, easily handled, and extremely cost-effective protein-expression system. rMAP30-KDEL was expressed in Nicotiana tobacum hairy roots, and its effect on different microorganisms was investigated. Analysis of the extracted total proteins of transgenic hairy roots showed that rMAP30-KDEL was expressed effectively and that this protein exhibited significant antibacterial activity in a dose-dependent manner. rMAP30-KDEL also possessed thermal and pH stability. Bioinformatic analysis of MAP30 and other RIPs regarding their conserved motifs, amino-acid contents, charge, aliphatic index, GRAVY value, and secondary structures demonstrated that these factors accounted for their thermophilicity. Therefore, RIPs such as MAP30 and its derived peptides might have promising applications as food preservatives, and their analysis might provide useful insights into designing clinically applicable antibiotic agents.

  8. Expression of a Recombinant Anti-HIV and Anti-Tumor Protein, MAP30, in Nicotiana tobacum Hairy Roots: A pH-Stable and Thermophilic Antimicrobial Protein.

    Directory of Open Access Journals (Sweden)

    Ali Moghadam

    Full Text Available In contrast to conventional antibiotics, which microorganisms can readily evade, it is nearly impossible for a microbial strain that is sensitive to antimicrobial proteins to convert to a resistant strain. Therefore, antimicrobial proteins and peptides that are promising alternative candidates for the control of bacterial infections are under investigation. The MAP30 protein of Momordica charantia is a valuable type I ribosome-inactivating protein (RIP with anti-HIV and anti-tumor activities. Whereas the antimicrobial activity of some type I RIPs has been confirmed, less attention has been paid to the antimicrobial activity of MAP30 produced in a stable, easily handled, and extremely cost-effective protein-expression system. rMAP30-KDEL was expressed in Nicotiana tobacum hairy roots, and its effect on different microorganisms was investigated. Analysis of the extracted total proteins of transgenic hairy roots showed that rMAP30-KDEL was expressed effectively and that this protein exhibited significant antibacterial activity in a dose-dependent manner. rMAP30-KDEL also possessed thermal and pH stability. Bioinformatic analysis of MAP30 and other RIPs regarding their conserved motifs, amino-acid contents, charge, aliphatic index, GRAVY value, and secondary structures demonstrated that these factors accounted for their thermophilicity. Therefore, RIPs such as MAP30 and its derived peptides might have promising applications as food preservatives, and their analysis might provide useful insights into designing clinically applicable antibiotic agents.

  9. Ectopic expression of Capsicum-specific cell wall protein Capsicum annuum senescence-delaying 1 (CaSD1) delays senescence and induces trichome formation in Nicotiana benthamiana.

    Science.gov (United States)

    Seo, Eunyoung; Yeom, Seon-In; Jo, Sunghwan; Jeong, Heejin; Kang, Byoung-Cheorl; Choi, Doil

    2012-04-01

    Secreted proteins are known to have multiple roles in plant development, metabolism, and stress response. In a previous study to understand the roles of secreted proteins, Capsicum annuum secreted proteins (CaS) were isolated by yeast secretion trap. Among the secreted proteins, we further characterized Capsicum annuum senescence-delaying 1 (CaSD1), a gene encoding a novel secreted protein that is present only in the genus Capsicum. The deduced CaSD1 contains multiple repeats of the amino acid sequence KPPIHNHKPTDYDRS. Interestingly, the number of repeats varied among cultivars and species in the Capsicum genus. CaSD1 is constitutively expressed in roots, and Agrobacterium-mediated transient overexpression of CaSD1 in Nicotiana benthamiana leaves resulted in delayed senescence with a dramatically increased number of trichomes and enlarged epidermal cells. Furthermore, senescence- and cell division-related genes were differentially regulated by CaSD1-overexpressing plants. These observations imply that the pepper-specific cell wall protein CaSD1 plays roles in plant growth and development by regulating cell division and differentiation.

  10. A three-dimensional RNA motif in Potato spindle tuber viroid mediates trafficking from palisade mesophyll to spongy mesophyll in Nicotiana benthamiana.

    Science.gov (United States)

    Takeda, Ryuta; Petrov, Anton I; Leontis, Neocles B; Ding, Biao

    2011-01-01

    Cell-to-cell trafficking of RNA is an emerging biological principle that integrates systemic gene regulation, viral infection, antiviral response, and cell-to-cell communication. A key mechanistic question is how an RNA is specifically selected for trafficking from one type of cell into another type. Here, we report the identification of an RNA motif in Potato spindle tuber viroid (PSTVd) required for trafficking from palisade mesophyll to spongy mesophyll in Nicotiana benthamiana leaves. This motif, called loop 6, has the sequence 5'-CGA-3'...5'-GAC-3' flanked on both sides by cis Watson-Crick G/C and G/U wobble base pairs. We present a three-dimensional (3D) structural model of loop 6 that specifies all non-Watson-Crick base pair interactions, derived by isostericity-based sequence comparisons with 3D RNA motifs from the RNA x-ray crystal structure database. The model is supported by available chemical modification patterns, natural sequence conservation/variations in PSTVd isolates and related species, and functional characterization of all possible mutants for each of the loop 6 base pairs. Our findings and approaches have broad implications for studying the 3D RNA structural motifs mediating trafficking of diverse RNA species across specific cellular boundaries and for studying the structure-function relationships of RNA motifs in other biological processes.

  11. Simultaneous determination of shikimic acid, salicylic acid and jasmonic acid in wild and transgenic Nicotiana langsdorffii plants exposed to abiotic stresses.

    Science.gov (United States)

    Scalabrin, Elisa; Radaelli, Marta; Capodaglio, Gabriele

    2016-06-01

    The presence and relative concentration of phytohormones may be regarded as a good indicator of an organism's physiological state. The integration of the rolC gene from Agrobacterium rhizogenes and of the rat glucocorticoid receptor (gr) in Nicotiana langsdorffii Weinmann plants has shown to determine various physiological and metabolic effects. The analysis of wild and transgenic N. langsdorffii plants, exposed to different abiotic stresses (high temperature, water deficit, and high chromium concentrations) was conducted, in order to investigate the metabolic effects of the inserted genes in response to the applied stresses. The development of a new analytical procedure was necessary, in order to assure the simultaneous determination of analytes and to obtain an adequately low limit of quantification. For the first time, a sensitive HPLC-HRMS quantitative method for the simultaneous determination of salicylic acid, jasmonic acid and shikimic acid was developed and validated. The method was applied to 80 plant samples, permitting the evaluation of plant stress responses and highlighting some metabolic mechanisms. Salicylic, jasmonic and shikimic acids proved to be suitable for the comprehension of plant stress responses. Chemical and heat stresses showed to induce the highest changes in plant hormonal status, differently affecting plant response. The potential of each genetic modification toward the applied stresses was marked and particularly the resistance of the gr modified plants was evidenced. This work provides new information in the study of N. langsdorffii and transgenic organisms, which could be useful for the further application of these transgenes.

  12. Ser/Thr kinase-like protein of Nicotiana benthamiana is involved in the cell-to-cell movement of Bamboo mosaic virus.

    Directory of Open Access Journals (Sweden)

    Shun-Fang Cheng

    Full Text Available To investigate the plant genes affected by Bamboo mosaic virus (BaMV infection, we applied a cDNA-amplified fragment length polymorphism technique to screen genes with differential expression. A serine/threonine kinase-like (NbSTKL gene of Nicotiana benthamiana is upregulated after BaMV infection. NbSTKL contains the homologous domain of Ser/Thr kinase. Knocking down the expression of NbSTKL by virus-induced gene silencing reduced the accumulation of BaMV in the inoculated leaves but not in the protoplasts. The spread of GFP-expressing BaMV in the inoculated leaves is also impeded by a reduced expression of NbSTKL. These data imply that NbSTKL facilitates the cell-to-cell movement of BaMV. The subcellular localization of NbSTKL is mainly on the cell membrane, which has been confirmed by mutagenesis and fractionation experiments. Combined with the results showing that active site mutation of NbSTKL does not change its subcellular localization but significantly affects BaMV accumulation, we conclude that NbSTKL may regulate BaMV movement on the cell membrane by its kinase-like activity. Moreover, the transient expression of NbSTKL does not significantly affect the accumulation of Cucumber mosaic virus (CMV and Potato virus X (PVX; thus, NbSTKL might be a specific protein facilitating BaMV movement.

  13. Enhancement of RNA-directed DNA methylation of a transgene by simultaneously downregulating a ROS1 orthologue using a virus vector in Nicotiana benthamiana

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    Shungo eOtagaki

    2013-04-01

    Full Text Available Cytosine methylation can be induced by double-stranded RNAs through the RNA-directed DNA methylation (RdDM pathway. A DNA glycosylase REPRESSOR OF SILENCING 1 (ROS1 participates in DNA demethylation in Arabidopsis and may possibly counteract RdDM. Here, we isolated an orthologue of ROS1 (NbROS1 from Nicotiana benthamiana and examined the antagonistic activity of NbROS1 against virus-induced RdDM by simultaneously inducing RdDM and NbROS1 knockdown using a vector based on Cucumber mosaic virus. Plants were inoculated with a virus that contained a portion of the CaMV 35S promoter, which induced RdDM of the promoter integrated in the plant genome and transcriptional silencing of the green fluorescent protein gene driven by the promoter. Plants were also inoculated with a virus that contained a portion of NbROS1, which induced downregulation of NbROS1. Simultaneous induction of RdDM and NbROS1 knockdown resulted in an increase in the level of cytosine methylation of the target promoter. These results provide evidence for the presence of antagonistic activity of NbROS1 against virus-induced RdDM and suggest that the simultaneous induction of promoter-targeting RdDM and NbROS1 knockdown by a virus vector is useful as a tool to enhance targeted DNA methylation.

