Pertumbuhan Plantlet Anggrek Cattleya sp. dengan Perlakuan Benzyl Amino Purine pada Media Dasar Pupuk Daun Modifikasi

Directory of Open Access Journals (Sweden)

HESTIN YUSWANTI

2015-09-01

Full Text Available Growth of Plantlets of Cattleya Orchid on The Fooliar Fertilizer-Based Medium added with Benzyl Amino Purine. The aim of the current research was to investigate the appropriate concentrationof plant growth regulator BAP on the growth of Cattleya plantlet. The experiment was utilized a Randomized Completely Design with five treatments and six replications. The basal medium used was modification of foliar fertilizer of Growmore (trade mark with addition of fish emulsion, Vitamin B1 and active charcoal. BAP concentration used as treatment were 0 ppm, 0.5 ppm, 1.0 ppm, 1.5 ppm and 2.0 ppm. Variables observed were height, leaf number, root number, root length, fresh weight and dry weight of plantlets. The result showed that concentration of 1 ppm BAP resulted in the highest value for plantlet height (5.67 cm, leaf number (4.67, root length (2.07 cm, fresh weight (0.36 g and dry weight (0.043 g.

Plant defense induced in in vitro propagated banana (Musa paradisiaca) plantlets by Fusarium derived elicitors.

Science.gov (United States)

Patel, Miral; Kothari, I L; Mohan, J S S

2004-07-01

Perception of microbial signal molecules is part of the strategy evolved by plants to survive attacks by potential pathogens. To gain a more complete understanding of the early signaling events involved in these responses, we used fungal components of Fusarium under in vitro condition and checked the rise in signal molecule, salicylic acid (SA), and marker enzymes in defense reactions against the pathogen. SA level increased by 21 folds in elicitor treated plantlets as compared to that of control plantlets and there was marked increase in phenylalanine ammonia-lyase(PAL), peroxidase(POX), polyphenol oxidase(PPO) along with higher total phenolic content. Present results indicated that use of fungal components had successfully induced systemic resistance in in vitro cultured banana plantlets.

Effects of CO 2 concentration and moisture content of sugar-free media on the tissue-cultured plantlets in a large growth chamber

Science.gov (United States)

Qu, Y. H.; Lin, C.; Zhou, W.; Li, Y.; Chen, B.; Chen, G. Q.

2009-01-01

The dynamic fluctuations of CO 2 concentration in the tissue culture growth chamber after transplantation of petunia, chrysanthemum and tomato plantlets were recorded with a real-time control system to determine the critical CO 2 concentration levels of 35 μl l -1 at which CO 2 enrichment is needed. The experimental data showed that the tissue-cultured plantlets of petunia, chrysanthemum and tomato had the same CO 2 concentration dynamics. The results indicated that CO 2 enrichment was proper on the second day after transplantation. Petunia plantlets were used to conduct experiments under PPFD of 80 μmol m -2 s -1, and CO 2 concentrations of 350 ± 50 μl l -1, 650 ± 50 μl l -1 and 950 ± 50 μl l -1 as well as medium moisture contents of 60%, 70% and 80%, with the result that plantlets grew better under CO 2 concentration of 650 ± 50 μl l -1 than under the other two concentrations with all the different media water contents. Three media water contents under the same CO 2 concentration produced plantlets with the same quality. The impacts of CO 2 concentrations on plantlets are more important than those of the media water contents. Sugar-free tissue culture, as compared with the conventional culture, showed that CO 2 enrichment to 350 ± 50 μl l -1 can promote the growth of the cultured plantlets. Sugar-free tissue culture produced healthy plantlets with thick roots, almost equivalent to the common plantlets.

Comparative proteomic analysis of off-type and normal phenotype somatic plantlets derived from somatic embryos of Feijoa (Acca sellowiana (O. Berg) Burret).

Science.gov (United States)

Fraga, Hugo Pacheco de Freitas; Agapito-Tenfen, Sarah Zanon; Caprestano, Clarissa Alves; Nodari, Rubens Onofre; Guerra, Miguel Pedro

2013-09-01

Morphological disorders in a relevant portion of emerged somatic embryos have been a limiting factor in the true-to-type plantlet formation in Acca sellowiana. In this sense, the present study undertook a comparison between normal phenotype and off-type somatic plantlets protein profiles by means of the 2-D DIGE proteomics approach. Off-type and normal phenotype somatic plantlets obtained at 10 and 20 days conversion were evaluated. Results indicated 12 exclusive spots between normal and off-type plantlets at 10 days conversion, and 17 exclusive spots at 20 days conversion. Also at 20 days conversion, 4 spots were differentially expressed, up- or down-regulated. Two proteins related to carbohydrate metabolism were only expressed in off-types at 10 days conversion, suggesting a more active respiratory pathway. A vicilin-like storage protein was only found in off-types at 20 days conversion, indicating that plantlets may present an abnormality in the mobilization of storage compounds, causing reduced vigor in the development of derived plantlets. The presence of heat shock proteins were only observed during formation of normal phenotype somatic plantlets, indicating that these proteins may be involved in normal morphogenesis of plantlets formed. These new findings shed light on possible genetic or epigenetic mechanisms governing A. sellowiana morphogenesis. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

In vitro regeneration of hybrid plantlets of cashew (Anacardium ...

African Journals Online (AJOL)

Embryos from immature nuts of cashew (Anacardium occidentale L.) were cultured in vitro to regenerate improved hybrid plantlets. Explants (embryo) were excised from developing F1 hybrid immature nuts derived from diallel cross and harvested at 2-, 4-, 6- and 8-weeks after pollination (WAPo) for in vitro culture.

Eucalyptus plantlet growth in relation to foliar application with ...

African Journals Online (AJOL)

In one experiment at greenhouse, condition was established to determine the effects of complete fertilizers on growth of plantlet Eucalyptus camaldulensis in Southeast of Iran. This experiment was conducted in the Agricultural Research Center, University of Zabol, in autumn year of 2008. This study was designed as a ...

Effects of gamma radiation on in vitro plantlets of Alpinia purpurata

International Nuclear Information System (INIS)

Fereol, L.; Louis, S.; Luce, L.

1996-01-01

Two cultivars of Alpinia purpurata were propagated by in vitro culture to provide numerous convenient plantlets that were exposed to 0, 15, 30, or 45 Gy of gamma rays to study their radiosensitivity. The plants of the C1 and C2 clonal generations showed decreases in plant survival rate, fresh weight gain and micropropagation rate with increasing doses of gamma radiation. On the basis of survival, LD 50 was approximately 30 Gy. The 45 Gy dose drastically affects the different responses studied. It proved lethal or induced dwarf plants. Some plantlets, produced in vitro from treated plants, showed morphological abnormalities of leaves or of whole plants. This technique has potential for increasing the genetic variation in Alpinia purpurata. (author)

Use of slow-release fertilizer on the production of sweet potatoes plantlets in tray

Directory of Open Access Journals (Sweden)

Amarílis Beraldo Rós

2013-12-01

Full Text Available The production of plantlets in containers generally requires the use of fertilizers. The aim of this study was to evaluate the production of sweet potato in styrofoam trays using slow-release fertilizer. The experiment was carried out, under a screen-protected nursery, in a factorial scheme 5x5, with five doses of slow-release fertilizer NPK 19-06-10 (0, 50, 100, 150 e 200 g per 25 kg de substrate and five times of plantlets permanence in tray (14, 28, 42, 56 and 70 days after planting the cuttings. The number and dry matter of roots and leaves were evaluated. The number of roots was not influenced by fertilizer addition. In general, there is not damage to plantlets growth until the highest dose used. Therefore, the fertilizer addition increases the sweet potato plants growth and the dose of 200 g per 25 kg of substrate is responsible for the best results.

Ultrastructural and physiological responses of potato (Solanum tuberosum L.) plantlets to gradient saline stress

Science.gov (United States)

Gao, Hui-Juan; Yang, Hong-Yu; Bai, Jiang-Ping; Liang, Xin-Yue; Lou, Yan; Zhang, Jun-Lian; Wang, Di; Zhang, Jin-Lin; Niu, Shu-Qi; Chen, Ying-Long

2015-01-01

Salinity is one of the major abiotic stresses that impacts plant growth and reduces the productivity of field crops. Compared to field plants, test tube plantlets offer a direct and fast approach to investigate the mechanism of salt tolerance. Here we examined the ultrastructural and physiological responses of potato (Solanum tuberosum L. c.v. “Longshu No. 3”) plantlets to gradient saline stress (0, 25, 50, 100, and 200 mM NaCl) with two consequent observations (2 and 6 weeks, respectively). The results showed that, with the increase of external NaCl concentration and the duration of treatments, (1) the number of chloroplasts and cell intercellular spaces markedly decreased, (2) cell walls were thickened and even ruptured, (3) mesophyll cells and chloroplasts were gradually damaged to a complete disorganization containing more starch, (4) leaf Na and Cl contents increased while leaf K content decreased, (5) leaf proline content and the activities of catalase (CAT) and superoxide dismutase (SOD) increased significantly, and (6) leaf malondialdehyde (MDA) content increased significantly and stomatal area and chlorophyll content decline were also detected. Severe salt stress (200 mM NaCl) inhibited plantlet growth. These results indicated that potato plantlets adapt to salt stress to some extent through accumulating osmoprotectants, such as proline, increasing the activities of antioxidant enzymes, such as CAT and SOD. The outcomes of this study provide ultrastructural and physiological insights into characterizing potential damages induced by salt stress for selecting salt-tolerant potato cultivars. PMID:25628634

Plantlets from encapsulated shoot buds of Catalpa ovata G. Don

Directory of Open Access Journals (Sweden)

Halina Wysokińska

2014-01-01

Full Text Available Shoot buds isolated from in vitro shoot cultures of Catalpa ovata G. Don were encapsulated using 3% sodium alginate with sucrose (3% and 50 mM calcium chloride. The morphogenic response of encapsulated buds was affected by such factors, like composition of the media and the presence of growth regulators. The highest frequency of plantlet germination from encapsulated buds (70% within 4 weeks was obtained on Woody Plant medium (WP (Lloyd and McCown 1980 containing indole-3-butyric acid (IBA (1 mg/l. The process was substantially inhibited by cold-storage (4oC of encapsulated buds. In this case, the frequency response ranged from 3% to 22% dependent on storage period (28 or 42 days and the presence of the paraffin coat covering the alginate capsules. The plantlets developed from both unstored and stored encapsulated buds of C. ovata were transplanted to soil and grew in pots to phenotypically normal plants.

Modification of the 137Cs, 90Sr, and 60Co transfer to wheat plantlets by NH4+ fertilizers.

Science.gov (United States)

Guillén, J; Muñoz-Muñoz, G; Baeza, A; Salas, A; Mocanu, N

2017-03-01

Inorganic fertilizers are used as agricultural countermeasures intended to inhibit the soil to plant transfer of radionuclides after a radioactive fallout. Two NH 4 + fertilizers, diammonium phosphate (DAP) and NPK, were applied to soil contaminated with a mixture of radionuclides to analyze whether they modify the transfer of 137 Cs, 90 Sr, and 60 Co and stable elements (K, Na, Ca, and Mg) to wheat plantlets grown under controlled laboratory conditions. DAP introduced NH 4 + in the soil, which can increase 137 Cs transfer, while NPK also introduced K + , which can decrease it. The application of DAP increased the accumulation of 137 Cs in wheat plantlets with increasing application rate, so did the 137 Cs/K in plantlets. Regarding the NPK application, the 137 Cs increased in all treatments, but at maximum rate, the available K introduced by the fertilizer was probably able to partially satisfy the nutritional requirements of the wheat plantlet and the 137 Cs decreased relative to the recommended rate. The 137 Cs/K ratio in plantlet decreased with increasing NPK rates. The transfer of 90 Sr increased with increasing DAP rate and only at the maximum NPK rate. The 60 Co transfer only increased at the maximum application rates for DAP and NPK. These modifications should be considered when using these fertilizers as agricultural countermeasures.

Diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple plantlets during acclimatization

Directory of Open Access Journals (Sweden)

Aurilena de Aviz Silva

Full Text Available ABSTRACT: This study examines the effect of inoculation with diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple cv. 'Vitória' plantlets during the acclimatization period. The experiment was carried out in a greenhouse in Campos dos Goytacazes, in randomized blocks, using a 2x5x5 factorial scheme, with the factors being two types of inocula (absence or presence of a mixture of diazotrophic bacteria that contained Burkholderia sp. UENF 114111, Burkholderia silvatlantica UENF 11711, and Herbaspirillum seropedicae HRC 54, five levels of urea (0, 2, 5, 7, and 10g L-1, and five evaluation periods (30, 60, 90, 120, and 150 days following the planting of plantlets in trays, and with four replications of each treatment. An increase in the dried weight of shoots and roots of micropropagated pineapple cv. 'Vitória' plants depended on inoculation with diazotrophic bacteria, and plantlets acclimatization could be reduced by up to 57 days, depending on the concentration of urea fertilizer.

Ultrastructural and physiological responses of potato (Solanum tuberosum L. plantlets to gradient saline stress

Directory of Open Access Journals (Sweden)

Hui-Juan eGao

2015-01-01

Full Text Available Salinity is one of the major abiotic stresses that impacts plant growth and reduces the productivity of field crops. Compared to field plants, test tube plantlets offer a direct and fast approach to investigate the mechanism of salt tolerance. Here we examined the ultrastructural and physiological responses of potato (Solanum tuberosum L. c.v. ‘Longshu No. 3’ plantlets to gradient saline stress (0, 25, 50, 100 and 200 mM NaCl with two consequent observations (two and six weeks, respectively. The results showed that, with the increase of external NaCl concentration and the duration of treatments, (1 the number of chloroplasts and cell intercellular spaces markedly decreased, (2 cell walls were thickened and even ruptured, (3 mesophyll cells and chloroplasts were gradually damaged to a complete disorganization containing more starch, (4 leaf Na and Cl contents increased while leaf K content decreased, (5 leaf proline content and the activities of catalase (CAT and superoxide dismutase (SOD increased significantly, and (6 leaf malondialdehyde (MDA content increased significantly and stomatal area and chlorophyll content decline were also detected. Severe salt stress (200 mM NaCl inhibited plantlet growth. These results indicated that potato plantlets adapt to salt stress to some extent through accumulating osmoprotectants, such as proline, increasing the activities of antioxidant enzymes, such as CAT and SOD. The outcomes of this study provide ultrastructural and physiological insights into characterizing potential damages induced by salt stress for selecting salt-tolerant potato cultivars.

Competitive-IgY- Enzyme Linked Immuno Sorbent Assay (CIgY-ELISA to detect the cytokinins in Gerbera jamesonii plantlets

Directory of Open Access Journals (Sweden)

Cleiton Mateus Sousa

2011-08-01

Full Text Available A competitive hyper-immune yolk Immunoglobulin Y - Enzyme Linked Immune Sorbent Assay (CIgY-ELISA, was developed as an alternative method to detect zeatin and 2ip in plantlets of gerbera. The endogenous level of hormones in the plantlets in vitro of gerbera with one or six weeks after replication was determined with competitive IgY-ELISA set to detect between 1 and 100 pmoles of plant hormone for each 1.0 g tissue. The plantlets of six weeks presented sprouts and root, while the plantlets of one week presented only sprouts. The CIgY-ELISA was set with high independent variables values of sensitivity/specificity of 96/89 % for zeatin and 94/78 % for 2ip, with high values of reproducibility (up to 90 % for both the cytokinins. Zeatin content varied from 2.2 to 2.8 pmoles.g-1 and from 2.7 to 3.3 pmoles.g-1 on the plantlet with one and six weeks, respectively. The 2ip content did not vary and was detected near the detection limit in all the assays. It was concluded that the observed capabilities of CIgY-ELISA were putative and the competitive assay was a highly robust and stable method, which could be used for the studies on plant physiology for endogenous cytokinins.

Molecular and biochemical characterization in Rauvolfia tetraphylla plantlets grown from synthetic seeds following in vitro cold storage.

Science.gov (United States)

Faisal, Mohammad; Alatar, Abdularhaman A; Hegazy, Ahmad K

2013-01-01

Synseed technology is one of the most important applications of plant biotechnology for in vitro conservation and regeneration of medicinal and aromatic plants. In the present investigation, synseeds of Rauvolfia tetraphylla were produced using in vitro-proliferated shoots upon complexation of 3 % sodium alginate and 100 mM CaCl(2). The encapsulated buds were stored at 4, 8, 12, and 16 °C and high conversion was observed in synseeds stored at 4 °C for 4 weeks. The effect of different medium strength on in vitro conversion response of synseed was evaluated and the maximum conversion (80.6 %) into plantlets was recorded on half-strength woody plant medium supplemented with 7.5 μM 6-benzyladenine and 2.5 μM α-naphthalene acetic acid after 8 weeks of culture. Plantlets with well-developed shoot and roots were hardened and successfully transplanted in field condition. After 4 weeks of transfer to ex vitro conditions, the performance of synseed-derived plantlets was evaluated on the basis of some physiological and biochemical parameters and compared with the in vivo-grown plants. Short-term storage of synthetic seeds at low temperature had no negative impact on physiological and biochemical profile of the plants that survived the storage process. Furthermore, clonal fidelity of synseed-derived plantlets was also assessed and compared with mother plant using rapid amplified polymorphic DNA and inter-simple sequence repeats analysis. No changes in molecular profiles were found among the regenerated plantlets and comparable to mother plant, which confirm the genetic stability among clones. This synseed protocol could be useful for in vitro clonal multiplication, conservation, and short-term storage and exchange of germplasm of this antihypertensive drug-producing plant.

Proteomic Analysis of Responsive Proteins Induced in Japanese Birch Plantlet Treated with Salicylic Acid

Directory of Open Access Journals (Sweden)

Hiromu Suzuki

2014-07-01

Full Text Available The present study was performed to unravel the mechanisms of systemic acquired resistance (SAR establishment and resistance signaling pathways against the canker-rot fungus (Inonotus obliquus strain IO-U1 infection in Japanese birch plantlet No.8. Modulation of protein-profile induced by salicylic acid (SA-administration was analyzed, and SA-responsive proteins were identified. In total, 5 specifically expressed, 3 significantly increased, and 3 significantly decreased protein spots were identified using liquid chromatography/tandem mass spectrometry (LC/MS/MS and the sequence tag method. These proteins were malate dehydrogenase, succinate dehydrogenase, phosphoglycerate kinase, diaminopimalate decarboxylase, arginase, chorismate mutase, cyclophilin, aminopeptidase, and unknown function proteins. These proteins are considered to be involved in SAR-establishment mechanisms in the Japanese birch plantlet No 8.

Reversion of hyperhydricity in pink (Dianthus chinensis L.) plantlets by AgNO3 and its associated mechanism during in vitro culture.

Science.gov (United States)

Gao, Hongyang; Xia, Xiuying; An, Lijia; Xin, Xin; Liang, Yuan

2017-01-01

Hyperhydricity occurs frequently in plant tissue culture and can severely affect commercial micropropagation and genetic improvement of the cultured plantlets. Hyperhydric shoots are charaterzized by high water content, but how this occurs is still a subject of investigation. Silver ion (Ag + ) can reduce the extent of hyperhydricity in plants, but its effect on the reversion of hyperhydric plantlets and the underlying mechanism of reversion has not been clarified. In this study, about 67% of the hyperhydric Dianthus chinensis L. plantlets were found to revert to normal condition when the plantlets were cultured in medium supplemented with 29.4μmolL -1 AgNO 3 . Water content and hydrogen peroxide (H 2 O 2 ) content in the guard cells of these plantlets were reduced, while stomatal aperture and water loss rate were increased. AgNO 3 also reduced the content of endogenous ethylene and expression of ethylene synthesis and ethylene signal transduction-associated genes. Reduced accumulation of ethylene consequently led to an increase in stomatal aperture mediated by decreased H 2 O 2 content in the guard cells. These results adequately verified the role of AgNO 3 in the reversion of hyperhydricity in D. chinensis L. and also provided clues for exploring the cause of excessive water accumulation in hyperhydric plants. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Procedure for identifying and prevent risks in in vitro sugarcane plantlets marketing

Directory of Open Access Journals (Sweden)

Ana Rosa Hernández Freire

2014-10-01

Full Text Available Seed Banks and agricultural farms benefit greatly from plant in vitro Culture techniques, for their rapid propagation processes and the production of desease-free plantlets. However, the production and commercialization process can fail and it is not as successful l as it was supposed to. The objective of this study is to identify potential failure modes at the acclimatiztion stage of plantlets to ex vitro conditions at Estación Territorial de Investigaciones de la Caña de Azúcar de Villa Clara (ETICA. A methodology of Failure Mode and Effects Analysis (FMEA was applied. A value of 80 or higher was determined to be the Risk Priority Number (RPN and the more relevant failure modes were defined a t the acclimatization and commercialization stages and ground rules and assumptions were established in order to prevent and control failure modes of the process. The study and implementation of the methodology to other processes at ETICA is strongly recommended.

Signal mediators at induction of heat resistance of wheat plantlets by short-term heating

Directory of Open Access Journals (Sweden)

Yu. V. Karpets

2015-12-01

Full Text Available The effects of functional interplay of calcium ions, reactive oxygen species (ROS and nitric oxide (NO in the cells of wheat plantlets roots (Triticum aestivum L. at the induction of their heat resistance by a short-term influence of hyperthermia (heating at the temperature of 42 °С during 1 minute have been investigated. The transitional increase of NO and H2O2 content, invoked by heating, was suppressed by the treatment of plantlets with the antagonists of calcium EGTA (chelator of exocellular calcium, lanthanum chloride (blocker of calcium channels of various types and neomycin (inhibitor of phosphatidylinositol-dependent phospholipase C. The rise of hydrogen peroxide content, caused by hardening, was partially suppressed by the action of inhibitors of nitrate reductase (sodium wolframate and NO-synthase (NG-nitro-L-arginine methyl ester – L-NAME, and the increasing of nitric oxide content was suppressed by the treatment of plants with the antioxidant ionol and with the scavenger of hydrogen peroxide (dimethylthiourea. These compounds and antagonists of calcium also partially removed the effect of the rise of plantlets’ heat resistance, invoked by hardening heating. The conclusion on calcium’s role in the activation of enzymatic systems, generating reactive oxygen species and nitric oxide, and on the functional interplay of these signal mediators at the induction of heat resistance of plantlets by hardening heating is made.

Using the "Kalanchoe daigremontiana" Plant To Show the Effects of Photoperiodism on Plantlet Formation.

Science.gov (United States)

Hershey, David R.

2002-01-01

Describes an activity demonstrating the importance of photoperiod on plant development. Uses the plant devil's backbone for the experiment and studies the details of photoperiodic requirement for plantlet formation. (Contains 12 references.) (YDS)

Gamma irradiation as activator of antioxidant activity and essential oil contents in lavender (Lavandula multifida) plantlets

International Nuclear Information System (INIS)

El-Naggar, H.A.; Atallah, R.K.; Aly, A.A.; Maraei, R.W.

2012-01-01

This study was conducted to evaluate the stimulation effect of γ-irradiation on the chemical composition of essential oils, total phenolic compounds, flavonoid contents and antioxidant activities in lavender plantlets (Lavandula multifida) at three multiplication stages. Lavender plantlets were irradiated using different γ- irradiation dose levels (0.0, 5, 15, 30, 45, 60 and 75 Gy). After irradiation; plantlets were sectioned to start the multiplication stage (three subcultures). Increasing irradiation dose levels at multiplication stages significantly increased the total phenolic content and reached to the maximum increment at the dose level of 75 Gy (26.88 g/100 g DW) in zero time stage in comparison with the untreated plantlets (7.250 g/100 g DW). The highest content of flavonoids (21.50 g/100 g DW) was detected at dose level of 75 Gy at zero time stage (M0). The highest applied irradiation dose of 75 Gy gave the highest reducing power activity compared with control at zero time stage (M0). Scavenging activity by DPPH was increased gradually by increasing irradiation dose levels in all multiplication stages until the high dose of 75 Gy which gave the maximum scavenging activity (91.05%) in zero time stage. Also, there was a significant increase in antioxidant activity on linoleic acid system with increasing the dose of γ-irradiation level. The application of γ-irradiation at dose level of 15 Gy and M3 stage produced the highest value of essential oil content (0.12%), followed by 5 Gy treatments (0.082%). The most increased volatile oil compounds by γ-irradiation were; limonene which increased from 4.87% to 5.37% at 0.0 and 5 Gy, respectively and linalool increased from 86.07% to 91.5% at 0.0 and 15 Gy respectively. The present study suggests that γ-irradiation led to increase antioxidant activities of lavender plantlets by increasing the availability of free polyphenolic compounds and also the content of volatile oil. This shows that lavender plants may be

Potato transformation and potato cyst nematode infection on potato plantlets in tissue culture

Science.gov (United States)

These two protocols describe the methods for generating transgenic potato plants and for evaluating potato cyst nematode (Globodera rostochiensis and G. pallida) infection on potato plantlets in tissue culture. These methods are useful tools that can be used in the study of the interactions between ...

Different exogenous sugars affect the hormone signal pathway and sugar metabolism in "Red Globe" (Vitis vinifera L.) plantlets grown in vitro as shown by transcriptomic analysis.

Science.gov (United States)

Mao, Juan; Li, Wenfang; Mi, Baoqin; Dawuda, Mohammed Mujitaba; Calderón-Urrea, Alejandro; Ma, Zonghuan; Zhang, Yongmei; Chen, Baihong

2017-09-01

Exogenously applied 2% fructose is the most appropriate carbon source that enhances photosynthesis and growth of grape plantlets compared with the same concentrations of sucrose and glucose. The role of the sugars was regulated by the expression of key candidate genes related to hormones, key metabolic enzymes, and sugar metabolism of grape plantlets ( Vitis vinifera L.) grown in vitro. The addition of sugars including sucrose, glucose, and fructose is known to be very helpful for the development of grape (V. vinifera L.) plantlets in vitro. However, the mechanisms by which these sugars regulate plant development and sugar metabolism are poorly understood. In grape plantlets, sugar metabolism and hormone synthesis undergo special regulation. In the present study, transcriptomic analyses were performed on grape (V. vinifera L., cv. Red Globe) plantlets in an in vitro system, in which the plantlets were grown in 2% each of sucrose (S20), glucose (G20), and fructose (F20). The sugar metabolism and hormone synthesis of the plantlets were analyzed. In addition, 95.72-97.29% high-quality 125 bp reads were further analyzed out of which 52.65-60.80% were mapped to exonic regions, 13.13-28.38% to intronic regions, and 11.59-28.99% to intergenic regions. The F20, G20, and S20 displayed elevated sucrose synthase (SS) activities; relative chlorophyll contents; Rubisco activity; and IAA and zeatin (ZT) contents. We found F20 improved the growth and development of the plantlets better than G20 and S20. Sugar metabolism was a complex process, which depended on the balanced expression of key potential candidate genes related to hormones (TCP15, LOG3, IPT3, ETR1, HK2, HK3, CKX7, SPY, GH3s, MYBH, AGB1, MKK2, PP2C, PYL, ABF, SnRK, etc.), key metabolic enzymes (SUS, SPS, A/V-INV, and G6PDH), and sugar metabolism (BETAFRUCT4 and AMY). Moreover, sugar and starch metabolism controls the generation of plant hormone transduction pathway signaling molecules. Our dataset advances our

Comparative physiological and proteomic analyses of poplar (Populus yunnanensis plantlets exposed to high temperature and drought.

Directory of Open Access Journals (Sweden)

Xiong Li

Full Text Available Plantlets of Populus yunnanensis Dode were examined in a greenhouse for 48 h to analyze their physiological and proteomic responses to sustained heat, drought, and combined heat and drought. Compared with the application of a single stress, simultaneous treatment with both stresses damaged the plantlets more heavily. The plantlets experienced two apparent response stages under sustained heat and drought. During the first stage, malondialdehyde and reactive oxygen species (ROS contents were induced by heat, but many protective substances, including antioxidant enzymes, proline, abscisic acid (ABA, dehydrin, and small heat shock proteins (sHSPs, were also stimulated. The plants thus actively defended themselves against stress and exhibited few pathological morphological features, most likely because a new cellular homeostasis was established through the collaborative operation of physiological and proteomic responses. During the second stage, ROS homeostasis was overwhelmed by substantial ROS production and a sharp decline in antioxidant enzyme activities, while the synthesis of some protective elements, such as proline and ABA, was suppressed. As a result, photosynthetic levels in P. yunnanensis decreased sharply and buds began to die, despite continued accumulation of sHSPs and dehydrin. This study supplies important information about the effects of extreme abiotic environments on woody plants.

Neubauer's plantlet method - an alternative procedure for evaluating the effectiveness of potassium based fertilizers in reducing radiocaesium from soil to plant

International Nuclear Information System (INIS)

Mocanu, N.; Cotarlea, M.; Boldor, O.

1996-01-01

Accidental releases of radiocaesium into the environment have necessitated the search for effective soil-based countermeasures to reduce its transfer along food-chains. As field experiments can be impractical and protracted to predict rapidly the effectiveness of chemical treatments of radioactive contaminated soils to reduce soil-to-plant transfer of radionuclides, laboratory experiments are a suitable alternative for rapid evaluation of the most appropriate countermeasure to apply under a range of different circumstances in the event of accidental radioactive contamination of agricultural lands. Taking into account these considerations our study focuses on the use of a laboratory experiment based on Neubauer's plantlet method to evaluate the effectiveness of potassic salt 30 % applied on two soil types (alluvial and brown-reddish forest type) contaminated with 137 Cs for reducing uptake of the radionuclide to wheat plants grown-up on these soils. The experimental results evidence diminished values for 137 Cs/K quotient in wheat plantlets grown-up on soils treated with potassic salt 30 %, compared to those registered for wheat plantlets grown-up on untreated soils. The diminished values of 137 Cs/K quotient result from the reduced uptake of 137 Cs to plantlets accompanied by the enhanced uptake of potassium. (author)

Cosmeceuticals based on Rhealba(®) Oat plantlet extract for the treatment of acne vulgaris.

Science.gov (United States)

Fabbrocini, G; Saint Aroman, M

2014-12-01

Recent evidence suggests that acne vulgaris begins as an inflammation in and around the sebaceous gland and alterations in the lipid content of sebum, which drive hyperproliferation and increased desquamation of keratinocytes within sebaceous follicles. This prevents sebum drainage, causing the formation of microcomedones, which spontaneously regress or become acne lesions when the pilosebaceous unit is further blocked by the accumulation of corneocytes. These conditions are favourable for the proliferation of Propionibacterium acnes, which further aggravates acne by enhancing abnormal desquamation, sebum production and inflammation. Also, skin fragility due to inflammation or irritation by anti-comedogenic agents can worsen the situation. Rhealba(®) Oat plantlet extract (Pierre Fabre Dermo Cosmetique) soothes and restores fragile skin in acne by reducing inflammation and inhibits bacterial adhesion of Propionibacterium acnes. Cosmeceuticals combining Rhealba(®) Oat plantlet extract and hydro-compensating actives, which are available with or without anti-comedogenic hydroxy acids, provide a balanced, multifaceted approach for acne patients. © 2014 European Academy of Dermatology and Venereology.

Assessment of genetic fidelity in Rauvolfia serpentina plantlets grown from synthetic (encapsulated) seeds following in vitro storage at 4 °C.

Science.gov (United States)

Faisal, Mohammad; Alatar, Abdulrahman A; Ahmad, Naseem; Anis, Mohammad; Hegazy, Ahmad K

2012-05-03

An efficient method was developed for plant regeneration and establishment from alginate encapsulated synthetic seeds of Rauvolfia serpentina. Synthetic seeds were produced using in vitro proliferated microshoots upon complexation of 3% sodium alginate prepared in Llyod and McCown woody plant medium (WPM) and 100 mM calcium chloride. Re-growth ability of encapsulated nodal segments was evaluated after storage at 4 °C for 0, 1, 2, 4, 6 and 8 weeks and compared with non-encapsulated buds. Effects of different media viz; Murashige and Skoog medium; Lloyd and McCown woody Plant medium, Gamborg’s B5 medium and Schenk and Hildebrandt medium was also investigated for conversion into plantlets. The maximum frequency of conversion into plantlets from encapsulated nodal segments stored at 4 °C for 4 weeks was achieved on woody plant medium supplement with 5.0 μM BA and 1.0 μM NAA. Rooting in plantlets was achieved in half-strength Murashige and Skoog liquid medium containing 0.5 μM indole-3-acetic acid (IAA) on filter paper bridges. Plantlets obtained from stored synseeds were hardened, established successfully ex vitro and were morphologically similar to each other as well as their mother plant. The genetic fidelity of Rauvolfia clones raised from synthetic seeds following four weeks of storage at 4 °C were assessed by using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. All the RAPD and ISSR profiles from generated plantlets were monomorphic and comparable to the mother plant, which confirms the genetic stability among the clones. This synseed protocol could be useful for establishing a particular system for conservation, short-term storage and production of genetically identical and stable plants before it is released for commercial purposes.

Influence of arbuscular mycorrhizas on the growth rate of mist-propagated olive plantlets

OpenAIRE

Porras Piedra, A.; Soriano Martín, M.L.; Porras Soriano, A.; Fernández Izquierdo, G.

2005-01-01

The aim of this work was to determine the influence of early inoculation of semi-woody olive cuttings (cv. Cornicabra), cultivated under mist propagation conditions, with arbuscular mycorrhizal fungi. A strongly positive response was seen to root inoculation with the three fungal species tested —Glomus mosseae, G. intraradices and G. claroideum— both in nursery phase plantlets, and in older plants grown in 5 L pots.

Acclimatization and leaf anatomy of micropropagated fig plantlets Aclimatização e anatomia foliar de plântulas de figo micropropagadas

Directory of Open Access Journals (Sweden)

Chrystiane Fráguas Chirinéa

2012-12-01

Full Text Available The survival of micropropagated plants during and after acclimatization is a limiting process to plant establishment. There is little information on how the anatomy of vegetative organs of Ficus carica can be affected by culture conditions and acclimatization. The present research aimed to study the effects of time on culture medium and substrates during the acclimatization of fig tree plantlets produced in vitro, characterizing some leaf anatomy aspects of plantlets cultured in vitro and of fig trees produced in field. Plantlets previously multiplied in vitro were separated and transferred into Wood Plant Medium (WPM where they were kept for 0, 15, 30, 45 and 60 days. Different substrates were tested and studies on leaf anatomy were performed in order to compare among plantlets grown in vitro, plantlets under 20, 40 and 60 days of acclimatization, and field grown plants. Keeping plantlets for 30 days in WPM allowed better development in Plantmax during acclimatization. Field grown plants presented higher number of stomata, greater epicuticular wax thickness and greater leaf tissue production compared to in vitro ones. The leaf tissues of in vitro plantlets show little differentiation and have great stomata number compared with acclimatized plants, which reduce the number of stomata during the acclimatization process.A sobrevivência de plantas micropropagadas durante e após a aclimatização é um processo limitante para o estabelecimento de plantas. Poucas informações são encontradas de como a anatomia de órgãos vegetativos de Ficus carica pode ser afetada pelas condições de cultivo e aclimatização. O presente trabalho teve por objetivo estudar os efeitos do tempo em meio de cultura e substratos durante a aclimatização de plântulas de figueira produzidas in vitro e cultivadas no campo. Plântulas multiplicadas in vitro foram separadas e transferidas para meio WPM, onde foram mantidas por 0; 15; 30; 45 e 60 dias. Diferentes

Effects of ventilation and sucrose concentrations on the growth and plantlet anatomy of micropropagated persian walnut plants

Science.gov (United States)

Plantlets grown in conventional tissue culture systems usually encounter physiological and anatomical abnormalities including inability to photosynthesize, low chlorophyll content, open stomata, lack of a cuticle layer in the leaf, abnormal xylem parenchyma etc. Photoautotrophic and photomixotrophic...