  14. Hydrophobin fusion of an influenza virus hemagglutinin allows high transient expression in Nicotiana benthamiana, easy purification and immune response with neutralizing activity.

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    Nicolas Jacquet

    Full Text Available The expression of recombinant hemagglutinin in plants is a promising alternative to the current egg-based production system for the influenza vaccines. Protein-stabilizing fusion partners have been developed to overcome the low production yields and the high downstream process costs associated with the plant expression system. In this context, we tested the fusion of hydrophobin I to the hemagglutinin ectodomain of the influenza A (H1N1pdm09 virus controlled by the hybrid En2PMA4 transcriptional promoter to rapidly produce high levels of recombinant antigen by transient expression in agro-infiltrated Nicotiana benthamiana leaves. The fusion increased the expression level by a factor of ∼ 2.5 compared to the unfused protein allowing a high accumulation level of 8.6% of the total soluble proteins. Hemagglutinin was located in ER-derived protein bodies and was successfully purified by combining an aqueous-two phase partition system and a salting out step. Hydrophobin interactions allowed the formation of high molecular weight hemagglutinin structures, while unfused proteins were produced as monomers. Purified protein was shown to be biologically active and to induce neutralizing antibodies after mice immunization. Hydrophobin fusion to influenza hemagglutinin might therefore be a promising approach for rapid, easy, and low cost production of seasonal or pandemic influenza vaccines in plants.

  15. Overexpression and self-assembly of virus-like particles in Nicotiana benthamiana by a single-vector DNA replicon system.

    Science.gov (United States)

    Moon, Ki-Beom; Lee, Jisu; Kang, Sebyung; Kim, Moonil; Mason, Hugh S; Jeon, Jae-Heung; Kim, Hyun-Soon

    2014-10-01

    Based on recent developments, virus-like particles (VLPs) are considered to be perfect candidates as nanoplatforms for applications in materials science and medicine. To succeed, mass production of VLPs and self-assembly into a correct form in plant systems are key factors. Here, we report expression of synthesized coat proteins of the three viruses, Brome mosaic virus, Cucumber mosaic virus, and Maize rayado fino virus, in Nicotiana benthamiana and production of self-assembled VLPs by transient expression system using agroinfiltration. Each coat protein was synthesized and cloned into a pBYR2fp single replicon vector. Target protein expression in cells containing p19 was fourfold higher than that of cells lacking p19. After agroinfiltration, protein expression was analyzed by SDS-PAGE and quantitative image analyzer. Quantitative analysis showed that BMVCP, CMVCP, and MRFVCP concentrations were 0.5, 1.0, and 0.8 mg · g(-1) leaf fresh weight, respectively. VLPs were purified by sucrose cushion ultracentrifugation and then analyzed by transmission electron microscopy. Our results suggested that BMVCP and CMVCP proteins expressed in N. benthamiana leaves were able to correctly self-assemble into particles. Moreover, we evaluated internal cavity accessibility of VLPs to load foreign molecules. Finally, plant growth conditions after agroinfiltration are critical for increasing heterologous protein expression levels in a transient expression system.

  16. A chimeric and truncated mitochondrial atpA gene is transcribed in alloplasmic cytoplasmic male-sterile tobacco with Nicotiana bigelovii mitochondria.

    Science.gov (United States)

    Bergman, P; Kofer, W; Håkansson, G; Glimelius, K

    1995-09-01

    Protoplast fusions were performed between two sexually produced alloplasmic male-sterile tobacco cultivars, with cytoplasms from Nicotiana bigelovii [Nta (big)S] and N. undulata[Nta(und)S], both of which exhibit homeotic-like phenotypes affecting the petal and stamen whorls. Among the fusion products obtained, both novel male-sterile and pollen-producing cybrid plants were identified. Of the pollen-producing cybrid plants, all of which were indehiscent, some had flowers with stamens that appeared normal when compared to male-fertile tobacco plants. Other hybrid plants were incompletely restored as they exhibited petaloid structures on the anther-bearing pollen-producing stamens. In this study, gel-blot analyses with mitochondrial geneprobes were conducted comparing the mitochondrial DNA of cybrids and male-sterile parents. It was found that the flower morphology typical of the Nta(big)S parental plants, as well as of the novel male-sterile cybrids, coincided with the presence of a chimeric atpA gene copy where an open reading frame of unknown origin was found to be linked in-frame to the 3'-end of a truncated atpA gene. RNA gel-blot hybridizations revealed the presence of atpA transcripts in the malesterile parent Nta(big)S and novel male-sterile cybrids, but which were absent in cybrids capable of pollen production.

  17. Downregulation of the NbNACa1 gene encoding a movement-protein-interacting protein reduces cell-to-cell movement of Brome mosaic virus in Nicotiana benthamiana.

    Science.gov (United States)

    Kaido, Masanori; Inoue, Yosuke; Takeda, Yoshika; Sugiyama, Kazuhiko; Takeda, Atsushi; Mori, Masashi; Tamai, Atsushi; Meshi, Tetsuo; Okuno, Tetsuro; Mise, Kazuyuki

    2007-06-01

    The 3a movement protein (MP) plays a central role in the movement of the RNA plant virus, Brome mosaic virus (BMV). To identify host factor genes involved in viral movement, a cDNA library of Nicotiana benthamiana, a systemic host for BMV, was screened with far-Western blotting using a recombinant BMV MP as probe. One positive clone encoded a protein with sequence similarity to the alpha chain of nascent-polypeptide-associated complex from various organisms, which is proposed to contribute to the fidelity of translocation of newly synthesized proteins. The orthologous gene from N. benthamiana was designated NbNACa1. The binding of NbNACa1 to BMV MP was confirmed in vivo with an agroinfiltration-immunoprecipitation assay. To investigate the involvement of NbNACa1 in BMV multiplication, NbNACa1-silenced (GSNAC) transgenic N. benthamiana plants were produced. Downregulation of NbNACa1 expression reduced virus accumulation in inoculated leaves but not in protoplasts. A microprojectile bombardment assay to monitor BMV-MP-assisted viral movement demonstrated reduced virus spread in GSNAC plants. The localization to the cell wall of BMV MP fused to green fluorescent protein was delayed in GSNAC plants. From these results, we propose that NbNACa1 is involved in BMV cell-to-cell movement through the regulation of BMV MP localization to the plasmodesmata.

  18. The Development of DNA Based Methods for the Reliable and Efficient Identification of Nicotiana tabacum in Tobacco and Its Derived Products

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    Sukumar Biswas

    2016-01-01

    Full Text Available Reliable methods are needed to detect the presence of tobacco components in tobacco products to effectively control smuggling and classify tariff and excise in tobacco industry to control illegal tobacco trade. In this study, two sensitive and specific DNA based methods, one quantitative real-time PCR (qPCR assay and the other loop-mediated isothermal amplification (LAMP assay, were developed for the reliable and efficient detection of the presence of tobacco (Nicotiana tabacum in various tobacco samples and commodities. Both assays targeted the same sequence of the uridine 5′-monophosphate synthase (UMPS, and their specificities and sensitivities were determined with various plant materials. Both qPCR and LAMP methods were reliable and accurate in the rapid detection of tobacco components in various practical samples, including customs samples, reconstituted tobacco samples, and locally purchased cigarettes, showing high potential for their application in tobacco identification, particularly in the special cases where the morphology or chemical compositions of tobacco have been disrupted. Therefore, combining both methods would facilitate not only the detection of tobacco smuggling control, but also the detection of tariff classification and of excise.

  19. iTRAQ-based quantitative proteomic analysis reveals proteomic changes in leaves of cultivated tobacco (Nicotiana tabacum) in response to drought stress.

    Science.gov (United States)

    Xie, He; Yang, Da-Hai; Yao, Heng; Bai, Ge; Zhang, Yi-Han; Xiao, Bing-Guang

    2016-01-15

    Drought is one of the most severe forms of abiotic stresses that threaten the survival of plants, including crops. In turn, plants dramatically change their physiology to increase drought tolerance, including reconfiguration of proteomes. Here, we studied drought-induced proteomic changes in leaves of cultivated tobacco (Nicotiana tabacum), a solanaceous plant, using the isobaric tags for relative and absolute quantitation (iTRAQ)-based protein labeling technology. Of identified 5570 proteins totally, drought treatment increased and decreased abundance of 260 and 206 proteins, respectively, compared with control condition. Most of these differentially regulated proteins are involved in photosynthesis, metabolism, and stress and defense. Although abscisic acid (ABA) levels greatly increased in drought-treated tobacco leaves, abundance of detected ABA biosynthetic enzymes showed no obvious changes. In contrast, heat shock proteins (HSPs), thioredoxins, ascorbate-, glutathione-, and hydrogen peroxide (H2O2)-related proteins were up- or down-regulated in drought-treated tobacco leaves, suggesting that chaperones and redox signaling are important for tobacco tolerance to drought, and it is likely that redox-induced posttranslational modifications play an important role in modulating protein activity. This study not only provides a comprehensive dataset on overall protein changes in drought-treated tobacco leaves, but also shed light on the mechanism by which solanaceous plants adapt to drought stress.

  20. Expression of a Recombinant Anti-HIV and Anti-Tumor Protein, MAP30, in Nicotiana tobacum Hairy Roots: A pH-Stable and Thermophilic Antimicrobial Protein

    Science.gov (United States)

    Moghadam, Ali; Niazi, Ali; Afsharifar, Alireza; Taghavi, Seyed Mohsen

    2016-01-01

    In contrast to conventional antibiotics, which microorganisms can readily evade, it is nearly impossible for a microbial strain that is sensitive to antimicrobial proteins to convert to a resistant strain. Therefore, antimicrobial proteins and peptides that are promising alternative candidates for the control of bacterial infections are under investigation. The MAP30 protein of Momordica charantia is a valuable type I ribosome-inactivating protein (RIP) with anti-HIV and anti-tumor activities. Whereas the antimicrobial activity of some type I RIPs has been confirmed, less attention has been paid to the antimicrobial activity of MAP30 produced in a stable, easily handled, and extremely cost-effective protein-expression system. rMAP30-KDEL was expressed in Nicotiana tobacum hairy roots, and its effect on different microorganisms was investigated. Analysis of the extracted total proteins of transgenic hairy roots showed that rMAP30-KDEL was expressed effectively and that this protein exhibited significant antibacterial activity in a dose-dependent manner. rMAP30-KDEL also possessed thermal and pH stability. Bioinformatic analysis of MAP30 and other RIPs regarding their conserved motifs, amino-acid contents, charge, aliphatic index, GRAVY value, and secondary structures demonstrated that these factors accounted for their thermophilicity. Therefore, RIPs such as MAP30 and its derived peptides might have promising applications as food preservatives, and their analysis might provide useful insights into designing clinically applicable antibiotic agents. PMID:27459300

  1. The Sesquiterpenes(E-ß-Farnesene and (E-α-Bergamotene Quench Ozone but Fail to Protect the Wild Tobacco Nicotiana attenuata from Ozone, UVB, and Drought Stresses.

    Directory of Open Access Journals (Sweden)

    Evan C Palmer-Young

    Full Text Available Among the terpenes, isoprene (C5 and monoterpene hydrocarbons (C10 have been shown to ameliorate abiotic stress in a number of plant species via two proposed mechanisms: membrane stabilization and direct antioxidant effects. Sesquiterpene hydrocarbons (C15 not only share the structural properties thought to lend protective qualities to isoprene and monoterpene hydrocarbons, but also react rapidly with ozone, suggesting that sesquiterpenes may similarly enhance tolerance of abiotic stresses. To test whether sesquiterpenes protect plants against ozone, UVB light, or drought, we used transgenic lines of the wild tobacco Nicotiana attenuata. The transgenic plants expressed a maize terpene synthase gene (ZmTPS10 which produced a blend of (E-ß-farnesene and (E-α-bergamotene, or a point mutant of the same gene (ZmTPS10M which produced (E-ß-farnesene alone,. (E-ß-farnesene exerted a local, external, and transient ozone-quenching effect in ozone-fumigated chambers, but we found no evidence that enhanced sesquiterpene production by the plant inhibited oxidative damage, or maintained photosynthetic function or plant fitness under acute or chronic stress. Although the sesquiterpenes (E-ß-farnesene and (E-α-bergamotene might confer benefits under intermittent heat stress, which was not tested, any roles in relieving abiotic stress may be secondary to their previously demonstrated functions in biotic interactions.

  2. Metabolism of methoxychlor by the P450-monooxygenase CYP6G1 involved in insecticide resistance of Drosophila melanogaster after expression in cell cultures of Nicotiana tabacum.

    Science.gov (United States)

    Joussen, Nicole; Schuphan, Ingolf; Schmidt, Burkhard

    2010-03-01

    Cytochrome P450 monooxygenase CYP6G1 of Drosophila melanogaster was heterologously expressed in a cell suspension culture of Nicotiana tabacum. This in vitro system was used to study the capability of CYP6G1 to metabolize the insecticide methoxychlor (=1,1,1-trichloro-2,2-bis(4-methoxyphenyl)ethane, 1) against the background of endogenous enzymes of the corresponding non-transgenic culture. The Cyp6g1-transgenic cell culture metabolized 96% of applied methoxychlor (45.8 microg per assay) within 24 h by demethylation and hydroxylation mainly to trishydroxy and catechol methoxychlor (16 and 17%, resp.). About 34% of the metabolism and the distinct formation of trishydroxy and catechol methoxychlor were due to foreign enzyme CYP6G1. Furthermore, methoxychlor metabolism was inhibited by 43% after simultaneous addition of piperonyl butoxide (458 microg), whereas inhibition in the non-transgenic culture amounted to 92%. Additionally, the rate of glycosylation was reduced in both cultures. These results were supported by the inhibition of the metabolism of the insecticide imidacloprid (6; 20 microg, 24 h) in the Cyp6g1-transgenic culture by 82% in the presence of piperonyl butoxide (200 microg). Due to CYP6G1 being responsible for imidacloprid resistance of Drosophila or being involved in DDT resistance, it is likely that CYP6G1 conveys resistance to methoxychlor (1). Furthermore, treating Drosophila with piperonyl butoxide could weaken the observed resistance phenomena.

  3. Jasmonate-Sensitivity-Assisted Screening and Characterization of Nicotine Synthetic Mutants from Activation-Tagged Population of Tobacco (Nicotiana tabacum L.)

    Science.gov (United States)

    Yin, Guoying; Wang, Wenjing; Niu, Haixia; Ding, Yongqiang; Zhang, Dingyu; Zhang, Jie; Liu, Guanshan; Wang, Sangen; Zhang, Hongbo

    2017-01-01

    Nicotine is a secondary metabolite that is important to the defense system and commercial quality of tobacco (Nicotiana tabacum L.). Jasmonate and its derivatives (JAs) are phytohormone regulators of nicotine formation; however, the underlying molecular mechanism of this process remains largely unclear. Owing to the amphitetraploid origin of N. tabacum, research on screening and identification of nicotine-synthetic mutants is relatively scarce. Here, we describe a method based on JA-sensitivity for screening nicotine mutants from an activation-tagged population of tobacco. In this approach, the mutants were first screened for abnormal JA responses in seed germination and root elongation, and then the levels of nicotine synthesis and expression of nicotine synthetic genes in the mutants with altered JA-response were measured to determine the nicotine-synthetic mutants. We successfully obtained five mutants that maintained stable nicotine contents and JA responses for three generations. This method is simple, effective and low-cost, and the finding of transcriptional changes of nicotine synthetic genes in the mutants shows potentials for identifying novel regulators involved in JA-regulated nicotine biosynthesis. PMID:28243248

  4. Isolation and characterization of an RNA-dependent RNA polymerase from Nicotiana clevelandii plants infected with red clover necrotic mosaic dianthovirus.

    Science.gov (United States)

    Bates, H J; Farjah, M; Osman, T A; Buck, K W

    1995-06-01

    A template-bound RNA polymerase was isolated from Nicotiana clevelandii plants infected with red clover necrotic mosaic dianthovirus (RCNMV) by differential centrifugation, solubilization with dodecyl beta-D-maltopyranoside, and chromatography on columns of Sephacryl S-400 and Q-Sepharose. Analysis of the purified polymerase by SDS-polyacrylamide gel electrophoresis, followed by silver staining or immunoblotting, showed that it contained virus-encoded proteins of molecular masses 27 kDa and 88 kDa together with several minor proteins possibly of host origin. After removal of endogenous RNA with micrococcal nuclease, the polymerase became template-dependent. It was also template-specific, being able to utilize as templates RNA of two strains of RCNMV, but not RNAs of three viruses in different taxonomic groups, namely cucumber mosaic cucumovirus, tomato bushy stunt tombusvirus and tomato mosaic tobamovirus. The products of RNA polymerase reactions were double-stranded RNAs corresponding to RCNMV RNAs 1 and 2. The ability of the template-dependent RNA polymerase to synthesize RNA was completely inhibited by antibodies to a peptide containing the GDD motif, whereas the activity of the template-bound enzyme was unaffected by these antibodies.

  5. The symbiosis between Nicotiana tabacum and the endomycorrhizal fungus Funneliformis mosseae increases the plant glutathione level and decreases leaf cadmium and root arsenic contents.

    Science.gov (United States)

    Degola, Francesca; Fattorini, Laura; Bona, Elisa; Sprimuto, Christian Triscari; Argese, Emanuele; Berta, Graziella; Sanità di Toppi, Luigi

    2015-07-01

    Over time, anthropogenic activities have led to severe cadmium (Cd) and arsenic (As) pollution in several environments. Plants inhabiting metal(loid)-contaminated areas should be able to sequester and detoxify these toxic elements as soon as they enter roots and leaves. We postulated here that an important role in protecting plants from excessive metal(loid) accumulation and toxicity might be played by arbuscular mycorrhizal (AM) fungi. In fact, human exploitation of plant material derived from Cd- and As-polluted environments may lead to a noxious intake of these toxic elements; in particular, a possible source of Cd and As for humans is given by cigarette and cigar smoke. We investigated the role of AM fungus Funneliformis mosseae (T.H. Nicolson & Gerd.) C. Walker & A. Schüßler in protecting Nicotiana tabacum L. (cv. Petit Havana) from the above-mentioned metal(loid) stress. Our findings proved that the AM symbiosis is effective in increasing the plant tissue content of the antioxidant glutathione (GSH), in influencing the amount of metal(loid)-induced chelators as phytochelatins, and in reducing the Cd and As content in leaves and roots of adult tobacco plants. These results might also prove useful in improving the quality of commercial tobacco, thus reducing the risks to human health due to inhalation of toxic elements contained in smoking products.