Effect of two vesicular-arbuscular mycorrhizal fungi on the growth of micropropagated potato plantlets and on the extent of disease caused by Rhizoctonia solani.

Science.gov (United States)

Yao, M K; Tweddell, R J; Désilets, H

2002-10-01

Two micropropagated potato cultivars, Goldrush and LP89221, were inoculated into sowing trays with either Glomus etunicatum or G. intraradices in a greenhouse. After 2 weeks, plantlets were transplanted into pots and roots were challenged 7 days later with Rhizoctonia solani. At different times after R. solani infection, disease severity, mortality rate, root colonization levels, various growth parameters, and shoot mineral content were evaluated. In Goldrush, only inoculation with G. etunicatum led to a significant reduction in disease severity, ranging between 60.2% and 71.2%, on both shoot and crown. This decrease was not observed in LP89221. Compared with the control plantlets, inoculation of Goldrush with G. etunicatum or G. intraradices reduced significantly the mortality rate by 77% and 26%, respectively, whereas vesicular-arbuscular mycorrhizal (VAM) fungi did not significantly influence the mortality rate in LP89221. In Goldrush, inoculation with G. etunicatum significantly increased shoot fresh weight, root dry weight and the number of tubers produced per plant, whereas G. intraradices only significantly increased the number of tubers. Tuber and root fresh weights of both potato cultivars were significantly reduced by R. solani infection. However, R. solani-infected plantlets of both Goldrush and LP89221, inoculated with G. etunicatum, produced significantly greater tuber fresh weight than non-VAM plantlets. In R. solani-infected plantlets of Goldrush but not LP89221, G. etunicatum and G. intraradices increased root fresh weight by approximately 140.3% and 76.5%, respectively, compared with non-VAM plants. The potato cultivars Goldrush and LP89221 responded differently to VAM fungal inoculation and to R. solani infection in terms of shoot mineral content.

Effect of cadmium on phenolic compounds, antioxidant enzyme activity and oxidative stress in blueberry (Vaccinium corymbosum L.) plantlets grown in vitro.

Science.gov (United States)

Manquián-Cerda, K; Escudey, M; Zúñiga, G; Arancibia-Miranda, N; Molina, M; Cruces, E

2016-11-01

Cadmium (Cd(2+)) can affect plant growth due to its mobility and toxicity. We evaluated the effects of Cd(2+) on the production of phenolic compounds and antioxidant response of Vaccinium corymbosum L. Plantlets were exposed to Cd(2+) at 50 and 100µM for 7, 14 and 21 days. Accumulation of malondialdehyde (MDA), hydrogen peroxide (H2O2) and the antioxidant enzyme SOD was determined. The profile of phenolic compounds was evaluated using LC-MS. The antioxidant activity was measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and the ferric reducing antioxidant power test (FRAP). Cd(2+) increased the content of MDA, with the highest increase at 14 days. The presence of Cd(2+) resulted in changes in phenolic compounds. The main phenolic compound found in blueberry plantlets was chlorogenic acid, whose abundance increased with the addition of Cd(2+) to the medium. The changes in the composition of phenolic compounds showed a positive correlation with the antioxidant activity measured using FRAP. Our results suggest that blueberry plantlets produced phenolic compounds with reducing capacity as a selective mechanism triggered by the highest activity of Cd(2+). Copyright © 2016 Elsevier Inc. All rights reserved.

LONG-TERM PRECONDITIONING OF PLANTLETS: A PRACTICAL METHOD FOR ENHANCING SURVIVAL OF PINEAPPLE (Ananas comosus Merr.) SHOOT TIPS CRYOPRESERVED USING VITRIFICATION.

Science.gov (United States)

Hu, W H; Liu, S F; Liaw, S I

2015-01-01

The purpose of this study was to develop an efficient cryopreservation protocol for pineapple (Ananas comosus Merr.) shoot tips. The optimal state of pineapple plantlets was investigated by using sucrose preconditioning to enhance survival after cryostorage. To achieve a suitable state of plantlets before cryopreservation, 0.2 M to 0.4 M sucrose concentrations combined with short- (0-7 days), medium- (15-30 days), and long-term (75-150 days) preconditioning periods were compared. The highest survival (100 %) was achieved using the following procedure: intact plantlets underwent long-term preconditioning with 0.2 M sucrose for 135 days, dissected shoot tips were treated with a loading solution containing 2.0 M glycerol + 0.4 M sucrose for 60 min at 25 degree and the shoot tips were dehydrated in PVS2 for 2h at 0 degree C before being plunged in liquid nitrogen. Rewarming was conducted in a water-bath for 30 s at 40 degree C and PVS2 was replaced with a 1.2 M sucrose solution for 30 min at 25 degree C. The shoot tips were transferred on semisolid medium and left in the dark for 1 week, then in dim light for 3 weeks.

EFFECTS INDUCED BY DIETHYL SULPHATE ON SOME CYTOGENETICAL PARAMETERS AND LENGTH GROWTH OF HEMP PLANTLETS

Directory of Open Access Journals (Sweden)

Elena Truta

2007-08-01

Full Text Available The hemp seeds were treated with diethyl sulphate, in four concentrations (0.1%, 0.25%, 0.5%, and 1% and in two variants of alkylant exposure (3 and 6 hours. The length growth of plantlets, mitotic index and frequency of chromosomal aberrations were the analyzed parameters. Significant modifications were obtained at the level of parameters in variants treated with DES, comparatively with control.

Diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple plantlets during acclimatization

OpenAIRE

Silva,Aurilena de Aviz; Carvalho,Almy Junior Cordeiro de; Freitas,Flávia Paiva de; Pessanha,Patrícia Gomes de Oliveira; Santos,Paulo Cesar dos; Silva,Mírian Peixoto Soares da; Vasconcelos,Tábatha de Souza; Olivares,Fábio Lopes

2016-01-01

ABSTRACT: This study examines the effect of inoculation with diazotrophic bacteria and nitrogen fertilization on the growth of micropropagated pineapple cv. 'Vitória' plantlets during the acclimatization period. The experiment was carried out in a greenhouse in Campos dos Goytacazes, in randomized blocks, using a 2x5x5 factorial scheme, with the factors being two types of inocula (absence or presence of a mixture of diazotrophic bacteria that contained Burkholderia sp. UENF 114111, Burkholder...

Salt stress encourages proline accumulation by regulating proline biosynthesis and degradation in Jerusalem artichoke plantlets.

Science.gov (United States)

Huang, Zengrong; Zhao, Long; Chen, Dandan; Liang, Mingxiang; Liu, Zhaopu; Shao, Hongbo; Long, Xiaohua

2013-01-01

Proline accumulation is an important mechanism for osmotic regulation under salt stress. In this study, we evaluated proline accumulation profiles in roots, stems and leaves of Jerusalem artichoke (Helianthus tuberosus L.) plantlets under NaCl stress. We also examined HtP5CS, HtOAT and HtPDH enzyme activities and gene expression patterns of putative HtP5CS1, HtP5CS2, HtOAT, HtPDH1, and HtPDH2 genes. The objective of our study was to characterize the proline regulation mechanisms of Jerusalem artichoke, a moderately salt tolerant species, under NaCl stress. Jerusalem artichoke plantlets were observed to accumulate proline in roots, stems and leaves during salt stress. HtP5CS enzyme activities were increased under NaCl stress, while HtOAT and HtPDH activities generally repressed. Transcript levels of HtP5CS2 increased while transcript levels of HtOAT, HtPDH1 and HtPDH2 generally decreased in response to NaCl stress. Our results supports that for Jerusalem artichoke, proline synthesis under salt stress is mainly through the Glu pathway, and HtP5CS2 is predominant in this process while HtOAT plays a less important role. Both HtPDH genes may function in proline degradation.

Biosynthesis of phenolic compounds and water soluble vitamins in culantro (Eryngium foetidum L. plantlets as affected by low doses of gamma irradiation

Directory of Open Access Journals (Sweden)

Amina Abd El-Hamid ALY

2010-11-01

Full Text Available Explants obtained from in-vitro propagated plantlets of Culantro (Eryngium foetidum L. were exposed to four dose levels of γ-irradiation (0.0, 10.0, 20.0 and 40.0 Gy to investigate the biosynthesis of phenolic compounds and water soluble vitamins in Culantro fresh plantlets. Among six identified phenolic compounds, the content of p-cumaric acid was the highest in the extracts, followed by caffeic acid, coumarin, benzoic acid, salicylic acid and apigenin. Significant increases were observed at dose 40.0 Gy (61.66 mg/g d.w. for flavonoids, 18.02 mg/g d.w, for flavonone and 5.06 mg/g d.w for anthocyanin compared to control. On the other hand, the flavonols were decreased by increasing the irradiation dose. Vitamin C was increased in irradiated samples and this increase was in correlation with irradiation dose level. Thiamin, riboflavin and nicotinic acid were enhanced by the applied dose level 10 Gy. In addition, folic acid was enhanced by the dose levels 20 and 40 Gy and not detected for the control and 10 Gy treatments. Meanwhile, pyridoxine was decreased by increasing the irradiation dose level. The results obtained suggested that both low doses of γ-irradiation and tissue culture technique could be used to produce plantlets with high amount of phenolic compounds and water soluble vitamins.

Salt stress encourages proline accumulation by regulating proline biosynthesis and degradation in Jerusalem artichoke plantlets.

Directory of Open Access Journals (Sweden)

Zengrong Huang

Full Text Available Proline accumulation is an important mechanism for osmotic regulation under salt stress. In this study, we evaluated proline accumulation profiles in roots, stems and leaves of Jerusalem artichoke (Helianthus tuberosus L. plantlets under NaCl stress. We also examined HtP5CS, HtOAT and HtPDH enzyme activities and gene expression patterns of putative HtP5CS1, HtP5CS2, HtOAT, HtPDH1, and HtPDH2 genes. The objective of our study was to characterize the proline regulation mechanisms of Jerusalem artichoke, a moderately salt tolerant species, under NaCl stress. Jerusalem artichoke plantlets were observed to accumulate proline in roots, stems and leaves during salt stress. HtP5CS enzyme activities were increased under NaCl stress, while HtOAT and HtPDH activities generally repressed. Transcript levels of HtP5CS2 increased while transcript levels of HtOAT, HtPDH1 and HtPDH2 generally decreased in response to NaCl stress. Our results supports that for Jerusalem artichoke, proline synthesis under salt stress is mainly through the Glu pathway, and HtP5CS2 is predominant in this process while HtOAT plays a less important role. Both HtPDH genes may function in proline degradation.

Germination and plantlet regeneration of encapsulated microshoots of aromatic rice (Oryza sativa L. Cv. MRQ 74).

Science.gov (United States)

Taha, Rosna Mat; Saleh, Azani; Mahmad, Noraini; Hasbullah, Nor Azlina; Mohajer, Sadegh

2012-01-01

Plant tissues such as somatic embryos, apical shoot tips, axillary shoot buds, embryogenic calli, and protocom-like bodies are potential micropropagules that have been considered for creating synthetic seeds. In the present study, 3-5 mm microshoots of Oryza sativa L. Cv. MRQ 74 were used as explant sources for obtaining synthetic seeds. Microshoots were induced from stem explants on Murashige and Skoog (MS) medium supplemented with 1.5 mg/L benzylaminopurine (BAP). They were encapsulated in 3% (w/v) sodium alginate, 3% sucrose, 0.1 mg/L BAP, and 0.1 mg/L α-Naphthalene acetic acid (NAA). Germination and plantlet regeneration of the encapsulated seeds were tested by culturing them on various germination media. The effect of storage period (15-30 days) was also investigated. The maximum germination and plantlet regeneration (100.0%) were recorded on MS media containing 3% sucrose and 0.8% agar with and without 0.1 mg/L BAP. However, a low germination rate (6.67%) was obtained using top soil as a sowing substrate. The germination rate of the encapsulated microshoots decreased from 93.33% to 3.33% after 30 days of storage at 4°C in the dark. Therefore, further research is being done to improve the germination rate of the synthetic seeds.

Silver Nitrate and Different Culture Vessels Influence High Frequency Microrhizome Induction In Vitro and Enhancement Growth of Turmeric Plantlet During Ex Vitro Acclimatization

Directory of Open Access Journals (Sweden)

Dikash Singh THINGBAIJAM

2012-11-01

Full Text Available Eleven cultivars of C. longa var. Lakadong were collected from Manipur having different topography. Curcumin content in different cultivars has been analyzed by using UV-Visible Spectrophotometer (100 Bio-Carry Spectrophotometer. The curcuminoids content were analyzed and quantified for identification of best quality cultivar. Thoubal Cultivar with highest curcumin content (9.44% was subjected for tissue culture technique using different culture vessels and silver nitrate for rapid multiplication and scaling up of microrhizome production. High multiplication rate of 27.40�0.47 were obtained in Murashige and Skoog�s medium supplemented with 3% sucrose + 1 mg L-1 ?-napthalene acetic acid, 4 mg L-1 6-benzyl-amino-purine and 11 ?M silver nitrate. Effect of different culture vessels and silver nitrate were studied for microrhizome and multiple shoots formation. Relatively higher rate of shoots along with microrhizome (17.5�0.32 can be seen in Growtek which was grown without any plant growth regulator. Growtek was used for scaling up of microrhizome production in vitro and utmost microrhizome was produced in liquid Murashige and Skoog�s medium supplemented with 8% sucrose, 1 mg L-1 ?-napthalene acetic acid, 4 mg L-1 6-benzyl-amino-purine and 11 ?M silver nitrate (36.25�0.27. Addition of silver nitrate in the medium resulted in improvement of microrhizome induction in vitro. Higher concentration of silver nitrate (33, 44, 66, 88 ?M negatively affected the microrhizome and shoot multiplication and shows inhibition of tissue response completely. Analysis of in vitro derived plantlets during acclimatization shows that the exogenous applied of silver nitrate shows superior growth as compared to control. 90-95% of plantlets with and 75-80% plantlets without silver nitrate treatment were successfully established under ex vitro acclimatization. The protocol could be utilized for large scale production of true-to-type plantlets and as alternative

Silver Nitrate and Different Culture Vessels Influence High Frequency Microrhizome Induction In Vitro and Enhancement Growth of Turmeric Plantlet During Ex Vitro Acclimatization

Directory of Open Access Journals (Sweden)

Dikash Singh THINGBAIJAM

2012-11-01

Full Text Available Eleven cultivars of C. longa var. Lakadong were collected from Manipur having different topography. Curcumin content in different cultivars has been analyzed by using UV-Visible Spectrophotometer (100 Bio-Carry Spectrophotometer. The curcuminoids content were analyzed and quantified for identification of best quality cultivar. Thoubal Cultivar with highest curcumin content (9.44% was subjected for tissue culture technique using different culture vessels and silver nitrate for rapid multiplication and scaling up of microrhizome production. High multiplication rate of 27.400.47 were obtained in Murashige and Skoogs medium supplemented with 3% sucrose + 1 mg L-1 ?-napthalene acetic acid, 4 mg L-1 6-benzyl-amino-purine and 11 ?M silver nitrate. Effect of different culture vessels and silver nitrate were studied for microrhizome and multiple shoots formation. Relatively higher rate of shoots along with microrhizome (17.50.32 can be seen in Growtek which was grown without any plant growth regulator. Growtek was used for scaling up of microrhizome production in vitro and utmost microrhizome was produced in liquid Murashige and Skoogs medium supplemented with 8% sucrose, 1 mg L-1 ?-napthalene acetic acid, 4 mg L-1 6-benzyl-amino-purine and 11 ?M silver nitrate (36.250.27. Addition of silver nitrate in the medium resulted in improvement of microrhizome induction in vitro. Higher concentration of silver nitrate (33, 44, 66, 88 ?M negatively affected the microrhizome and shoot multiplication and shows inhibition of tissue response completely. Analysis of in vitro derived plantlets during acclimatization shows that the exogenous applied of silver nitrate shows superior growth as compared to control. 90-95% of plantlets with and 75-80% plantlets without silver nitrate treatment were successfully established under ex vitro acclimatization. The protocol could be utilized for large scale production of true-to-type plantlets and as alternative method to step

The effects of varying concentrations of growth regulators benzyladenine and naphthalene acetic acid and gamma irradiation on the gross plantlet morphology of Garcinia mangostana L

International Nuclear Information System (INIS)

Bonsol, Ma. Fatima Nona M.; Lasiste, Jade Marie Edenvirg F.; Quinio, Ma. Franchesca S.

2004-02-01

Mangosteen (Garcinia mangostana L.) is regarded as one of the best flavored fruits in the world. Though this fruit yields high income and has great potential as an export, productivity in the Philippines is still limited due to its slow and difficult conventional propagation. To increase the plants' productivity, growth regulators can be added, the seeds can be exposed to irradiation to cause genetic mutations, and micropropagation techniques can be used. This study seeks to determine the effects of these three variables on the general morphological characteristics of the plantlets grown both in vitro and in vivo. The first set of mangosteen seeds was inoculated and grown in Murashige and Skoog (MS) basal culture medium with different treatment combinations of growth regulators, naphthalene acetic acid (NAA) and benzyladenine (BA). Treatment concentrations ranged from 0 parts per million (ppm) to 6 ppm. The treatment that best induced stem, leaf and root formation, and produced plantlets with long stems and roots was MS with 6 ppm NAA combined with 6 ppm BA. A second set of seeds was then exposed to varying levels of gamma radiation and propagated in vivo and in vitro. For in vivo, it was observed that the length of stem of the plantlet decreases with increasing radiation dose; length of leaves of the mangosteen plantlets was best noted in seeds exposed to 10 Gy gamma rays. In in vitro, the two treatment concentrations that best helped seeds to develop callus, which were 0 ppm BA with 4 pmm NAA and 4 ppm BA with 6 ppm NAA, were added to MS basal medium. Seeds exposed to 5 Gy and grown in basal medium with 0 ppm BA and 4 ppm NAA were observed to have developed the most callus. (Author)

Generation and multiplication of plantlets from callus derived from Haplopappus gracilus (Nutt.) Gray and their karyotype analysis

Science.gov (United States)

Kann, R. P.; O'Connor, S. A.; Levine, H. G.; Krikorian, A. D.

1991-01-01

Unopened flower heads of Haplopappus gracilis (2n = 4) provided primary explants for callus production and subsequent induction of organized growth. Callus was initiated from small (3-5 mm in length) floral buds with benzylaminopurine (BAP) (44.4 micromoles; 10 mg/l) and naphthalene acetic acid (NAA) (0.54 micromole; 0.1 mg/l). Lowering the BAP level to 4.44 micromoles (1 mg/l) but maintaining the NAA level, gave rise to organized but highly compressed shoot growing points from an otherwise undifferentiated callus mass. Shoots selected from such cultures were maintainable and could be proliferated by growing 1-1.5-cm stem tip cuttings on Murashige and Skoog basal medium (solidified with agar) containing 0.444 micromole (0.1 mg/l) BAP and 0.054 micromole (0.01 mg/l) NAA. The stem tip multiplication rates obtainable by these means permit reliable strategies for shoot multiplication or production of rooted plantlets. Prolonged subculture and maintenance of shoots on growth regulator-free medium leads to in vitro flowering and greatly reduces rooting capacity. Karyotype analysis of chromosomes from root tip cells at metaphase and chromosome measurements show that karyologically uniform plantlets (based on chromosome number and morphology) can be obtained.

Germination and Plantlet Regeneration of Encapsulated Microshoots of Aromatic Rice (Oryza sativa L. Cv. MRQ 74

Directory of Open Access Journals (Sweden)

Rosna Mat Taha

2012-01-01

Full Text Available Plant tissues such as somatic embryos, apical shoot tips, axillary shoot buds, embryogenic calli, and protocom-like bodies are potential micropropagules that have been considered for creating synthetic seeds. In the present study, 3–5 mm microshoots of Oryza sativa L. Cv. MRQ 74 were used as explant sources for obtaining synthetic seeds. Microshoots were induced from stem explants on Murashige and Skoog (MS medium supplemented with 1.5 mg/L benzylaminopurine (BAP. They were encapsulated in 3% (w/v sodium alginate, 3% sucrose, 0.1 mg/L BAP, and 0.1 mg/L α-Naphthalene acetic acid (NAA. Germination and plantlet regeneration of the encapsulated seeds were tested by culturing them on various germination media. The effect of storage period (15–30 days was also investigated. The maximum germination and plantlet regeneration (100.0% were recorded on MS media containing 3% sucrose and 0.8% agar with and without 0.1 mg/L BAP. However, a low germination rate (6.67% was obtained using top soil as a sowing substrate. The germination rate of the encapsulated microshoots decreased from 93.33% to 3.33% after 30 days of storage at 4°C in the dark. Therefore, further research is being done to improve the germination rate of the synthetic seeds.

Organ and plantlet regeneration of Menyanthes trifoliata through tissue culture

Directory of Open Access Journals (Sweden)

Urszula Adamczyk-Rogozińska

2014-01-01

Full Text Available The conditions for the regeneration of plants through organogenesis from callus tissues of Menyanthes trifoliata are described. The shoot multiplication rate was affected by basal culture media, the type and concentration of cytokinin and subculture number. The best response was obtained when caulogenic calli were cultured on the modified Schenk and Hildebrandt medium (SH-M containing indole-3-acetic acid (IAA 0,5 mg/l and 6-benzyladenine (BA 1 mg/l or zeatin (2 mg/l. Under these conditions ca 7 shoots (mostly 1 cm or more in length per culture in the 5th and 6th passages could be developed. In older cultures (after 11-12 passages there was a trend for more numerous but shorter shoot formation. All regenerated shoots could be rooted on the SH-M medium supplemented with 0.5 mg/l IAA within 6 weeks; 80% of in vitro rooted plantlets survived their transfer to soil.

Assessment of Genetic Fidelity in Rauvolfia serpentina Plantlets Grown from Synthetic (Encapsulated Seeds Following in Vitro Storage at 4 °C

Directory of Open Access Journals (Sweden)

Mohammad Anis

2012-05-01

Full Text Available An efficient method was developed for plant regeneration and establishment from alginate encapsulated synthetic seeds of Rauvolfia serpentina. Synthetic seeds were produced using in vitro proliferated microshoots upon complexation of 3% sodium alginate prepared in Llyod and McCown woody plant medium (WPM and 100 mM calcium chloride. Re-growth ability of encapsulated nodal segments was evaluated after storage at 4 °C for 0, 1, 2, 4, 6 and 8 weeks and compared with non-encapsulated buds. Effects of different media viz; Murashige and Skoog medium; Lloyd and McCown woody Plant medium, Gamborg’s B5 medium and Schenk and Hildebrandt medium was also investigated for conversion into plantlets. The maximum frequency of conversion into plantlets from encapsulated nodal segments stored at 4 °C for 4 weeks was achieved on woody plant medium supplement with 5.0 μM BA and 1.0 μM NAA. Rooting in plantlets was achieved in half-strength Murashige and Skoog liquid medium containing 0.5 μM indole-3-acetic acid (IAA on filter paper bridges. Plantlets obtained from stored synseeds were hardened, established successfully ex vitro and were morphologically similar to each other as well as their mother plant. The genetic fidelity of Rauvolfia clones raised from synthetic seeds following four weeks of storage at 4 °C were assessed by using random amplified polymorphic DNA (RAPD and inter-simple sequence repeat (ISSR markers. All the RAPD and ISSR profiles from generated plantlets were monomorphic and comparable to the mother plant, which confirms the genetic stability among the clones. This synseed protocol could be useful for establishing a particular system for conservation, short-term storage and production of genetically identical and stable plants before it is released for commercial purposes.

Produção de mudas de alface com biofertilizantes e substratos Production of lettuce plantlets using biofertilizer and substrates

Directory of Open Access Journals (Sweden)

Damiana Cleuma de Medeiros

2007-09-01

Full Text Available Com o objetivo de avaliar o desenvolvimento de mudas de alface que foram desenvolvidas em diferentes substratos e sob efeito de adubação foliar usando biofertilizantes, foi realizado o experimento em casa de vegetação da UFERSA. Utilizou-se o delineamento de blocos ao acaso com 4 repetições. Os tratamentos foram resultados de um fatorial 4 x 5 envolvendo tipos de substrato (composto orgânico, Gold Mix 11, areia lavada e húmus + areia lavada 1:1 e fertilizante foliar (organo-mineral comercial Fertamin, testemunha e três tipos de biofertilizantes. Foram feitas adubações foliares nas mudas, aos oito e aos 15 dias após a semeadura. O fertilizante organo-mineral comercial Fertamin apresentou os melhores resultados para massa seca da parte aérea, massa seca da raiz e número de folhas. Dentre os biofertilizantes, aquele à base de esterco de curral fresco, cinzas, leite e caldo de cana apresentou os menores valores para essas mesmas características avaliadas. O composto orgânico foi o que apresentou os maiores valores para todas as características avaliadas, exceto para comprimento da raiz.The development of lettuce plantlets was evaluated, employing different substrate types and applying foliar biofertilization. The research was done in a green house in Mossoró, Rio Grande do Norte State, Brazil. The experimental design was of randomized blocks with four replications. The treatments were composed of the interaction substract type (composed organic, Golden Mix 11, washed sand and humus + washed sand 1:1 and fertilizer (the commercial organo-mineral Fertamin, the witness and tree biofertilizer types. Plantlets were foliar fertilized 8 and 15 days after sowing date. Plantlets treated with Fertamim presented the best results in the dry matter of the aerial part, dry matter of roots and number of leaves. Among the biofertilizers, the one based on cattle manure, leached ash, milk and broth of sugar cane presented the lesser values for the

THE EFFICIENCY OF SHOOT AND PLANTLET FORMATION OF Cephaelis ipecacuanha AFTER THREE SUBCULTURES IN VITRO

Directory of Open Access Journals (Sweden)

Osmar Alves Lameira

1994-12-01

Full Text Available Multiple adventitious shoot formed from internodal segments of Cephaelis ipecacuanha cultured 25 days on Gamborg basal medium (GAMBORG et al., 1968 supplemented with 6.66mM 6-benzylaminopurine there was a maximum of nine shoots per segment and an average of five shoots per segment formed. The presence of gibberellic acid in the subculture media promoted shoot elongation in all treatments. The shoots attained 3cm in height and rooting of 100% after 35 days of culturing upon Murashige and Skoog's basal medium (MS, added with 4.92mM indole-3-butyric acid, 0.87m gibberellic acid and 0.1% activated charcoal. Further growth was accelerated after the transfer to 1/2 MS without growth regulators. Rooted plantlets transferred to potting soil could be successfully established.

Assessment of clonal fidelity of Tylophora indica (Burm. f.) Merrill "in vitro" plantlets by ISSR molecular markers.

Science.gov (United States)

Sharma, Madan Mohan; Verma, Roop Narayan; Singh, Abhijeet; Batra, Amla

2014-01-01

Tylophora indica Burm F. Merrill. is widely used against various diseases owing to the presence an array of medicinally important secondary metabolites. Its stem is bitter, stomachic, stimulates bile secretion, enriches the blood and cures diseases like diabetes, fever, flatulence, hypertension, jaundice, leucorrhoea, urinary disease and upper respiratory tract infection. It is neglected for tissue culture work because of deciduous nature of climbing shrub, facing problems for micropropagation. Hence, in vitro regeneration of complete plantlets was done through indirect organogenesis in Tylophora indica. Calli were produced from in vivo leaves of T. indica on MS medium supplemented with 6-Benzylaminopurine (BAP: 2.0 mg l(-1)) and Indole-3-butyric acid (IBA: 0.5 mg l(-1)). The multiple shoots (12.00 ± 1.50) emerged and elongated on MS medium fortified with Thidiazuron (TDZ: 0.1 mg l(-1)). They were rooted on half strength MS medium having IBA (0.5 mg l(-1)) (7.75 ± 0.25) after 20 days of sub-culturing followed by hardening and acclimatization. During indirect regeneration of plants, chances of somaclonal variations may arise. These variations should be identified to produce true to type plants. Plantlets raised through tissue culture were used to validate the clonal fidelity through Inter simple sequence repeat markers (ISSR). Clonal fidelity is a major consideration in commercial micropropagation using in vitro tissue culture methods. During the study, total 71 clear and distinct bands were produced using 6 primers. The banding pattern of each primer was uniform and comparable to mother plant and showed about 93% homology using un-weighted pair group method with arithmetic averaging (UPGMA). ISSR analysis confirmed the genetic stability of in vitro raised plants.

Micropropagation of herbal plants for mass production of in vitro plantlets

International Nuclear Information System (INIS)

Sobri Hussein; Rusli Ibrahim

2006-01-01

Malaysia herbal industry is considered to be one of the most dynamic enterprises with annual growth estimated at 20 % a year. The total import value of the medicinal and aromatic plants increased from RM 141 million in 1986 to RM 431 million in 1996. Species that have been identified in terms of current priority are Eurycoma longifolia (Tongkat Ali), Labisia pumila (Kacip Fatimah), Andrographis paniculata (Hempedu Bumi), Morinda citrifolia (Mengkudu), Centella asiatica (Pegaga), Orthosiphon aristatus (Misai Kucing) and Gynura procumbens (Sambung Nyawa). Herbal and medicinal plants have a larger pool of genetic resources for the production of compounds valuable to the industry and human well being such as in pharmaceuticals, nutraceuticals and health food. The use of tissue culture technology has long been considered as an attractive solution to the problem of limited supply of raw materials for the industries. Kacip Fatimah (Labisia pumila) has been well known to increase female hormone, overcome tiredness and regain overall health. Misai Kucing (Orthosiphon aristatus) has proven to be active against kidney stones, high blood pressure, diabetes and gout and Sambung Nyawa (Gynura procumbens) is active against hypertension, diabetes and cancer. Tissue culture protocols had been optimized for the mass production of in vitro plantlets of these three selected herbal plants. (Author)

Micropropagation of herbal plants for mass production of in vitro plantlets

Energy Technology Data Exchange (ETDEWEB)

Hussein, Sobri; Ibrahim, Rusli [Malaysian Institute for Nuclear Technology Research MINT, Bangi (Malaysia). Agrotechnology and Biosciences Div.

2006-09-15

Malaysia herbal industry is considered to be one of the most dynamic enterprises with annual growth estimated at 20 % a year. The total import value of the medicinal and aromatic plants increased from RM 141 million in 1986 to RM 431 million in 1996. Species that have been identified in terms of current priority are Eurycoma longifolia (Tongkat Ali), Labisia pumila (Kacip Fatimah), Andrographis paniculata (Hempedu Bumi), Morinda citrifolia (Mengkudu), Centella asiatica (Pegaga), Orthosiphon aristatus (Misai Kucing) and Gynura procumbens (Sambung Nyawa). Herbal and medicinal plants have a larger pool of genetic resources for the production of compounds valuable to the industry and human well being such as in pharmaceuticals, nutraceuticals and health food. The use of tissue culture technology has long been considered as an attractive solution to the problem of limited supply of raw materials for the industries. Kacip Fatimah (Labisia pumila) has been well known to increase female hormone, overcome tiredness and regain overall health. Misai Kucing (Orthosiphon aristatus) has proven to be active against kidney stones, high blood pressure, diabetes and gout and Sambung Nyawa (Gynura procumbens) is active against hypertension, diabetes and cancer. Tissue culture protocols had been optimized for the mass production of in vitro plantlets of these three selected herbal plants. (Author)

Asymbiotic germination, seedling development and plantlet propagation of Encyclia aff. oncidioides - an endangered orchid

Directory of Open Access Journals (Sweden)

Ewa Łojkowska

2011-01-01

Full Text Available In order to estimate the best germination conditions of Encyclia aff. oncidioides seeds, five different media (Fast, Knudson C modified by Vajrabhaya, Murashige and Skoog, PB2 and modified Vacin and Went with different concentrations of plant growth regulators such as benzyladenine (BA, naphthaleneacetic acid (NAA and gibberellic acid (GA3 were tested. No beneficial effect was observed when BA and NAA were applied to the germination medium and GA3 inhibited germination. The effect of light, activated charcoal, coconut water and casein hydrolysate on seed germination was also studied. The growth rate of seedlings on three different media supplemented with activated charcoal and plant growth regulators was checked. The applied plant growth regulators had no beneficial effect on the further growth of seedlings. Fast and PB2 media with 0.2% activated charcoal proved to be the best for E. aff. oncidioides seed germination, seedling development and plantlet propagation.

In Vitro Selection of Peanut Somatic Embryos on Medium Containing Culture Filtrate of Sclerotium rolfsii and Plantlet Regeneration

Directory of Open Access Journals (Sweden)

YUSNITA

2005-06-01

Full Text Available Attempts to identify somaclonal variants of peanut with resistance to Sclerotium stem rot disease due to infection of S. rolfsii were conducted. The objectives of this study were to develop in vitro selection method using culture filtrates of S. rolfsii, identify culture filtrate-insensitive somatic embryo (SE of peanut after in vitro selection and regenerate peanut R0 lines originated from culture filtrate-insensitive SE. To achieve these objectives, peanut embryogenic tissues were cultured on selective medium containing various concentrations of S. rolfsii culture filtrates and sublethal concentration of the filtrates. Medium containing sublethal level of S. rolfsii culture filtrates was used to identify culture filtrate-insensitive SE of peanut. Subsequently, the selected SEs were germinated, plantlets were regenerated and preliminary tested against S. rolfsii. Results of the experiments showed that addition of S. rolfsii culture filtrates into medium for inducing peanut somatic embryos drastically reduced their growth and proliferation. S. rolfsii culture filtrates at 10% concentration has significantly reduced the number of proliferated SE per explant. However, sublethal level was achieved at 30% of culture filtrates concentration. Responses of five peanut cultivars against 30% of culture filtrates were similar, indicating they were similar in their susceptibility against S. rolfsii. A number of culture filtrate-insensitive SE were identified after culturing 1500 clumps of embryogenic tissue of peanut cv. Kelinci for three consecutive passages on medium containing 30% of culture filtrates. Germination of selected SE and regeneration of plantlet from culture filtrate-insensitive SE resulted in 50 peanut R0 lines. These lines have been grown in the plastic house and produced normal seeds for further evaluation. Results of S. rolfsii inoculation indicated the existence of chimera for insensitivity against S. rolfsii.

Biomimetic synthesis of antimicrobial silver nanoparticles using in vitro-propagated plantlets of a medicinally important endangered species: Phlomis bracteosa.