  6. Biological and chemical induction of resistance to the Globodera tabacum solanacearum in oriental and flue-cured tobacco (Nicotiana tabacum L.).

    Science.gov (United States)

    Parkunan, Venkatesan; Johnson, Charles S; Eisenback, Jon D

    2009-09-01

    The effects of acibenzolar-S-methyl (ASM) and four combinations of plant growth-promoting rhizobacteria (PGPR) on the reproduction of a tobacco cyst nematode, Globodera tabacum solanacearum, and growth of Nicotiana tabacum (cv. K326 and Xanthi) were tested under greenhouse and field conditions. The PGPR included combinations of Bacillus subtilis A13 with B. pumilis INR7, B. pumilis SE34, B. licheniformis IN937b, or B. amyloliquefaciens IN937a, respectively. Among the four rhizobacterial combinations, IN937a + A13 exhibited the most consistent reduction in G. t. solanacearum cysts under greenhouse and field conditions. No undesirable effects of IN937a + A13 were observed on tobacco growth under greenhouse and field conditions. Use of INR7 + A13 reduced G. t. solanacearum reproduction on flue-cured tobacco cv. K326 but not on oriental tobacco cv. Xanthi. Application of ASM reduced final numbers of G. t. solanacearum cysts, but also resulted in phytotoxicity mainly under the greenhouse conditions. When oriental tobacco seedlings were pre-grown in a IN937a + A13-treated soil-less medium, a single application of ASM at 200 mg/L one week after transplanting significantly reduced G. t. solanacearum reproduction in the field.

  7. Transplastomic Nicotiana benthamiana plants expressing multiple defence genes encoding protease inhibitors and chitinase display broad-spectrum resistance against insects, pathogens and abiotic stresses.

    Science.gov (United States)

    Chen, Peng-Jen; Senthilkumar, Rajendran; Jane, Wann-Neng; He, Yong; Tian, Zhihong; Yeh, Kai-Wun

    2014-05-01

    Plastid engineering provides several advantages for the next generation of transgenic technology, including the convenient use of transgene stacking and the generation of high expression levels of foreign proteins. With the goal of generating transplastomic plants with multiresistance against both phytopathogens and insects, a construct containing a monocistronic patterned gene stack was transformed into Nicotiana benthamiana plastids harbouring sweet potato sporamin, taro cystatin and chitinase from Paecilomyces javanicus. Transplastomic lines were screened and characterized by Southern/Northern/Western blot analysis for the confirmation of transgene integration and respective expression level. Immunogold localization analyses confirmed the high level of accumulation proteins that were specifically expressed in leaf and root plastids. Subsequent functional bioassays confirmed that the gene stacks conferred a high level of resistance against both insects and phytopathogens. Specifically, larva of Spodoptera litura and Spodoptera exigua either died or exhibited growth retardation after ingesting transplastomic plant leaves. In addition, the inhibitory effects on both leaf spot diseases caused by Alternaria alternata and soft rot disease caused by Pectobacterium carotovorum subsp. carotovorum were markedly observed. Moreover, tolerance to abiotic stresses such as salt/osmotic stress was highly enhanced. The results confirmed that the simultaneous expression of sporamin, cystatin and chitinase conferred a broad spectrum of resistance. Conversely, the expression of single transgenes was not capable of conferring such resistance. To the best of our knowledge, this is the first study to demonstrate an efficacious stacked combination of plastid-expressed defence genes which resulted in an engineered tolerance to various abiotic and biotic stresses.

  8. A calmodulin-like protein suppresses RNA silencing and promotes geminivirus infection by degrading SGS3 via the autophagy pathway in Nicotiana benthamiana

    Science.gov (United States)

    Li, Fangfang; Zhao, Nan; Xu, Xiongbiao; Wang, Yaqin; Yang, Xiuling; Liu, Shu-Sheng; Wang, Aiming; Zhou, Xueping

    2017-01-01

    A recently characterized calmodulin-like protein is an endogenous RNA silencing suppressor that suppresses sense-RNA induced post-transcriptional gene silencing (S-PTGS) and enhances virus infection, but the mechanism underlying calmodulin-like protein-mediated S-PTGS suppression is obscure. Here, we show that a calmodulin-like protein from Nicotiana benthamiana (NbCaM) interacts with Suppressor of Gene Silencing 3 (NbSGS3). Deletion analyses showed that domains essential for the interaction between NbSGS3 and NbCaM are also required for the subcellular localization of NbSGS3 and NbCaM suppressor activity. Overexpression of NbCaM reduced the number of NbSGS3-associated granules by degrading NbSGS3 protein accumulation in the cytoplasm. This NbCaM-mediated NbSGS3 degradation was sensitive to the autophagy inhibitors 3-methyladenine and E64d, and was compromised when key autophagy genes of the phosphatidylinositol 3-kinase (PI3K) complex were knocked down. Meanwhile, silencing of key autophagy genes within the PI3K complex inhibited geminivirus infection. Taken together these data suggest that NbCaM acts as a suppressor of RNA silencing by degrading NbSGS3 through the autophagy pathway. PMID:28212430

  9. N-glycosylation of cholera toxin B subunit in Nicotiana benthamiana: impacts on host stress response, production yield and vaccine potential.

    Science.gov (United States)

    Hamorsky, Krystal Teasley; Kouokam, J Calvin; Jurkiewicz, Jessica M; Nelson, Bailey; Moore, Lauren J; Husk, Adam S; Kajiura, Hiroyuki; Fujiyama, Kazuhito; Matoba, Nobuyuki

    2015-01-23

    Plant-based transient overexpression systems enable rapid and scalable production of subunit vaccines. Previously, we have shown that cholera toxin B subunit (CTB), an oral cholera vaccine antigen, is N-glycosylated upon expression in transgenic Nicotiana benthamiana. Here, we found that overexpression of aglycosylated CTB by agroinfiltration of a tobamoviral vector causes massive tissue necrosis and poor accumulation unless retained in the endoplasmic reticulum (ER). However, the re-introduction of N-glycosylation to its original or an alternative site significantly relieved the necrosis and provided a high CTB yield without ER retention. Quantitative gene expression analysis of PDI, BiP, bZIP60, SKP1, 26Sα proteasome and PR1a, and the detection of ubiquitinated CTB polypeptides revealed that N-glycosylation significantly relieved ER stress and hypersensitive response, and facilitated the folding/assembly of CTB. The glycosylated CTB (gCTB) was characterized for potential vaccine use. Glycan profiling revealed that gCTB contained approximately 38% plant-specific glycans. gCTB retained nanomolar affinity to GM1-ganglioside with only marginal reduction of physicochemical stability and induced an anti-cholera holotoxin antibody response comparable to native CTB in a mouse oral immunization study. These findings demonstrated gCTB's potential as an oral immunogen and point to a potential role of N-glycosylation in increasing recombinant protein yields in plants.

  10. Effects of sublethal concentrations of tobacco (Nicotiana tobaccum leaf dust on some biochemical parameters of Hybrid catfish (Clarias gariepinus and Heterobranchus bidorsalis

    Directory of Open Access Journals (Sweden)

    Kabir Mohammed Adamu

    2011-02-01

    Full Text Available The aim of this work was to study the effect of tobacco (Nicotiana tobaccum leaf dust at sublethal concentrations (375.6, 187.8, 93.9 0.0mg/L on biochemical parameters of Hybrid catfish in a static semi-renewable bioassay system with the aim to ascertain its effect on the test fish after 14 days exposure period. The effects of the plant dust on the test fish was dose-dependent, revealing significant difference in albumin, glucose, triglyceride, cholesterol, urea, and uric acid of the serum, liver and kidney; total protein and globulin of serum and kidney and creatinine and calcium of the serum while insignificant difference were observed for calcium and creatinine in liver and kidney and total protein and globulin in liver compared to the control. The effect of the plant dust was most pronounced in the serum, less in the kidney and least in the liver. However, the monitored water quality parameters revealed that the plant dust had effects on primary productivity, thus the biodiversity of organisms.

  11. Transcriptomic responses of the aphid Myzus persicae nicotianae Blackman (Hemiptera: Aphididae to insecticides: Analyses in the single Chilean clone of the tobacco aphid

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    Marco Cabrera-Brandt

    2014-04-01

    Full Text Available The tobacco aphid Myzus persicae nicotianae Blackman is a subspecies of the highly polyphagous and agricultural pest Myzus persicae (Sulzer. For its control, insecticide applications are widely used, but resistance to numerous molecules has been reported, displaying at least three insecticide resistance mechanisms, including: (i elevated carboxylesterases (E-Carb, (ii modification of the acetylcholinesterase (MACE, and (iii kdr and super-kdr insensitivity mutations. In Chile, populations of the tobacco aphid are characterized by the presence of a single predominant clone, which is also present in high proportions in other countries of the Americas. This aphid clone exhibits low levels of carboxylesterase activity and is kdr susceptible, but the MACE mechanism of insecticide resistance has not been studied. In order to characterize the tobacco aphid in terms of the MACE mechanism and to identify a preliminary group of aphid genes putatively involved in insecticide resistance, a cDNA microarray was used to study the transcriptomic responses when aphids are sprayed with a carbamate insecticide. The single Chilean clone of the tobacco aphid was characterized as MACE susceptible, but we found 38 transcripts significantly regulated by insecticide exposure (13 up- and 25 down-regulated genes. The expression of six of them was validated by qRT-PCR experiments at several time points (6, 12, 18, 24, 30, 36, and 42 h after insecticide application. This mutational and transcriptomic characterization of the tobacco aphid responding to insecticide spray opens new hypotheses in the understanding of the molecular mechanisms underlying insecticide resistance.