Science.gov (United States)

Anjum, Sumaira; Abbasi, Bilal Haider

2016-01-01

In vitro-derived cultures of plants offer a great potential for rapid biosynthesis of chemical-free antimicrobial silver nanoparticles (AgNPs) by enhancing their phytochemical reducing potential. Here, we developed an efficient protocol for in vitro micropropagation of a high-value endangered medicinal plant species, Phlomis bracteosa, in order to explore its biogenic potential in biomimetic synthesis of antimicrobial AgNPs. Murashige and Skoog medium supplemented with 2.0 mg/L thidiazuron was found to be more efficient in inducing optimum in vitro shoot regeneration (78%±4.09%), and 2.0 mg/L indole-3-butyric acid was used for maximum root induction (86%±4.457%). Antimicrobial AgNPs were successfully synthesized by using aqueous extract (rich in total phenolics and flavonoids content) of in vitro derived plantlets of P. bracteosa. Ultraviolet-visible spectroscopy of synthesized AgNPs showed characteristic surface plasmon band in the range of 420-429 nm. The crystallinity, size, and shape of the AgNPs were characterized by X-ray diffraction and scanning electron microscopy. Face-centered cubic AgNPs of almost uniform spherical size (22.41 nm) were synthesized within a short time (1 hour) at room temperature. Fourier-transform infrared spectroscopy revealed that the polyphenols were mainly responsible for reduction and capping of synthesized AgNPs. Energy dispersive X-ray analysis further endorsed the presence of elemental silver in synthesized AgNPs. These biosynthesized AgNPs displayed significantly higher bactericidal activity against multiple drug-resistant human pathogens. The present work highlighted the potent role of in vitro-derived plantlets of P. bracteosa for feasible biosynthesis of antimicrobial AgNPs, which can be used as nanomedicines in many biomedical applications.

Physiological responses in the banana plantlets treateds with strobilurinsRespostas fisiológicas em mudas de banananeira tratadas com estrobilurinas

Directory of Open Access Journals (Sweden)

Sivia Helena Modenese-Gorla da Silva

2012-04-01

Full Text Available There are reports that strobilurin besides having a fungicide effect can promote physiologic benefits to the plants. However, this effect on banana plants was not studied yet. The objective of the present study was to evaluate the effect of strobirulins on the physiology of banana plantlets. For this purpose, cultivar Grand Naine banana plantlets were transferred to pots containing substrate and kept in a nursery with 50% shading. The experimental design was a completely randomized design with three treatments (water, azoxystrobin and pyraclostrobin and five replications. The treatments were applied at 15, 30, 45, 60 and 75 days after transplanting at a dose 100 g a. i. ha-1 with manual spray. Plant height, pseudostem diameter, shoot dry matter in strobilurin treated plants were higher than the untreated plants, however, the effect of fungicide treatment was different, being the most pronounced effect of pyraclostrobin compared to azoxystrobin. Plants treated with pyraclostrobin had higher leaf area, nitrate reductase activity and chlorophyll content of leaf total nitrogen than the plants treated with azoxystrobin and water, which did not differ. Strobilurins affect the physiology of the banana plantlets differently, the effect being more pronounced by pyraclostrobin.Há relatos de que estrobilurinas, além de atuarem como fungicida, promovem benefícios fisiológicos às plantas. Assim sendo, o presente trabalho teve como objetivo avaliar o efeito de estrobirulinas na fisiologia de mudas de bananeira. Para tal, mudas micropropagadas da cultivar Grand Naine foram repicadas para vasos contendo substrato e mantidas em viveiro a 50% de sombreamento. O delineamento experimental foi inteiramente casualizado, com três tratamentos (água, azoxistrobina e piraclostrobina e cinco repetições. As estrobirulinas foram aplicadas com pulverizador manual aos 15, 30, 45, 60 e 75 dias após a repicagem, na dose de 100 g i. a. ha-1. A altura da planta, di

Comparative studies about the influence of salicylic and acetylsalicilic acid on content of assimilatory pigments in the primary leaves of wheat (Triticum aestivum plantlets

Directory of Open Access Journals (Sweden)

Cornelia PURCAREA

2007-05-01

Full Text Available Salicylic acid (SA and some of its derivates are phenolic compounds recently recognized as plant growth regulators involved in many physiological processes including photosynthesis. One of the important derivates of Salicylic Acid is the Acetylsalicylic Acid. In the present investigation we studied the influence of exogenous Acetylsalicylic and Salicylic acid with different concentrations on the assimilatory pigments contents of the primary leaves of wheat seedlings in comparison with the same parameters of the control lots which were treated with water. The wheat seedlings were soaked for 6 hours in 0.01mM; 0.1mM; 0.5mM and 1 mM SA or ASA solutions and in water for the control lot, germinated for 7 days on filter paper moistened with water. After that, we planted the plantlets in sand and sprayed their coleoptiles and primary leaves, each day for an additional 7 days, with water. In the 14th days of germination we determined the content of assimilatory pigments extracted with N,N-dimethylformamide (DMF. The results showed that exogenous 0.01 mM, 0.1mM, 0.5 mM or 1.0 mM SA solution treatments cause more significant increases in the assimilatory pigments contents in leaves of wheat plantlets than treatments with ASA solutions of the same concentrations do.

Crescimento de mudas micropropagadas de bananeira micorrizadas em diferentes recipientes Growth of mycorrhizal micropropagated banana plantlets in different recipients

Directory of Open Access Journals (Sweden)

Patrícia Lopes Leal

2005-04-01

Full Text Available Realizou-se um experimento em casa de vegetação, com o objetivo de avaliar a utilização de diferentes recipientes e a eficiência da inoculação com fungo micorrízico arbuscular (FMA, Glomus clarum, no crescimento de mudas micropropagadas de bananeira. O delineamento experimental foi o de blocos casualizados, no esquema fatorial 2x2, sendo 2 tratamentos microbiológicos: Glomus clarum e controle; e 2 recipientes: blocos prensados e tubetes, com 6 repetições. O substrato utilizado para a confecção dos blocos e para o enchimento dos tubetes foi constituído por uma mistura de materiais orgânicos (bagaço de cana + torta de filtro de usina açucareira e vermiculita. Mudas de bananeira produzidas em blocos prensados e inoculadas com o FMA apresentaram melhores resultados quando comparadas com as produzidas nos tubetes, com incrementos na altura, na produção de matéria seca da parte aérea e no acúmulo de N, P e K de 90%, 829%, 2774%, 249% e 403%, respectivamente.An experiment under greenhouse was carried out in order to evaluate different recipients and the inoculation with arbuscular mycorrhizal fungus (AMF, Glomus clarum, on the growth of micropropagated banana plantlets. A random block design in a factorial scheme 2x2 (two microbiological treatments: Glomus clarum and control; two types of recipients: pressed blocks and plastic tubes, with six repetitions, was used. The substrate utilized to construct the pressed blocks and to fill the tubes was constituted by a mixture of organic residue (sugarcane bagasse and filter cake and vermiculite. Banana plantlets produced in pressed blocks and inoculated with AMF let to better results than their counterparts grown in tubes, increasing the height, shoot dry weight, contents of N, P and K in 90%, 829%, 2774%, 249% and 403%, respectively.

Growing Tomato Plantlets on Various Mixtures of Peat and Sand or Peat and Perlite. Note 1

Directory of Open Access Journals (Sweden)

Antonia Patruno

Full Text Available Given the considerable interest in use of substrates derived from various mixtures in the nursery sector and in light of the enormous variety of possibilities offered by this technique, in contrast with the still small number of researches dedicated to this theme, this study was set out to examine in-depth the growing of tomato plantlets on peat-based substrates. Two series of peat mixtures were produced, one with sand and the other with perlite, with a volume ratio of the other two components with respect to the peat of 1:0, 2.5:1, 1:1 and 1:2.5. Tomato seedlings were cultivated for 30 or 25 days in small perforated pots containing these mixtures. The irrigation was calculated by weighing each pot daily, measuring the water lost by evaporation-transpiration, then just past an established lower threshold value bringing the water back up to a defined upper threshold. Two water regimes were compared in the sand series and three in the perlite series.

Interaction of Ulocladium atrum, a Potential Biological Control Agent, with Botrytis cinerea and Grapevine Plantlets

Directory of Open Access Journals (Sweden)

Sébastien Ronseaux

2013-09-01

Full Text Available The effectiveness of biological control agent, Ulocladium atrum (isolates U13 and U16 in protecting Vitis vinifera L. cv. Chardonnay against gray mold disease caused by Botrytis cinerea, and simulation of the foliar defense responses was investigated. A degraded mycelium structure during cultural assay on potato dextrose agar revealed that U. atrum isolates U13 and U16 were both antagonistic to B. cinerea, mainly when isolates were inoculated two days before Botrytis. Under in vitro conditions, foliar application of U. atrum protected grapevine leaves against gray mold disease. An increase in chitinase activity was induced by the presence of U. atrum isolates indicating that the biological control agents triggered plant defense mechanisms. Moreover, U13 has the potential to colonize the grapevine plantlets and to improve their growth. The ability of U. atrum isolates to exhibit an antagonistic effect against B. cinerea in addition to their aptitude to induce plant resistance and to promote grapevine growth may explain a part of their biological activity. Hence, this study suggests that U. atrum provides a suitable biocontrol agent against gray mold in grapevines.

Resposta de plantas micropropagadas de abacaxizeiro à inoculação de bactérias diazotróficas em casa de vegetação Response of pineapple plantlets to inoculation with diazotrophic bacteria in greenhouse

Directory of Open Access Journals (Sweden)

Olmar Baller Weber

2003-12-01

Full Text Available O objetivo deste trabalho foi selecionar e avaliar bactérias diazotróficas isoladas de abacaxizeiro (Ananas comosus (L. Merril no desenvolvimento de cultivares micropropagadas da mesma espécie em casa de vegetação. Plantas da cultivar Perolera foram submetidas à inoculação com Asaia bogorensis (AB219 e cultivadas em tubetes, durante 145 dias, com as misturas: casca de arroz carbonizada, folha de carnaubeira triturada e vermicomposto; casca de arroz carbonizada, pó da casca do coco maduro e vermicomposto; casca de arroz carbonizada, vermiculita e vermicomposto. Plantas da cultivar Primavera receberam inóculos com o AB219 e bactérias relacionadas a Burkholderia cepacia (AB202 e AB213, enquanto plantas das cultivares Pérola e Smooth Cayenne receberam AB219 e AB213, sendo cultivadas, por 140 dias, em tubetes com a mistura de vermicomposto e vermiculita. A colonização dos abacaxizeiros pelas bactérias diazotróficas foi confirmada. As plantas da cultivar Perolera cresceram melhor em casca de arroz carbonizada, vermiculita e vermicomposto e responderam positivamente ao AB219. Já as plantas da cultivar Primavera não apresentaram resposta significativa à inoculação com AB219, AB202 e AB213. Houve incremento de 23,1% a 38,5% na matéria seca de raízes das plantas da cultivar Pérola na presença de AB213 e AB219, respectivamente. A presença de AB213 incrementou em 15,2% a matéria seca da parte aérea das plantas da cultivar Smooth Cayenne. Os resultados revelam a eficiência de bactérias diazotróficas na promoção do crescimento de abacaxizeiros.The objective of this work was to select and to evaluate isolates of diazotrophic bacteria associated to pineapple (Ananas comosus (L. Merril plants in the development of micropropagated plantlets of cultivars pineapple in greenhouse. Plantlets from the Perolera cultivar have been submitted to inoculation with Asaia bogorensis (AB219 and were cultivated during 145 days on the following

Micropropagation of an Exotic Ornamental Plant, Calathea crotalifera, for Production of High Quality Plantlets

Directory of Open Access Journals (Sweden)

Shahril Efzueni Rozali

2014-01-01

Full Text Available A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera. The effects of different sterilization techniques, explant type, and the combination and concentration of plant growth regulators on shoots induction were studied. The axillary shoot buds explants sprouted from rhizomes in soil free conditions showed high induction rate of shoots with lowest contamination percentage when treated with combination of 30% (v/v NaOCl, 70% (v/v ethanol, and 0.3% (w/v HgCl2. In the present study, the highest number of multiple shoots was obtained in MS basal medium supplemented with 3.5 mg/L 6-Benzylaminopurine (BAP, 1.0 mg/L 1-Naphthaleneacetic acid (NAA, 3% sucrose, and 6 g/L plant agar for both varieties and was used as multiplication medium. Microshoots were highly induced when the young shoot bud explants were incised longitudinally prior subculture. Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem. The maximum roots induction was recorded on MS medium supplemented with 1.0 mg/L 1-Naphthaleneacetic acid (NAA compared to indolebutyric acid (IBA. The complete regenerated plantlets were successfully acclimatized in the soilless medium under greenhouse condition. This is the first report of rapid mass propagation for C. crotalifera.

Micropropagation of an exotic ornamental plant, Calathea crotalifera, for production of high quality plantlets.

Science.gov (United States)

Rozali, Shahril Efzueni; Rashid, Kamaludin A; Taha, Rosna Mat

2014-01-01

A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera. The effects of different sterilization techniques, explant type, and the combination and concentration of plant growth regulators on shoots induction were studied. The axillary shoot buds explants sprouted from rhizomes in soil free conditions showed high induction rate of shoots with lowest contamination percentage when treated with combination of 30% (v/v) NaOCl, 70% (v/v) ethanol, and 0.3% (w/v) HgCl2. In the present study, the highest number of multiple shoots was obtained in MS basal medium supplemented with 3.5 mg/L 6-Benzylaminopurine (BAP), 1.0 mg/L 1-Naphthaleneacetic acid (NAA), 3% sucrose, and 6 g/L plant agar for both varieties and was used as multiplication medium. Microshoots were highly induced when the young shoot bud explants were incised longitudinally prior subculture. Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem. The maximum roots induction was recorded on MS medium supplemented with 1.0 mg/L 1-Naphthaleneacetic acid (NAA) compared to indolebutyric acid (IBA). The complete regenerated plantlets were successfully acclimatized in the soilless medium under greenhouse condition. This is the first report of rapid mass propagation for C. crotalifera.

Cryopreservation of chayote (Sechium edule JACQ. SW.) zygotic embryos and shoot-tips from in vitro plantlets.

Science.gov (United States)

Abdelnour-Esquivel, Ana; Engelmann, Florent

2002-01-01

This paper presents the development of cryopreservation protocols for zygotic embryos and apices of chayote (Sechium edule Jacq. Sw.), a tropical plant species with recalcitrant seeds. Zygotic embryos of two cultivars, Ccocro negro (CN) and Claudio (Cl) could withstand cryopreservation, with survival percentages of 10 and 30 %, after desiccation to 23 and 19 % moisture content (fresh weight basis), respectively. Apices sampled on in vitro plantlets of cultivars Cl, 13 and JM were successfully cryopreserved using a vitrification technique. Optimal conditions included the culture of mother-plants for 22 days on medium containing 0.3 M sucrose, culture of excised apices on the same medium for 1 day, loading of apices for 20 min with 2M glycerol + 0.4M glycerol, treatment with a series of diluted PVS2 solution (60 % PVS2 followed by 80 % PVS2 solution for 15 min (cultivar Cocoro Blanco [CB]) or 30 min (cultivars CN and Cl) at each concentration), rapid freezing and thawing, washing of shoot-tips with a 1.2 M sucrose solution, followed by recovery on media with progressively decreasing sucrose concentrations until the standard concentration of 0.1 M was reached. The highest survival percentages achieved ranged between 17 and 38 %, depending on the cultivar.

Selection of suitable propagation method for consistent plantlets production in Stevia rebaudiana (Bertoni).

Science.gov (United States)

Khalil, Shahid Akbar; Zamir, Roshan; Ahmad, Nisar

2014-12-01

Stevia rebaudiana (Bert.) is an emerging sugar alternative and anti-diabetic plant in Pakistan. That is why people did not know the exact time of propagation. The main objective of the present study was to establish feasible propagation methods for healthy biomass production. In the present study, seed germination, stem cuttings and micropropagation were investigated for higher productivity. Fresh seeds showed better germination (25.51-40%) but lost viability after a few days of storage. In order to improve the germination percentage, seeds were irradiated with 2.5, 5.0, 7.5 and 10 Gy gamma doses. But gamma irradiation did not show any significant change in seed germination. A great variation in survival of stem cutting was observed in each month of 2012. October and November were found the most suitable months for stem cutting survival (60%). In order to enhance survival, stem cuttings were also dipped in different plant growth regulators (PGRs) solution. Only indole butyric acid (IBA; 1000 ppm) treated cutting showed a higher survival (33%) than control (11.1%). Furthermore, simple and feasible indirect regeneration system was established from leaf explants. Best callus induction (84.6%) was observed on MS-medium augmented with 6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D; 2.0 mg l(-1)). For the first time, we obtained the highest number of shoots (106) on a medium containing BA (1.5 mg l(-1)) and gibberellic acid (GA3; 0.5 mg l(-1)). Plantlets were successfully acclimatized in plastic pots. The current results preferred micropropagation (85%) over seed germination (25.51-40%) and stem cutting (60%).

Selection of suitable propagation method for consistent plantlets production in Stevia rebaudiana (Bertoni)

Science.gov (United States)

Khalil, Shahid Akbar; Zamir, Roshan; Ahmad, Nisar

2014-01-01

Stevia rebaudiana (Bert.) is an emerging sugar alternative and anti-diabetic plant in Pakistan. That is why people did not know the exact time of propagation. The main objective of the present study was to establish feasible propagation methods for healthy biomass production. In the present study, seed germination, stem cuttings and micropropagation were investigated for higher productivity. Fresh seeds showed better germination (25.51–40%) but lost viability after a few days of storage. In order to improve the germination percentage, seeds were irradiated with 2.5, 5.0, 7.5 and 10 Gy gamma doses. But gamma irradiation did not show any significant change in seed germination. A great variation in survival of stem cutting was observed in each month of 2012. October and November were found the most suitable months for stem cutting survival (60%). In order to enhance survival, stem cuttings were also dipped in different plant growth regulators (PGRs) solution. Only indole butyric acid (IBA; 1000 ppm) treated cutting showed a higher survival (33%) than control (11.1%). Furthermore, simple and feasible indirect regeneration system was established from leaf explants. Best callus induction (84.6%) was observed on MS-medium augmented with 6-benzyladenine (BA) and 2,4-dichlorophenoxyacetic acid (2,4-D; 2.0 mg l−1). For the first time, we obtained the highest number of shoots (106) on a medium containing BA (1.5 mg l−1) and gibberellic acid (GA3; 0.5 mg l−1). Plantlets were successfully acclimatized in plastic pots. The current results preferred micropropagation (85%) over seed germination (25.51–40%) and stem cutting (60%). PMID:25473365

RAPD and ISSR based evaluation of genetic stability of micropropagated plantlets of Morus alba L. variety S-1

Science.gov (United States)

Saha, Soumen; Adhikari, Sinchan; Dey, Tulsi; Ghosh, Parthadeb

2015-01-01

Plant regeneration through rapid in vitro clonal propagation of nodal explants of Morus alba L. variety S-1 was established along with genetic stability analysis of regenerates. Axillary shoot bud proliferation was achieved on Murashige and Skoog (MS) medium in various culture regimes. Highest number of shoots (5.62 ± 0.01), with average length 4.19 ± 0.01 cm, was initially achieved with medium containing 0.5 mg/l N6-benzyladenine (BA) and 3% sucrose. Repeated subculturing of newly formed nodal parts after each harvest up to sixth passage, yielded highest number of shoots (about 32.27) per explants was obtained after fourth passage. Rooting of shoots occurred on 1/2 MS medium supplemented with 1.0 mg/1 Indole-3-butyric acid (IBA). About 90% (89.16) of the plantlets transferred to the mixture of sand:soil:organic manure (2:2:1) in small plastic pots acclimatized successfully. Genetic stability of the discussed protocol was confirmed by two DNA-based fingerprinting techniques i.e. RAPD (random amplified polymorphic DNA) and ISSR (inter-simple sequence repeat). This protocol can be used for commercial propagation and for future genetic improvement studies. PMID:26693403

Comparative Digital Gene Expression Analysis of Tissue-Cultured Plantlets of Highly Resistant and Susceptible Banana Cultivarsin Response to Fusarium oxysporum

Directory of Open Access Journals (Sweden)

Yuqing Niu

2018-01-01

Full Text Available Banana Fusarium wilt caused by Fusarium oxysporum f. sp. cubense (Foc is one of the most destructive soil-borne diseases. In this study, young tissue-cultured plantlets of banana (Musa spp. AAA cultivars differing in Foc susceptibility were used to reveal their differential responses to this pathogen using digital gene expression (DGE. Data were evaluated by various bioinformatic tools (Venn diagrams, gene ontology (GO annotation and Kyoto encyclopedia of genes and genomes (KEGG pathway analyses and immunofluorescence labelling method to support the identification of gene candidates determining the resistance of banana against Foc. Interestingly, we have identified MaWRKY50 as an important gene involved in both constitutive and induced resistance. We also identified new genes involved in the resistance of banana to Foc, including several other transcription factors (TFs, pathogenesis-related (PR genes and some genes related to the plant cell wall biosynthesis or degradation (e.g., pectinesterases, β-glucosidases, xyloglucan endotransglucosylase/hydrolase and endoglucanase. The resistant banana cultivar shows activation of PR-3 and PR-4 genes as well as formation of different constitutive cell barriers to restrict spreading of the pathogen. These data suggest new mechanisms of banana resistance to Foc.

Direct plantlet inoculation with soil or insect-associated fungi may control cabbage root fly maggots.

Science.gov (United States)

Razinger, Jaka; Lutz, Matthias; Schroers, Hans-Josef; Palmisano, Marilena; Wohler, Christian; Urek, Gregor; Grunder, Jürg

2014-07-01

A potential Delia radicum biological control strategy involving cauliflower plantlet inoculation with various fungi was investigated in a series of laboratory and glasshouse experiments. In addition to entomopathogenic fungi, fungi with a high rhizosphere competence and fungi with the ability to survive as saprotrophs in soil were tested. The following fungal species were evaluated in the experiments: Trichoderma atroviride, T. koningiopsis, T. gamsii, Beauveria bassiana, Metharhizium anisopliae, M. brunneum and Clonostachys solani. A commercial carbosulfan-based insecticide was used as a positive control. Additionally, two commercial products, one based on B. bassiana (Naturalis) and one on Bacillus thuringiensis (Delfin) were used as reference biocontrol agents. The aims were (i) to assess the pathogenicity of the selected fungal isolates to Delia radicum, (ii) to evaluate the fungal isolates' rhizosphere competence, with the emphasis on the persistence of the original inoculum on the growing roots, (iii) to assess possible endophytic plant tissue colonization, and (iv) to evaluate potential plant growth stimulating effects of the added inoculi. Significant pathogenicity of tested fungi against Delia radicum was confirmed in in vitro and glasshouse experiments. All tested fungi persisted on cauliflower rhizoplane. More importantly, the added fungi were found on thoroughly washed roots outside the original point of inoculation. This provided us with evidence that our tested fungi could be transferred via or grow with the elongating roots. In addition to colonizing the rhizoplane, some fungi were found inside the plant root or stem tissue, thus exhibiting endophytic characteristics. The importance of fungal ecology as a criterion in appropriate biological control agent selection is discussed. Copyright © 2014 Elsevier Inc. All rights reserved.

Structural Interaction Between GFP-Labeled Diazotrophic Endophytic Bacterium Herbaspirillum seropedicae RAM10 and Pineapple Plantlets 'VitóRia'.

Science.gov (United States)

Estrela Borges Baldotto, Lílian; Lopes Olivares, Fábio; Bressan-Smith, Ricardo

2011-01-01

The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets 'Vitória' were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae.

Structural interaction between GFP-labeled diazotrophic endophytic bacterium Herbaspirillum seropedicae RAM10 and pineapple plantlets 'Vitória'

Directory of Open Access Journals (Sweden)

Lílian Estrela Borges Baldotto

2011-03-01

Full Text Available The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets 'Vitória' were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae.

Genotypic differences in proembryoid development and green plantlets regeneration through androgenesis in barley varieties Diferenças genotípicas no desenvolvimento de pró-embrióides e regeneração de plântulas verdes via androgênese em genótipo de cevada

Directory of Open Access Journals (Sweden)

Paula Wiethölter

2008-02-01

Full Text Available The development of in vitro haploid plants followed by spontaneous or induced genome duplication allows to achieve, in one generation, the recovery of total homozygosis. The efficiency of the haplodiploidization process through in vitro anther culture of barley is variable among genotypes. This study was aimed at determining the androgenetic response of nine barley genotypes from the breeding program of Embrapa Trigo, analysing proembryoid development and green plantlets regeneration in anthers cultivated in vitro. Cultivar 'BR2' presented the highest average of proembryoids (104/anther and 'MN698' presented the highest average of green plantlets (0,41/anther. There was a significant variation among the average values of barley genotypes for embryo formation and green plantlets regeneration, making possible the selection to combine androgenetic capacity and good agronomic traits.A obtenção, na cevada, de plantas haplóides in vitro e a posterior duplicação natural ou artificial do genoma permitem alcançar a homozigose completa, em uma geração. A eficiência da haplodiploidização pela cultura de anteras é variável entre os genótipos. Foi avaliada a resposta androgenética através da formação de pró-embrióides e da regeneração de plântulas verdes em nove cultivares do programa de melhoramento de cevada da Embrapa Trigo, em anteras cultivadas in vitro. A cultivar "BR2" apresentou maior média de pró-embrióides (104/antera, enquanto "MN698" mostrou a maior média de plântulas verdes (0,41/antera. Houve variação significativa entre os valores médios dos genótipos em relação à formação de pró-embrióides e à regeneração de plântulas verdes, indicando a possibilidade de seleção para combinar a capacidade androgenética com boas características agronômicas.

Induction of embryogenic callus and plantlet regeneration from young leaves of high yielding mature oil palm

Directory of Open Access Journals (Sweden)

Yeedum, I.

2004-09-01

Full Text Available Callus induction and plantlet regeneration from young leaves of high-yielding mature oil palm were carried out using 10-year and 20-year-old trees from Thepa Research Station, Faculty of Natural Resources,Prince of Songkla University, Hat Yai, and Trang Agricultural College, respectively. Culture media used in this experiment were Murashige and Skoog (1962 and Oil Palm supplemented with various concentrations of α-naphthaleneacetic acid (NAA or 2,4- dichlorophenoxy acetic acid (2,4-D or dicamba (Di and antioxidants.Young leaves from 6th to 11st frond were excised, sterilized, cut into 5x5 mm pieces and cultured in the dark at 26±4ºC or 28±0.5ºC for 3 months. The results revealed that MS medium with 200 mg/l ascorbic acid (As and 1 mg/l Di (MS-AsDi gave the highest callus induction percentage (7.93 after culture for 3 months at 28±0.5ºC. Leaf segments from 6th - 8th frond yielded callus forming percentage at 10% (averaged from 1, 2.5 and 5 mg/l Di containing MS medium. Ascorbic acid as an antioxidant at concentration of 200 mg/l supplemented in MS medium in the presence of 2.5 mg/l Di produced the highest callus induction percentage (11.2 and number of nodules (7.06. A high percentage of embryogenic callus formation (66.67 was obtained when the calli were transferred to the same medium component supplemented with 0.5 mg/l Di and 1,000 mg/l casein hydrolysate (CH (MS-AsDiCH. Haustorial-staged embryos were observed to be isolated as an individual embryo and germinated on MS medium without plant growth regulator (MS-free. Development of root could be classified into two distinct types, fibrous and tap root.

Structural Interaction Between GFP-Labeled Diazotrophic Endophytic Bacterium Herbaspirillum seropedicae RAM10 and Pineapple Plantlets ‘VitóRia’

Science.gov (United States)

Estrela Borges Baldotto, Lílian; Lopes Olivares, Fábio; Bressan-Smith, Ricardo

2011-01-01

The events involved in the structural interaction between the diazotrophic endophytic bacterium Herbaspirillum seropedicae, strain RAM10, labeled with green fluorescent protein, and pineapple plantlets ‘Vitória’ were evaluated by means of bright-field and fluorescence microscopy, combined with scanning electron microscopy for 28 days after inoculation. After 6 hours of inoculation, H. seropedicae was already adhered to the roots, colonizing mainly root hair surface and bases, followed by epidermal cell wall junctions. Bacteria adherence in the initial periods occurred mainly in the form of solitary cells and small aggregates with pleomorphic cells. Bacteria infection of root tissue occurred through the cavities caused by the disruption of epidermal cells during the emergence of lateral roots and the endophytic establishment by the colonization of intercellular spaces of the cortical parenchyma. Moreover, within 1 day after inoculation the bacteria were colonizing the shoots. In this region, the preferred sites of epiphytic colonization were epidermal cell wall junctions, peltate scutiform trichomes and non-glandular trichomes. Subsequently, the bacteria occupied the outer periclinal walls of epidermal cells and stomata. The penetration into the shoot occurred passively through stoma aperture followed by the endophytic establishment on the substomatal chambers and spread to the intercellular spaces of spongy chlorenchyma. After 21 days of inoculation, bacterial biofilm were seen at the root hair base and on epidermal cell wall surface of root and leaf, also confirming the epiphytic nature of H. seropedicae. PMID:24031612

Influência de porta-enxertos na resistência de mudas de cajueiro ao estresse salino Influence of rootstocks on the resistance of cashew plantlets to salt stress

Directory of Open Access Journals (Sweden)

Sérgio Luiz Ferreira-Silva

2009-04-01

Full Text Available O objetivo deste trabalho foi avaliar a influência de porta-enxertos na resistência de mudas de cajueiro (Anacardium occidentale L. à salinidade. As mudas foram obtidas pela enxertia do clone BRS 226 sobre os porta-enxertos CAPI 4, CCP 09 e BRS 226. Foram expostas a meio hidropônico sem NaCl (controle ou com NaCl 200 mM (tratamento salino, sob condições controladas de temperatura, umidade e luminosidade, durante 12 dias. O delineamento foi o inteiramente casualizado, em esquema fatorial 3x2 (três combinações de enxerto/porta-enxerto e duas concentrações de NaCl, com quatro repetições. Foram determinados a concentração de Na+, Cl-, K+ e solutos orgânicos e os sintomas visuais de toxicidade nas folhas. Os conteúdos de Na+ e Cl-, a relação K+/Na+ e as concentrações de aminoácidos e de prolina livres nas folhas tiveram relação direta com os sintomas visuais de toxicidade. Os porta-enxertos CAPI 4, CCP 09 e BRS 226 foram classificados como sensível, intermediário e resistente à salinidade elevada, respectivamente. Essa variação foi decorrente da influência do porta-enxerto na partição do Na+ e do Cl-.The aim of this work was to evaluate the influence of cashew (Anacardium occidentale L. rootstocks on salt resistance in grafted plantlets. The plantlets were obtained by grafting the BRS 226 scion on CAPI 4, CCP 09 and BRS 226 rootstocks, and were exposed to nutrient solution without NaCl (control or with 200 mM NaCl (salt treatment in hydroponics under controlled temperature, humidity, and luminosity conditions during 12 days. The experimental design was a completely randomized in factorial arrangement 3x2 (three scion/rootstock combinations, two NaCl concentrations with four replicates. Na+, Cl-, K+ and organic solute concentrations and toxicity symptoms on leaves were determined. Na+ and Cl- contents, the K+/Na+ ratio and the free amino acid and proline concentrations in leaves were directly related to the visual

Resistance selection on banana CV. Ambon Kuning Against Fusarium Wilt

International Nuclear Information System (INIS)

Sutarto, Ismiyarti; Meldia, Yeni; Jumjunidang

1998-01-01

This research was conducted in order to study the occurrence of mutation on irradiated plantlets and their resistance of plants of banana cv. Ambon Kuning against Fusarium wilt. Plantlets of banana cv. Ambon Kuning sized 5 cm were exposed to gamma rays at the doses 5 - 35 Gy intervals, then were subcultured for obtained M 1 V 5 plantlets. More over, the planlets were acclimatized and were planted in the field was already infected by Fasarium (f).culbense (FOC). The result indicated that irradiated plantlets of the doses 20 - 35 Gy were not able to survive up to 6 months after exposing to gamma rays. Abnormalities of M 1 V 5 plantlets originated from irradiated plantlets at the doses 10 and 15 Gy were shown on rossette plantlets with rigid and dark green leaves, and the formation of smooth mass morphologically shaped like calculus. The appearance of plant height and number of suckers of suckers of M 1 V 5 plants in the field was quite various. The number of survival plants after 8 moths planting was 8, 7, 15, and 28, respectively originated from untreated plants and irradiated plantlets at the doses 5, 10, and 15 Gy. After one year planting , only 2 plants were able to survive from irradiated plantlet at the dose 15 Gy. The plants could produce 27 plantlets obtained from culturing their shoot tips. Further study of these plantlets was needed in order create the stability of their resistance to FOC. (author)

Salt Tolerant Mutants in Potato sp. Cara Induced by Gamma Irradiation and Tissue Culture Technique

International Nuclear Information System (INIS)

Sharabash, M.T.; Mohamed, A.A.; Ahmed, F.A.; Afifi, A.M.

2003-01-01

Sprouts of 2-3 cm length were cut off from potatoes cv. Cara tubers and sterilized. Apical meristemic tips were sown to produce virus free plantlets. After 6-8 weeks, the new plantlets became at 5-7cm heights. Micro-propagation was begun to obtain sufficient number of plantlets. Plantlets obtained from micro-propagation stage were divided into 3 groups to be exposed to 0.0,20 or 40Gy gamma rays. The dose rate was 27.7rad/sec. Irradiated and unirradiated plantlets were trimmed to be with 5-6 nodes and without leaves. They were transplanted onto 1/2MS [1] liquid medium supplemented with sodium chloride at different concentrations, i.e.,0.0, 2000 or 4000 ppm. Healthy plantlets were chosen, and all unfit ones were excluded. The plantlets were sub-cultured six times on the same fresh medium with the same concentrations of NaCI, with insisting to exclude all unfit plantlets. All maintained healthy plantlets, in the sixth vegetative generation (M 1 V 6 ), grown under salinity stress, whatever the concentration of NaCI, were considered as mutant lines

Morpho-physiological changes in pineapple plantlets [Ananas comosus (L. merr.] during acclimatization Mudanças morfofisiológicas em plântulas de abacaxi [Ananas comosus (L. Merr.] durante a aclimatização

Directory of Open Access Journals (Sweden)

Ariel Villalobo

2012-12-01

Full Text Available Pineapple producing countries lack good quality propagation material to expand cultivars into new areas. Previously, research protocols may increase the offer of high quality plantlets and speed the introduction of new pineapple cultivars. The present work is to evaluate the morpho-physiological changes in plantlets of pineapple [Ananas comosus (L. Merr. 'MD-2'] during the acclimatization phase. Plantlets were acclimatized under 80% relative humidity, 25.5 °C temperature and photosynthetic photon flux of 400-500 µmol m-2 s-1 as average for 45 d under natural photoperiods. All measurements (plant length, number of leaves and roots, fresh weight, width and length of leaf 'D', net photosynthesis and total transpiration rate were carried out at the end of in vitro rooting phase coincident with 0 d of acclimatization and at 15, 30 and 45 d thereafter. Photosynthetic activity of in vitro plantlets did not increase during the first 30 d of the acclimatization phase. After 30 d, photosynthetic activity ranged from 5.72 to 9.36 µmol CO2 m-2 s-1 while total transpiration ranged from 6.0 to 1.42 mmol H2O m-2 s-1. During the first 30 days there were no significant differences in number of leaves, length or width of the longest ('D' leaf (cm or plant length (cm. However, after 45 days plant fresh weight (g, length and width of the 'D' leaf (cm and root number all increased significantly, while transpiration (mmol H2O m-2 s-1 declined. There were small but significant decreases in chlorophyll a and b (µg g-1 mf.. Increased photosynthetic activity after 30 d shows that the increase in light intensity and the reduction of relative humidity during acclimatization did not constitute inhibitory factors.Os países produtores do abacaxi precisam de mudas de boa qualidade para a introdução de novas culturas nas áreas de cultivo. Os protocolos de pesquisa podem aumentar a oferta de mudas de alta qualidade e acelerar a introdução de novas culturas do abacaxi

Mass propagation of in vitro plantlets and extraction of bioactive compounds from Eurycoma Longifolia Jack (Tongkat Ali) using cell and organ cultures

International Nuclear Information System (INIS)

Sobri Hussein; Rusli Ibrahim

2006-01-01

Eurycoma longifolia Jack or better known in Malaysia as Tongkat Ali is an important herbal plant that owes its popularity locally and worldwide to its aphrodisiac claim and has been sought after as an essential component for the treatment of anti-malarial, anti-ulcer, anti-tumour promoting and anti-parasitic agent and also as health supplements. Quassinoids and canthin-6-one alkaloids contained in Tongkat Ali roots are the main substances having active functions for human body. Propagation is commonly done through seeds or some vegetative means such as cuttings of stems and root, grafting or layering. However, propagation by conventional method is inadequate to meet the demand due to low viability, low germination rate, delayed rooting and long life cycle. Tissue culture has been successfully applied for mass production of plantlets and extraction of many useful secondary metabolites, including pharmaceuticals, pigments and other fine chemicals. Tongkat Ali active substances have also been derived through cell culture technology but its processes need to be optimized before the product can be commercialized. Cell and organ cultures have been proven to be an efficient alternative method for biomass production due to its fast growth and stable metabolite production. In view of its potential commercial value as a plantation crop as well as to conserve its germplasm, micropropagation technique using cell culture system is an attractive method for regeneration and mass propagation. (Author)

Tanggapan Planlet Vanili yang Diradiasi dengan Sinar Gamma dan Keragaannya Setelah Diinokulasi dengan Jamur Fusarium oxysporum f.sp. vanillae

Directory of Open Access Journals (Sweden)

Woerjono Mangoendidjojo

2000-12-01

initiation. The untreated plantlets showed better responses as compared to those treated. Evaluation on their resistance after inoculation indicated that the survival plantlets irradiated at 3 and 1 krad showed lower infection intensity, eventhough variation within a treatment was observed. Evaluation on the individual plantlet basis seemed to be more helpful in order to identify mutant plantlets with better resistance.