  12. Spermine either delays or promotes cell death in Nicotiana tabacum L. corolla depending on the floral developmental stage and affects the distribution of transglutaminase.

    Science.gov (United States)

    Cai, Giampiero; Della Mea, Massimiliano; Faleri, Claudia; Fattorini, Laura; Aloisi, Iris; Serafini-Fracassini, Donatella; Del Duca, Stefano

    2015-12-01

    The role of spermine (SM) was studied to verify if SM supplied to Nicotiana tabacum flower can modulate programmed cell death (PCD) of the corolla. SM has strong effects on the development and senescence of excised flowers despite its low physiological levels. The timing and duration of SM treatment is a key factor; SM counteracts PCD (verified by morphological observations, pigment contents and DNA laddering) only in the narrow developmental window of corolla expansion. Before and after, SM promotes PCD. SM exerts its pro-survival role by delaying fresh weight loss, by inhibiting reduction of pigments and finally by preventing DNA degradation. Moreover, SM deeply alters the distribution of the PA-conjugating enzyme transglutaminase (TGase). TGase is present in the epidermis during development, but it sprays also in the cell walls of inner parenchyma at senescence. After SM treatment, parenchyma cells accumulate TGase, increase in size and their cell walls do not undergo stiffening contrarily to control cells. The subcellular localization of TGase has been validated by biolistic-transformation of onion epidermal cells. Results indicated that SM is a critical factor in the senescence of N. tabacum corolla by controlling biochemical and morphological parameters; the lasts are probably interconnected with the action of TGase.

  13. Phenylpropanoid Defences in Nicotiana tabacum Cells: Overlapping Metabolomes Indicate Common Aspects to Priming Responses Induced by Lipopolysaccharides, Chitosan and Flagellin-22.

    Directory of Open Access Journals (Sweden)

    Msizi I Mhlongo

    Full Text Available Plants have evolved both constitutive and inducible defence strategies to cope with different biotic stimuli and stresses. Exposure of a plant to a challenging stress can lead to a primed state that allows it to launch a more rapid and stronger defence. Here we applied a metabolomic approach to study and compare the responses induced in Nicotiana tabacum cells by microbe-associated molecular pattern (MAMP molecules, namely lipopolysaccharides (LPS, chitosan (CHT and flagellin-22 (FLG22. Early response metabolites, extracted with methanol, were analysed by UHPLC-MS/MS. Using multivariate statistical tools the metabolic profiles induced by these elicitors were analysed. In the metabolic fingerprint of these agents a total of 19 cinnamic acid derivatives conjugated to quinic acids (chlorogenic acids, shikimic acid, tyramine, polyamines or glucose were found as discriminant biomarkers. In addition, treatment with the phytohormones salicylic acid (SA, methyljasmonic acid (MJ and abscisic acid (ABA resulted in differentially-induced phenylpropanoid pathway metabolites. The results indicate that the phenylpropanoid pathway is activated by these elicitors while hydroxycinnamic acid derivatives are commonly associated with the metabolic response to the MAMPs, and that the activated responses are modulated by both SA and MJ, with ABA not playing a role.

  14. An Accessory Protease Inhibitor to Increase the Yield and Quality of a Tumour-Targeting mAb in Nicotiana benthamiana Leaves.

    Science.gov (United States)

    Jutras, Philippe V; Marusic, Carla; Lonoce, Chiara; Deflers, Carole; Goulet, Marie-Claire; Benvenuto, Eugenio; Michaud, Dominique; Donini, Marcello

    2016-01-01

    The overall quality of recombinant IgG antibodies in plants is dramatically compromised by host endogenous proteases. Different approaches have been developed to reduce the impact of endogenous proteolysis on IgGs, notably involving site-directed mutagenesis to eliminate protease-susceptible sites or the in situ mitigation of host protease activities to minimize antibody processing in the cell secretory pathway. We here characterized the degradation profile of H10, a human tumour-targeting monoclonal IgG, in leaves of Nicotiana benthamiana also expressing the human serine protease inhibitor α1-antichymotrypsin or the cysteine protease inhibitor tomato cystatin SlCYS8. Leaf extracts revealed consistent fragmentation patterns for the recombinant antibody regardless of leaf age and a strong protective effect of SlCYS8 in specific regions of the heavy chain domains. As shown using an antigen-binding ELISA and LC-MS/MS analysis of antibody fragments, SlCYS8 had positive effects on both the amount of fully-assembled antibody purified from leaf tissue and the stability of biologically active antibody fragments containing the heavy chain Fc domain. Our data confirm the potential of Cys protease inhibitors as convenient antibody-stabilizing expression partners to increase the quality of therapeutic antibodies in plant protein biofactories.

  15. SGS3 Cooperates with RDR6 in Triggering Geminivirus-Induced Gene Silencing and in Suppressing Geminivirus Infection in Nicotiana Benthamiana

    Directory of Open Access Journals (Sweden)

    Fangfang Li

    2017-09-01

    Full Text Available RNA silencing has an important role in defending against virus infection in plants. Plants with the deficiency of RNA silencing components often show enhanced susceptibility to viral infections. RNA-dependent RNA polymerase (RDRs mediated-antiviral defense has a pivotal role in resistance to many plant viruses. In RDR6-mediated defense against viral infection, a plant-specific RNA binding protein, Suppressor of Gene Silencing 3 (SGS3, was also found to fight against some viruses in Arabidopsis. In this study, we showed that SGS3 from Nicotiana benthamiana (NbSGS3 is required for sense-RNA induced post-transcriptional gene silencing (S-PTGS and initiating sense-RNA-triggered systemic silencing. Further, the deficiency of NbSGS3 inhibited geminivirus-induced endogenous gene silencing (GIEGS and promoted geminivirus infection. During TRV-mediated NbSGS3 or N. benthamiana RDR6 (NbRDR6 silencing process, we found that their expression can be effectively fine-tuned. Plants with the knock-down of both NbSGS3 and NbRDR6 almost totally blocked GIEGS, and were more susceptible to geminivirus infection. These data suggest that NbSGS3 cooperates with NbRDR6 against GIEGS and geminivirus infection in N. benthamiana, which provides valuable information for breeding geminivirus-resistant plants.

  16. NaJAZh Regulates a Subset of Defense Responses against Herbivores and Spontaneous Leaf Necrosis in Nicotiana attenuata Plants[C][W][OA

    Science.gov (United States)

    Oh, Youngjoo; Baldwin, Ian T.; Gális, Ivan

    2012-01-01

    The JASMONATE ZIM DOMAIN (JAZ) proteins function as negative regulators of jasmonic acid signaling in plants. We cloned 12 JAZ genes from native tobacco (Nicotiana attenuata), including nine novel JAZs in tobacco, and examined their expression in plants that had leaves elicited by wounding or simulated herbivory. Most JAZ genes showed strong expression in the elicited leaves, but NaJAZg was mainly expressed in roots. Another novel herbivory-elicited gene, NaJAZh, was analyzed in detail. RNA interference suppression of this gene in inverted-repeat (ir)JAZh plants deregulated a specific branch of jasmonic acid-dependent direct and indirect defenses: irJAZh plants showed greater trypsin protease inhibitor activity, 17-hydroxygeranyllinalool diterpene glycosides accumulation, and emission of volatile organic compounds from leaves. Silencing of NaJAZh also revealed a novel cross talk in JAZ-regulated secondary metabolism, as irJAZh plants had significantly reduced nicotine levels. In addition, irJAZh spontaneously developed leaf necrosis during the transition to flowering. Because the lesions closely correlated with the elevated expression of programmed cell death genes and the accumulations of salicylic acid and hydrogen peroxide in the leaves, we propose a novel role of the NaJAZh protein as a repressor of necrosis and/or programmed cell death during plant development. PMID:22496510

  17. Tobacco plants transformed with the bean. alpha. ai gene express an inhibitor of insect. alpha. -amylase in their seeds. [Nicotiana tabacum; Tenebrio molitor

    Energy Technology Data Exchange (ETDEWEB)

    Altabella, T.; Chrispeels, M.J. (Univ. of California, San Diego, La Jolla (USA))

    1990-06-01

    Bean (Phaseolus vulgaris L.) seeds contain a putative plant defense protein that inhibits insect and mammalian but not plant {alpha}-amylases. We recently presented strong circumstantial evidence that this {alpha}-amylase inhibitor ({alpha}Al) is encoded by an already-identified lectin gene whose product is referred to as lectin-like-protein (LLP). We have now made a chimeric gene consisting of the coding sequence of the lectin gene that encodes LLP and the 5{prime} and 3{prime} flanking sequences of the lectin gene that encodes phytohemagglutinin-L. When this chimeric gene was expressed in transgenic tobacco (Nicotiana tabacum), we observed in the seeds a series of polypeptides (M{sub r} 10,000-18,000) that cross-react with antibodies to the bean {alpha}-amylase inhibitor. Most of these polypeptides bind to a pig pancreas {alpha}-amylase affinity column. An extract of the seeds of the transformed tobacco plants inhibits pig pancreas {alpha}-amylase activity as well as the {alpha}-amylase present in the midgut of Tenebrio molitor. We suggest that introduction of this lectin gene (to be called {alpha}ai) into other leguminous plants may be a strategy to protect the seeds from the seed-eating larvae of Coleoptera.