Impact Of Thermotherapy And Chlorothalonil On Plantlets Production Of Some Genotypes Of Cassava Manihot Esculenta Crantz Produce In Benin

Directory of Open Access Journals (Sweden)

J.A.

2017-01-01

Full Text Available Cassava Manihot esculenta is a starchy root plant of great economic importance in sub-Saharan Africa and particularly in Benin. Its production is confronted to virus diseases which cause a considerable losses of yield. This work aims to determine the impact of thermotherapy and chlorothalonil in the production of cassava material of plantation. Cuttings of four varieties RB89509 BEN86052 9102319 92B0057 are cultivated under two conditions of thermotherapy and a control under greenhouse during 4 weeks. These different conditions are a closed drying oven with 16 hours photoperiod at 40 C the day and 36C the night a drying oven Binder with photoperiod of 12 hours at 38C the day and 28C the night and the control carried out under the conditions of the greenhouse. The media used was Murashige and Skoog MS added with various amounts of chlorothalonil 0.6 gl and 2gl and control without chlorothalonil. Both techniques of thermotherapy eliminate the virus symptoms of cassava at the rate of 0 seedling infected in thermotherapy against 16 seedlings in natural condition. The technique of closed drying oven significantly favors the production of nodes at 5 level p0.000 and shoots p0.02 on the other hand Binder drying oven has no significant effect on the production of shoots p0.68. The chlorothalonil had a positive effect on in vitro infestations elimination of cassava p0.05 but influenced the growth and development of cassava explants by reducing of nodes production p0.01 without a lethal effect on the plantlets until the dose of 2gl.

Adubação foliar com macro e micronutrientes no crescimento de mudas micropropagadas do abacaxizeiro cv. Gold [Ananas comosus (L. Merrill] em diferentes recipientes Foliar fertilization with macro and micronutrients in the growth of plantlets micropropagated of pineapple cv. Gold [Ananas comosus (L. Merrill] in different containers

Directory of Open Access Journals (Sweden)

Izaias dos Santos Bregonci

2008-06-01

Full Text Available Objetivou-se com este trabalho avaliar o efeito da adubação foliar com macro e micronutrientes no crescimento das mudas micropropagadas do abacaxizeiro cv. Gold [Ananas comosus (L. Merrill], em diferentes recipientes. O experimento foi em esquema fatorial 8x3, adubação foliar em 8 níveis e recipientes em 3 níveis, através de um delineamento inteiramente casualizado com 5 repetições. As mudas foram padronizadas com altura média de 7,12 cm. As adubações foliares foram feitas com uréia, cloreto de potássio, ácido bórico, um formulado comercial com macro e micronutrientes e testemunha (pulverização com água e os recipientes: bandeja de isopor com 200 células; tubete pequeno de 115 cm³; e tubete grande com 300 cm³. O substrato utilizado foi o plantmax hortaliças®. Avaliaram-se as características área foliar, altura de planta e massa seca da parte aérea e da raiz, aos 140 dias do transplantio. Os adubos foliares proporcionaram maior crescimento em área foliar, altura e massa seca da parte aérea às mudas do abacaxizeiro, embora com resultados diferentes. Os adubos foliares não aumentaram a massa seca do sistema radicular. A bandeja de isopor apresentou as menores médias, com todos os adubos foliares para área foliar, altura e massa seca da parte aérea das mudas do abacaxizeiro. O tubete pequeno e o tubete grande apresentaram resultados semelhantes com a maioria dos adubos foliares utilizados.The objective of this work was to evaluate the effect of the foliar fertilization with macro and micronutrients on the growth of the plantlets micropropagated of pineapple cv. Gold [Ananas comosus (L. Merrill] in different containers. The experiment was mounted in factorial arrangement 8x3, with foliar fertilization in 8 levels and containers in 3 levels, through a completely randomized design with five repetitions. The plantlets were standardized with average height of 7,12 cm. The foliar fertilization was used in the urea

Use of radiation for inducing mutants in potatoes through tissue culture technique

International Nuclear Information System (INIS)

Sharabash, M.T.

1997-01-01

Meristem-tips obtained from sprouts of potato tubers, cv. 'Diamant' (Solanum tuberosum) were cultured on MS-medium and multiplied into plantlets through micropropagation. After 2-3 weeks, the micropropagated plantlets had 5-6 nodes each. The plantlets were irradiated with 20 to 40 Gy gamma rays at 27.7 rad/sec. Irradiated plantlets were cut into single nodes and cultured on MS-medium supplemented with 2 g NaCl. Salt resistant plantlets were transferred to MS-liquid medium supplemented with 2g NaCl/l 5.2, and microtubers were collected after 6 weeks. Minitubers were produced under the same level of salinity. (author). 2 tabs

Use of radiation for inducing mutants in potatoes through tissue culture technique

Energy Technology Data Exchange (ETDEWEB)

Sharabash, M T [National Centre for Radiation Research and Technology, Cairo (Egypt)

1997-07-01

Meristem-tips obtained from sprouts of potato tubers, cv. `Diamant` (Solanum tuberosum) were cultured on MS-medium and multiplied into plantlets through micropropagation. After 2-3 weeks, the micropropagated plantlets had 5-6 nodes each. The plantlets were irradiated with 20 to 40 Gy gamma rays at 27.7 rad/sec. Irradiated plantlets were cut into single nodes and cultured on MS-medium supplemented with 2 g NaCl. Salt resistant plantlets were transferred to MS-liquid medium supplemented with 2g NaCl/l 5.2, and microtubers were collected after 6 weeks. Minitubers were produced under the same level of salinity. (author). 2 tabs.

In vitro propagation and assessment of genetic stability of acclimated plantlets of Cornus alba L. using RAPD and ISSR markers.

Science.gov (United States)

Ilczuk, Agnieszka; Jacygrad, Ewelina

2016-01-01

Cornus alba L. (white dogwood) is an important ornamental shrub having a wide range of applications such as reforestation programs and soil retention systems. The vegetative propagation of dogwood by cuttings may be slow, difficult, and cultivar dependent; therefore, an improved micropropagation method was developed. Nodal stem segments of C. alba cultivars 'Aurea' and 'Elegantissima' were cultured on media enriched with six different sources of macronutrients. Media were supplemented with either N 6 -benzyladenine (BA) or thidiazuron (TDZ) in combination with 1-naphthaleneacetic acid (NAA). Regardless of the cultivar, the best shoot proliferation was observed on Lloyd and McCown medium (woody plant medium (WPM)) at pH 6.2, containing 1.0 mg L -1 BA, 0.1 mg L -1 NAA, and 20-30 g L -1 sucrose. Rooting of regenerated shoots was achieved by an in vitro method when different concentrations of NAA or indole-3-butyric acid (IBA) were tested. Microcuttings were rooted for 8 wk on medium enriched with 0.25 mg L -1 NAA and potted into P9 containers in the greenhouse. The final survival rate of the plants after 20 wk was 80% for 'Aurea' and 90% for 'Elegantissima'. Genetic stability of the micropropagated plants was confirmed by using two DNA-based molecular marker techniques. A total of 30 random amplified polymorphic DNA (RAPD) and 20 inter-simple sequence repeat (ISSR) primers resulted in 197-199 and 184-187 distinct and reproducible band classes, respectively, in 'Aurea' and 'Elegantissima' plantlets. All of the RAPD and ISSR profiles were monomorphic and comparable with the mother plant.

Molecular Diversity Analysis of Two in vitro and Irradiated Potato Varieties Expressed by Random Amplified Polymorphic DNA

Directory of Open Access Journals (Sweden)

Ayman El-FIKI

2018-03-01

Full Text Available Potato buds cvs. ҅Valor’ and ‘Spunta’ were cultured in vitro on MS solid medium with 0.2 mg -1 BAP. The resulting plantlets were irradiated with gamma radiation doses 10, 20, 30, 40 and 50 Gy. Irradiated segments were transferred onto fresh MS with BAP and plantlets survival percentage was calculated after eight weeks. Gamma radiation caused the death of 3.8% to 81% in cv. ҅Spunta’ and 2.9% to 83.9% in cv. ҅Valor’. Microtubers produced from irradiated plantlets were decreased with increasing gamma radiation doses, with notable changes in shape, size and numbers. The proline contents in irradiated plantlets were steady increase with gamma radiation doses. The genomic DNA of the two cultivars and ten radiation treatments was amplified with 10 RAPD primers that generated 53 polymorphic bands. The highest number of genetic identity was 0.9672 showed between irradiated plantlets with 20 and 30 Gy in cv. ҅Valor’. However, the highest genetic distance was 0.3995 observed between irradiated plantlets with dose 20 Gy in cv. ҅Valor’ and 30 Gy in cv. ҅Spunta’. The dendrogram generated by cluster analysis distinguished the irradiated plantlets genetically.

Morfologi dan Pertumbuhan Planlet Hasil Induksi Poliploidi melalui Perlakuan Kolkisin pada Kuncup Bunga Anggrek Bulan (Phalaenopsis amabilis (L. Blume

Directory of Open Access Journals (Sweden)

Tubagus Kiki Kawakibi Azmi

2016-06-01

Full Text Available Induction of polyploid gametes is one of useful plant polyploidization methods. Some of its benefits are to obtain triploid and tetraploid progenies at the same time by cross and self pollination. Previous research showed that some morphological characters which could be the indications of polyploidy plantlets before the analysis of chromosome number. Colchicine treatment on flower bud of diploid Phalaenopsis amabilis was conducted to determine the effect of colchicine on flower bud development, plantlets morphology and growth, and potential of polyploidy induction based on plantlets morphology. Colchicine concentrations in the experiment were 0, 50, 500, 1,000, and 2,000 mg L-1, with three days duration of treatments with aluminium foil wraps on flower buds. The results showed that high colchicine concentrations (2,000 mg L-1 inhibited flowers blooming of treated flower buds. Based on morphological characters, plantlets were classified into normal and putative polyploid plantlets. Putative polyploid plantlets from colchicine with the concentration of 50, 500, and 1,000 mg L-1 were 71.2, 86.4, and 100.0% respectively.Keywords: colchicine concentration, morphological characters, normal plantlets, putative polyploidy, reproductive organ

Study On In Vitro Variation Generation Of Cymbidium Labell Anna Belle By Gamma Co-60 Irradiation

International Nuclear Information System (INIS)

Nguyen Huynh Phuong Uyen; Vo Thi Thu Ha; Le Quang Luan

2011-01-01

Radiation ionization is one of useful methods for mutation breeding of plants. In this study, γ-ray (Co-60) was used for generating morphological variations of Cymbidium Labell Anna Belle. The lethal doses of 50% samples (LD 50 ) after 4 months cultivation were determined at 35.0 Gy for protocorm like bodies (PLBs), 37.4 Gy for in vitro shoot bubs and 70.1 Gy for plantlets. PLBs were found as suitable samples for generation of variation of mentioned Cymbidium. The dose range from 20 to 50 Gy was determined with a high ability of variation on C. Labell Anna Belle. The highest frequency of variation was obtained ca. 3.82 x 10 -3 at the dose of 30 Gy. 157 in vitro variant plantlets were selected in M 1 V 1 generation, among variations, the variations were mainly found with 5 following types: removal of chlorophyll (8 plantlets), the leaves become shorter (64 plantlets), the leaves become longer (64 plantlets), increase of leaf number (11 plantlets) and the colour of pericardium was changed from green into violet (10 plantlets). Twenty one variant lines were selected in M 1 V 2 generation. (author)

Response of Musa species to macro-propagation. I: Genetic and ...

African Journals Online (AJOL)

Days to emergence of the first and fifth plantlet were shortest in 'FHIA 17\\' (a dessert banana hybrid) and longest in 'PITA 25\\' (a plantain hybrid). Emergence of the first three plantlets in landrace plantain ('Agbagba\\') was earlier than in dessert banana landrace ('Nsukka Local\\'). A higher proportion of plantlets excised from ...

Study of the effects of Cobalt 60 gamma radiations on the seeds of two textile hibiscus species. Influence of seed water content on their radiosensitivity. Mutagenic effects on two plantlet generation

International Nuclear Information System (INIS)

Mahama, Adamou.

1980-10-01

This work on textile hibiscus was undertaken for two reasons: to contribute by physical mutagenesis methods to the genetic improvement of these species and especially to obtain the genetic diversity which is seriously lacking in the species sabdariffa var. altissima; to add to research on the effects of mutagenic seed treatments and thus to help towards a better definition of optimum seed irradiation conditions. These two purposes have been fulfilled. The main results may be summed up as follows. - Concerning the improvement of textile hibiscus. Many strains with various new features were selected at the second generation and may lead either to new varieties or at least to interesting precursors for selection programmes. In the second generation derived from Hibiscus saddariffa var. THS 22 some plants were observed with one or more of the following characteristics specific to the edible form: ramified bearing - fleshy calix - smooth leaf. - Concerning the study of mutagenic seed treatment effects. Our experiments showed up the importance of the seed water content and its effect on the response of first-generation plantlets to cobalt 60 γ-ray treatments [fr

Interactive effects of aluminum and cadmium on phenolic compounds, antioxidant enzyme activity and oxidative stress in blueberry (Vaccinium corymbosum L.) plantlets cultivated in vitro.

Science.gov (United States)

Manquián-Cerda, K; Cruces, E; Escudey, M; Zúñiga, G; Calderón, R

2018-04-15

To evaluate the potential role of phenolic compounds in Al and Cd stress tolerance mechanisms, Vaccinium corymbosum cv. Legacy plantlets were exposed to different metal concentrations. The present study used an in vitro plant model to test the effects of the following treatments: 100μM Al; 100μMAl + 50μMCd; and 100μMAl + 100μMCd during periods of 7, 14, 21 and 30 days. The oxidative damage was determined by the accumulation of malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ). The antioxidant activity values were determined using 1,1-diphenyl-2-picrylhydrazine (DPPH) and the ferric reducing antioxidant power test (FRAP). Additionally, the phenolic compound concentrations were determined using HPLC-DAD. The exposure to Al and Cd increased the MDA and H 2 O 2 contents differentially, while the antioxidant capacity values showed differences between DPPH and FRAP with the largest changes in FRAP relative to Cd. SOD had the highest activity in the first 7 days, leading to a significant increase in phenolic compounds observed after 14 days, and chlorogenic acid was the major compound identified. Our results revealed that phenolic compounds seem to play an important role in the response to ROS. Therefore, the mechanisms of tolerance to Al and Cd in V. corymbosum will be determined by the type of metal and time of exposure. Copyright © 2017 Elsevier Inc. All rights reserved.

THE BIOTECHNOLOGY OF EMBRYOGENIC CELL LINES OBTAINING AND PLANTLETS OF CONIFEROUS SPECIES IN SIBERIA IN CULTURE IN VITRO

Directory of Open Access Journals (Sweden)

Tretiakova I.

2012-08-01

Full Text Available Experiments of culturing the immature isolated embryos and megagamethophytes of Siberian coniferous species were carried out on different modified media: ½ LV medium for Pinus sibirica and Pinus pumila, MSG and AI media (patent № 2431651 for Larix sibirica and Larix gmelinii, DCR medium for Picea ajanensis. For induction of embryogenic callus every species needs the optimized medium supplemented with L-glutamine, casein hydrolysate, ascorbic acid and hormones with different concentrations and their different proportions. The active proliferation of embryonal masses is observed on the same medium with reduced concentration of cytokinins. The proliferation of embryonal masses was significantly improved when they were subcultured after dispersing in liquid medium. The somatic embryos from embryonal masses are matured on basal medium with ABA (60-120 mM and PEG. In spite of species specificity the embryogenesis of morphogenic structures had the same scheme: elongation and asymmetric division of somatic cells, formation of initial cells and embryonal tubes, development of globular, torpedo and bipolar somatic embryos, embryos maturation and germination. However, not all donor-plants of coniferous species can form the embryogenic cell lines and somatic embryos. The active development of embryonal masses and formation of somatic embryos are observed from zygotic embryo of hybrid seeds of P. sibirica and L. sibirica. The obtained embryogenic lines were characterized by different proliferative activity. During 10 months cultivation the value of embryonal masses in different lines was 140-570 g. The number of somatic embryos varies from 210 to 410 per 100 mg of callus fresh weight. Decreasing proliferation activity did not observed during 24-45 months cultivation. However, development of somatic embryos in long cultivated lines decreased. Maturation of somatic embryos and development of plantlets were established in L. sibirica and P. pumila 50

Germinação de sementes e crescimento de plântulas de soja (Glycine max L. Merrill sob cobertura vegetal = Soybean (Glycine max L. Merrill seed germination and plantlet growth under vegetation cover

Directory of Open Access Journals (Sweden)

Lúcia Helena Pereira Nóbrega

2009-07-01

Full Text Available A eficiência do sistema de rotação de cultura requer, entre outrostratamentos, a escolha adequada das espécies a serem instaladas. Culturas vegetais podem apresentar compostos aleloquímicos, os quais são liberados por meio de pelos radiculares, sementes, raízes, colmos e folhas, em quantidades variáveis, capazes de interferir nas culturas subsequentes, comprometendo a produção. Assim, este estudoanalisou o potencial alelopático de aveia-preta (Avena strigosa Schreb (AP, nabo forrageiro (Raphanus sativus L. (NF, ervilhaca (Vicia sativa L. (ER, azevém (Lolium multiflorum Lam. (AZ e consórcio (CO - AP+ER+NF na germinação de sementes e crescimento de plântulas de soja. O experimento foi em laboratório, com substrato deareia, onde foram cultivadas plantas de cobertura, por 30 dias, mantendo os restos radiculares das plantas com e sem restos de parte aérea. Observou-se redução na emergência de plântulas de soja sob CO, AZ e AP. O índice de velocidade de emergência (IVE, a porcentagem de emergência em areia (EA e a massa fresca de hipocótilo (MFH foram afetados negativamente pelas plantas de cobertura.In order to be efficient, the crop rotation system requires, among other factors, an adequate choice of species to be installed. Vegetalcultures can feature allelochemical compounds, released by root hairs, roots, stems and leaves in variable amounts, which are able to interfere on subsequent cultures, as well as cause a delay in their production. Thus, this study analyzed the allelopathic potentialof black oat (Avena strigosa Schreb (BO, forage turnip (Raphanus sativus L. (FT, vetch (Vicia sativa L. (V, ryegrass (Lolium multiflorum Lam. (RG and consortium (CO - BO+V+FT on soybean seed germination and plantlet growth. This trial was carried out in a laboratory, with sand substrate, where coverage plants were grown, during 30 days. The remaining root portions of plants were kept with and without the remaining aerial part portions

Mutation induction of blechnum gibium a fern species through in vitro propagation

International Nuclear Information System (INIS)

Azhar Mohamad; Amran Abd Halim; Rusli Ibrahim

2002-01-01

Blechnum gibum is a fern with beutiful symmetry and unique palm-like appearance. Mutation induction is an alternative way to increase its variability in characteristics. Information on the radiosensitivity test is an important factor in deciding the approach to be taken for genetic changes of the fern in mutation breeding. Plantlets of B. gibum were irradiated with doses of 0, 10, 20, 40, 60, 80, 100 and 150 Gy using a gamma cell with a 60 Co source at dose rate of 0.25 Gy s -1 . . Irradiated plantlets were cultured on semi solid modified hormone free Murashige and Skoog (MS) media and incubated at 24 degree C with a photoperiod of 16 hours (3500 lux). Radiosensitivity was assessed by the survival rate 7 weeks after the treatment. Increasing gamma ray doses caused a reduction of survival rate as well as the number of fronds produced per plantlet. LD 50 for B. gibum was 54 Gy as estimated from the dose rate vs survival curve. Regenerated plantlets from treated plantlets showed a stunted and slow recovery growth especially for higher doses as compared to control plantlets. (Author)

Acclimatization of in Vitro-derived Dendrobium

Directory of Open Access Journals (Sweden)

Jaime A. Teixeira da Silva

2017-05-01

Full Text Available The successful ex vitro establishment of Dendrobium plantlets raised in vitro determines the quality of the end product (cut flowers or potted plants in commercial production for economic gain. When in vitro Dendrobium plantlets are transplanted from the culture room to greenhouse conditions, they may desiccate or wilt rapidly and can die as a result of changes in the environment, unless substantial precautions are taken to adapt plantlets to a new environment. The acclimatization of in vitro-grown Dendrobium plantlets to an ex vitro environment by gradually weaning them towards ambient relative humidity and light levels facilitates better survival of young and physiologically sensitive plantlets. Dendrobium plantlets raised in vitro must thus undergo a period of acclimatization or transitional development to correct anatomical abnormalities and to enhance their physiological performance to ensure survival under ex vitro conditions. The most common approach to improve the survival of Dendrobium plantlets upon transfer to an ex vitro environment is their gradual adaptation to that environment. Under such conditions, plants convert rapidly from a heterotrophic or photomixotrophic state to an autotrophic growth, develop a fully functional root system, and better control their stomatal and cuticular transpiration. Gradual adaptation is carried out in a greenhouse by decreasing relative humidity using fog or mist chambers and by increasing light intensity using shading techniques. This review details the acclimatization and ex vitro survival of Dendrobium plants produced in vitro. This advice is also useful for other orchids.

Effect of indole-3-butyric acid (IBA) on in vitro root induction in ...

African Journals Online (AJOL)

Root induction pre-developed in vitro plantlets of orchid was carried out using indole-3-butyric acid (IBA) (0, 0.5, 1.0, 1.5, 2.0, 2.5 and 3 mM) on basal Murashige and Skoog (MS) medium. Among the concentrations of IBA, the number of roots per plantlet with 1 mM IBA was found to be the highest (2.25 roots per plantlet) ...

IMPROVEMENTS OF CLONAL MICROPROPAGATION TECHNOLOGY OF APPLE ROOTSTOCKS AT THE STAGE OF INTRODUCTION TO IN VITRO CULTURE

OpenAIRE

Besedina E. N.; Buntsevich L. L.

2015-01-01

The stage of introduction of plantlets to in vitro culture is an important stage of technology of clonal micropropagation of plants. For the purpose of decrease in a share of the plantlets that were lost from an infection and increase of their regeneration ability, sterilizers and antibiotics, effective and safe for apple rootstocks’ plantlets wee allocated, and also favorable terms of introduction to in vitro culture were allocated. As a result of the conducted researches, we have establishe...

Enhanced resistance to blister blight in transgenic tea (Camellia sinensis [L.] O. Kuntze) by overexpression of class I chitinase gene from potato (Solanum tuberosum).

Science.gov (United States)

Singh, H Ranjit; Deka, Manab; Das, Sudripta

2015-07-01

Tea is the second most consumed beverage in the world. A crop loss of up to 43 % has been reported due to blister blight disease of tea caused by a fungus, Exobasidium vexans. Thus, it directly affects the tea industry qualitatively and quantitatively. Solanum tuberosum class I chitinase gene (AF153195) is a plant pathogenesis-related gene. It was introduced into tea genome via Agrobacterium-mediated transformation with hygromycin phosphotransferase (hpt) gene conferring hygromycin resistance as plant selectable marker. A total of 41 hygromycin resistant plantlets were obtained, and PCR analysis established 12 plantlets confirming about the stable integration of transgene in the plant genome. Real-time PCR detected transgene expression in four transgenic plantlets (T28, C57, C9, and T31). Resistance to biotrophic fungal pathogen, E. vexans, was tested by detached leaf infection assay of greenhouse acclimated plantlets. An inhibitory activity against the fungal pathogen was evident from the detached leaves from the transformants compared with the control. Fungal lesion formed on control plantlet whereas the transgenic plantlets showed resistance to inoculated fungal pathogen by the formation of hypersensitivity reaction area. This result suggests that constitutive expression of the potato class I chitinase gene can be exploited to improve resistance to fungal pathogen, E. vexans, in economical perennial plantation crop like tea.

Detection of Methylobacterium radiotolerans IMBG290 in potato plants by in situ hybridization

Directory of Open Access Journals (Sweden)

Pirttila A. M.

2009-04-01

Full Text Available A new bacterial strain of pink-pigmented facultative methylotroph (M. radiotolerans IMBG290 which was previously isolated from in vitro grown potato plantlets after their inoculation with Pseudomonas fluorescens IMBG163 was detected in tissues by in situ hybridization method (ISH/FISH. The presence of Methylobacterium rRNA was observed in leaves and stems of potato plantlets, whereas no signal was detected in potato roots. The signal was less abundant in the untreated plants than in the plantlets infected with M. radiotolerans IMBG290.

Dynamics of Short-Term Phosphorus Uptake by Intact Mycorrhizal and Non-mycorrhizal Maize Plants Grown in a Circulatory Semi-Hydroponic Cultivation System.

Science.gov (United States)

Garcés-Ruiz, Mónica; Calonne-Salmon, Maryline; Plouznikoff, Katia; Misson, Coralie; Navarrete-Mier, Micaela; Cranenbrouck, Sylvie; Declerck, Stéphane

2017-01-01

A non-destructive cultivation system was developed to study the dynamics of phosphorus (Pi) uptake by mycorrhizal and non-mycorrhizal maize plantlets. The system consisted of a plant container connected via silicon tubes to a glass bottle containing a nutrient solution supplemented with Pi. The nutrient solution is pumped with a peristaltic pump to the upper part of the container via the silicon tubes and the solution percolate through the plantlet container back into the glass bottle. Pi is sampled from the glass bottle at regular intervals and concentration evaluated. Maize plantlets were colonized by the AMF Rhizophagus irregularis MUCL 41833 and Pi uptake quantified at fixed intervals (9, 21, and 42 h) from the depletion of the Pi in the nutrient solution flowing through the plantlets containers. Plants and fungus grew well in the perlite substrate. The concentration of Pi in the bottles followed an almost linear decrease over time, demonstrating a depletion of Pi in the circulating solution and a concomitant uptake/immobilization by the plantlet-AMF associates in the containers. The Pi uptake rate was significantly increased in the AMF-colonized plantlets (at 9 and 21 h) as compared to non-colonized plantlets, although no correlation was noticed with plant growth or P accumulation in shoots. The circulatory semi-hydroponic cultivation system developed was adequate for measuring Pi depletion in a nutrient solution and by corollary Pi uptake/immobilization by the plant-AMF associates. The measurements were non-destructive so that the time course of Pi uptake could be monitored without disturbing the growth of the plant and its fungal associate. The system further opens the door to study the dynamics of other micro and macro-nutrients as well as their uptake under stressed growth conditions such as salinity, pollution by hydrocarbon contaminants or potential toxic elements.

DNA methylation in sugarcane somaclonal variants assessed through methylation-sensitive amplified polymorphism.

Science.gov (United States)

Francischini, J H M B; Kemper, E L; Costa, J B; Manechini, J R V; Pinto, L R

2017-05-04

Micropropagation is an important tool for large-scale multiplication of plant superior genotypes. However, somaclonal variation is one of the drawbacks of this process. Changes in DNA methylation have been widely reported as one of the main causes of somaclonal variations in plants. In order to investigate the occurrence of changes in the methylation pattern of sugarcane somaclonal variants, the MSAP (methylation-sensitive amplified polymorphism) technique was applied to micro-propagated plantlets sampled at the third subculture phase. The mother plant, in vitro normal plantlets, and in vitro abnormal plantlets (somaclonal variants) of four sugarcane clones were screened against 16 MSAP selective primers for EcoRI/MspI and EcoRI/HpaII restriction enzymes. A total of 1005 and 1200 MSAP-derived markers with polymorphism percentages of 28.36 and 40.67 were obtained for EcoRI/HpaII and EcoRI/MspI restriction enzyme combinations, respectively. The genetic similarity between the mother plant and the somaclonal variants ranged from 0.877 to 0.911 (EcoRI/MspI) and from 0.928 to 0.955 (EcoRI/HpaII). Most of the MASPs among mother plant and micro-propagated plantlets were derived from EcoRI/MspI restriction enzymes suggesting alteration due to gain or loss of internal cytosine methylation. A higher rate of loss of methylation (hypomethylation) than gain of methylation (hypermethylation) was observed in the abnormal in vitro sugarcane plantlets. Although changes in the methylation pattern were also observed in the in vitro normal plantlets, they were lower than those observed for the in vitro abnormal plantlets. The MASP technique proved to be a promising tool to early assessment of genetic fidelity of micro-propagated sugarcane plants.

Dynamics of Short-Term Phosphorus Uptake by Intact Mycorrhizal and Non-mycorrhizal Maize Plants Grown in a Circulatory Semi-Hydroponic Cultivation System

Directory of Open Access Journals (Sweden)

Mónica Garcés-Ruiz

2017-08-01

Full Text Available A non-destructive cultivation system was developed to study the dynamics of phosphorus (Pi uptake by mycorrhizal and non-mycorrhizal maize plantlets. The system consisted of a plant container connected via silicon tubes to a glass bottle containing a nutrient solution supplemented with Pi. The nutrient solution is pumped with a peristaltic pump to the upper part of the container via the silicon tubes and the solution percolate through the plantlet container back into the glass bottle. Pi is sampled from the glass bottle at regular intervals and concentration evaluated. Maize plantlets were colonized by the AMF Rhizophagus irregularis MUCL 41833 and Pi uptake quantified at fixed intervals (9, 21, and 42 h from the depletion of the Pi in the nutrient solution flowing through the plantlets containers. Plants and fungus grew well in the perlite substrate. The concentration of Pi in the bottles followed an almost linear decrease over time, demonstrating a depletion of Pi in the circulating solution and a concomitant uptake/immobilization by the plantlet-AMF associates in the containers. The Pi uptake rate was significantly increased in the AMF-colonized plantlets (at 9 and 21 h as compared to non-colonized plantlets, although no correlation was noticed with plant growth or P accumulation in shoots. The circulatory semi-hydroponic cultivation system developed was adequate for measuring Pi depletion in a nutrient solution and by corollary Pi uptake/immobilization by the plant-AMF associates. The measurements were non-destructive so that the time course of Pi uptake could be monitored without disturbing the growth of the plant and its fungal associate. The system further opens the door to study the dynamics of other micro and macro-nutrients as well as their uptake under stressed growth conditions such as salinity, pollution by hydrocarbon contaminants or potential toxic elements.

Effect of applied synthetic auxin on root growth in plantlet propagation by cuttage and tissue culture; Sashiki to soshiki baiyo ni okeru gosei auxin rui no shiyo koka

Energy Technology Data Exchange (ETDEWEB)

Shoji, K; Yoshihara, T [Central Research Institute of Electric Power Industry, Tokyo (Japan)

1996-12-01

The effect of synthetic plant hormone 4-C1-IAA and TFIBA on root growth in plantlet propagation was clarified by the cuttage and the issue culture of strawberry seedling production. A periwinkle, vine, and azalea are the effect of 4-C1-IAA on root growth, and a promotion effect was recognized for rooting and root elongation. The concentration of 4-C1-IAA in which the growth promotion effect of a root most appears varies depending on the species of a plant. The concentration of a periwinkle was 20 ppm, and that of an azalea was 2000 ppm. The growth promotion effect of a root in 4-C1-IAA and TFIBA was compared with IBA for an azalea. The result showed that 4-C1-IAA is the same in the effect as IBA and that TFIBA is higher than for IBA. The growth of a vine`s terminal bud was promoted by the effect of TFIBA on root growth, and the callus occurring when IBA was treated was not formed. The rooting of a strawberry was promoted by the effect of TFIBA on root growth. The combined use of TFIBA and BA promotes the growth of a side bud and forms a multi-bud plant. However, rooting was inhibited. The callus caused by the effect of BA on root growth could be suppressed through the combined use with TFIBA. 6 refs., 7 figs., 2 tabs.

The Occurrence of Hyperhydricity on Several Carnations (Dianthus carryophyllus L. Cultivars during Low Temperature Storage

Directory of Open Access Journals (Sweden)

KURNIAWAN BUDIARTO

2009-04-01

Full Text Available The incident of hyperhydricity was a common problem in propagated carnation (Dianthus carryophyllus L. during in vitro culture. Due to its possible relations with the decrease in phenotypic performance of plantlets, the observation on the occurrence of hyperhydricity was conducted on several in vitro conserved carnation cultivars. The research was conducted from July 2007 to August 2008 at The Indonesian Ornamental Crops Research Institute, Cianjur, West Java. A complete factorial experiment with 24 replications was designed to accomplish the combination of two factors. The first factor was six commercial carnation cultivars, namely light pink candy, malaga, opera, white candy, liberty and pink maladi, while the seconds dealt with type of conservation media, i.e. ½MS+DMSO 3%, ½MS+DMSO 3%+3% sucrose and control (½MS+3% sucrose. The results showed that the percentage of hyperhydric plantlet and plantlet viability after in vitro conservation were varied among carnation cultivars. Single treatment of sucrose had the least capacity in inducing plantlet resistance to low temperature conditions during in vitro conservation. Supplemental DMSO postponed the occurrence of hyperhydricity and with the existence of sucrose, higher plantlet viability were achieved.

Induksi Poliploidi Phalaenopsis amabilis (L. Blume dan Phalaenopsis amboinensis J. J. Smith dengan Kolkisin dalam Kultur In Vitro

Directory of Open Access Journals (Sweden)

Eka Martha Della Rahayu

2016-02-01

Full Text Available ABSTRACTPhalaenopsis amabilis (L. Blume and Phalaenopsis amboinensis J.J. Smith (diploid are important in Phalaenopsis breeding. Polyploid species are needed for crossing with polyploid hybrid varities of Phalaenopsis. The objectives of this study were to obtain effective concentration of colchicine to induce polyploidy and to produce polyploid plantlets of P. amabilis and P. amboinensis. Experiment was arranged in randomized complete block design with one factor, the colchicine concentration. Protocorms of P. amabilis and P. amboinensis were immersed in half strength of Murashige-Skoog (1/2 MS liquid media added with colchicine (0; 0.5; 5; 25; 50, and 75 mg L-1 for 10 days. The results showed that higher concentration of colchicine on both species did not have significant effect on the survival of the plantlets at 24 weeks after treatment. The average number of leaves and roots of colchicine treated planlets from both species were less than the control plantlets. Immersing protocorm in colchicine at concentration of 50 mg L-1 for 10 days was effective in inducing polyploid plantlets of P. amabilis and P. amboinensis with the frequency of 33.3% and 40%, respectively. Polyploid plantlet has larger stomata size and lower stomata density than the diploid ones.Keywords: chromosome number, colchicine, polyploid, protocorm, stomatal density, stomatal size

Methylobacterium sp. resides in unculturable state in potato tissues in vitro and becomes culturable after induction by Pseudomonas fluorescens IMGB163.

Science.gov (United States)

Podolich, O; Laschevskyy, V; Ovcharenko, L; Kozyrovska, N; Pirttilä, A M

2009-03-01

To induce growth of endophytic bacteria residing in an unculturable state in tissues of in vitro-grown potato plantlets. To isolate and identify the induced bacteria and to localize the strains in tissues of in vitro-grown potato plantlets. The inoculation of in vitro-grown potato plants with Pseudomonas fluorescens IMBG163 led to induction of another bacterium, a pink-pigmented facultative methylotroph that was identified as Methylobacterium sp. using phylogenetic 16S rDNA approach. Two molecular methods were used for localizing methylobacteria in potato plantlets: PCR and in situ hybridization (ISH/FISH). A PCR product specific for the Methylobacterium genus was found in DNA isolated from the surface-sterilized plantlet leaves. Presence of Methylobacterium rRNA was detected by ISH/FISH in leaves and stems of inoculated as well as axenic potato plantlets although the bacterium cannot be isolated from the axenic plants. Methylobacterium sp. resides in unculturable state within tissues of in vitro-grown potato plants and becomes culturable after inoculation with P. fluorescens IMBG163. In order to develop endophytic biofertilizers and biocontrol agents, a detailed knowledge of the life-style of endophytes is essential. To our knowledge, this is the first report on increase of the culturability of endophytes in response to inoculation by nonpathogenic bacteria.

How Planting Density Affects Number and Yield of Potato Minitubers in a Commercial Glasshouse Production System

NARCIS (Netherlands)

Veeken, van der A.J.H.; Lommen, W.J.M.

2009-01-01

Commercial potato minituber production systems aim at high tuber numbers per plant. This study investigated by which mechanisms planting density (25.0, 62.5 and 145.8 plants/m2) of in vitro derived plantlets affected minituber yield and minituber number per plantlet. Lowering planting density

Mutant in tobacco anther culture induced by 60Co γ-rays

International Nuclear Information System (INIS)

Tong Daoru; Jia Xinghua

1991-01-01

The tobacco anthers at uninucleate eccentric stage were irradiated by 60 Co γ-rays for the purpose of inducing desirable mutants. The results showed that the induction frequency of plantlets increased following 1kR of 60 Co γ-rays treatment. However, the time of plantlet induction was delayed and the percentage of responding anthers as well as the number of plantlets induced per anther significantly decreased after 3kR of 60 Co γ-ray radiation which was considered as a semilethal exposure. The plantlet numbers induced per anther were extremely low following 6kR of 60 Co γ-ray radiation. A white flower mutant appeared in the induced progenies. The tobacco leaf quality of this mutant were significantly improved as compared with its parental line. The mutant line has been tested and proved to have commercial value though the resistance to the black shank of tobacco slightly decreased as compared with the parental line

Indução de metabólitos secundários em plântulas de Hypericum brasiliense Choisy crescendo in vitro Induction of secondary metabolites in plantlets of Hypericum brasiliense Choisy in vitro

Directory of Open Access Journals (Sweden)

Mariella Araújo Luna Velloso

2009-01-01

Full Text Available A produção de rutina, quercetina, 1,5-diidroxixantona e ácido betulínico foi investigada em plântulas de H. brasiliense crescendo in vitro, sob a influência de ácido salicílico, polietilenoglicol, NaCl, 24-epibrassinolídeo, benzotiadiazole (BION, metiljasmonato e concentrações aumentadas de boro e nitrogênio no meio líquido de cultura. As avaliações foram feitas após 5 e 10 dias do início dos tratamentos. Os maiores aumentos de conteúdo foram observados com quercetina para boro e ácido salicílico aos 5 dias, e 24-epibrassinolídeo e BION aos 10 dias.The production of rutin, quercetin, 1,5-dihydroxyxanthone and betulinic acid was investigated in plantlets of H. brasieliense in vitro, and exposed to salycilic acid, poliethylene glycol, NaCl, 24-epibrassinolide, benzothiadiazole (BION, methyljasmonate and increased concentrations of boron and nitrogen in the liquid culture medium. Evaluations of the contents were carried out after 5 and 10 days of treatments. The highest increase was observed in quercetin in the salycilic acid and B treatments after 5 days of exposure, and in 24-epibrassinolide and BION after 10 days.

Improvement of cassava for resistance to insect pests and diseases

International Nuclear Information System (INIS)

Mwanga, R.O.M.

1997-01-01

The African cassava mosaic virus and cassava mealybug are devastating the cassava crop in Uganda. Because of the severe widespread occurrence of the virus and mealybug, in vitro cultured cassava plantlets instead of stem cuttings will be irradiated. In addition, the project has incorporated sweet potato. Installation of tissue culture laboratory at Namulonge was completed in early 1993. Work is in progress to establish efficient in vitro culture micropropagation techniques for the two crops. Small numbers of cassava plantlets of varieties 'TMS 30337' and 'TMS 4(2)1425' and sweet potato entry 30 are in vitro culture. Mass irradiation of plantlets is planned in future. (author). 4 refs

Improvement of cassava for resistance to insect pests and diseases

Energy Technology Data Exchange (ETDEWEB)

Mwanga, R O.M. [Namulonge Agricultural and Animal Production Research Inst., Kampala (Uganda)

1997-12-01

The African cassava mosaic virus and cassava mealybug are devastating the cassava crop in Uganda. Because of the severe widespread occurrence of the virus and mealybug, in vitro cultured cassava plantlets instead of stem cuttings will be irradiated. In addition, the project has incorporated sweet potato. Installation of tissue culture laboratory at Namulonge was completed in early 1993. Work is in progress to establish efficient in vitro culture micropropagation techniques for the two crops. Small numbers of cassava plantlets of varieties `TMS 30337` and `TMS 4(2)1425` and sweet potato entry 30 are in vitro culture. Mass irradiation of plantlets is planned in future. (author). 4 refs.

Effect of nodal positions, seasonal variations, shoot clump and ...

African Journals Online (AJOL)

SAM

2014-05-07

May 7, 2014 ... neck in the micro propagation of many plants (Hazarika,. 2003). Plantlets that were grown in vitro had continuously exposed to a unique microenvironment with high level of humidity, aseptic conditions, on a medium containing ample sugar and nutrients. Thereafter, in vitro raised plantlets of the species ...

Effects of adenine sulphate, glutamine and casein hydrolysate on in ...

African Journals Online (AJOL)

The in vitro raised plantlets were acclimatized under culture room conditions in different potting mixture, of which the combination of garden soil, sand and vermiculite mixture in 1:1:2 ratio was found most supportive. After 30 days of acclimatization, plantlets were transferred to soil, where established plants showed more ...

In vitro propagation of Irvingia gabonensis

African Journals Online (AJOL)

GREGO

2007-04-16

Apr 16, 2007 ... Full-grown plantlets were obtained and work is in progress on mass propagation. ... subsequent mass propagation to produce seedlings for farmers, and to improve food security and ... Shooting and rooting were observed, and full grown plantlets were obtained. ¼ MS +0.2 mg KIN. +0.1 mg NAA. Rooting ...

Use of plastic films for weed control during field establishment of micropropagated hardwoods

Science.gov (United States)

J. W. Van Sambeek; John E. Preece; Carl A. Huetteman; Paul L. Roth

1995-01-01

This study compares the use of plastic films to conventional methods for establishing hardwoods on a recently cultivated old field site using 1-year-old micropropagated plantlets of white ash (Fraxinus americana L.) and silver maple (Acer saccharinum L.). After one growing season in the field, height of plantlets with all weed...

Influence of salicylic and succinic acids on antioxidant enzymes activity, heat resistance and productivity of Panicum miliaceum L.

Directory of Open Access Journals (Sweden)

Miroshnichenko N.N.

2011-05-01

Full Text Available The influence of treatment of millet (Panicum miliaceum L. seeds with the solutions of salicylic and succinic acids on the heat resistance of plantlets and activity of antioxidant enzymes – superoxide dismutase (SOD, catalase and peroxidase – in them have been investigated. In the micro-field experiment the influence of these acids on the millet yield was estimated. The action of salicylic (10 μM and succinic (1 mM acids caused the increase of plantlets resistance to the damaging heating that expressed in the rise of relative quantity of survived plantlets in 5 days after heating at the temperature of 47°С and in the reduced content of lipid peroxidation product malonic dialdehyde during the poststress period. The increase of activity of SOD, catalase and peroxidase took place in millet plantlets under the influence of salicylic and succinic acids. The increase of productivity of millet grain under the action of salicylic and succinic acids on 13,3-52,0 and 6,4-38,8% respectively depending on weather conditions in the field experiments was noted.

Radiation induced variation in potato for tolerance to salinity using tissue culture technique

International Nuclear Information System (INIS)

Sharabash, M.T.

2001-01-01

Meristem-tips of potato (Solanum tuberosum) cv. 'Diamant', obtained from tuber sprouts, were cultured on MS medium, and multiplied into plantlets through micropropagation. To induce variation for salt tolerance, the obtained plantlets were irradiated with 0, 20, and 40 Gy gamma rays at 27.7 rad/sec. Irradiated plantlets were cut into single nodes and cultured on MS medium, supplemented with 2000 and 4000 ppm NaCI. Salt tolerant plantlets were transferred for tuberization on MS liquid medium supplemented with the same concentration of NaCI. Micro-tubers, collected after 6 weeks of culture, had fresh weight between 0.03 to 0.3 g. Mini-tubers were obtained by planting micro-tubers in 25 cm pots under insect proof greenhouse. Mini-tuber number per plant ranged from 3 to 6, and the mini-tuber weight ranged from 0.5-3.0 g, depending upon the treatment. Further studies are in progress to produce conventional tubers under salinity stress from the promising variants, specially those tolerant to 4000 ppm, and to assure the stability of the obtained variants. (author)

A dual bacterial culture augments Kalanchoe spp. photosynthesis under extreme conditions

Science.gov (United States)

Burlak, Olexii; Rogutskyy, Ivan; Danilchenko, Boris; Mikheev, Olexander; Zaetz, Iryna; Lorek, Andreas; Koncz, Alexander; de Vera, Jean-Pierre; Foing, Bernard H.; Kozyrovska, Natalia

In consistence with conception of using microbial technology for plant growing/protosoil for-mation for Lunar/Martian greenhouses (Kozyrovska et al., 2004-2010), we anticipate microbes to alleviate impact of the environmental stressors on plant development. Bacteria can augment physiological processes in plants, for example, photosynthesis, by regulating a hormone level and decreasing glucose sensing in planta (Zhang et al., 2008). The study aimed to examine impact of consortium of well-defined bacteria Klebsiella oxytoca IMBG26 and Paenibacillus sp. IMBG150 on the CAM-plantlets Kalanhoe diagramontiana and Kalanhoe tubiflora pho-tosynthetic activity after acute action of gamma radiation (60Co), Near Martian ultraviolet radiation, low pressure (100 mbar), and high concentrations of CO2 (95Plantlets of K. tubi-flora were exposed to harmful doses of Near Martian UV radiation for 3 hours (26.53 J/cm2). A week before experiment kalanchoe plantlets were subjected to acute effects of ionizing radiation at doses of 30 and 70 Gy. In noninoculated plantlets after 30 Gy the photosynthetic activity fell to 71

Respon Pertumbuhan Planlet Anggrek Dendrobium sp. Melalui Aplikasi Ekstrak Tauge dan Air Kelapa pada Media Organik

Directory of Open Access Journals (Sweden)

HESTIN YUSWANTI

2015-09-01

Full Text Available The Growth Response of Orchid Plantlets of Dendrobium sp. by Aplicación of Bean Sprouts Extract and Coconut Water in Organic Medium The aim of this researh is to obtain the effect of coconut water and bean sprouts extract and the best combination added to organic medium on growth of orchid planlets in vitro. The experiment was conducted at Tissue Culture Laboratory of Agriculture Faculty, Udayana University, Pegok Denpasar from Mei until Oktober 2012. The research design was Completely Randomized Design (RAL with ten treatments and six replications. The results of this ezperiment showed that addition of coconut water at concentration of 150 ml/l and bean sprouts extract of 50 ml/l resulted in the best growth of orchid planlet which includes the height of plantlets (4,10 cm, the number of leaf (3,33 leaves, length of root (1,07 cm. The total fresh weight of plantlet was 0,20 g and total oven dry-weight of plantlet was 0,021 g.

Effectiveness of gamma ray irradiation and ethyl methane ...

African Journals Online (AJOL)

Survival rate and plantlet performance of DNKW001 in gamma ray + EMS 7uM treatment declined profoundly with increasing doses and LD50 was lower (104 Gy) than LD50 in gamma ray irradiation (177 Gy) alone. Variants of plantlets were detected in pre (white streaked leaf and bigger petiole with distorted leaf) and post ...

Seismomorphogenesis: a novel approach to acclimatization of tissue culture regenerated plants.

Science.gov (United States)

Sarmast, Mostafa Khoshhal; Salehi, Hassan; Khosh-Khui, Morteza

2014-12-01

Plantlets under in vitro conditions transferred to ex vivo conditions are exposed to biotic and abiotic stresses. Furthermore, in vitro regenerated plants are typically frail and sometimes difficult to handle subsequently increasing their risk to damage and disease; hence acclimatization of these plantlets is the most important step in tissue culture techniques. An experiment was conducted under in vitro conditions to study the effects of shaking duration (twice daily at 6:00 a.m. and 9:00 p.m. for 2, 4, 8, and 16 min at 250 rpm for 14 days) on Sansevieria trifasciata L. as a model plant. Results showed that shaking improved handling, total plant height, and leaf characteristics of the model plant. Forty-eight hours after 14 days of shaking treatments with increasing shaking time, leaf length decreased but proline content of leaf increased. However, 6 months after starting the experiment different results were observed. In explants that received 16 min of shaking treatment, leaf length and area and photosynthesis rate were increased compared with control plantlets. Six months after starting the experiment, control plantlets had 12.5 % mortality; however, no mortality was observed in other treated explants. The results demonstrated that shaking improved the explants' root length and number and as a simple, cost-effective, and non-chemical novel approach may be substituted for other prevalent acclimatization techniques used for tissue culture regenerated plantlets. Further studies with sensitive plants are needed to establish this hypothesis.

Correlation of epiphyllous bud differentiation with foliar senescence in crassulacean succulent Kalanchoe pinnata as revealed by thidiazuron and ethrel application.

Science.gov (United States)

Jaiswal, Sarita; Sawhney, Sudhir

2006-05-01

Leaves of Kalanchoe pinnata have crenate margins with each notch bearing a dormant bud competent to develop into a healthy plantlet. Leaf detachment is a common signal for inducing two contrastingly different leaf-based processes, i.e. epiphyllous bud development into plantlet and foliar senescence. To investigate differentiation of bud and its correlation, if any, with foliar senescence, thidiazuron (TDZ), having cytokinin activity and ethrel (ETH), an ethylene releasing compound, were employed. The experimental system was comprised of marginal leaf discs, each harbouring an epiphyllous bud. Most of the growth characteristics of plantlet developing from the epiphyllous bud were significantly inhibited by TDZ but promoted by ETH. The two regulators modulated senescence in a manner different for leaf discs and plantlet leaves. Thus, TDZ caused a complete retention whereas ETH a complete loss of chlorophyll in the leaf discs. In contrast, the former resulted in a complete depletion of chlorophyll from the plantlet leaves producing an albino effect, while the latter reduced it by 50% only. In combined dispensation of the two regulators, the effect of TDZ was expressed in majority of responses studied. The results presented in this investigation clearly show that the foliar processes of epiphyllous bud differentiation and senescence are interlinked as TDZ that delayed senescence inhibited epiphyllous bud differentiation and ETH that hastened senescence promoted it. A working hypothesis to interpret responsiveness of the disc-bud composite on lines of a source-sink duo, has been proposed.

EFFECT OF COMPLEX ORGANIC COMPOUNDS ON GROWTH PLANLET OF DENDROBIUM ORCHID

Directory of Open Access Journals (Sweden)

Sitti Raodah Garuda

2015-01-01

Full Text Available Uniqueness of stunning Dendrobium variety such as shapes, colors, and sizes are main attraction of this plant. Germination oforchid seeds can be carried out in a laboratory with in vitro techniques.Medium used for germination of orchid seeds are Vacin and Went medium. Researcher stried to add other substances that may increase growth explants, such as complex organic compounds. Study aims to determine effect of complex organic compounds into growth medium VW Dendrobium plantlets. Research used complete randomized design consist five treatment:VW medium without extract (control, VW medium+banana extract, VW medium+ melon extrac, VW medium+guava extract and VW medium+pepaya extract, with three replications, each replication consist two culture bottles.. Each culture bottle planted four planlets. Addition of complex organic compounds such as melon extract gave best vegetative growth of leaves quantity, roots quantity, root length and fresh weight. While guava extract provide best results to plantlet high and saplings. Plant lets with melon extract treatment showed appearance of muscular orchid plantlets is characteristic of plants that can survive during acclimatization. While both guava extract is best used for purpose of orchid plantlets regeneration.

Investigation about selecting strong type of melons by using melon paleness factor fusarium oxysporum f.sp.melonis and mutation techniques

International Nuclear Information System (INIS)

Kantoglu, Y.; Secer, E.; Kunter, B.; Erzurum, K.; Maden, S.; Yanmaz, R.

2009-01-01

Fusarium wilt is a vascular disease of the Cucurbitaceae family, especially in muskmelon (Cucumis melo L.), caused by the soil fungus Fusarium oxysporum f. sp. melonis (FOM). This pathogen persists in the soil for extended periods of time, and the only effective control is the use of resistant varieties. Fusarium oxysporum f. sp. melonis is a very serious disease factor for farmers in Turkey. In this research, we show a method for mass-selection of melon mutants resistant to Fusarium wilt. In vitro selection of resistant cells, which are come from irradiated and non-irradiated explants, is done using culture filtrates of different FOM races. According to our results we determined effective irradiation doses and filtrate treatment dose by Linear Regression Analysis. According to our results 21.75 Gy is effective dose for in vitro Yuva cv. explants to induce mutation and for filtrate treatment 6.73% is the proper dose to select survive calluses and plantlets. We recommended using 10 and 20 Gy gamma ray doses for in vitro melon plantlets to induce mutation by our results. We succeed to regenerate 6% plantlets which were obtained and selected from irradiated plantlets and regenerated in in vitro medias which were include 6.73 % filtrate. Although 16.7% of resistant or tolerant plantlets can continue their viability in greenhouse conditions after disease inoculation treatment, we observed 4 plants had a surviving capability in a limited time. That is very important for breeding cycle and this research can lead to the development of new melon cultivars that will be resistant to Fusarium wilt.

Piper nigrum: micropropagation, antioxidative enzyme activities, and chromatographic fingerprint analysis for quality control.

Science.gov (United States)

Ahmad, Nisar; Abbasi, Bilal Haider; Rahman, Inayat ur; Fazal, Hina

2013-04-01

A reliable in vitro regeneration system for the economical and medicinally important Piper nigrum L. has been established. Callus and shoot regeneration was encouraged from leaf portions on Murashige and Skoog (MS) medium augmented with varied concentrations of plant growth regulators. A higher callus production (90 %) was observed in explants incubated on MS medium incorporated with 1.0 mg L(-1) 6-benzyladenine (BA) along with 0.5 mg L(-1) gibberellic acid after 4 weeks of culture. Moreover, a callogenic response of 85 % was also recorded for 1.0 mg L(-1) BA in combination with 0.25 mg L(-1) α-naphthalene acetic acid (NAA) and 0.25 mg L(-1) 2,4-dichlorophenoxyacetic acid or 0.5 mg L(-1) indole butyric acid (IBA) along with 0.25 mg L(-1) NAA and indole acetic acid. Subsequent sub-culturing of callus after 4 weeks of culture onto MS medium supplemented with 1.5 mg L(-1) thiodiazoran or 1.5 mg L(-1) IBA induced 100 % shoot response. Rooted plantlets were achieved on medium containing varied concentrations of auxins. The antioxidative enzyme activities [superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX)] revealed that significantly higher SOD was observed in regenerated plantlets than in other tissues. However, POD, CAT, and APX were higher in callus than in other tissues. A high-performance liquid chromatography (HPLC) fingerprint analysis protocol was established for quality control in different in vitro-regenerated tissues of P. nigrum L. During analysis, most of the common peaks represent the active principle "piperine." The chemical contents, especially piperine, showed variation from callus culture to whole plantlet regeneration. Based on the deviation in chromatographic peaks, the in vitro-regenerated plantlets exhibit a nearly similar piperine profile to acclimated plantlets. The in vitro regeneration system and HPLC fingerprint analysis established here brought a novel approach to the quality control of in vitro

Induction of mutations in garlic by combined use of gamma-rays and tissue culture

International Nuclear Information System (INIS)

Zhen Hai Rou

1997-01-01

Callus cultures were initiated from leaf explants of garlic on MS medium supplemented with 1.0 mg/l KIN + 1.0 mg/l IAA + 2.0 mg/l 2,4-D. Plantlets were induced from leaf calli on MS medium with 2.0 mg/l BA + 2.0 mg/l IAA. Bulblets were induced from plantlets on MS medium containing 3.0 mg/l ITA. Callus growth and plantlets induction were remarkably inhibited with irradiation doses of 8 and 10 Gy. It was found that doses of 3 and 5 Gy were suitable to induce variation. Somatic embryos were induced on MS medium supplemented with 2.0 mg/l 2,4-D + 500 mg/l casein, 1000 mg/l yeast extract and 3 to 5% sucrose. (author). 11 tabs

Induction of mutations in garlic by combined use of gamma-rays and tissue culture

Energy Technology Data Exchange (ETDEWEB)

Rou, Zhen Hai [Horticultural Research Inst., Shanghai Academy of Agricultural Sciences, Shanghai (China)

1997-07-01

Callus cultures were initiated from leaf explants of garlic on MS medium supplemented with 1.0 mg/l KIN + 1.0 mg/l IAA + 2.0 mg/l 2,4-D. Plantlets were induced from leaf calli on MS medium with 2.0 mg/l BA + 2.0 mg/l IAA. Bulblets were induced from plantlets on MS medium containing 3.0 mg/l ITA. Callus growth and plantlets induction were remarkably inhibited with irradiation doses of 8 and 10 Gy. It was found that doses of 3 and 5 Gy were suitable to induce variation. Somatic embryos were induced on MS medium supplemented with 2.0 mg/l 2,4-D + 500 mg/l casein, 1000 mg/l yeast extract and 3 to 5% sucrose. (author). 11 tabs.

Using Gamma Irradiation To Induce New Mutants In Potatoes Cv. Diamant Through Tissue Culture Technique

International Nuclear Information System (INIS)

Sharabash, M.T.; Ali, Amina A. M.; Ahmed, F. A.; Afifi, Abd El-Moneim M.

2004-01-01

The excess salt, usually NaCl, inhibits potato plant growth and decreases tubers yield. The use of gamma irradiation to induce new mutants in potato cv. Diamant through tissue culture technique was the main task of this study. Sterilized meristemic tips of potato tubers were cultured on aseptic solid MS-medium, pH 5.7, and were incubated at 20 ± 2 d eg C and 16 hrs day length of 3000-Lux light intensity, to produce virus-free plantlets. Micro-propagation started after 6-8 weeks and plantlets were sub-cultured every 3-4 weeks to increase plantlets population. Plantlets were exposed to 0, or 40 Gy, dose rate 27.7 rad / sec., using Co 60 source at the National Center for Research and Radiation Technology, Cairo, Egypt. Irradiated and unirradiated plantlets were transplanted into 60 ml liquid 1/2MS-medium, pH 5.7, and supplemented with 0, 2000 or 4000 ppm NaCl. And, they were incubated for 2 weeks under the same conditions of temperature and light till the new plantlets were grown up. Healthy plantlets were selected, and micro-propagated up to the sixth vegetative generation (M 1 V 6 ), under the same conditions of salinity and incubation conditions Thereafter, the plantlets were transferred to tuberization liquid 1/2MS-medium, supplemented with the same mentioned concentrations of NaCl, to obtain microtubers. The microtubers were collected after 6-8 weeks and preserved at 10 deg C for 3 months approximately, to break the dormancy. Sprouted microtubers were sown to obtain minitubers, and subsequently macrotubers. All cultures were performed in 30-cm pots in a protected greenhouse, and were irrigated with the same concentrations of NaCl. It could be elicited that cv. Diamant is salinity sensitive. This was evidenced by the decrease in the average number of tubers per plant and average fresh weight of tuber under salinity stress up 4000 ppm NaCl, comparing to unsaline control treatment. Potato plants, which still healthy and produced tubers under salinity stress up to

Irradiation effect on in vitro organogenesis, callus growth and plantlet development of Gerbera jamesonii Efeito da irradiação na organogênese in vitro, crescimento de calos e desenvolvimento de plântulas de gerbera

Directory of Open Access Journals (Sweden)

Nor A Hasbullah

2012-06-01

Full Text Available The present work was carried out to study the effects of gamma irradiation on in vitro growth of explants, callus and the formation of shoots and plantlets. Irradiation is known to exhibit or inhibit the differentiation of cells and growth of plants in vitro, which helps in producing new plant varieties. Gamma irradiation is one of the physical mutagens that are widely used for mutation breeding. A gradual decline was observed in the number of shoots regenerated from irradiated petiole explants compared to control. Numbers of shoots regenerated from irradiated petiole explant cultured on Murashige & Skoog medium supplemented with 2.0 mg L-1 BAP and 0.5 mg L-1 NAA was reduced to 6.6±0.9 from 7.5±0.4 (control when explants were exposed to 20 Gray of irradiation dose. Similar observation was reported on effects of gamma irradiation on in vitro propagated plantlets. Gradual decline was observed based on plant height as the dose of gamma irradiation increased. A significant decline was observed in the fresh weight of irradiated callus compared to control. In this case, growth responses of callus were strongly influenced by the radiation dose. The fresh weight of callus was reduced to 76.4±2.2% compared to 89.7±0.5% of control when callus tissues were exposed to 20 Gy.O presente trabalho foi realizado para estudar os efeitos da radiação gama no crescimento in vitro de explantes de calos, e a formação de brotos e mudas. A irradiação é conhecida por induzir ou inibir a diferenciação de células e o crescimento das plantas in vitro, o que ajuda na produção de novas variedades vegetais. Radiação gama é um dos agentes mutagenicos que são amplamente utilizados para o melhoramento através da mutação. Um declínio gradual foi observado no número de brotos regenerados a partir de explantes de pecíolos irradiados comparado com o controle. O número de brotações regeneradas de explantes de pecíolos irradiados, cultivados em meio

Preparation and Low Temperature Short-term Storage for Synthetic Seeds of Caladium bicolor

Directory of Open Access Journals (Sweden)

Mehpara MAQSOOD

2015-03-01

Full Text Available An efficient somatic embryo encapsulation and in vitro plant regeneration technique were established with Caladium bicolor, an important ornamental plant.Tuber derived embryogenic callus (95.50% was obtained on Murashige and Skoog (MS medium amended with 0.5 mg L-1 α-Naphthalene acetic acid (NAA + 0.5 mg L-1 6-Benzyladenine (BA. The embryogenic callus later differentiated into somatic embryos in the same plant growth regulators (PGRs added medium (NAA and BA. The induced embryos matured and developed into plantlets in NAA and BA added media; maximum plantlets development was observed at 1.0 mg L-1 NAA + 1.0 mg L-1 BA supplemented medium. Synthetic seeds were produced by encapsulating embryos in gel containing 3.0% sucrose + 3.0% sodium alginate and 100 mM of calcium chloride.The highest synthetic seed germination (97.6% was observed on medium supplemented with 1.0 mg L-1 NAA + 1.0 mg L-1 BA. The synthetic seeds were kept at low temperatures for storage; the encapsulated beads were viable and demonstrated good germination even after 12 weeks of storage at 4 °C. The plantlet recovery frequency was however declined with time. The synthetic seed derived plantlets were morphologically similar to the mother plant.

Preparation and Low Temperature Short-term Storage for Synthetic Seeds of Caladium bicolor

Directory of Open Access Journals (Sweden)

Mehpara MAQSOOD

2015-03-01

Full Text Available An efficient somatic embryo encapsulation and in vitro plant regeneration technique were established with Caladium bicolor, an important ornamental plant.Tuber derived embryogenic callus (95.50% was obtained on Murashige and Skoog (MS medium amended with 0.5 mg L-1 α-Naphthalene acetic acid (NAA + 0.5 mg L-1 6-Benzyladenine (BA. The embryogenic callus later differentiated into somatic embryos in the same plant growth regulators (PGRs added medium (NAA and BA. The induced embryos matured and developed into plantlets in NAA and BA added media; maximum plantlets development was observed at 1.0 mg L-1 NAA + 1.0 mg L-1 BA supplemented medium. Synthetic seeds were produced by encapsulating embryos in gel containing 3.0% sucrose + 3.0% sodium alginate and 100 mM of calcium chloride.The highest synthetic seed germination (97.6% was observed on medium supplemented with 1.0 mg L-1 NAA + 1.0 mg L-1 BA. The synthetic seeds were kept at low temperatures for storage; the encapsulated beads were viable and demonstrated good germination even after 12 weeks of storage at 4 °C. The plantlet recovery frequency was however declined with time. The synthetic seed derived plantlets were morphologically similar to the mother plant.

Global Reprogramming of Transcription in Chinese Fir (Cunninghamia lanceolata during Progressive Drought Stress and after Rewatering

Directory of Open Access Journals (Sweden)

Ruiyang Hu

2015-07-01

Full Text Available Chinese fir (Cunninghamia lanceolata, an evergreen conifer, is the most commonly grown afforestation species in southeast China due to its rapid growth and good wood qualities. To gain a better understanding of the drought-signalling pathway and the molecular metabolic reactions involved in the drought response, we performed a genome-wide transcription analysis using RNA sequence data. In this study, Chinese fir plantlets were subjected to progressively prolonged drought stress, up to 15 d, followed by rewatering under controlled environmental conditions. Based on observed morphological changes, plantlets experienced mild, moderate, or severe water stress before rehydration. Transcriptome analysis of plantlets, representing control and mild, moderate, and severe drought-stress treatments, and the rewatered plantlets, identified several thousand genes whose expression was altered in response to drought stress. Many genes whose expression was tightly coupled to the levels of drought stress were identified, suggesting involvement in Chinese fir drought adaptation responses. These genes were associated with transcription factors, signal transport, stress kinases, phytohormone signalling, and defence/stress response. The present study provides the most comprehensive transcriptome resource and the first dynamic transcriptome profiles of Chinese fir under drought stress. The drought-responsive genes identified in this study could provide further information for understanding the mechanisms of drought tolerance in Chinese fir.

The basic design and requirement for plant tissue culture laboratory in MINT

International Nuclear Information System (INIS)

Azraf Azman; Rosli Darmawan; Rusli Ibrahim; Mohd Nazir Basiran; Azhar Mohamad; Mohamed Najli Mohamed Yasin; Shuhaimi Shamsuddin

2005-01-01

The production of multiple species plantlets involves a relatively complex process and it is a highly specialized operation. Tissue culture technology is rapidly becoming a commercialized method for propagating new cultivars, rare species and difficult-to-propagate plant. Not only are skills and knowledge essential but the laboratory itself also plays an important role to ensure the successful growth of the plantlets. To produce quality plantlets, plant tissue culture laboratories should fulfill the basic requirements. The laboratory should have proper building and layout which comprise of media preparation and washing room, sterilization or autoclave room, transfer room and culture or growth room. The scope of this paper is to compare these fundamental requirements with the plant tissue culture laboratory in MINT. All the basic needs and differences will be discussed and the proposal for corrective actions will be presented. (Author)

Pineapple (Ananas comosus cv. pérola ex vitro growth and mycorrhizal colonization affected by in vitro sucrose concentration

Directory of Open Access Journals (Sweden)

Alceu Kunze

2014-09-01

Full Text Available The aim of this study was to evaluate the effect of sucrose concentration in the culture medium on growth and on the establishment of mycorrhizas during the acclimatization of pineapple cv. Pérola. The plantlets were micropropagated in MS culture medium with 0, 10, 20 and 30 g L-1 of sucrose and then they were acclimatized during 12 weeks under greenhouse conditions, in a sandy soil - compost mixture, uninoculated or inoculated with a Rhizophagus clarus isolate. Plantlets from the culture medium with 20 g and 30 g of sucrose L-1 showed higher shoot and root biomass than those from sugar-free medium. Mycorrhizal colonization was lower in plantlets micropropagated in sucrose-free medium, but the intensity of arbuscules did not differ among treatments. In the 12-week period of acclimatization, mycorrhizal colonization had no effect on plant biomass.

Pengaruh Eksplan Dan Zpt Terhadap Pertumbuhan Nepenthes Albomarginata Secara in Vitro

OpenAIRE

Sukamto, Lazarus Agus

2011-01-01

Nepenthes albomarginata Lobb ex Lindl. is a carnivorous plant, distributes in several regions in Indonesia. The plant population decreases drastically because of over exploitation and ruining nature habitat. Plant propagation by nature and cutting are not enough to rehabilitation its population. In vitro culture of N. albomarginata was carried out using plantlets grown from the seeds in vitro. Plantlets were cut to became two part explants, consisted of shoot tip and under-shoot tip cuttings....

In vitro rescue of interspecific embryos from Elaeis guineensis x E. oleifera (Arecaceae).