  18. The effect of methyl jasmonate and different chloride concentrations on photosynthetic pigments and proline content in Nicotiana tabacum L. cv. Cooker 347

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    Mahyar Moshtaghi

    2014-12-01

    Full Text Available Jasmonic acid and its methylated ester (methyl jasmonate, as natural plant growth regulators, are widely found in plants. In this study, the effect of methyl jasmonate (30 μM, in the early stages of Nicotiana tabacum L. cv. Cooker 347 growth, on photosynthetic pigments and proline was studied at high concentration of chloride. This experiment was performed in randomized complete block design with 4 replications, during 2011 crop year at Guilan Tobacco Research Center, outside greenhouse and under pot conditions. After transplanting similar seedlings to pots in rapid growth phase, the plants were irrigated with three levels chloride (50, 150 and 300 mg/L as CaCl2 both in presence and absence of methyl jasmonate for 4 weeks. Data analysis showed that by increasing Cl- concentrations up to 300 (mg/L, chloride and calcium accumulation of leaves increased, whilst photosynthetic pigments and proline decreased. Exogenous application of methyl jasmonate (30 μM significantly increased photosynthetic pigments in all treatments and proline in the 300 mg/L treatment. It seemed that the application of external methyl jasmonate (30 µM could improve tobacco plant injuries at high chloride concentration.

  19. Multivariate optimization of a headspace solid-phase microextraction method followed by gas chromatography with mass spectrometry for the determination of terpenes in Nicotiana langsdorffii.

    Science.gov (United States)

    Ardini, Francisco; Carro, Marina Di; Abelmoschi, Maria Luisa; Grotti, Marco; Magi, Emanuele

    2014-07-01

    A simple and sensitive procedure based on headspace solid-phase microextraction and gas chromatography with mass spectrometry was developed for the determination of five terpenes (α-pinene, limonene, linalool, α-terpineol, and geraniol) in the leaves of Nicotiana langsdorffii. The microextraction conditions (extraction temperature, equilibration time, and extraction time) were optimized by means of a Doehlert design. The experimental design showed that, for α-pinene and limonene, a low temperature and a long extraction time were needed for optimal extraction, while linalool, α-terpineol, and geraniol required a high temperature and a long extraction time. The chosen compromise conditions were temperature 60°C, equilibration time 15 min and extraction time 50 min. The main analytical figures of the optimized method were evaluated; LODs ranged from 0.07 ng/g (α-pinene) to 8.0 ng/g (geraniol), while intraday and interday repeatability were in the range 10-17% and 9-13%, respectively. Finally, the procedure was applied to in vitro wild-type and transgenic specimens of N. langsdorffii subjected to abiotic stresses (chemical and heat stress). With the exception of geraniol (75-374 ng/g), low concentration levels of terpenes were measured (ng/g level or lower); some interesting variations in terpene concentration induced by abiotic stress were observed.

  20. In vitro Anthelmintic Activity of Oily Extracts of Azadirachta indica and Aqueous Extracts of Nicotiana tabacum on Gastrointestinal Nematodes in Goats

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    Richard Zapata Salas

    2013-12-01

    Full Text Available Gastrointestinal nematodes are the most common parasites found in ruminants in the world. These parasites cause parasitic gastroenteritis and have a negative effect on productivity. Nematode control has been based on the use of anthelmintic chemicals—against which nematodes have developed a certain degree of resistance—which have been rated as residual substances in goat products that may cause adverse effects on the final consumer. As a result, the study of plant components has been proposed as a sustainable alternative to control nematodosis in goats. The anthelmintic potential of aqueous extracts of Nicotiana tabacum and oily extracts of Azadirachta indica on gastrointestinal nematodes affecting the goat chain was evaluated in vitro in this study. Nematicidal activity tests were performed on stool samples from goats with a high parasite load (trichostrongyles, by performing dose/ response curves. Percent inhibition in egg hatch for the aqueous extract of N. tabacum and the oily extract of A. indica was 99% and 80%, respectively. Extracts showed an effect on larva 3 (infective stage, with a mean lethal time of 8 ± 1 minutes for extracts of N. Tabacum, and of 8 ± 1 minutes for extract of A. indica. The in vitro results of the nematicidal activity show that N. tabacum and A. indica extracts can be a promising alternative for controlling nematodes in ruminants.

  1. Heterologous Expression Screens in Nicotiana benthamiana Identify a Candidate Effector of the Wheat Yellow Rust Pathogen that Associates with Processing Bodies

    Science.gov (United States)

    Petre, Benjamin; Saunders, Diane G. O.; Sklenar, Jan; Lorrain, Cécile; Krasileva, Ksenia V.; Win, Joe; Duplessis, Sébastien; Kamoun, Sophien

    2016-01-01

    Rust fungal pathogens of wheat (Triticum spp.) affect crop yields worldwide. The molecular mechanisms underlying the virulence of these pathogens remain elusive, due to the limited availability of suitable molecular genetic research tools. Notably, the inability to perform high-throughput analyses of candidate virulence proteins (also known as effectors) impairs progress. We previously established a pipeline for the fast-forward screens of rust fungal candidate effectors in the model plant Nicotiana benthamiana. This pipeline involves selecting candidate effectors in silico and performing cell biology and protein-protein interaction assays in planta to gain insight into the putative functions of candidate effectors. In this study, we used this pipeline to identify and characterize sixteen candidate effectors from the wheat yellow rust fungal pathogen Puccinia striiformis f sp tritici. Nine candidate effectors targeted a specific plant subcellular compartment or protein complex, providing valuable information on their putative functions in plant cells. One candidate effector, PST02549, accumulated in processing bodies (P-bodies), protein complexes involved in mRNA decapping, degradation, and storage. PST02549 also associates with the P-body-resident ENHANCER OF mRNA DECAPPING PROTEIN 4 (EDC4) from N. benthamiana and wheat. We propose that P-bodies are a novel plant cell compartment targeted by pathogen effectors. PMID:26863009

  2. Heterologous Expression Screens in Nicotiana benthamiana Identify a Candidate Effector of the Wheat Yellow Rust Pathogen that Associates with Processing Bodies.

    Directory of Open Access Journals (Sweden)

    Benjamin Petre

    Full Text Available Rust fungal pathogens of wheat (Triticum spp. affect crop yields worldwide. The molecular mechanisms underlying the virulence of these pathogens remain elusive, due to the limited availability of suitable molecular genetic research tools. Notably, the inability to perform high-throughput analyses of candidate virulence proteins (also known as effectors impairs progress. We previously established a pipeline for the fast-forward screens of rust fungal candidate effectors in the model plant Nicotiana benthamiana. This pipeline involves selecting candidate effectors in silico and performing cell biology and protein-protein interaction assays in planta to gain insight into the putative functions of candidate effectors. In this study, we used this pipeline to identify and characterize sixteen candidate effectors from the wheat yellow rust fungal pathogen Puccinia striiformis f sp tritici. Nine candidate effectors targeted a specific plant subcellular compartment or protein complex, providing valuable information on their putative functions in plant cells. One candidate effector, PST02549, accumulated in processing bodies (P-bodies, protein complexes involved in mRNA decapping, degradation, and storage. PST02549 also associates with the P-body-resident ENHANCER OF mRNA DECAPPING PROTEIN 4 (EDC4 from N. benthamiana and wheat. We propose that P-bodies are a novel plant cell compartment targeted by pathogen effectors.

  3. Facetas em dentes anteriores

    OpenAIRE

    Veloso, Helena Rafaela Lourenço Martins

    2015-01-01

    Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Medicina Dentária A presente revisão bibliográfica aborda as facetas estéticas em dentes anteriores, pela crescente valorização de um sorriso esteticamente agradável, facto que faz com que as pessoas procurem cada vez mais alternativas de tratamento para melhorar a aparência do seu sorriso. Os dentes anteriores são decisivos na aparência estética e, c...

  4. EMS & the DEA.

    Science.gov (United States)

    Beeson, Jeff; Ayres, Chris

    2010-01-01

    It's clear that EMS medical directors and management staff must be vigilant in their oversight of implementation, administration and monitoring of controlled substances within their agencies to best serve the public and avoid running afoul of investigation and incurring significant penalties. Those potentially affected by the need for individual registrations of both emergency vehicles and central inventory systems should carefully monitor upcoming developments in the interpretation od DEA regulations.