Science.gov (United States)

Angelo, Paula Cristina da Silva; Moraes, Larissa Alexandra Cardoso; Lopes, Ricardo; Sousa, Nelcimar Reis; da Cunha, Raimundo Nonato Vieira; Quisen, Regina Caetano

2011-09-01

The African oil palm (Elaeis guineensis) is the most effective oil producer in tons per hectare. Nevertheless, its increasing cultivation in Latin America is harmed by the "lethal yellowing". Genetic resistance to this anomaly can be found in the germplasm of American oil palm or caiaué (E. oleifera), a native species from the Amazon rainforest. However, the procedures adopted to induce seeds of E. guineensis to germination frequently result mild for interespecific hybrids. Embryo in vitro cultivation can be a viable option. This work was aimed initially to test liquid MS medium supplemented with different glucose or sucrose concentrations for the in vitro cultivation of zygotic embryos from E. guineensis x E. oleifera controlled pollinations. Additionally we investigated different compost mixtures to acclimatize the regenerated hybrid plantlets. Concentrations of 10, 20 and 30g/L of both sugars were tested on flasks containing five mature zygotic embryos, with 15 repetitions per treatment in a total of 450 explants. The number of embryos displaying shoots and radicles at least 2mm in length per experimental unit was evaluated during phase one of in vitro cultivation. Plantlets displaying shoots and radicles were transferred to phase two of in vitro cultivation and subsequently to acclimatization, under 70% shading with manual water supply. The experiments of acclimatization were conducted with 130 plantlets randomly distributed in pure horticultural compost, 3:1 or 1:1 compost:sand mixtures and each plantlet was defined as an experimental unit. Data were submitted to ANOVA, t test and analyzes of correlation (p < or = 0.05). Highest emergence rates were 97% for shoots and 73% for radicles, observed in MS medium supplemented with 20g/L (110mM) of glucose. This sugar in concentrations of 20 or 30g/L provided balanced shoot/root development, and this was considered one of the reasons for the higher frequency of plantlet establishment. The survival percentage was 55

Detection of Methylobacterium radiotolerans IMBG290 in potato plants by in situ hybridization

OpenAIRE

Pirttila A. M.; Kozyrovska N. O.; Ovcharenko L. P.; Podolich O. V.

2009-01-01

A new bacterial strain of pink-pigmented facultative methylotroph (M. radiotolerans IMBG290) which was previously isolated from in vitro grown potato plantlets after their inoculation with Pseudomonas fluorescens IMBG163 was detected in tissues by in situ hybridization method (ISH/FISH). The presence of Methylobacterium rRNA was observed in leaves and stems of potato plantlets, whereas no signal was detected in potato roots. The signal was less abundant in the untreated plants than in the pla...

Lipid peroxidation and seed emergency in progenies of the yellow passion fruit plant

Directory of Open Access Journals (Sweden)

João Paulo Bestete de Oliveira

2012-09-01

Full Text Available The objective was to evaluate the percentage of emergency plantlets and lipid peroxidation in seeds of 29 half-sib progenies of yellow passion fruit (Passiflora edulis Sims. after 24 months under storage. The experimental design was completely randomized, with four replications of 50 seeds each, from which the treatments were the progenies (1-29. The evaluation of the percent plantlet emergency was accomplished at 14 and 28 days after sowing. The lipid peroxidation of the seeds was expressed as malondialdehyde (MDA content that was determined by the TBARS method. Approximately 21% of those half-sib progenies maintained the viability of their seeds for twenty-four months under storage. The results point out a remarkable genetic variability for vigor and emergency of the yellow passion fruit plantlets, with occurrence of individuals with high and other ones with low capacity to maintaining the physiologic quality of their seeds after storage.

Effect of Different Concentrations of Growth Regulators on Gardenia jasminoides cv. Veitchii Micropropagation by Tissue Culture Technique

Directory of Open Access Journals (Sweden)

G. R. Abdullah

2003-01-01

Full Text Available Micropropagation techniques were set up for Gardenia jasminoides c.v. veitchi. Many plantlets were obtained by culturing shoot cuttings in MS nutrient media, 30 g/L Sucrose, 7 g/L Agar Agar, and different concentrations of BAP and IAA. The best concentration was 1mg /L BAP with 0.5 mg/L IAA. This concentration gave the best sprout growth suitable for rooting in primary and secondary culture by reculturing the stuck cutting every 6 weeks and for many times. We also obtained a high rooting percentage up to 98 % of natural rooting in rooting media different from propagation media by reducing mineral salt concentration to half, Sucrose to 20gm/L, and 2gm/L active charcoal, and 1mg/L IAA. Plantlets were transferred to greenhous and subjected for hardening. This technique gave 22 plantlets from one cutting in one year.

Morphogenesis and tissue culture of sweet orange (Citrus sinensis (L.) Osb.): effect of temperature and photosynthetic radiation

International Nuclear Information System (INIS)

Duran-Vila, N.; Gogorcena, Y.; Ortega, V.; Ortiz, J.; Navarro, L.

1992-01-01

Both incubation temperature and photosynthetic radiation affected morphogenesis, callus culture and plantlet culture of sweet orange (Citrus sinensis) cultured in vitro. Bud culture from nodal stem segments, regeneration of shoots and buds from internode stem segments and induction of primary callus were near optimal at incubation temperatures between 21–30°C. The optimal temperature for root formation was 27°C with temperatures above and below being clearly deleterious. Incubation in the dark or under low photosynthetic photon flux density (PPFD) was beneficial for callus induction and growth and also favored the production of rooted plantlets from bud cultures. Incubation in the dark improved considerably the regeneration of shoots and buds from internode segments and the recovery of whole plants. No off-types, as determined by protein and isoenzyme analysis, were observed among plantlets recovered from bud cultures or from regeneration of shoots from internode stem segments

Genetic Changes in Stevia rebaudiana after Gamma Irradiation

International Nuclear Information System (INIS)

Aly, A.A.

2005-01-01

In vitro propagated plantlets of Stevia rebaudiana Bertoni J.were irradiated with doses 0, 5, 10 and 20 Gy. Irradiated plantlets exhibited changes in electrophoretic profile of proteins, there were some new bands induced with molecular weight of 100, 45, 32, and 30 kDa. In some treatments and some other bands disappeared such as the 205 and 100 kDa bands form plantlets treated with 10 Gy. Isoenzyme were also examined, esterase isozyme, isopolyphenol oxidase, alkaline phosphatase, catalse, acid phosphatase and peroxidase isozyme also altered by treatments. RAPD analysis was performed to determine the effect of gamma-irradiation on DNA changes. Polymorphisms between regenerates from non-irradiated and irradiated plantlets were found. The scope of variation spectrum by gamma-irradiation was larger than that by tissue culture. All the primers used produced polymorphic bands. Six primers generated 129.0 RAPD markers, among which 49.0 (37.98%) were polymorphic, with a mean of 8.17 pol morphisms per primer. The results showed that gamma-irradiation induced changes in plantlets that can be detected by molecular and biochemical markers. Stevia rebaudiana Bertoni. Stevia is a member of the Compositae family and native to the valley of the Rio Monday in the highlands of Paraguay, where it has been used by aboriginal people as a sweetener for centuries. It is one of 154 members of the genus Stevia and one of only two that produce sweet steviol glycosides. Stevioside has a sweetening potency of 200-300 times that of sucrose and it is stable to heat (Soejarto et a/., 1982 and 1983 and Lewis, 1992). The leaves were used either to sweeten mate or as a general sweetening agent. Currently Stevia production is centred in China and there is a major market in Japan (Kinghorn and Soejarto 1985)

In vitro induction, isolation, and selection of potato mutants tolerant to salinity

International Nuclear Information System (INIS)

Al-Safadi, B.; Arabi, M. I. E.

2008-01-01

A mutation breeding program was conducted to improve potato (Solanum tuberosum) tolerance to salinity. In vitro cultured explants from potato cvs. Draga, Diamant, and Spunta were irradiated with gamma ray doses of 25, 30, and 35 Gy. Growing plantlets were subsequently propagated to obtain enough explants for in vitro selection of plants tolerant to salinity. Around 1300 MV 4 plantlets from the three cultivars were subjected to selection pressure. MV 4 explants were cultured on an MS medium supplemented with NaCl in varying concentrations ranging from 50 to 200 mM. Surviving plantlets were propagated and re-cultured on a similar medium to insure their tolerance to salinity. Salt tolerant plantlets were acclimatized and transferred to pots and grown under greenhouse conditions. Mutant and control plants were later subjected to a second selection pressure by irrigating them with water containing NaCl in concentrations ranging from 50 to 250 mM. Cultivar Spunta produced the highest number of tolerant plants. Four plants of Spunta appeared to be tolerant to salinity whereas only one plant from Diamant was tolerant and no plants from cultivar Draga were tolerant. The average number of produced minitubers per plant varied in the mutant plants from eight to 14. Also, weight of these minitubers varied from less than 1 to 31 grams. (author)

In vitro induction, isolation and selection of potato mutants resistant to late blight

International Nuclear Information System (INIS)

Al-Safadi, B.; Arabi, M.I.E.

2003-01-01

A mutation breeding program was conducted to improve potato resistance to late blight disease caused by Phytophthora infestans. In vitro cultured explants from cvs Draga, Diamant, Spunta were irradiated with gamma ray doses 25, 30, and 35 Gy. Growing shoots were cut and re-cultured every 2 weeks until the 4 th generation (MV 4 ) to make sure no chimeral tissues still existed in the mutant material. Plantlets were subsequently propagated to obtain enough explants for in vitro selection pressure. Around 3,000 plantlets from the 3 cultivars were subjected to selection pressure using co-culture technique. MV 4 explants were incubated in jars, containing MS medium, with mycelia of P. infestans. Surviving plantlets were propagated and re-incubated with the pathogen for 3 consecutive generations. Resistant plantlets were acclimatized and transferred to pots and grown under glasshouse conditions. Plants were later inoculated, at the adult stage, with sporangial suspension. Cv Draga produced the highest number of resistant plants. Ten plants of Draga appeared to be resistant to late blight, whereas only one plant from each of the other 2 cvs was resistant. Mutant plants varied in number of produced minitubers from 13 to 70. Also, weight of these minitubers varied from less than 1 to 35 grams. Selected mutant lines will undergo further testing under field conditions for P. infestans resistance and other agronomic characteristics

The use of tissue culture techniques with irradiation to improve potato resistance to late blight

International Nuclear Information System (INIS)

Al-Safadi, B.; Arabi, M.I.E.

2004-01-01

A mutation breeding program was conducted to improve potato (Solanum tuberosum) resistance to late blight disease caused by Phytophthora infestans. In vitro cultured explants from potato cvs. Draga, Diamant, Spunta were irradiated with gamma ray doses 25, 30, and 35 Gy. Growing shoots were cut and re-cultured every 2 weeks until the 4 t h generation (MV 4 ) to make sure no chimeral tissues still existed in the mutant material. Plantlets were subsequently propagated to obtain enough explants for in vitro selection pressure. Around 3000 plantlets from the three cultivars were subjected to selection pressure using co-culture technique. MV 4 explants were incubated in jars, containing MS medium, with mycelia of P. infestans. Surviving plantlets were propagated and re-incubated with the pathogen for three consecutive generations. Resistant plantlets were acclimatized and transferred to pots and grown under glasshouse conditions. Plants were later inoculated, at the adult stage, with sporangial suspension. Cultivar Draga produced the highest number of resistant plants. Ten plants of Draga appeared to be resistant to late blight whereas only one plant from each of the other 2 cultivars was resistant. Mutant plants varied in number of produced minitubers from 13 to 70, Also, weight of these minitubers varied from less than 1 to 35 grams. Selected mutant lines will undergo further testing under field conditions for P. infestans resistance and other agronomic characteristics. (author)

Somatic embryogenesis for efficient micropropagation of guava (Psidium guajava L.).

Science.gov (United States)

Akhtar, Nasim

2013-01-01

Guava (Psidium guajava L.) is well known for edible fruit, environment friendly pharmaceutical and commercial products for both national and international market. The conventional propagation and in vitro organogenesis do not meet the demand for the good quality planting materials. Somatic embryogenesis for efficient micropropagation of guava (P. guajava L.) has been developed to fill up the gap. Somatic embryogenesis and plantlets regeneration are achieved from 10-week post-anthesis zygotic embryo explants by 8-day inductive treatment with different concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D) on MS agar medium containing 5% sucrose. Subsequent development and maturation of somatic embryos occur after 8 days on MS basal medium supplemented with 5% sucrose without plant growth regulator. The process of somatic embryogenesis shows the highest relative efficiency in 8-day treatment of zygotic embryo explants with 1.0 mg L(-1) 2,4-D. High efficiency germination of somatic embryos and plantlet regeneration takes place on half strength semisolid MS medium amended with 3% sucrose within 2 weeks of subculture. Somatic plantlets are grown for additional 2 weeks by subculturing in MS liquid growth medium containing 3% sucrose. Well-grown plantlets from liquid medium have survived very well following 2-4 week hardening process. The protocol of somatic embryogenesis is optimized for high efficiency micropropagation of guava species.

PENGARUH EKSPLAN DAN ZPT TERHADAP PERTUMBUHAN nEPEnTHES ALBOmARGInATA SECARA In VITRO

OpenAIRE

Sukamto, Lazarus Agus

2016-01-01

Nepenthes albomarginata Lobb ex Lindl. is a carnivorous plant, distributes in several regions in Indonesia. The plant population decreases drastically because of over exploitation and ruining nature habitat. Plant propagation by nature and cutting are not enough to rehabilitation its population. In vitro culture of N. albomarginata was carried out using plantlets grown from the seeds in vitro. Plantlets were cut to became two part explants, consisted of shoot tip and under-shoot tip cuttings....

Evaluation on the effectiveness of 2-deoxyglucose-6-phosphate phosphatase (DOGR1) gene as a selectable marker for oil palm (Elaeis guineensis Jacq.) embryogenic calli transformation mediated by Agrobacterium tumefaciens

Science.gov (United States)

Izawati, Abang Masli Dayang; Masani, Mat Yunus Abdul; Ismanizan, Ismail; Parveez, Ghulam Kadir Ahmad

2015-01-01

DOGR1, which encodes 2-deoxyglucose-6-phosphate phosphatase, has been used as a selectable marker gene to produce transgenic plants. In this study, a transformation vector, pBIDOG, which contains the DOGR1 gene, was transformed into oil palm embryogenic calli (EC) mediated by Agrobacterium tumefaciens strain LBA4404. Transformed EC were exposed to 400 mg l-1 2-deoxyglucose (2-DOG) as the selection agent. 2-DOG resistant tissues were regenerated into whole plantlets on various regeneration media containing the same concentration of 2-DOG. The plantlets were later transferred into soil and grown in a biosafety screenhouse. PCR and subsequently Southern blot analyses were carried out to confirm the integration of the transgene in the plantlets. A transformation efficiency of about 1.0% was obtained using DOGR1 gene into the genome of oil palm. This result demonstrates the potential of using combination of DOGR1 gene and 2-DOG for regenerating transgenic oil palm. PMID:26442041

In vitro micropropagation of Dracaena sanderiana Sander ex Mast: An important indoor ornamental plant.

Science.gov (United States)

Aslam, Junaid; Mujib, Abdul; Sharma, Maheshwar Prasad

2013-01-01

A protocol has been developed for in vitro plant regeneration from a nodal explant of Dracaena sanderiana Sander ex Mast. Nodal explant showed high callus induction potentiality on MS medium supplemented with 6.78 μM 2,4-dichlorophenoxyacetic acid (2,4-D) followed by 46.5 μM chlorophenoxy acetic acid (CPA). The highest frequency of shoot regeneration (85%) and number of shoots per explant (5.6) were obtained on medium supplemented with 7.84 μM N(6)-benzylaminopurine (BA). Rooting was high on MS solid compared to liquid medium when added with 7.38 μM indole-3-butyric acid (IBA). Fifty percent of the roots were also directly rooted as microcuttings on soil rite, sand and peat mixture (1:1:1). In vitro and ex vitro raised plantlets were used for acclimatization. More than 90% of the plantlets was successfully acclimatized and established in plastic pots. Ex vitro transferred plantlets were normal without any phenotypic aberrations.

Rhizophagus irregularis MUCL 41833 transitorily reduces tomato bacterial wilt incidence caused by Ralstonia solanacearum under in vitro conditions.

Science.gov (United States)

Chave, Marie; Crozilhac, Patrice; Deberdt, Péninna; Plouznikoff, Katia; Declerck, Stéphane

2017-10-01

Bacterial wilt caused by Ralstonia solanacearum is one of the world's most important soil-borne plant diseases. In Martinique, French West Indies, a highly virulent new pathogenic variant of this bacterium (phylotype IIB/4NPB) severely impacts tomato production. Here we report on the effect of R. solanacearum CFBP 6783, classified in phytotype IIB/4NPB, on tomato plantlets grown under strict in vitro culture conditions in the presence or absence of the arbuscular mycorrhizal fungus Rhizophagus irregularis MUCL 41833. A mycelium donor plant (i.e. Crotalaria spectabilis) was used for rapid, uniform mycorrhization of tomato plantlets that were subsequently infected by the bacterium. Bacterial wilt was significantly delayed and the incidence of the disease consequently reduced in the mycorrhizal tomato plantlets. Conversely, R. solanacearum did not affect root colonization by the AMF within the 16 days of the experiment. These results suggested that the mycorrhizal fungus was able to reduce bacterial wilt symptoms, probably by eliciting defence mechanisms in the plant.

Chemical evaluation of strawberry plants produced by tissue culturing of gamma irradiated seedlings

International Nuclear Information System (INIS)

Maraei, R.W.

2007-01-01

studies were conducted to evaluate the influence of gamma irradiation as a supplementary factor precedes tissue culture application on strawberry seedlings (c.v.Rosa Linda). the strawberry seedling were irradiated using 8 doses of co 60 gamma rays 50.75.100.125 ,150,250, 350 and 500 gray. tissue culture technique was applied on irradiated and unirradiated strawberry seedling. different characteristics of plantlets, plant and fruit of strawberry produced from the double treatment (irradiation followed by tissue culture) were studied as well as the early, total and exportable fruit yields. data indicated that, low radiation doses 50,75 and 100 gray increased all morphological and chemical characteristics of the plantlets, plant and fruit of strawberry, whereas radiation doses higher than 100 gray decreased them significantly. moreover 350 and gray were lethal doses. radiation dose 50 gray increased the survival percentage and the length of plantlets by 1.5% and 50% respectively more than the unirradiated treatment in all multiplication stages

Mutant induction through adventitious buds of Kohleria

International Nuclear Information System (INIS)

Parliman, B.J.; Stushnoff, C.

1979-01-01

Freshly cut leaves of Kohleria eriantha and K. x 'Longwood' were exposed in a wide range of gamma irradiation doses and allowed to root and form adventitious buds. K. Eriantha could not be successfully propagated from leaf half cuttings. 'Longwood' produced a small number of adventitious plantlets as compared to other Gesneriads. Colchicine treatments reduced leaf half survival in 'Longwood' by more than 50%. Leaf halves exposed to low and moderate doses of gamma irradiation showed increased overall plantlet production compared to nonirradiated leaf halves. (Auth.)

In vitro rescue of interspecific embryos from Elaeis guineensis x E. oleifera (Arecaceae

Directory of Open Access Journals (Sweden)

Paula Cristina da Silva Angelo

2011-09-01

Full Text Available The African oil palm (Elaeis guineensis is the most effective oil producer in tons per hectare. Nevertheless, its increasing cultivation in Latin America is harmed by the “lethal yellowing”. Genetic resistance to this anomaly can be found in the germplasm of American oil palm or caiaué (E. oleifera, a native species from the Amazon rainforest. However, the procedures adopted to induce seeds of E. guineensis to germination frequently result mild for interespecific hybrids. Embryo in vitro cultivation can be a viable option. This work was aimed initially to test liquid MS medium supplemented with different glucose or sucrose concentrations for the in vitro cultivation of zygotic embryos from E. guineensis x E. oleifera controlled pollinations. Additionally we investigated different compost mixtures to acclimatize the regenerated hybrid plantlets. Concentrations of 10, 20 and 30g/L of both sugars were tested on flasks containing five mature zygotic embryos, with 15 repetitions per treatment in a total of 450 explants. The number of embryos displaying shoots and radicles at least 2mm in length per experimental unit was evaluated during phase one of in vitro cultivation. Plantlets displaying shoots and radicles were transferred to phase two of in vitro cultivation and subsequently to acclimatization, under 70% shading with manual water supply. The experiments of acclimatization were conducted with 130 plantlets randomly distributed in pure horticultural compost, 3:1 or 1:1 compost:sand mixtures and each plantlet was defined as an experimental unit. Data were submitted to ANOVA, t test and analyzes of correlation (p≤0.05. Highest emergence rates were 97% for shoots and 73% for radicles, observed in MS medium supplemented with 20g/L (110mM of glucose. This sugar in concentrations of 20 or 30g/L provided balanced shoot/root development, and this was considered one of the reasons for the higher frequency of plantlet establishment. The survival

Improvement of potato tolerance to salinity using tissue culture techniques and irradiation with in vitro selection

International Nuclear Information System (INIS)

Al-Safadi, B.; Arabi, M. I. E.

2005-06-01

A mutation breeding program was conducted to improve potato (Solanum tuberosum) tolerance to salinity. In vitro cultured explants from potato cvs. Draga, Diamant, Spunta were irradiated with gamma doses 25, 30, and 35 Gy.Growing shoots were cut and re-cultured every 2 weeks until the 4th generation (MV 4 ) to make sure no chimeral tissues still existed in the mutant material. Plantlets were subsequently propagated to obtain enough explants for in vitro selection pressure. Around 3000 plantlets from the three cultivars were subjected to selection pressure. MV 4 explants were cultured on MS medium supplemented with the NaCl in varying concentrations ranging between 50 to 200 mM. Surviving plantlets were propagated and re-cultured on a similar medium to insure their tolerance to salinity. Tolerant plantlets were acclimatized and transferred to pots and grown under glasshouse conditions. Plants were later subjected to another selection pressure, by irrigating them using water containing NaCl in concentrations ranging between 50-250 mM in addition to controls irrigated with normal water. Cultivar Spunta produced the highest number of tolerant plants. Four plants of Spunta appeared to be tolerant to salinity whereas only one plant from Diamant and was tolerant and no plants from cultivar Draga were tolerant. Mutant plants varied in number of produced minitubers from 8 - 14. Also, weight of these minitubers varied from less than 1 to 31 grams. (author)

Thidiazuron enhanced regeneration and silymarin content in silybum marianum

International Nuclear Information System (INIS)

Khan, M.A.; Abbasi, B.H.; Shinwari, Z.K.

2014-01-01

Silybum marianum, of family Asteraceae is renowned for production of biologically important silymarin, which has shown multi-dimensional medicinal properties. It has a high protective role against jaundice and hepatitis C worldwide. We hereby established a feasible and efficient method for indirect regeneration of S. marianum for production of consistent plantlets. Calli were induced from leaf explants of seed-derived plantlets on Murashige and Skoog (MS) medium supplemented with several concentrations of different plant growth regulators (PGRs). Highest callogenic response (89%) was recorded for 4.4 meu M Thidiazuron (TDZ) in combination with 6.6 meu M Kinetin (Kn). Subsequent sub-culturing of callus after 4 weeks of culture, on medium with similar compositions of PGRs induced shoot organogenesis. Highest shoot induction frequency (86%) with maximum mean multiple shoots (26 shoots per explant) were recorded for 11meu M TDZ after 4 weeks of transfer. Longest shoots (4.1 cm) were recorded for MS medium augmented with 6.6 meu M TDZ and 4.4 meu M naphthalene acetic acid (NAA). Furthermore, rooted plantlets were developed on MS medium containing different concentrations of indole acetic acid (IAA). Silymarin was determined by High performance liquid chromatography (HPLC) and 8.47 mg/g DW silymarin was detected in the regenerated plantlets. This study contributes to a better understanding of the different mechanisms involved in morphogenesis and production of biologically active principle in Silybum marianum. (author)

The effect of in vitro mycorrhization on growth characteristics, changes in endogenous hormones and performance of microplants in potato (Solanum tuberosum L.

Directory of Open Access Journals (Sweden)

Khosro PARVIZI

2015-12-01

Full Text Available Potato (Solanum tuberosum L. plantlets were inoculated in vitro with an arbuscular mycorrhizal fungus (Glomus intraradices and their growth response, performance and endogenous hormonal status evaluated. A factorial experiment based on a completely randomized design consisting of two potato cultivars (Agria and Sante and four culture media was conducted with four replications. The culture media included non-inoculated Murashige and Skoog (MS medium (control, inoculated MS medium, inoculated half-strength MS medium and inoculated MS medium without vitamins and sugar. To do inoculation, germinated spores of fungus were transferred to the root zone of plantlets. Colonization percentage, total chlorophyll content, internodes and stolon length, shoot diameter, shoot and root fresh and dry weight, leaf area and the level of three endogenous hormones (total auxins, gibberellins and cytokinins were determined. In addition, mini-tuber production was assayed quantitatively and qualitatively. Results demonstrated that the effect of two factors on all characteristics including mini-tuber production was different significantly. Interaction between cultivar and inoculation systems was significantly associated with endogen auxins as well as all range of mini-tuber production. The inoculated MS medium showed the better results, but it did not have significant difference to half-strength MS in terms of plantlet performance and growing parameters. Responses of both cultivars to inoculation were very conspicuous in the production of endogen hormones. Higher endogen hormone levels were associated with elevated growth parameters, greater biomass production and better plantlet performance.

Desenvolvimento de mudas de bromélia (Neoregelia cruenta (R. Graham L. B. Smith cultivadas em diferentes substartos e adubação foliar Development of plantlets of bromeliad (Neoregelia cruenta cultivated in different substrates and leaf fertilizers

Directory of Open Access Journals (Sweden)

Clarissa Alves Ferreira

2007-06-01

extraction. This situation is also a consequence of the reduced technical information regarding its propagation and development. One of the limitations is the lack of knowledge regarding the type of substrate and adequate fertilizer for its cultivation. The objective of this study was to evaluate the development of Neoregelia cultivated in different substrates and leaf fertilizers through the variables height, leaf number and fresh and dry matter of shoots and roots. The plantlets used were produced by tissue culture. After a period of pre-acclimatization, the plantlets were transferred to a green house without nebulization. The substrates consisted of different proportions of soil, sand and carbonized rice husk and a commercial substrate composed of vermiculite. Leaf fertilizers were applied to the substrates combining urea and sucrose in intervals of fifteen days. The results showed that the interactions of the substrates and the sucrose and urea dosages had no effect on height and leaf number. The use of sucrose also showed no influence on plant development. The commercial substrate composed of vermiculite, independent of the leaf fertilizer application, showed the best results for plant height and leaf number. The use of urea presented a crescent linear effect during the evaluated period.

Crescimento de plântulas do milho 'Saracura' e atividade de alfa-amilase e invertases associados ao aumento da tolerância ao alagamento exercido pelo cálcio exógeno Growth of maize 'Saracura' (BRS-4154 and alpha-amylase and invertases activity associated with the increase of flooding tolerance caused by exogenous calcium

Directory of Open Access Journals (Sweden)

Daniela Deitos Fries

2007-01-01

Full Text Available Objetivou-se avaliar o crescimento da plântula e o metabolismo de carboidratos associados ao aumento da tolerância à hipoxia exercido pela presença do cálcio no período de germinação e/ou alagamento de plântulas de milho com diferentes idades. O experimento foi desenvolvido na Universidade Federal de Lavras, MG, em 2002. Cariopses de milho da variedade 'Saracura' (BRS-4154 foram germinadas em água ou solução de CaCl2. Após dois e quatro dias, as plântulas foram submetidas ao alagamento em tubos de PVC com tampão (com ou sem CaCl2 por três dias, sendo então avaliadas a sobrevivência, massa seca e as características bioquímicas. O cálcio aumentou a sobrevivência ao alagamento de plântulas com quatro dias, entretanto, não influenciou naquelas com dois dias. O alagamento reduziu a massa seca das plântulas sem prejudicar sua recuperação pós-estresse, entretanto, o cálcio influenciou no acúmulo de massa seca pós-hipoxia somente em plântulas com quatro dias, mostrando uma recuperação mais rápida dos danos causados no período de alagamento, o que ocorre em plântulas de dois dias de germinação, independentemente desse elemento. Em plântulas mais jovens também houve maior mobilização das reservas de amido, assim como, maior translocação e utilização dessas reservas. O alagamento reduziu drasticamente a atividade da invertase ácida do vacúolo, independentemente da presença do cálcio, comprovando sua não participação nos processos comandados por essa enzima. A redução na atividade das invertases favorece menor concentração de hexoses, diminuindo a respiração anaeróbica e, conseqüentemente, a produção de substâncias tóxicas, aumentando a sobrevida das plântulas em condições de hipoxia.This research evaluated the plantlet growth and carbohydrate metabolism associated with the increase in hypoxia tolerance caused by calcium presence during germination and/or flooding of maize plantlets with

Rice improvement, involving altered flower structure more suitable to cross-pollination, using in vitro culture in combination with mutagenesis

International Nuclear Information System (INIS)

Min, S.K.

1998-01-01

Anther and somatic tissue culture in combination with mutagenesis were carried out to evaluate the efficiency of different mutagenic treatments of various in vitro culture materials, and to obtain some promising variants for rice improvement. Results indicated that in japonica rice radiation treatment of dry seeds and young panicles influenced the percentage of green plantlets regeneration from anther culture. Both treatments increased significantly the percentage of regenerated green plantlets in comparison with the control. Irradiation with 30 Gy of rice callus increased also the percentage of regenerated green plantlets. For indica rice, the combination of the suitable dose of gamma rays irradiation on seeds and an improved medium, increased the percentage of callus induction. This approach made it possible to use anther culture in indica rice breeding. Somatic tissue cultures combined with radiation-induced mutagenesis led to the development of a number of promising mutants including some new cytoplasm-nucleus interacting male-sterile lines with almost 100% stigma exertion. Their development would be of practical significance for increasing the genetic diversity for production of hybrid rice. (author)

Somatic embryogenesis and plant regeneration from cell suspension cultures of Cucumis sativus L.

Science.gov (United States)

Chee, P P; Tricoli, D M

1988-06-01

A procedure for the regeneration of whole cucumber plants (Cucumis sativus L. cv. Poinsett 76) by embryogenesis from cell suspension cultures is described. Embryogenic callus was initiated from the primary leaves of 14-17 day old plants. Suspension cultures of embryogenic cells were grown in liquid Murashige and Skoog basal medium containing 5 uM 2,4,5-trichlorophenoxyacetic acid and 4 uM 6-benzylaminopurine. Suspension cultures were composed of a population of cells that were densely cytoplasmic and potentially embryogenic. Differentiation of embryos was enhanced by washing the suspension culture cells with MS basal medium containing 0.5% activated charcoal and twice with MS basal medium followed by liquid shake cultures in MS basal medium. Sixty to 70 percent of the embryos prewashed with activated charcoal germinated into plantlets with normal morphology. Embryos obtained from suspension cultured cells without prewashing with activated charcoal organized into plantlets with abnormal primary leaves. Morphologically normal plantlets were obtained by excising the shoot tips and transferring them to fresh medium.

In Vitro Seeds Germination and Seedling Growth of Bambara Groundnut (Vigna subterranea (L.) Verdc. (Fabaceae)).

Science.gov (United States)

Koné, Mongomaké; Koné, Tchoa; Silué, Nakpalo; Soumahoro, André Brahima; Kouakou, Tanoh Hilaire

2015-01-01

Bambara groundnut (Vigna subterranea (L.) Verdc.) is an indigenous grain legume. It occupies a prominent place in the strategies to ensure food security in sub-Saharan Africa. Development of an efficient in vitro regeneration system, a prerequisite for genetic transformation application, requires the establishment of optimal conditions for seeds germination and plantlets development. Three types of seeds were inoculated on different basal media devoid of growth regulators. Various strengths of the medium of choice and the type and concentration of carbon source were also investigated. Responses to germination varied with the type of seed. Embryonic axis (EA) followed by seeds without coat (SWtC) germinated rapidly and expressed a high rate of germination. The growth performances of plantlets varied with the basal medium composition and the seeds type. The optimal growth performances of plants were displayed on half strength MS basal medium with SWtC and EA as source of seeds. Addition of 3% sucrose in the culture medium was more suitable for a maximum growth of plantlets derived from EA.

Plant Growth-Promoting Rhizobacteria Stimulate Vegetative Growth and Asexual Reproduction of Kalanchoe daigremontiana.

Science.gov (United States)

Park, Yong-Soon; Park, Kyungseok; Kloepper, Joseph W; Ryu, Choong-Min

2015-09-01

Certain bacterial species associate with plant roots in soil. The plant growth-promoting rhizobacteria (PGPR) stimulate plant growth and yield in greenhouse and field. Here, we examined whether application of known bacilli PGPR strains stimulated growth and asexual reproduction in the succulent plant Kalanchoe daigremontiana. Four PGPR strains B. amyloliquefaciens IN937a, B. cereus BS107, B. pumilus INR7, and B. subtilis GB03 were applied to young plantlets by soil-drenching, and plant growth and development was monitored for three months. Aerial growth was significantly stimulated in PGPR-inoculated plants, which was observed as increases in plant height, shoot weight, and stem width. The stimulated growth influenced plant development by increasing the total number of leaves per plant. Treatment with bacilli also increased the total root biomass compared with that of control plants, and led to a 2-fold increase in asexual reproduction and plantlet formation on the leaf. Collectively, our results firstly demonstrate that Bacillus spp. promote vegetative development of K. daigremontiana, and the enhanced growth stimulates asexual reproduction and plantlet formation.

Plant Growth-Promoting Rhizobacteria Stimulate Vegetative Growth and Asexual Reproduction of Kalanchoe daigremontiana

Directory of Open Access Journals (Sweden)

Yong-Soon Park

2015-09-01

Full Text Available Certain bacterial species associate with plant roots in soil. The plant growth-promoting rhizobacteria (PGPR stimulate plant growth and yield in greenhouse and field. Here, we examined whether application of known bacilli PGPR strains stimulated growth and asexual reproduction in the succulent plant Kalanchoe daigremontiana. Four PGPR strains B. amyloliquefaciens IN937a, B. cereus BS107, B. pumilus INR7, and B. subtilis GB03 were applied to young plantlets by soil-drenching, and plant growth and development was monitored for three months. Aerial growth was significantly stimulated in PGPR-inoculated plants, which was observed as increases in plant height, shoot weight, and stem width. The stimulated growth influenced plant development by increasing the total number of leaves per plant. Treatment with bacilli also increased the total root biomass compared with that of control plants, and led to a 2-fold increase in asexual reproduction and plantlet formation on the leaf. Collectively, our results firstly demonstrate that Bacillus spp. promote vegetative development of K. daigremontiana, and the enhanced growth stimulates asexual reproduction and plantlet formation.

Detection of Variation in Long-Term Micropropagated Mature Pistachio via DNA-Based Molecular Markers.

Science.gov (United States)

Akdemir, Hülya; Suzerer, Veysel; Tilkat, Engin; Onay, Ahmet; Çiftçi, Yelda Ozden

2016-12-01

Determination of genetic stability of in vitro-grown plantlets is needed for safe and large-scale production of mature trees. In this study, genetic variation of long-term micropropagated mature pistachio developed through direct shoot bud regeneration using apical buds (protocol A) and in vitro-derived leaves (protocol B) was assessed via DNA-based molecular markers. Randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), and amplified fragment length polymorphism (AFLP) were employed, and the obtained PIC values from RAPD (0.226), ISSR (0.220), and AFLP (0.241) showed that micropropagation of pistachio for different periods of time resulted in "reasonable polymorphism" among donor plant and its 18 clones. Mantel's test showed a consistence polymorphism level between marker systems based on similarity matrices. In conclusion, this is the first study on occurrence of genetic variability in long-term micropropagated mature pistachio plantlets. The obtained results clearly indicated that different marker approaches used in this study are reliable for assessing tissue culture-induced variations in long-term cultured pistachio plantlets.

Adição de torta de mamona em substratos na aclimatação de mudas micropropagadas de bananeira Effect of addition of castor bean pie in substrates in acclimatization of micropropagated banana plantlets

Directory of Open Access Journals (Sweden)

Adriana Novais Martins

2011-03-01

Full Text Available A utilização de mudas micropropagadas de bananeira que oferecem qualidade genética e fitossanitária, favorecendo o desenvolvimento, instalação e uniformidade do pomar, é importante para a exploração comercial da bananicultura. O objetivo deste trabalho foi avaliar a influência da adição da torta de mamona nos substratos na aclimatação de mudas micropropagadas de bananeira da cv. Willians. O delineamento estatístico utilizado foi o inteiramente casualizado, com 10 tratamentos, sendo 2 substratos e 5 dosagens de torta de mamona (0; 6; 12; 18 e 24 g vaso-1. O substrato Vivatto Slim Plus® possibilitou o melhor desenvolvimento das plantas na aclimatação. Não são recomendadas doses superiores a 12 g planta-1 de torta de mamona misturadas ao substrato na aclimatação de mudas de bananeira.The use of micropropagated banana tree seedlings offer genetic and phytosanitary qualities, favoring the development, installation and uniformity of the yield and it is very important for the commercial exploration of banana fruits. The objective of this study was to evaluate the influence of castor oil plant pie added to the substrate in the acclimatization of micropropagated banana plantlets cv. 'Willians'. It was use the completely randomized, with 10 treatments, being 2 substrates and 5 dosages of castor oil plant pie (0; 6; 12; 18 and 24 g for each recipient. The substrate Vivatto Slim Plus® showed the best development of the plants in acclimatization processes. Dosages above 12 g of castor oil plant pie for each plant are not recommended mixed to the substrate in the acclimatization of banana tree seedlings

In vitro propagation and reintroduction of the endangered Renanthera imschootiana Rolfe.

Science.gov (United States)

Wu, Kunlin; Zeng, Songjun; Lin, Danni; Teixeira da Silva, Jaime A; Bu, Zhaoyang; Zhang, Jianxia; Duan, Jun

2014-01-01

Renanthera imschootiana Rolfe is an endangered tropical epiphytic orchid that is threatened with extinction due to over-collection and the loss of suitable habitats. In vitro propagation is a useful way to mass produce plants for re-establishment in the wild and for commercial propagation. Seeds collected 150 days after pollination (DAP) were the optimum stage for in vitro culture. Seed germination reached 93.1% on quarter-strength MS (i.e., MS containing a quarter of macro- and micronutrients) medium containing 0.5 mg l(-1) α-naphthaleneacetic acid (NAA), 20% coconut water (CW), 1.0 g l(-1) peptone, 10 g l(-1) sucrose and 1.0 g l(-1) activated charcoal (AC). Quarter-strength MS medium supplemented with 1.0 mg l(-1) BA, 0.5 mg l(-1) NAA, 1.0 g l(-1) peptone, 10 g l(-1) sucrose and 20% CW was suitable for the sub-culture of protocorm-like bodies (PLBs) in which the PLB proliferation ratio was 2.88. Quarter-strength MS medium containing 1.0 mg l(-1) NAA, 1.0 g l(-1) peptone, 100 g l(-1) banana homogenate (BH), and 1.0 g l(-1) AC was suitable for plantlet formation and 95.67% of plantlets developed from PLBs within 60 days of culture. Hyponex N016 medium supplemented with 0.5 mg l(-1) NAA, 1.0 g l(-1) peptone, 20 g l(-1) sucrose, 150 g l(-1) BH, and 1.0 g l(-1) AC was suitable for the in vitro growth of plantlets about 2-cm in height. Plantlets 3-cm in height or taller were transplanted to Chilean sphagnum moss, and 95% of plantlets survived after 60 days in a greenhouse. Three hundred transplanted of seedlings 360-days old were reintroduced into three natural habitats. Highest percentage survival (79.67%) was observed in Yuanjiang Nature Reserve two years after reintroduction, followed by Huolu Mountain forest park (71.33%). This protocol is an efficient means for the large-scale propagation and in vitro and in vivo germplasm conservation of R. imschootiana.

Influence of 2,4-D and picloram on embryogenic competence of three cassava (manihot esculenta crantz) accessions

International Nuclear Information System (INIS)

Elegba, W.

2008-06-01

The influence of 2,4-D and picloram on embryogenic competence of three local cassava accessions namely, Nkabom, Wenchi and ADI 001 was studied. Young leaf lobes of cassava cultured on MS medium supplemented with 4 to 20 mg/L 2,4-D or picloram (initiation medium) resulted in 90 to 100% embryogenic calli formation depending on the accession. The transfer of embryogenic calli to an MS medium supplemented with 0.1 mg/L BAP produced primary embryos. The duration and number of primary embryos produced on the maturation medium depended on the auxin as well as the genotype. picloram decreased the number of days to maturation and produced comparatively more embryos at both the primary and cyclic stages than 2,4-D. However, the optimal concentration required for increased embryo production in picloram was higher (16 mg/L) than 2,4-D ( 8 mg/L). Fragmentation of matured primary somatic embryos and reculture on initiation medium led to cyclic embryo production. Cyclic embryogenesis doubled (with 2,4-D) or tripled (with picloram) the number of embryos produced at the primary stage. However, recycling of embryos through three successive cycles resulted in a slight reduction in the number of embryos produced. The presence of lower concentration of ABA (1 mg/L) in the maturation medium enhanced embryo maturation and plantlet regeneration while increased concentration of ABA (2 or 4 mg/L) decreased embryo production and plantlet regeneration indicating inhibition or dormancy effect of the growth regulator on germination of embryos. Desiccation significantly (P ≤ 0.05) improved plantlet regeneration from both 2,4-D and picloram-induced somatic embryos in all the accessions. However, comparatively high somatic embryos derived from picloram germinated into plantlets than 2,4-D.This study has shown that all three accessions, are embryogenically competent and picloram is a superior auxin to 2,4-D in somatic embryo production and successful regeneration of plantlets. This holds great

Reproduction at the extremes: pseudovivipary, hybridization and genetic mosaicism in Posidonia australis (Posidoniaceae).

Science.gov (United States)

Sinclair, Elizabeth A; Statton, John; Hovey, Renae; Anthony, Janet M; Dixon, Kingsley W; Kendrick, Gary A

2016-02-01

Organisms occupying the edges of natural geographical ranges usually survive at the extreme limits of their innate physiological tolerances. Extreme and prolonged fluctuations in environmental conditions, often associated with climate change and exacerbated at species' geographical range edges, are known to trigger alternative responses in reproduction. This study reports the first observations of adventitious inflorescence-derived plantlet formation in the marine angiosperm Posidonia australis, growing at the northern range edge (upper thermal and salinity tolerance) in Shark Bay, Western Australia. These novel plantlets are described and a combination of microsatellite DNA markers and flow cytometry is used to determine their origin. Polymorphic microsatellite DNA markers were used to generate multilocus genotypes to determine the origin of the adventitious inflorescence-derived plantlets. Ploidy and genome size were estimated using flow cytometry. All adventitious plantlets were genetically identical to the maternal plant and were therefore the product of a novel pseudoviviparous reproductive event. It was found that 87 % of the multilocus genotypes contained three alleles in at least one locus. Ploidy was identical in all sampled plants. The genome size (2 C value) for samples from Shark Bay and from a separate site much further south was not significantly different, implying they are the same ploidy level and ruling out a complete genome duplication (polyploidy). Survival at range edges often sees the development of novel responses in the struggle for survival and reproduction. This study documents a physiological response at the trailing edge, whereby reproductive strategy can adapt to fluctuating conditions and suggests that the lower-than-usual water temperature triggered unfertilized inflorescences to 'switch' to growing plantlets that were adventitious clones of their maternal parent. This may have important long-term implications as both genetic and

Radiosensibility curve with 60 Co in guinea grass (Panicum maximum Jacq.) cv. 'K-249'

International Nuclear Information System (INIS)

Lajonchere, G.; Mesa, A.R.; Prieto, M.; Sánchez-Quiroz, E.

1995-01-01

In order to select the adequate radiosensibility criterion and useful dosis were studied the effects of gamma radiation on the embryogenic callus of guinea cv. K-249. The application dosages were 0, 10, 20, 40, 60, 80, 100, 150, 200, 250, 300 and 400 Gy and the measures evaluated were: increase of fresh mass of the callus in fresh medium, number of shoots, plantlets and total by callus and by gram of callus in regeneration medium. It was determined that the curves shaped with the number of shoots, plantlets and total by callus and by gram of callus were more radiosensible than that shoped with increase of fresh mass of the callus; as criterion of radiosensibility in this cultivar the total number of shoots and plantlets from gram of callus was selected for it's simplicity and veracity as much as in the evaluation and in the results. The useful dosis that determined one GR 10, 20, 50 and 70 percent were 16, 20, 29 and 160 Gy respectively. Albino shoots were also observed whose values were randomizely showed and it was observed albinism in 0 dosis. (author) [es

Substratos alternativos ao xaxim e adubação de plantas de orquídea na fase de aclimatização Alternative substrates to xaxim and fertilization on orchids plantlets acclimatization phase

Directory of Open Access Journals (Sweden)

Aparecida Gomes de Araujo

2007-04-01

Full Text Available Objetivou-se estudar substratos e adubação em plantas de orquídea da espécie C. loddgesii "Alba" x C. loddgesii "Atibaia". Plantas oriundas de cultivo in vitro foram transplantadas para bandejas coletivas contendo pó de xaxim e aclimatizadas em casa de vegetação. Após seis meses, quando as plantas tinham de 3,0 a 5,0cm de comprimento, foram transferidas para bandejas de plástico com 24 células de 150cm³ cada contendo os substratos brita no 0, casca de arroz carbonizada (CAC, xaxim desfibrado e fibra de piaçava e submetidas a adubações semanais com Biofert Plus® (5,0mL L-1, Dyna-Grow® (2,5mL L-1 e formulação elaborada. Após 12 meses, verificou-se que os melhores substratos para plantas da orquídea C. loddgesii "Alba" x C. loddgesii "Atibaia" são casca de arroz carbonizada e fibra de piaçava, e melhores respostas à adubação são obtidas com o adubo foliar Biofert Plus®.This paper was aimed at studing different substrates and fertilization for the development of C. loddgesii 'Alba' x C. loddgesii 'Atibaia' orchid species. Seedlings from in vitro culture were transplanted to plastic trays containing xaxim powder. After 6-months of acclimatization inside a greenhouse, plantles measuring beetwen 3.0 to 5.0cm length were transferred to 24 cell trays with a volume of 150cm³. Four substrates were tested (broken stone number 0, carbonized rice hulls, defibered xaxim and piassava fiber with tree different fertilizations (Biofert Plus®, Dyna Grow® and a specific formulation developed for this work. Pure water was tested as control. After 12-months of growth the best treatments for the acclimatization of in vitro produced C. loddgesii 'Alba' x C. loddgesii 'Atibaia' orchid plantlets were carbonized rice hulls or piassava fiber substrates combined with leaf fertilizer Biofert Plus®.

Obtaining a new variety of rape by biotechnology

Energy Technology Data Exchange (ETDEWEB)

Chen, Z H; Cun, S X; Zhang, Z Z; Wang, W F; Zhang, T H; Li, W B; Zhang, L H [Institute of Genetics, Chinese Academy of Sciences, Beijing (China); [Laboratory of Genetic Engineering, Yunnan Academy of Agricultural Sciences, Kunming (China)

1990-01-01

Full text: High-frequency induction of pollen embryoids and plantlets of rape was obtained by stepped float anther culture. In addition, somatic embryoids and plantlets were induced with a high frequency from several species by cell suspension culture. The erucic acid (EA) content of embryoids was analysed by a micro-analysis technique and a semi-micro-analysis of glucosinolates (GS) content in culture was used. A new variety 'H86-166' with low EA and low GS content was selected by these techniques. It gave a yield of 3169 kg/ha and has been released for commercial production on about 1000 ha in Yunnan Province. (author)

Gamma ray induced effects in in vitro mammillaria san angelensis (CACTACEAE)

International Nuclear Information System (INIS)

Brunner, I.; Rubluo, A.; De la Cruz, E.; Gonzalez, J.

2001-01-01

The nearly extinct Mammillaria san angelensis has been recovered in vitro and large quantities of individuals are growing successfully in the greenhouse, however due to its axillary shoot proliferation origin the genetic make-up of this species is extremely narrow. For this reason is urgently required to increase its genetic variability. The induction of point mutations through irradiation of in vitro plantlets can favour the survival of this plant. Radiosensibility experiments using gamma rays were performed in order to determine the primary effects of radiation in in vitro plantlets of this species as a necessary preliminary for the use of the mutation induction strategy

Genetic fidelity in micropropagated plantlets of Ochreinauclea ...

African Journals Online (AJOL)

STORAGESEVER

2009-07-06

Jul 6, 2009 ... show the genetic similarity among the 22 plants (21 micropropagated plants and 1 donor mother plant). In the present study, ..... Determination of genetic stability in long-term micropropagated shoots of Pinus thunbergii Parl. using. RAPD markers. Plant Cell Rep. 18: 193-197. Guo WL, Gong L, Ding ZF, ...

Somatic embryogenesis and plantlet regeneration from protoplast ...

African Journals Online (AJOL)

After 4 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads. Growth of microcalli in the medium with nurse cells (33.3%) was much better than those in the medium without nurse cells (6.5%). Transferring beads onto MS medium supplemented with 0.1 mg/l BA increased the growth of embryogenic ...

Photosynthetic and biochemical mechanisms of an EMS-mutagenized cowpea associated with its resistance to cowpea severe mosaic virus.

Science.gov (United States)

Souza, Pedro F N; Silva, Fredy D A; Carvalho, Fabricio E L; Silveira, Joaquim A G; Vasconcelos, Ilka M; Oliveira, Jose T A

2017-01-01

The seed treatment of a CPSMV-susceptible cowpea genotype with the mutagenic agent EMS generated mutagenized resistant plantlets that respond to the virus challenge by activating biochemical and physiological defense mechanisms. Cowpea is an important crop that makes major nutritional contributions particularly to the diet of the poor population worldwide. However, its production is low, because cowpea is naturally exposed to several abiotic and biotic stresses, including viral agents. Cowpea severe mosaic virus (CPSMV) drastically affects cowpea grain production. This study was conducted to compare photosynthetic and biochemical parameters of a CPSMV-susceptible cowpea (CE-31 genotype) and its derived ethyl methanesulfonate-mutagenized resistant plantlets, both challenged with CPSMV, to shed light on the mechanisms of virus resistance. CPSMV inoculation was done in the fully expanded secondary leaves, 15 days after planting. At 7 days post-inoculation, in vivo photosynthetic parameters were measured and leaves collected for biochemical analysis. CPSMV-inoculated mutagenized-resistant cowpea plantlets (MCPI) maintained higher photosynthesis index, chlorophyll, and carotenoid contents in relation to the susceptible (CE-31) CPSMV-inoculated cowpea (CPI). Visually, the MCPI leaves did not exhibit any viral symptoms neither the presence of the virus as examined by RT-PCR. In addition, MCPI showed higher SOD, GPOX, chitinase, and phenylalanine ammonia lyase activities, H 2 O 2 , phenolic contents, and cell wall lignifications, but lower CAT and APX activities in comparison to CPI. All together, these photosynthetic and biochemical changes might have contributed for the CPSMS resistance of MCPI. Contrarily, CPI plantlets showed CPSMV accumulation, severe disease symptoms, reduction in the photosynthesis-related parameters, chlorophyll, carotenoid, phenolic compound, and H 2 O 2 contents, in addition to increased β-1,3-glucanase, and catalase activities that might have

Nuclear DNA content of the pigeon orchid (Dendrobium crumenatum Sw. with the analysis of flow cytometry

Directory of Open Access Journals (Sweden)

Upatham Meesawat

2008-05-01

Full Text Available Nuclear DNA content for the adult plants grown in a greenhouse and in vitro young plantlets of the pigeon orchid (Dendrobium crumenatum Sw. was analyzed using flow cytometry. The resulting 2C DNA values ranged from 2.30±0.14 pgto 2.43±0.06 pg. However, nuclear DNA ploidy levels of long-term in vitro plantlets were found to be triploid and tetraploid.These ploidy levels were confirmed by chromosome counting. Tetraploid individuals (2n = 4x = 76 had approximately two times DNA content than diploid (2n = 2x = 38 individuals. This variation may be due to prolonged cultivation and thepresence of exogenous plant growth regulators.

THE EFFECT OF SPRUCE BARK POLYPHENOLS EXTRACT IN COMBINATION WITH DEUTERIUM DEPLETED WATER (DDW ON GLYCINE MAX L. AND HELIANTHUS ANNUUS L. DEVELOPMENT

Directory of Open Access Journals (Sweden)

Corneliu Tanase

2010-09-01

Full Text Available The aim of this study was to evaluate the effect of spruce bark aqueous extract and deuterium depleted water (DDW as bioregulators on the plant growth Glycine max L. and Helianthus annuus. The following specific parameteres were closely monitorised: germination energy and germination capacity, plants vegetative organelles growth and development and photoassimilatory pigments concentrations. The results have shown that DDW presents different effects depending on tested plant species. In the case of soybean, DDW presented stimulatory effects on both germination energy and capacity, radicles elongation, primary leaves growth and development but inhibitory effects on photoassimilatory pigments. Spruce bark extract reduced the germination capacity of soybean seeds, but accelerated the germination process of sunflower seeds and present stimulatory effects on plantlets biomass accumulation. The combination of DDW with Picea abies polyphenolic extract promoted soybean plantlet elongation, especially the rootlets ones and stimulated green biomass accumulation for both soybean and sunflower plantlets. Analyzing the photoassimilatory pigments concentration for sunflower, it can be observed an increasing trend (almost 100% comparing with control when introduce into the growth medium DDW and P. abies polyphenolic extract. DDW and P. abies bark extract have shown an important role in plant growth and development, improving photoassimiliation process.

Somatic Embryogenesis in Peach-Palm (Bactris gasipaes) Using Different Explant Sources.

Science.gov (United States)

Steinmacher, Douglas A; Heringer, Angelo Schuabb; Jiménez, Víctor M; Quoirin, Marguerite G G; Guerra, Miguel P

2016-01-01

Peach palm (Bactris gasipaes Kunth) is a member of the family Arecaceae and is a multipurpose but underutilized species. Nowadays, fruit production for subsistence and local markets, and heart-of-palm production for local, national, and international markets are the most important uses of this plant. Conventional breeding programs in peach palm are long-term efforts due to the prolonged generation time, large plant size, difficulties with controlled pollination and other factors. Although it is a caespitose palm, its propagation is currently based on seeds, as off-shoots are difficult to root. Hence, tissue culture techniques are considered to be the most likely strategy for efficient clonal plantlet regeneration of this species. Among various techniques, somatic embryogenesis offers the advantages of potential automated large-scale production and putative genetic stability of the regenerated plantlets. The induction of somatic embryogenesis in peach palm can be achieved by using different explant sources including zygotic embryos, immature inflorescences and thin cell layers from the young leaves and shoot meristems. The choice of a particular explant depends on whether clonal propagation is desired or not, as well as on the plant conditions and availability of explants. Protocols to induce and express somatic embryogenesis from different peach palm explants, up to acclimatization of plantlets, are described in this chapter.

Identification of genes related to Paulownia witches' broom by AFLP and MSAP.

Science.gov (United States)

Cao, Xibing; Fan, Guoqiang; Deng, Minjie; Zhao, Zhenli; Dong, Yanpeng

2014-08-21

DNA methylation is believed to play important roles in regulating gene expression in plant growth and development. Paulownia witches' broom (PaWB) infection has been reported to be related to gene expression changes in paulownia plantlets. To determine whether DNA methylation is associated with gene expression changes in response to phytoplasma, we investigated variations in genomic DNA sequence and methylation in PaWB plantlets treated with methyl methane sulfonate (MMS) using amplified fragment length polymorphism (AFLP) and methylation-sensitive amplification polymorphism (MSAP) techniques, respectively. The results indicated that PaWB seedings recovered a normal morphology after treatment with more than 15 mg·L(-1) MMS. PaWB infection did not cause changes of the paulownia DNA sequence at the AFLP level; However, DNA methylation levels and patterns were altered. Quantitative real-time PCR (qRT-PCR) showed that three of the methylated genes were up-regulated and three were down-regulated in the MMS-treated PaWB plantlets that had regained healthy morphology. These six genes might be involved in transcriptional regulation, plant defense, signal transduction and energy. The possible roles of these genes in PaWB are discussed. The results showed that changes of DNA methylation altered gene expression levels, and that MSAP might help identify genes related to PaWB.

Identification of Genes Related to Paulownia Witches’ Broom by AFLP and MSAP

Science.gov (United States)

Cao, Xibing; Fan, Guoqiang; Deng, Minjie; Zhao, Zhenli; Dong, Yanpeng

2014-01-01

DNA methylation is believed to play important roles in regulating gene expression in plant growth and development. Paulownia witches’ broom (PaWB) infection has been reported to be related to gene expression changes in paulownia plantlets. To determine whether DNA methylation is associated with gene expression changes in response to phytoplasma, we investigated variations in genomic DNA sequence and methylation in PaWB plantlets treated with methyl methane sulfonate (MMS) using amplified fragment length polymorphism (AFLP) and methylation-sensitive amplification polymorphism (MSAP) techniques, respectively. The results indicated that PaWB seedings recovered a normal morphology after treatment with more than 15 mg·L−1 MMS. PaWB infection did not cause changes of the paulownia DNA sequence at the AFLP level; However, DNA methylation levels and patterns were altered. Quantitative real-time PCR (qRT-PCR) showed that three of the methylated genes were up-regulated and three were down-regulated in the MMS-treated PaWB plantlets that had regained healthy morphology. These six genes might be involved in transcriptional regulation, plant defense, signal transduction and energy. The possible roles of these genes in PaWB are discussed. The results showed that changes of DNA methylation altered gene expression levels, and that MSAP might help identify genes related to PaWB. PMID:25196603

Banana and coconut in-vitro cultures for induced mutations studies

International Nuclear Information System (INIS)

Guzman, E.V. de; Ubalde, E.M.; Rosario, A.G. del

1976-01-01

Explants obtained from shoot tips of developing suckers of banana were found to be suitable materials for plantlet formation in vitro. Subculture of the shoots formed from the original explant can lead to regeneration of numerous shoots from a single sucker. The technique can be a practical method of propagation. Though a more rapid rate of multiplication is still to be desired the present rate is much faster than the maximum rate of sucker production obtained under natural conditions. The conditions which favor shoot formation were the addition of benzyladenine, the use of dextrose at 2% instead of 4%, and the use of either Murashige and Skoog or of Vacin and Went salt solution. Viable explants can be obtained from suckers irradiated up to 10 Kr. Survival and shoot formation are sometimes not adversely affected by irradiation. Chlorosis and reduction in plantlet size and unusual seedling characters were among the features observed in plantlets regenerated from irradiated tissues. Proliferations derived from the cotyledonary sheath in coconut embryos could be successfully subcultured. The initial and subsequent proliferations may appear as nodular growths. In continuous subculture a nodular growth may change over to a large mass with smooth surface. Cellular characteristics of the two types of growth are markedly different. (author)

Radiation and chemical mutagen induced somaclonal variations through in vitro organogenesis of cotton (Gossypium hirsutum L.).

Science.gov (United States)

Muthusamy, Annamalai; Jayabalan, Narayanasamy

2014-12-01

The purpose of the investigation was to induce somaclonal variations by gamma rays (GR), ethylmethane sulphonate (EMS) and sodium azide (SA) during in vitro organogenesis of cotton. The shoot tip explants were irradiated with 5-50 Gray (Gy) GR (Cobalt 60), 0.5-5.0 mM EMS and SA separately, and inoculated on Murashige and Skoog (MS) medium fortified with plant growth regulator (PGR) for organogenesis. The plantlets with well-developed root systems were acclimatized and transferred into the experimental field to screen the somaclonal variations during growth and development. The number of somaclonal variations was observed in growth of irradiated/treated shoot tips, multiplication, plantlet regeneration and growth in vitro and ex vitro. The lower doses/concentrations of mutagenic treatments showed significant enhancement in selected agronomical characters and they showed decreased trends with increasing doses/concentrations of mutagenic agents. The results of the present study revealed the influence of lower doses/concentrations of mutagenic treatments on in vitro and ex vitro growth of cotton plantlets and their significant improvement in agronomical characters which needs further imperative stability analysis. The present observations showed the platform to use lower doses/concentrations of mutagenic agents to induce variability for enhanced agronomical characters, resistant and tolerant cotton varieties.

Effect of leaf harvesting on reproduction and natural populations of Indian Wild Banana Ensete superbum (Roxb. Cheesman (Zingiberales: Musaceae

Directory of Open Access Journals (Sweden)

Mahendra R. Bhise

2015-04-01

Full Text Available Ensete superbum (Roxb. Cheesman an important taxon in India is threatened in Maharashtra. It is sporadically distributed on high altitude slopes and rocky cliffs in the Western Ghats. It is an important medicinal and economic plant utilized by people living in rural areas, while the leaves are also utilized in urban areas. The leaves are harvested for commercial purposes. The effect of leaf harvest on natural population with respect to regeneration of new plantlets was evaluated. The results revealed that, non-scientific leaf harvesting resulted in significantly reduced flowering and fruiting, less number of new plantlets in the population, and population degradation. Therefore, leaf harvesting should be practiced in a controlled manner to maintain the population health of this highly potential species. 

Investigation of the effects of monochromatic lights combination on Phalaenopsis orchid and chrysanthemum in vitro plant growth

International Nuclear Information System (INIS)

Le Tien Thanh; Nguyen Tuong Mien; Huynh Thi Trung; Vu Thi Trac; Pham Van Nhi

2016-01-01

Eleven independent monochromatic light combination systems were established using super blue and red light LEDs. These are adjustable for mixing ratio of red and blue lights, the intensity and time for illumination. A fluorescent light illumination system was included as control. These were used to implement experiments for investigation effects of monochromatic lights combination in in vitro plantlet fully-forming stage on in vitro plantlet fully-forming and on the development of plant in nursery of chrysanthemum and Phalaenopsis orchid. Results from these experiments showed that illumination with intensity of 750 lx and light mixture of 20% blue-80% red is suitable for Phalaenopsis orchid; illumination with intensity of 1100 lx and light mixture of 90% red light-10% blue light is suitable for chrysanthemum. (author)

Callus induction, regeneration and transformation of sugarcane ...

African Journals Online (AJOL)

Yomi

2012-03-22

Mar 22, 2012 ... dichlorophenoxyacetic acid (2,4-D), naphthalene acetic acid (NAA), yeast extract and coconut water .... transformed (control) plantlets using CTAB method (Doyle and .... Embryogenic Sugarcane Callus by Image Analysis.

Somatic embryogenesis, scanning electron microscopy, histology and biochemical analysis at different developing stages of embryogenesis in six date palm (Phoenix dactylifera L.) cultivars.

Science.gov (United States)

Aslam, Junaid; Khan, Saeed Ahmad; Cheruth, Abdul Jaleel; Mujib, Abdul; Sharma, Maheshwar Pershad; Srivastava, Prem Shanker

2011-10-01

An efficient somatic embryogenesis system has been established in six date palm (Phoenix dactylifera L.) cultivars (Barhee, Zardai, Khalasah, Muzati, Shishi and Zart). Somatic embryogenesis (SE) was growth regulators and cultivars dependent. Friable embryogenic callus was induced from excised shoot tips on MS medium supplemented with various auxins particularly 2,4-dichlorophenoxyacetic acid (2,4-D, 1.5 mg 1(-l)). Suspension culture increased embryogenesis potentiality. Only a-naphthaleneacetic acid (NAA, 0.5 mg 1(-1)) produced somatic embryos in culture. Somatic embryos germinated and converted into plantlets in N(6)-benzyladenine (BAP, 0.75 mg 1(-l)) added medium following a treatment with thidiazuron (TDZ, 1.0 mg 1(-l)) for maturation. Scanning electron microscopy showed early stages of somatic embryo particularly, globular types, and was in masses. Different developing stages of embryogenesis (heart, torpedo and cotyledonary) were observed under histological preparation of embryogenic callus. Biochemical screening at various stages of somatic embryogenesis (embryogenic callus, somatic embryos, matured, germinated embryos and converted plantlets) of date palm cultivars has been conducted and discussed in detail. The result discussed in this paper indicates that somatic embryos were produced in numbers and converted plantlets can be used as a good source of alternative propagation. Genetic modification to the embryo precursor cell may improve the fruit quality and yield further.

Identification of Genes Related to Paulownia Witches’ Broom by AFLP and MSAP

Directory of Open Access Journals (Sweden)

Xibing Cao

2014-08-01

Full Text Available DNA methylation is believed to play important roles in regulating gene expression in plant growth and development. Paulownia witches’ broom (PaWB infection has been reported to be related to gene expression changes in paulownia plantlets. To determine whether DNA methylation is associated with gene expression changes in response to phytoplasma, we investigated variations in genomic DNA sequence and methylation in PaWB plantlets treated with methyl methane sulfonate (MMS using amplified fragment length polymorphism (AFLP and methylation-sensitive amplification polymorphism (MSAP techniques, respectively. The results indicated that PaWB seedings recovered a normal morphology after treatment with more than 15 mg·L−1 MMS. PaWB infection did not cause changes of the paulownia DNA sequence at the AFLP level; However, DNA methylation levels and patterns were altered. Quantitative real-time PCR (qRT-PCR showed that three of the methylated genes were up-regulated and three were down-regulated in the MMS-treated PaWB plantlets that had regained healthy morphology. These six genes might be involved in transcriptional regulation, plant defense, signal transduction and energy. The possible roles of these genes in PaWB are discussed. The results showed that changes of DNA methylation altered gene expression levels, and that MSAP might help identify genes related to PaWB.

Physiological differences and changes in global DNA methylation levels in Agave angustifolia Haw. albino variant somaclones during the micropropagation process.

Science.gov (United States)

Duarte-Aké, Fátima; Castillo-Castro, Eduardo; Pool, Felipe Barredo; Espadas, Francisco; Santamaría, Jorge M; Robert, Manuel L; De-la-Peña, Clelia

2016-12-01

Global DNA methylation changes caused by in vitro conditions are associated with the subculturing and phenotypic variation in Agave angustifolia Haw. While the relationship between the development of albinism and in vitro culture is well documented, the role of epigenetic processes in this development leaves some important questions unanswered. During the micropropagation of Agave angustifolia Haw., we found three different phenotypes, green (G), variegated (V) and albino (A). To understand the physiological and epigenetic differences among the somaclones, we analyzed several morphophysiological parameters and changes in the DNA methylation patterns in the three phenotypes during their in vitro development. We found that under in vitro conditions, the V plantlets maintained their CAM photosynthetic capacity, while the A variant showed no pigments and lost its CAM photosynthetic ability. Epigenetic analysis revealed that global DNA methylation increased in the G phenotype during the first two subcultures. However, after that time, DNA methylation levels declined. This hypomethylation correlated with the appearance of V shoots in the G plantlets. A similar correlation occurred in the V phenotype, where an increase of 2 % in the global DNA methylation levels was correlated with the generation of A shoots in the V plantlets. This suggests that an "epigenetic stress memory" during in vitro conditions causes a chromatin shift that favors the generation of variegated and albino shoots.

Phytotoxicity and cytogenotoxicity of hydroalcoholic extracts from Solanum muricatum Ait. and Solanum betaceum Cav. (Solanaceae) in the plant model Lactuca sativa.

Science.gov (United States)

Dos Santos, Fabio Eduardo; Carvalho, Marcos Schleiden Sousa; Silveira, Graciele Lurdes; Correa, Felipe Folgaroli; Cardoso, Maria das Graças; Andrade-Vieira, Larissa Fonseca; Vilela, Luciane Resende

2018-03-05

Plants are rich in biologically active compounds. They can be explored for the production of bioherbicides. In this context, the present work aimed to evaluate the allelopathic effect of hydroalcoholic extracts from two Solanaceae species: Solanum muricatum Ait. and Solanum betaceum Cav. For this end, we conducted phytochemical screening and biological assays, determining the effects of the extracts on germination, early development, cell cycle, and DNA fragmentation in plantlets and meristematic cells of the plant model Lactuca sativa L. (lettuce). The percentage of seeds germinated under effect of S. muricatum extract did not differ from the control, but plantlet growth was reduced at the highest concentrations. For S. betaceum extract, dose dependence was observed for both germination and plantlet development, with the highest concentrations inhibiting germination. The growth curves revealed the concentrations of 2.06 and 1.93 g/L for S. muricatum and S. betaceum extracts, respectively, as those reducing 50% of root growth (RG). At these concentrations, both extracts presented mitodepressive effect, besides inducing significant increase in the frequency of condensed nuclei, associated to DNA fragmentation and cytoplasmic shrinkage. The frequency of chromosome alterations was not significant. We further discuss the mechanisms of action related to the chemical composition of the extracts, which presented organic acids, reducing sugars, proteins, amino acids, and tannins, besides catechins and flavonoids, only found in the extract of S. betaceum.

Micropropagation and in vitro conservation of the rare and threatened plants Ramonda serbica and Ramonda nathaliae.

Science.gov (United States)

Gashi, Bekim; Abdullai, Kasamedin; Sota, Valbona; Kongjika, Efigjeni

2015-01-01

Ramonda serbica and Ramonda nathaliae are rare and endemo relict plant species from Balkan Peninsula. An efficient micro propagation and in vitro conservation method via direct and indirect organogenesis from seed and leaf explants, respectively, was established in this study. The seed of both Ramonda species were collected from different populations in Kosovo, and were germinated in nutrient media JG-B without any phytohormone. The highest number of shoots and multiplication rate was observed on JG-B medium supplemented with BAP and IAA (0.5 mg l(-1) each), whereas the highest number of leaves per plantlets was found on WPM and RA medium supplemented with BAP and IAA (0.1 mg l(-1) each). During this stage of micro propagation some significant differences were observed in plantlets from different populations. The indirect organogenesis from parts of leaves of natural plants was not successful due to unavailability of established protocol for disinfections of the plant material. On other hand, parts of leaves from micro propagated plantlets, cultured on MS medium supplemented with different ratio of BAP and NAA, resulted in the highest efficiency for shoot regeneration. In vitro conservation of micro propagated plants at the lower temperature (4 °C) had a significantly positive effect for storage of more than 12 months.

INFLUENCIA DE DIFERENTES SUBSTRATOS E FERTILIZANTES NA ACLIMATIZAÇÃO DE PLANTAS DE FIGUEIRA (FICUS CARICA L.

Directory of Open Access Journals (Sweden)

Ester Alice Ferreira

2008-01-01

Full Text Available Due to a great amount of environmental differences between in vitro and ex vitro conditions, the acclimatization phase of plantlets is one of the essential points to guarantee the success in obtaining micropropagated plants. Using a substrate with chemical and organic composition is important on this phase, because it will influence in the nutritional state of the seedlings. The present paper aimed to evaluate the effects of different substrate on acclimatization of fig plants 'Roxo de Valinhos'. It were selected uniform plantlets within a 5 cm of height which were transferred to the greenhouse, with an intermittent mist system, with 50% mesh of shadow by sombrite® planted in stereo foam trays where were submitted to the following treatments: Plantmax®; carbonized rice husk; soil in combination with Osmocote®; manure (3:1:1; NPKMg. The experimental design adopted was randomized blocks in factorial scheme and the evaluation was done one hundred and twenty days after the acclimatization by the following characteristics: plant high (cm, number of leaves/plant, dry weight of aerial part and roots and total weight (g/plant. As results, it was verified that on the acclimatization of fig plantlets 'Roxo de Valinhos' using of substrate soil + carbonized rice husk+ manure promotted highest number of leaves and largest weight of aerial dry matter were obtained Plantmax® substrate without any addition of fertilizers.