  5. CORANTES ARTIFICIAIS EM ALIMENTOS

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    Marcelo Alexandre PRADO

    2009-07-01

    Full Text Available

    O emprego de aditivos químicos é, sem dúvida, um dos mais polêmicos avanços alcançados pela indústria de alimentos. Os corantes artificiais pertencem a uma dessas classes de aditivos alimentares e têm sido objeto de muitas críticas, já que seu uso em muitos alimentos justifica-se apenas por questões de hábitos alimentares. Ainda existem diferentes opiniões quanto à inocuidade dos diversos corantes artificiais. Visando, principalmente, o controle no uso dos corantes sintéticos, mas tendo em vista que produtos coloridos artificialmente são exportados e importados, a análise desses aditivos requer métodos eficientes e rápidos para a detecção, identificação e quantificação. A cromatografia em papel e em camada delgada, apesar de serem técnicas relativamente rápidas, apresentam dados com baixa exatidão e precisão. Já na cromatografia líquida de alta eficiência (CLAE as maiores dificuldades encontram-se nas etapas de extração, mas principalmente no alto custo do equipamento. A eletroforese capilar apresenta os mesmos problemas da CLAE, aliados ao fato de se tratar de uma técnica relativamente recente para a análise desse tipo de substância e, portanto, existem poucos estudos a cerca da determinação e quantificação. PALAVRAS-CHAVE: Corantes artificiais; análise; legislação; CLAE; EC

  6. Bacharelado em Psicopedagogia

    OpenAIRE

    Ramos, Maria Beatriz Jacques; PUCRS

    2007-01-01

    Este texto mostra a história da implantação do Curso de Bacharelado em Psicopedagogia na FACED, PUCRS. Neste texto são apresentados os referenciais teóricos e dados sobre a atuação do psicopedagogo nos campos preventivo e terapêutico explicitando a relevância da formação de profissionais nesta área. Descritores – Psicopedagogia; aprendizagem; problema de aprendizagem.

  7. Ampliação da distribuição geográfica de Triatoma deaneorum Galvão, Souza & Lima 1967, nova denominação para Triatoma deanei (Hemiptera, Reduviidae New geographical distribution of Triatoma deaneorum Galvão, Souza & Lima 1967, new designation for Triatoma deanei (Hemiptera, Reduviidae

    Directory of Open Access Journals (Sweden)

    Cleber Galvão

    2001-12-01

    Full Text Available A distribuição geográfica de Triatoma deaneorum Galvão, Souza & Lima, 1967, nova denominação para Triatoma deanei, é ampliada baseada em um espécime recentemente encontrado no município de Chapada de Guimarães, próximo a Cuiabá, Mato Grosso, Brasil. A nova denominação é baseada no Código Internacional de Nomenclatura Zoológica.Authors extend the geographical distribution of Triatoma deaneorum Galvão, Souza & Lima, 1967, new designation for Triatoma deanei, based on a specimen recently found in Chapada dos Guimarães, near to Cuiabá, Mato Grosso, Brazil. The new designation is based on the International Code of Zoological Nomenclature.

  8. On subspecific taxonomy of <em>Microtus saviiem> (Rodentia, Arvicolidae

    Directory of Open Access Journals (Sweden)

    Longino Contoli

    2003-10-01

    Full Text Available Riassunto Sulla tassonomia sottospecifica di <em>Microtus saviiem> (Rodentia, Arvicolidae Viene riveduta e riassunta la situazione tassonomica sottospecifica di <em>Microtus (Terricola saviiem>, anche tramite la descrizione di due nuovi taxa: <em>Microtus (Terricola savii tolfetanusem>, dei Monti della Tolfa e <em>Microtus (Terricola savii niethammericusem>, del Gargano.

  9. How <em>Varroa> Parasitism Affects the Immunological and Nutritional Status of the Honey Bee, <em>Apis melliferaem>

    Directory of Open Access Journals (Sweden)

    Katherine A. Aronstein

    2012-06-01

    Full Text Available We investigated the effect of the parasitic mite <em>Varroa destructorem> on the immunological and nutritional condition of honey bees, <em>Apis melliferaem>, from the perspective of the individual bee and the colony. Pupae, newly-emerged adults and foraging adults were sampled from honey bee colonies at one site in S. Texas, USA. <em>Varroa>‑infested bees displayed elevated titer of Deformed Wing Virus (DWV, suggestive of depressed capacity to limit viral replication. Expression of genes coding three anti-microbial peptides (<em>defensin1, abaecin, hymenoptaecinem> was either not significantly different between <em>Varroa>-infested and uninfested bees or was significantly elevated in <em>Varroa>-infested bees, varying with sampling date and bee developmental age. The effect of <em>Varroa> on nutritional indices of the bees was complex, with protein, triglyceride, glycogen and sugar levels strongly influenced by life-stage of the bee and individual colony. Protein content was depressed and free amino acid content elevated in <em>Varroa>-infested pupae, suggesting that protein synthesis, and consequently growth, may be limited in these insects. No simple relationship between the values of nutritional and immune-related indices was observed, and colony-scale effects were indicated by the reduced weight of pupae in colonies with high <em>Varroa> abundance, irrespective of whether the individual pupa bore <em>Varroa>.

  10. <em>Lotus japonicusem> nodulation requires two GRAS-domain regulators, <em>NSP1em> and <em>NSP2em>

    DEFF Research Database (Denmark)

    Heckmann, Anne Birgitte Lau; Lombardo, Fabien; Miwa, Hiroki

    Signalling Pathway 1) gene (Ljnsp1-1) resulting in loss of the C-terminal 23 amino acids (aa) and we recently identified another mutant in NSP1 (Ljnsp1-2) with a truncated protein of 341 aa. Additionally, we have sequenced and isolated a mutant in LjNSP2 (Ljnsp2-3) that has a premature stop codon showing...... a similar phenotype to the Ljnsp1 mutants, whereas another mutant (Ljnsp2-4) showed increased nodulation. Both LjNSP1 and LjNSP2 are predicted GRAS (GAI, RGA, SCR) domain transcriptional regulators. Transcript steady-state levels of LjNSP1 and LjNSP2 initially decreased and then increased following...... infection by M. loti. In hairy root transformations, LjNSP1 and MtNSP1 complemented both Mtnsp1-1 and Ljnsp1-1 mutants, demonstrating that these orthologous proteins have a conserved biochemical function. A Nicotiana benthamiana NSP1-like gene (NbNSP1) was shown to restore nodule formation in both Ljnsp1...

  11. Synthesis and Spectroscopic Analysis of Novel 1<em>H-Benzo[d>]imidazoles Phenyl Sulfonylpiperazines

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    Amjad M. Qandil

    2012-05-01

    Full Text Available A group of benzimidazole analogs of sildenafil, 3-benzimidazolyl-4-methoxy-phenylsulfonylpiperazines 2–4 and 3-benzimidazolyl-4-methoxy-<em>N,N>-dimethyl- benzenesulfonamide (5, were efficiently synthesized. Compounds 2–5 were characterized by NMR and MS and contrary to the reported mass spectra of sildenafil, the spectra of the piperazine-containing compounds 2–4 showed a novel fragmentation pattern leading to an <em>m/z> = 316. A mechanism for the formation of this fragment was proposed.

  12. Isolation and Characterization of Microsatellite Markers in Brown Planthopper (<em>Nilaparvata em>lugens> Stål

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    Guangcun He

    2012-07-01

    Full Text Available Brown planthopper (<em>Nilaparvata lugensem> Stål (Homoptera: Delphacidae is an economically important pest on rice. In this study, 30 polymorphic microsatellite markers were developed from <em>N. lugensem> genomic libraries using the method of Fast Isolation by AFLP of Sequence Containing Repeats (FIASCO. Polymorphism of each locus was detected in 48 individuals from two natural populations. These microsatellite loci revealed 2 to 18 alleles, and the expected and observed heterozygosities ranged from 0.042 to 0.937 and from 0.042 to 0.958, respectively. These markers will be useful for the future study of this agricultural pest in population genetics and molecular genetics.

  13. Negative-Pressure Cavitation Extraction of Four Main Vinca Alkaloids from <em>Catharanthus em>roseus> Leaves

    OpenAIRE

    Yuangang Zu; Xiaorui Guo; Yujie Fu; Meng Luo; Wei Wang; Liuqing Yang; Fansong Mu

    2012-01-01

    In the present study, an improved method termed negative-pressure cavitation extraction (NPCE) followed by reverse phase high-performance liquid chromatography (RP-HPLC) was developed for the extraction and quantification of vindoline (VDL), catharanthine (CTR), vincristine (VCR) and vinblastine (VLB) from <em>Catharanthus roseus em>leaves. The optimized method employed 60-mesh particles, 80% ethanol, a negative pressure of −0.075 MPa, a solid to liquid ratio of 1:20, 30 min of ...

  14. Proximate Composition, Nutritional Attributes and Mineral Composition of <em>Peperomia> <em>pellucida> L. (Ketumpangan Air Grown in Malaysia

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    Maznah Ismail

    2012-09-01

    Full Text Available This study presents the proximate and mineral composition of <em>Peperomia> <em>pellucida> L., an underexploited weed plant in Malaysia. Proximate analysis was performed using standard AOAC methods and mineral contents were determined using atomic absorption spectrometry. The results indicated <em>Peperomia> <em>pellucida> to be rich in crude protein, carbohydrate and total ash contents. The high amount of total ash (31.22% suggests a high-value mineral composition comprising potassium, calcium and iron as the main elements. The present study inferred that <em>Peperomia> <em>pellucida> would serve as a good source of protein and energy as well as micronutrients in the form of a leafy vegetable for human consumption.

  15. Casca de soja em dietas para coelhos em crescimento

    Directory of Open Access Journals (Sweden)

    Ana Carolina Kohlrausch Klinger

    2015-01-01

    Full Text Available O presente trabalho teve como objetivo avaliar efeitos da inclusão de casca de soja em substituição ao feno de alfafa sobre o desempenho, características da carcaça e trato gastrointestinal e parâmetros bioquímicos de coelhos em fase de crescimento. Foram utilizados 30 coelhos da raça Nova Zelândia Branca, divididos igualmente em três tratamentos, compostos pela dieta experimental padrão (sem inclusão de casca de soja; e pelas dietas com 50% e 100% de substituição de alfafa por casca de soja nas respectivas rações experimentais. Concluiu-se que a casca de soja pode ser incluída em dietas para coelhos em crescimento, em substituição ao feno de alfafa em até 100% sem causar danos no desempenho nas características de carcaça e do trato gastrointestinal e parâmetros bioquímicos dos animais, durante a fase de crescimento. Porém, a utilização deste co-produto exige a necessidade de estudos mais aprofundados em virtude da escassez de dados.

  16. Antioxidant Profile of <em>Trifolium pratenseem> L.

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    Heidy Schwartsova

    2012-09-01

    Full Text Available In order to examine the antioxidant properties of five different extracts of <em>Trifolium pratenseem> L. (Leguminosae leaves, various assays which measure free radical scavenging ability were carried out: 1,1-diphenyl-2-picrylhydrazyl, hydroxyl, superoxide anion and nitric oxide radical scavenger capacity tests and lipid peroxidation assay. In all of the tests, only the H2O and (to some extent the EtOAc extracts showed a potent antioxidant effect compared with BHT and BHA, well-known synthetic antioxidants. In addition, <em>in vivo em>experiments were conducted with antioxidant systems (activities of GSHPx, GSHR, Px, CAT, XOD, GSH content and intensity of LPx in liver homogenate and blood of mice after their treatment with extracts of <em>T. pratenseem> leaves, or in combination with CCl4. Besides, in the extracts examined the total phenolic and flavonoid amounts were also determined, together with presence of the selected flavonoids: quercetin, luteolin, apigenin, naringenin and kaempferol, which were studied using a HPLC-DAD technique. HPLC-DAD analysis showed a noticeable content of natural products according to which the examined <em>Trifolium pratenseem> species could well be regarded as a promising new source of bioactive natural compounds, which can be used both as a food supplement and a remedy.

  17. Identification and Characterization of a <em>Bursaphelenchus xylophilusem> (Aphelenchida: Aphelenchoididae Thermotolerance-Related Gene: <em>Bx-HSP90em>

    Directory of Open Access Journals (Sweden)

    Danlei Li

    2012-07-01

    Full Text Available Temperatures directly influence the distribution and intensity of pine wilt disease caused by the pine wood nematode, <em>Bursaphelenchus xylophilusem>. To date, however, little is known about the causation and mechanism of this influence. The molecular chaperone HSP90 is a key component that contributes to survival in the abiotic stress response. In this study, we investigated the relationship between the survival of <em>B. xylophilusem> and the functionality of the HSP90 gene<em>.> <em>Bx-HSP90 em>was cloned from a suppression subtractive hybridization library. <em>In situem> mRNA hybridization showed that <em>Bx-HSP90em> was constitutively expressed in response to all of the temperatures tested, and RT-PCR indicated that all of the temperatures could induce <em>Bx-HSP90 em>transcription, with the highest transcript level detected at 30 °C. The suppression of the <em>Bx-HSP90em> transcript by RNA interference led to a 25% reduction in the number of nematodes at 30 °C after 44 h. Sharp declines in the survival of the RNAi-treated nematodes were observed after 8 days at 25 °C, 48 h at 30 °C and 24 h at 35 °C. Both heat shock and the knockdown of <em>Bx-HSP90 em>hindered the growth of the <em>B. xylophilusem> populations. The results indicate that <em>Bx-HSP90em> is essential for the survival of <em>B. xylophilusem>, confirming the thermoregulatory function of the gene, and delineate the timeframe and temperature range within which the gene function occurs.

  18. Effects of <em>Citrus aurantiumem> (Bitter Orange Fruit Extracts and <em>p>-Synephrine on Metabolic Fluxes in the Rat Liver

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    Rosane Marina Peralta

    2012-05-01

    Full Text Available The fruit extracts of <em>Citrus aurantiumem> (bitter orange are traditionally used as weight-loss products and as appetite supressants. An important fruit component is <em>p>-synephrine, which is structurally similar to the adrenergic agents. Weight-loss and adrenergic actions are always related to metabolic changes and this work was designed to investigate a possible action of the <em>C. aurantiumem> extract on liver metabolism. The isolated perfused rat liver was used to measure catabolic and anabolic pathways, including oxygen uptake and perfusion pressure. The <em>C. aurantiumem> extract and <em>p>-synephrine increased glycogenolysis, glycolysis, oxygen uptake and perfusion pressure. These changes were partly sensitive to a- and b-adrenergic antagonists. <em>p>-Synephrine (200 mM produced an increase in glucose output that was only 15% smaller than the increment caused by the extract containing 196 mM <em>p>-synephrine. At low concentrations the <em>C. aurantiumem> extract tended to increase gluconeogenesis, but at high concentrations it was inhibitory, opposite to what happened with <em>p>-synephrine. The action of the <em>C. aurantiumem> extract on liver metabolism is similar to the well known actions of adrenergic agents and can be partly attributed to its content in <em>p>-synephrine. Many of these actions are catabolic and compatible with the weight-loss effects usually attributed to <em>C. aurantiumem>.

  19. Nanodispositivos baseados em grafeno

    OpenAIRE

    José Eduardo Padilha de Sousa

    2012-01-01

    Nesta tese investigamos a partir de cálculos de primeiros princípios, dispositivos e componentes de dispositivos baseados em grafeno. Abordamos os campos da nanoeletrônica e da spintrônica. Dentro da nanoeletrônica investigamos: (i) propriedades de transporte de um nanotransistor de bicamada de grafeno na presença de um gate duplo. Demonstramos que sobre a ação de um campo elétrico externo, mesmo utilizando um gate da ordem de 10 nm, à temperatura ambiente e 4.5K uma corrente nula nunca é exi...

  20. Sade em Lacan

    OpenAIRE

    Riaviz, Eduardo

    2000-01-01

    Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Comunicação e Expressão. Esta dissertação analisa o lugar do Marquês de Sade no ensino de Lacan. Começamos por reconstruir a recepção de Sade pela vanguarda francesa, particularmente no movimento surrealista e nos seus dissidentes. Em seguida, acompanhamos as modificações do lugar de Sade na obra de Lacan. Para isto nos orientamos nos seis paradigmas do gozo através dos quais J.-A. Miller ordena a doutrina lacaniana...

  1. Flashes em Circuito

    Directory of Open Access Journals (Sweden)

    Ivete Lara Camargos Walty

    2002-12-01

    Full Text Available Reflexão sobre o papel da narrativa urbana no tratamento do tema da exclusão social, tendo como ponto de partida uma leitura do livro Passaporte, de Fernando Bonassi, que envolve a discussão do próprio gênero, a partir, sobretudo, da teoria do conto de Ricardo Piglia e de sua retomada das Seis propostas para o próximo milênio, de Ítalo Calvino, em artigo recente.

  2. Perspectives on EM metamaterials

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    Christophe Caloz

    2009-03-01

    Full Text Available Electromagnetic (EM metamaterials have become a field of intense research activities. This paper presents a critical perspective of the field, with emphasis on fundamental concepts and practical applications. Metamaterials are explained in the general context of periodic structures. Resonant particle type and transmission line type metamaterials are compared, and their fundamental connection is established. Exotic phenomena recently reported and associated challenges are briefly reviewed. Practical applications are enumerated and illustrated by an example. The paper concludes with an optimistic outlook regarding the future of metamaterials.

  3. Anestesia Diploica em Endodontia

    OpenAIRE

    Macedo, Ricardo Ribeiro Veiga de

    2013-01-01

    Trabalho final do 5º ano com vista à atribuição do grau de mestre no âmbito do ciclo de estudos de Mestrado Integrado em Medicina Dentária apresentado à Faculdade de Medicina da Universidade de Coimbra. Objetivos Comparar a eficácia das técnicas de anestesia convencionais, a anestesia infiltrativa periapical, com a anestesia diploica. Metodologia Foram selecionados 32 voluntários, saudáveis, aos quais foram administradas ambas as técnicas anestésicas no dente 1.4. Numa primeira fase os...

  4. Perossomus elumbis em bovino

    OpenAIRE

    1986-01-01

    Relato de um caso de distocia fetal em uma fêmea bovina mestiça Holandesa)Gir, provocado pela presença do monstro fetal Perossomus elumbis. Foi observado que o monstro fetal apresentava agenesia de vértebras lombares e coccígeas, com hi-poplasia da medula vertebral, terminando na região das últimas vértebras torácicas. A pelve apresentava-se hipoplásica com estreitamento do canal pélvico e anquilose dos membros posteriores e articulações da bacia, bem como atrofia muscular.  

  5. Monitoramento da suscetibilidade de populações de Triatoma sordida Stål, 1859 (Hemiptera: Reduviidae ao inseticida deltametrina, na região Centro-Oeste do Brasil Monitoring the susceptibility of Triatoma sordida Stål, 1859 (Hemiptera: Reduviidae to deltamethrin insecticide, in Central-Western Brazil

    Directory of Open Access Journals (Sweden)

    Marcos Takashi Obara

    2011-04-01

    Full Text Available INTRODUÇÃO: Populações de Triatoma sordida Stål, 1859 foram investigadas quanto à suscetibilidade à deltametrina. MÉTODOS: Análise por meio de bioensaios por aplicação tópica em 11 populações de T. sordida procedentes dos Estados de Goiás, Mato Grosso e Mato Grosso do Sul. RESULTADOS: As estimativas de DL50 e