Acclimatization of the endangered Mexican epiphytic orchid, Laelia speciosa (H.B.K. Schltr

Directory of Open Access Journals (Sweden)

Martha Mireya Ortega-Loeza

2012-02-01

Full Text Available In vitro propagation could be an alternative for the conservation of the endemic and endangered Mexican epiphytic orchid Laelia speciosa (H.B.K. Schltr. The goal of this study was to develop a protocol that would enhance acclimatization of in vitro – derived L. speciosa plantlets – a critical stage in propagation and subsequent conservation. Observations of stomata opening during ex vitro acclimatization, and the time of in vitro culture (0, 5, 10, 15, 20, 25, 30 days in greenhouse conditions (pre-acclimatization, on the survival and development of seedlings during the ex vitro acclimatization were carried out. In addition, the effect of different levels of nutrients (100%, 75%, 50%, 25% and 0%-strength salts and sucrose (0, 10, 20, 30, 40 g l−1 in the Murashige and Skoog medium (MS on the same parameters were measured. Plantlets incubated 20 days in greenhouse conditions before ex vitro acclimatization also displayed the best growth with a survival rate of 97.5%, related with high stomata opening. Plantlets on MS containing 100%-strength salts (with 20 days of pre-acclimatization, 40 g l−1 sucrose had the highest rate (97.5–100% of survival and vigor when acclimatized. By improving micropropagation through acclimatization, the sustainable management of L. speciosa now more likely, benefitting the conservation of this endangered species.

Field Performance of Cassava (Manihot esculenta Crantz ...

African Journals Online (AJOL)

Field Performance of Cassava (Manihot esculenta. Crantz) ... Keywords: Tissue culture-derived plantlets, Field plant growth, Yield, Root tuber characteristics,. Cassava ..... Micro-propagation of ... Roca, W.M.; Henry, G., Angel, F. and Sarria, R.

Callus regeneration from stem explants of Pseudarthira viscida (L ...

African Journals Online (AJOL)

STORAGESEVER

2009-09-01

Sep 1, 2009 ... regeneration frequency have come from tissue culture work done in ... Table 1. In vitro responses from stem callus of Psudarthria viscida Wight & Arn. Growth .... plantlets regeneration from cotyledonary callus of Tomato.

In vitropropagation in Temporary Immersion System of sugarcane plants variety `RB 872552' derived from somatic embryos

Directory of Open Access Journals (Sweden)

Marina Medeiros de Araújo Silva

2015-07-01

Full Text Available In this study, we used a temporary immersion system (TIS to multiply sugarcane (Saccharum spp. plants obtained by somatic embryogenesis (SE. SE was induced from immature leaf segments that were grown in culture medium supplemented with 2,4-D and BAP. Embryo formation occurred in 81% of the inoculated explants and 254 plants were regenerated. Ninety plants were transferred to TIS and cultured in medium supplemented with BAP. After three subcultures, 60 000 plantlets were obtained and transferred to rooting media. After 30 days of acclimatization period plantlets were well developed and exhibited a 96% survival. The results demonstrate the feasibility of the combined use of two important techniques of in vitro culture (SE and shoot multiplication in TIS to sugarcane in vitro propagation. Key words: acclimatization, 6-benzylaminopurine, Saccharum spp.

Studies on the tissue culture of Stevia rebaudiana and its components; (II). Induction of shoot primordia.

Science.gov (United States)

Miyagawa, H; Fujioka, N; Kohda, H; Yamasaki, K; Taniguchi, K; Tanaka, R

1986-08-01

Shoot primordia, which were able to propagate vegetatively with a very high rate and to redifferentiate easily to new plants, were induced from shoot tips of Stevia rebaudiana Bertoni on Gamborg B5 medium containing 6-benzylaminopurine (BAP) and alpha-naphthaleneacetic acid (NAA) under light. The propagation of the shoot primordia of Stevia rebaudiana is rapid, and they are highly stable in chromosome number and karyotype. The shoot primordia can propagate at a high rate for a long time without differentiation. At any time, the shoot primordia readily developed into plantlets with shoots and roots within 2 or 3 weeks in static culture on B5 medium containing 0.02 mg/l BAP and 2% sucrose. The plantlets were transplanted to sterilized soil to grow to normal adult plants.

Regeneration of begonia plantlets by direct organogenesis | Mendi ...

African Journals Online (AJOL)

African Journal of Biotechnology. Journal Home · ABOUT THIS JOURNAL · Advanced Search · Current Issue · Archives · Journal Home > Vol 8, No 9 (2009) >. Log in or Register to get access to full text downloads.

Responses of macropropagated Musa plantlets to transient moisture ...

African Journals Online (AJOL)

Moisture stress causes serious devastating effects on crops ranging from retarded growth, tissue desiccation, and severe yield drop to outright death. In this study, the efficacy of six nursery substrates formulated in varying volume ratios of composted plantain fruit/bunch wastes (PFW), sawdust (SD), poultry manure (PM) and ...

Regeneration of plantlets from unpollinated ovary cultures of ...

African Journals Online (AJOL)

Tsega

2013-09-25

Sep 25, 2013 ... 3Ethiopian Institute of Agricultural Research (EIAR): Holetta ... cultured unpollinated ovaries, 1,100 embryonic tissues (7.6 %) and 75 regenerants were obtained. The ... viral diseases, insect pests and increasing human popu-.

High frequency induction of somatic embryos and plantlet ...

African Journals Online (AJOL)

PRECIOUS

2009-11-16

Nov 16, 2009 ... plant, Hygrophila spinosa through direct somatic embryogenesis from nodal explants excised from 4 week old ... medicinal purposes further curbs propagation via seed. Plant tissue ... buted a great deal of information for the genetic, morpho- ..... analysis of peroxidase in cultured lettuce (Lactuca sativa L.).

Assessment of genetic stability in micropropagules of Jatropha curcas genotypes by RAPD and AFLP analysis

KAUST Repository

Sharma, Sweta K.; Pamidimarri, D. V N N Sudheer; Vijay Anand, K. G.; Reddy, Muppala P.

2011-01-01

of high yielding clones and their large scale multiplication by vegetative propagation to obtain true to type plants. In the current investigation plantlets of J. curcas generated by axillary bud proliferation (micropropagation) using nodal segments

Studies on seed germination and in vitro shoot multiplication of ...

African Journals Online (AJOL)

Yomi

2011-12-21

Dec 21, 2011 ... vitro seed germination and plantlet regeneration of this plant. ... Key words: Germination, gibberellic acid, growth regulators, node explants, Satureja ..... Abscisic Acid: A. Seed Maturation and Antistress Signal, 3rd ed. Sinauer ...

Systemic induced tolerance against root-knot nematodes in rice

African Journals Online (AJOL)

user

Rice plantlets were randomly grown in laboratory, sprayed with hormones and riboflavin .... between riboflavin, SA, JA and ET pathways in rice-nematodes interaction, ..... oxidative damage caused by aging as well as biotic and abiotic stress.

In vitro embryo rescue and plant regeneration following self ...

African Journals Online (AJOL)

EJIRO

2015-07-08

Jul 8, 2015 ... calculated using the simple matching Euclidean distance (DE) followed by cluster analysis ..... we plan to fully characterize these plantlets and to establish whether or not .... Tesis de Grado, Universidad. ICESI, Cali. Colombia.

A review of somaclonal variation in plantain (Musa spp ...

African Journals Online (AJOL)

SARAH

2013-07-30

Jul 30, 2013 ... Plant tissue has contributed immensely in large-scale production of resistant and pest free plantlets, .... Potato is a native of South America (Peru) where considerable ..... An efficient, short and cost-effective regeneration ...

Effect of different combinations of 6-benzyl amino purine and ...

African Journals Online (AJOL)

SNAP CUSTOMER

2013-02-13

Feb 13, 2013 ... The culture meristem grew into callus after 28 days then adventitious plantlets ... plantain is greatly reduced by virus disease (Lepoivre,. 2000). ..... for Production of Quality Banana Planting Material in Asia-Pacific. Asia-Pacific ...

UNIVERSIDADE ESTADUAL DE MONTES CLAROS

African Journals Online (AJOL)

OF DO MICRO

2013-01-30

Jan 30, 2013 ... the mesocarp and seed were determined, and lipids and proteins were identified through histochemical analyses of the .... ghts of their structures did not vary greatly in plantlets obtained from .... A classification system for seed.

An improved embryo-rescue protocol for hybrid progeny from seedless Vitis vinifera grapes × wild Chinese Vitis species.

Science.gov (United States)

Li, Gui Rong; Ji, Wei; Wang, Gang; Zhang, Jian Xia; Wang, Yue Jin

A highly efficient technique of embryo rescue is critical when using stenospermocarpic Vitis vinifera cultivars (female parents) to breed novel, disease-resistant, seedless grape cultivars by hybridizing with wild Chinese Vitis species (male parents) having many disease-resistance alleles. The effects of various factors on the improvement of embryo formation, germination, and plantlet development for seven hybrid combinations were studied. The results indicated that Beichun and Shuangyou were the best male parents. The best sampling time for ovule inoculation differed among the female parents. When hybrid ovules were cultured on a double-phase medium with five different solid medium types, percent embryo formation was highest (11.3-28.3%) on a modified MM3 medium. Percentages of embryo germination (15.4-55.4%) and plantlet development (11.15-44.6%) were all highest when embryos were cultured on Woody Plant Medium + 5.7 μM indole-3-acetic acid + 4.4 μM 6-benzylaminopurine + 1.4 μM gibberellic acid + 2% sucrose + 0.05% casein hydrolysate + 0.3% activated charcoal + 0.7% agar. In the absence of other amino acids, the addition of proline significantly increased embryo formation (36.1%), embryo germination (64.6%), and plantlet development (90.5%). A highly efficient protocol has been developed for hybrid embryo rescue from seedless V. vinifera grapes × wild Chinese Vitis species that results in a significant improvement in breeding efficiency for new disease-resistant seedless grapes.

Monitoring the colonization of sugarcane and rice plants by the endophytic diazotrophic bacterium Gluconacetobacter diazotrophicus marked with gfp and gusA reporter genes.

Science.gov (United States)

Rouws, L F M; Meneses, C H S G; Guedes, H V; Vidal, M S; Baldani, J I; Schwab, S

2010-09-01

To evaluate the colonization process of sugarcane plantlets and hydroponically grown rice seedlings by Gluconacetobacter diazotrophicus strain PAL5 marked with the gusA and gfp reporter genes. Sugarcane plantlets inoculated in vitro with PAL5 carrying the gfp::gusA plasmid pHRGFPGUS did not present green fluorescence, but beta-glucuronidase (GUS)-stained bacteria could be observed inside sugarcane roots. To complement this existing inoculation methodology for micropropagated sugarcane with a more rapid colonization assay, we employed hydroponically grown gnotobiotic rice seedlings to study PAL5-plant interaction. PAL5 could be isolated from the root surface (10(8) CFU g(-1)) and from surface-disinfected root and stem tissues (10(4) CFU g(-1)) of inoculated plants, suggesting that PAL5 colonized the internal plant tissues. Light microscopy confirmed the presence of bacteria inside the root tissue. After inoculation of rice plantlets with PAL5 marked with the gfp plasmid pHRGFPTC, bright green fluorescent bacteria could be seen colonizing the rice root surface, mainly at the sites of lateral root emergence, at root caps and on root hairs. The plasmids pHRGFPGUS and pHRGFPTC are valid tools to mark PAL5 and monitor the colonization of micropropagated sugarcane and hydroponic rice seedlings. These tools are of use to: (i) study PAL5 mutants affected in bacteria-plant interactions, (ii) monitor plant colonization in real time and (iii) distinguish PAL5 from other bacteria during the study of mixed inoculants.

PENGARUH IRADIASI SINAR GAMMA TERHADAP VARIASI PERTUMBUHAN ANGGREK Brachypeza indusiata (Reichb.f Garay SECARA IN VITRO

Directory of Open Access Journals (Sweden)

Maria Utami Dewi Wardhani

2007-01-01

Full Text Available A research was carried out in Orchid Laboratory of Bogor Botanical Gardens to observe the effect of gamma ray irradiation on the growth variation of Brachypezo indusiata (Reichb.f Garray and suitable culture medium for propagating the plant. The research was arranged in a factorial experiment with a Completely Randomized Design. The first factor was gamma ray irradiation on protocorm-like­bodies (plb of germinated seeds of B. indusiata in which four levels of irradiation was applied, i.e. 0, 10, 20, and 30 Gy. The second factor was the use of two different culture media, i.e. Vacin & Went medium supplemented with either banana pulp or a mixture of tomato and sprout bean as the essential organic compound of the medium. The result showed that the irradiation treatment improved the survival of B. indusiata explants, most significantly at 20 Gy level of irradiation, and could modify the growth of the resulted plantlets by reducing the height of the plant and the length and width of the leaves. The higher the level of irradiation the smaller the plantlets produced. Meanwhile, the addition of organic compound derived from banana fruit into the basal medium of Vacin & Went enhanced the growth of the plantlets better than that derived from a mixture of tomato fruit and bean sprout. However, there was no significant interaction effect detected between the irradiation treatments and the use of different culture media.

Development of a new wheat germplasm with high anther culture ability by using a combination of gamma-ray irradiation and anther culture.

Science.gov (United States)

Zhao, Linshu; Liu, Luxiang; Wang, Jing; Guo, Huijun; Gu, Jiayu; Zhao, Shirong; Li, Junhui; Xie, Yongdun

2015-01-01

Wheat with high anther culture ability would be beneficial for breeding. We aimed to screen a wheat germplasm to with high anther culture ability as well as good agronomic characteristics. The F1 young spikes of winter wheat cross combination Yanfu188/Jimai37 were irradiated with gamma rays at a dose of 1.5 Gy to develop a new germplasm H307 with high anther culture ability. The proportion of green plantlets per 100 anthers (GP/100A) of H307 was 14.50% which was higher than other H2 lines (P green plantlet regeneration ability of H307 remained high in all 3 years. Reciprocal crosses between H307 and Nongda3308 showed no significant differences in their values for calli per 100 anthers (CA/100A), green plantlets per 100 calli (GP/100C) and GP/100A (P > 0.05). Five main wheat varieties used in production, namely Yumai68, Yanzhan4110, Bainongaikang58, Zhoumai18 and Xinmai18, were selected to cross with the new H307. CA/100A, GP/100C and GP/100A were used to assess the anther culture ability of F1 hybrids, demonstrating that the anther culture ability of H307 was heritable. H307 possessed high anther culture ability that was heritable, which would be potential germplasm for improving wheat anther breeding ability. © 2014 Society of Chemical Industry.

In vitro micropropagation of orchid, Oncidium sp. (Dancing Dolls)

African Journals Online (AJOL)

Administrator

2007-05-16

May 16, 2007 ... international market. ... dipped quickly in 70% alcohol and flamed over a spirit lamp. ... Table 1. Effect of different concentrations of plant growth regulators on in vitro development of plantlets from immature seeds of Oncidium ...

Effect of benzylaminopurine on in vivo multiplication of French plantain

African Journals Online (AJOL)

SARAH

2014-02-28

Feb 28, 2014 ... of French plantain (Musa spp. AAB) cv. ... Figure 1: Plant and fruit appearance of French plantain cv. "Itoke Sege" .... Control of lethal browning of tissue culture plantlets of ... Research Workshop on Food Security,. Morogoro ...

In vitro propagation of cordyline plant using electromagnetic waves

International Nuclear Information System (INIS)

EL-Sharnouby, M.E.

2007-01-01

Shoot tips of cordyline terminals were cultured in vitro on different media (MS. NN and A) to select the best culture medium. Explants were subjected to different cytokines, microwaves periods, auxins and agriculture media. The results showed that MS was the best culture medium. The maximum number of shoots as well as per plant and number of leaves per plant were produced on MS medium supplemented with 4 mg/l 2ip. Exposing the plantlets to microwave irradiation for one minute caused the best proliferation rate, comparing control plantlets. The addition of auxin IBA at concentrate 2 mg/I to culture medium gave a significant increasing in number of roots and root length. Acclimatization culture medium containing peatmoss, sand and loam at equal volume parts gave a significant increasing in plant growth and greening than other acclimatization treatments

Pollen-gamma irradiation on pineapple and pollination using irradiated pollen

International Nuclear Information System (INIS)

Benega, R.; Cisneros, A.; Martinez, J.; Arias, E.; Yabor, L.; Isidron, M.; Castillo, E.; Fernandez, J.

1997-01-01

In order to try to induce haploid plants, pineapple (Ananas comosus (L.) ;err.) pollen grains were gamma-rays irradiated with a Co 60 source at rates of doses from 0 to 300 Gy. The effect of gamma-rays on the generative-nucleus division and vitro pollen viability as well as, seed contents and plantlets obtained after pollinations was analysed. The were reducing in the division frequencies of generative nucleus at all assayed doses. The LD50 was achieved among 200 and 250 Gy. There were not differences on pollen viability in the rates of assayed doses. Seed contents were dose-dependent. The percentages of seed full and seed containing only embryos decreases at increasing of irradiation doses. Contrary effect was observed on the seed empty. Some plantlets regenerated from the different irradiation doses showed phenotypes with small and leaves and short inter nodes

Micropropagation of six Paulownia genotypes through tissue culture

Directory of Open Access Journals (Sweden)

Lydia Shtereva

2014-12-01

Full Text Available We investigated the effect of genotype and culture medium on the in vitro germination and development of plantlets from seeds of 6 different Paulownia genotypes (P. tomentosa, hybrid lines P. tomentosa P. fortunei (Mega, Ganter and Caroline, P. elongata and hybrid line P. elongata P. fortunei. Nodal and shoot tip explants were used for micropropagation of Paulownia genotypes by manipulating plant growth regulators. The highest germination percentage for all genotypes was obtained for seeds inoculated on medium supplemented with 50 mg*L GA3 (MSG2. On Thidiazuron containing media, the explants of hybrid line P. elongata P. fortunei exhibited the highest frequency of axillary shoot proliferation following by P. tomentosa P. fortunei. The results are discussed with the perspective of applying an improved protocol for in vitro seed germination and plantlet formation in several economically valuable Paulownia genotypes.

Microsatellite loci and peroxidase alleles correlation in somaclonal ...

African Journals Online (AJOL)

A dimer locus, a tetramer locus and two epigenetic bands were observed. Genome variation among somaclonal plantlets were investigated using microsatellite markers. SSR (Simple Sequence Repeat) markers revealed polymorphism among the studied population. Nonparametric statistical analysis showed significant ...

African Journal of Biotechnology - Vol 8, No 9 (2009)

African Journals Online (AJOL)

High frequency callus induction and plantlet regeneration from different explants of Picrorhiza kurroa - a medicinal herb of Himalayas · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. H Sood, RS Chauhan ...

(Tongkat Ali) for callus

African Journals Online (AJOL)

hope&shola

2010-12-06

Dec 6, 2010 ... The induction of callus cultures using leaf, petiole, rachis, stem, tap root, fibrous root, cotyledon and ... other countries of the South-East Asia region and widely distributed ..... Indian ginseng plantlets from stem callus. Plant Cell ...

PRODUCTION OF THE NEW ANTIMALARIAL DRUG ARTEMISININ IN SHOOT CULTURES OF ARTEMISIA-ANNUA L

NARCIS (Netherlands)

WOERDENBAG, HJ; LUERS, JFJ; VANUDEN, W; PRAS, N; MALINGRE, TM; ALFERMANN, AW

From aseptically grown Artemisia annua plantlets, shoot cultures were initiated. Using different concentrations of auxine, cytokinine and sucrose, a suitable culture medium was developed, with respect to the growth of the shoots and their artemisinin accumulation. Nitrate concentration and

Ac-rolC transgenic p

African Journals Online (AJOL)

mfladung

urea derivative (Yip and Yang, 1986) are described as prerequisite for future transformation experiments. Evi- dence is provided here for the first ..... as a growth regulator offers two options: either high effi- ciency of plantlet regeneration for 40 ...

Influence of different cytokinins used in in vitro culture on the stoma ...

African Journals Online (AJOL)

GREGO

2007-03-05

Mar 5, 2007 ... ture result in plantlets of abnormal growth and develop- ment. After ex ... The anatomical changes seen in in ... and North Africa, the Middle East, China and California. ... export product, pistachio nut is a very important fruit for.

In vitro multiplication of banana (Musa sp.) cv. Grand Naine

African Journals Online (AJOL)

SAM

2014-07-02

Jul 2, 2014 ... with IAA 2.00 mg/l resulted in maximum establishment of cultures in lesser time. MS medium + BAP. 4.00 mg/l + ... is, Musa acuminata (Malaysia) and Musa balbsiana. (India) (Georget et al., 2000). Banana plantlets produced.

Journal of Biosciences | Indian Academy of Sciences

Indian Academy of Sciences (India)

However, expression of megatransgenes impacted the growth of the transplastomic plantlets. By combining two exogenous pathways, artemisinic acid was produced in transplastomic plants, which can be improved further using better metabolic engineering strategies for commercially viable yield of desirable isoprenoid ...

Establishment of efficient in vitro culture protocol for wheat land ...

African Journals Online (AJOL)

ADNAN

2012-02-07

Feb 7, 2012 ... etc. MATERIALS AND METHODS. The proposed research was carried out at the Center of ... contained in 25 ml capacity Pyrex test tubes (18 × 150 mm). The cultures were ... After 15 to 20 days of hardening, the plantlets were ...

Screening of endophytic fungi that promote the growth of Euphorbia ...

African Journals Online (AJOL)

STORAGESEVER

2008-10-06

Oct 6, 2008 ... host plantlets. Indole acetic acid (IAA) and gibberellin (GA) were found in the fermented broth of E4 and. E5. ... plant has been hampered by its slow maturation of this perennial ..... µmol L-1 abscisic acid. .... grown from seeds.

Evaluación y selección de un protocolo para la regeneración in vitro de la variedad de tomate Unapal-Arreboles Evaluation and selection of a protocol for in vitro regeneration of tomato variety Unapal-Arreboles

Directory of Open Access Journals (Sweden)

Hernando Ramírez

2009-01-01

Full Text Available Se compararon tres medios de cultivo para seleccionar el apropiado para la regeneración in vitro de la variedad de tomate UNAPAL-Arreboles. Se evaluó la producción de callos/explante, brotes/explante y plántulas /explante. El análisis de varianza y la prueba de Duncan permitieron concluir que el medio M3 presentó los mejores promedios para las variables evaluadas.The main objective of this research was to carried out the comparison of three culture medium of major use for in vitro regeneration of tomato, to select the more appropriate medium for in vitro regeneration of tomato, variety UNAPAL-Arreboles. The analysis of variance and media comparison was made based on the callus, shoots and plantlets production. The highest efficiency of callus, shoots and plantlets production was obtained with the M3 media.

Sauvetage d'embryons chez les légumineuses alimentaires en général et dans le genre Phaseolus en particulier (synthèse bibliographique

Directory of Open Access Journals (Sweden)

Barikissou, E.

2012-01-01

Full Text Available A review of embryos rescue in food legumes in general and in the genus Phaseolus in particular. Genetic improvement of Phaseolus vulgaris L. by interspecific hybridization with Phaseolus coccineus L. and Phaseolus polyanthus Greenm., used as female parents, often gives rise to embryo abortion at globular developmental stage. In vitro culture of embryos at cotyledonary and torpedo shaped stages, leads to hybrid plants, but with very low percentages of success. Several investigations of in vitro culture in selfed genotypes of Phaseolus and from embryos at globular or young heart shaped stages have allowed to regenerate some young plantlets. However, problems of rooting and stopping of growth restrict the number of developing plantlets. Analysis of the results achieved from interspecific embryo rescue in others food legumes of the genus Lupinus, Cajanus, Cicer, Lens and Trifolium, helped to identify some solutions to resolve incompatibility problems in Phaseolus.

Somatic embryogenesis and plant regeneration of Capsicum baccatum L.

Directory of Open Access Journals (Sweden)

Peddaboina Venkataiah

2016-06-01

Full Text Available A plant regeneration protocol via somatic embryogenesis was achieved in cotyledon and leaf explants of Capsicum baccatum, when cultured on MS medium supplemented with various concentrations of 2,4-dichlorophenoxy acetic acid (2,4-D, 0.5–5.0 mg l−1 in combination with Kinetin (Kn, 0.5 mg l−1 and 3% sucrose. Various stages were observed during the development of somatic embryos, including globular, heart, and torpedo-stages. Torpedo stage embryos were separated from the explants and subcultured on medium supplemented with various concentrations of different plant growth regulators for maturation. Maximum percentage (55% of somatic embryo germination and plantlet formation was found at 1.0 mg l−1 BA. Finally, about 68% of plantlets were successfully established under field conditions. The regenerated plants were morphologically normal, fertile and able to set viable seeds.

In vitro regeneration of some Iranian alfalfa (Medicago sativa L. genotypes via somatic embryogenesis

Directory of Open Access Journals (Sweden)

Majid Shokrpour

2014-12-01

Full Text Available An effective in vitro regeneration system is one of the prerequisites for genetic manipulation of alfalfa (Medicago sativa L. varieties and genotypes. In this research, somatic embryogenesis of four alfalfa genotypes, 6-18 (synthetic, 4-14 (Kara Yonje- Karakozlu, 3-27 (Kara Yonje Maraghe and y-6 (Regen-SY, were investigated using leaf and petiole explants. Formation of callus and somatic embryogenesis was significantly influenced by the explant type and interaction of genotype and culture medium. Petiole explants of genotype 4-14 produced the highest yield of callus (0.406 gr fresh weight of callus. Percentage of somatic embryogenesis and the number of embryos per callus in petiole explants of genotype 4-14 was higher than those of other genotypes and explants. In genotype 6-18, the highest percentage of somatic embryogenesis was achieved on MS medium containing 5 mg/L 2,4-D and 2 mg/L kinetin. There was no significant differences between genotypes and explants in terms of embryo conversion to plantlet, and on average, 58% of somatic embryos converted to plantlet on MS medium. The petiole explants of genotype 6-18 did not exhibit somatic embryogenesis response in medium containing low ratio of 2,4-D:Kinetin (5 mg/L 2,4-D and 2 mg/L kinetin. While, these explants showed somatic embryogenesis in higher ratio of 2,4-D:Kinetin (5:1. The plantlet conversion efficiency of somatic embryos produced through this study was relatively higher and therefore, the method presented in this study could be used in alfalfa genetic manipulation and molecular studies.

In vitro co-cultures of Pinus pinaster with Bursaphelenchus xylophilus: a biotechnological approach to study pine wilt disease.

Science.gov (United States)

Faria, Jorge M S; Sena, Inês; Vieira da Silva, Inês; Ribeiro, Bruno; Barbosa, Pedro; Ascensão, Lia; Bennett, Richard N; Mota, Manuel; Figueiredo, A Cristina

2015-06-01

Co-cultures of Pinus pinaster with Bursaphelenchus xylophilus were established as a biotechnological tool to evaluate the effect of nematotoxics addition in a host/parasite culture system. The pinewood nematode (PWN), Bursaphelenchus xylophilus, the causal agent of pine wilt disease (PWD), was detected for the first time in Europe in 1999 spreading throughout the pine forests in Portugal and recently in Spain. Plant in vitro cultures may be a useful experimental system to investigate the plant/nematode relationships in loco, thus avoiding the difficulties of field assays. In this study, Pinus pinaster in vitro cultures were established and compared to in vivo 1 year-old plantlets by analyzing shoot structure and volatiles production. In vitro co-cultures were established with the PWN and the effect of the phytoparasite on in vitro shoot structure, water content and volatiles production was evaluated. In vitro shoots showed similar structure and volatiles production to in vivo maritime pine plantlets. The first macroscopic symptoms of PWD were observed about 4 weeks after in vitro co-culture establishment. Nematode population in the culture medium increased and PWNs were detected in gaps of the callus tissue and in cavities developed from the degradation of cambial cells. In terms of volatiles main components, plantlets, P. pinaster cultures, and P. pinaster with B. xylophilus co-cultures were all β- and α-pinene rich. Co-cultures may be an easy-to-handle biotechnological approach to study this pathology, envisioning the understanding of and finding ways to restrain this highly devastating nematode.

Induksi Embriogenesis Somatik dari Jaringan Endosperma Jeruk Siam (Citrus nobilis Lour. cv Simadu

Directory of Open Access Journals (Sweden)

Mia Kosmiatin

2014-07-01

Full Text Available ABSTRACTTriploid plants can be obtained from endosperm tissues through somatic embryogenesis regeneration. This research aimed to obtain somatic embryogenesis regeneration technique of tangerine endosperm. There were 3 experiments conducted in this research: 1 Embryogenic callus induction of tangerine endosperm. Endosperms isolated from fruits that were harvested from mother plants 11-13 weeks after anthesis and cultured on Murashige and Skoog (MS medium by modified vitamin Morel and Wetmore (MW which treated by 0.1 mg L-1 biotin, 500 mg L-1 malt extract (ME, 500 mg L-1 Casein hydrolisate (CH, 500 mg L-1 ME + 0.1 mg L-1 biotin, and 500 mg L-1CH + 0.1 mg L-1 biotin, 2 Maturation and germination of somatic embryos conducted by embryogenic callus cultured on MS medium by vitamin MW modified with addition of ABA, glutamine, and biotin, and 3 Plantlet elongation conducted on MS medium modified by MW vitamin with addition of GA3 and Kinetin. The best induction medium for embryogenic callus was modified MS enriched with 3 mg L-1 BA and 500 L-1 CH or ME, in succession 84.0 and 80.0%. The best medium for somatic embryos maturation with normal morphological plantlets (54.8% was modified MS medium without plant growth regulator with higher rate of solidified agent (from 2.5 to 3 g L-1 Phytagel. Plantlets elongation was highly (0.9 cm occurred on modified MS with enriched of 2.5 mg L-1 GA3. Keywords: Citrus nobilis (Lour., endosperm culture, in vitro, Simadu tangerine

Assessment of genetic stability in micropropagules of Jatropha curcas genotypes by RAPD and AFLP analysis

KAUST Repository

Sharma, Sweta K.

2011-07-01

Jatropha curcas (Euphorbiaceae), a drought resistant non edible oil yielding plant, has acquired significant importance as an alternative renewable energy source. Low and inconsistent yields found in field plantations prompted for identification of high yielding clones and their large scale multiplication by vegetative propagation to obtain true to type plants. In the current investigation plantlets of J. curcas generated by axillary bud proliferation (micropropagation) using nodal segments obtained from selected high yielding genotypes were assessed for their genetic stability using Randomly Amplified Polymorphic DNA (RAPD) and Amplified Fragment Length Polymorphism (AFLP) analyses. For RAPD analysis, 21 out of 52 arbitrary decamer primers screened gave clear reproducible bands. In the micropropagated plantlets obtained from the 2nd sub-culture, 4 out of a total of 177 bands scored were polymorphic, but in the 8th and 16th sub-cultures (culture cycle) no polymorphisms were detected. AFLP analysis revealed 0.63%, 0% and 0% polymorphism in the 2nd, 8th and 16th generations, respectively. When different genotypes, viz. IC 56557 16, IC 56557 34 and IC 56557 13, were assessed by AFLP, 0%, 0.31% and 0.47% polymorphisms were found, respectively, indicating a difference in genetic stability among the different genotypes. To the best of our knowledge this is the first report on assessment of genetic stability of micropropagated plantlets in J. curcas and suggests that axillary shoot proliferation can safely be used as an efficient micropropagation method for mass propagation of J. curcas. © 2011 Elsevier B.V.

Effect Of Gamma Rays And Growth Regulators On Explants Excised From In Vitro Shoots And Greenhouse Seedlings, Of Pepper (Capsicum Annum L.)

International Nuclear Information System (INIS)

Maarouf, A. A.; Kassem, M.

2004-01-01

This experiment was conducted on pepper (Capsicum annum L.) to compare the ability of the in vitro explants with those of greenhouse grown seedlings on shoot proliferation and callus formation and their ability to form plantlets and the effect of gamma irradiation and growth regulators on the shoot tip, hypocotyls and leaf tissue was used as laboratory explants, leaf tissue nodes and internodes were taken from greenhouse seedlings. 6- benzyla-minopurine (BAP) in different concentrations was combined with Indoleacertic acid (IAA) to know their effect on shoot proliferation, 2,4 - Dichlorophenoxy acetic acid (2,4- D) was used for callus formation, and use stimulation effect of gamma irradiation, potassium nitrat (KNO 3 ), Thidaiazurom (TDZ) and casine hydrolysate (CH) for plantlet formation. The results showed that the highest percentage of callus was obtained by in vitro hypocotyls and greenhouse grown nodes followed by in vitro leaf tissue thereafter greenhouse leaf tissue. The shoot tips were the lowest efficient explants in producing callus in both in vitro and greenhouse ones. The highest percentage of shooting resulted from shoot tip, hypocotyls and leaf tissue of in vitro explants, followed by shoot tip, nodes and internodes of greenhouse grown explants and the lowest percentage was recorded by leaf tissue. Highest percentage of shoot number was obtained form greenhouse grown shoot tip followed by in vitro shoot tip, hypocotyls and leaf tissue of greenhouse grown seedlings the internodes were the lowest efficient in producing shoots. The highest success in plantlet formation was caused by TDZ followed by gamma irradiation and the other treatments were equaled. (Authors)

Use in plant breeding of acute, chronic or fractionated doses of X-rays or fast neutrons as illustrated with leaves of Saintpaulia

NARCIS (Netherlands)

Broertjes, C.

1971-01-01

The parameters used were survival of irradiated leaves of S. ionantha , production of adventitious plantlets at the base of the petiole and mutation frequency.

The differences between unfiltered X-rays from a 250/25 deep therapy apparatus and fast neutrons

Micropropagation of Helianthemum lippii L. var Sessiliforuim ...

African Journals Online (AJOL)

HAMZA

2013-11-13

Nov 13, 2013 ... or the compress of the aerial part is used to treat coetaneous lesion. ... plastic pots (150 ml) containing a mixture of peat and perlite (2:1, v/v). Potted plantlets were ... wrap and the shoots begin to be less vigorous. However,.

Efficient in vitro regeneration protocol of Centella asiatica (L.) Urban ...

African Journals Online (AJOL)

The present communication reports an efficient in vitro plantlet regeneration protocol for endemic umbellifer Centella asiatica (L.) urban via callus mediated organogenesis from leaf and stem explants. The plant is pharmacologically very important and its consumption as underutilized green leafy vegetable affluent in ...

Shoot organogenesis in oleaster (Elaeagnus angustifolia L.)

African Journals Online (AJOL)

STORAGESEVER

2009-02-04

Feb 4, 2009 ... Regenerated plantlets were acclimatized and successfully transplanted to soil. Key words: shoot organogenesis, callus, ... saline and alkaline soils (Economou and Maloupa, 1995). Its fruits have been used as a .... cally inert compounds as reported by Kaminek (1992). Direct shoot organogenesis without ...

Profound improvements of isolated microspores culture techniques ...

African Journals Online (AJOL)

We studied the effects of sampling stages, physical conditions (like temperature), culture conditions, embryo long-distance transportation methodology and plantlet regeneration on isolated microspores from donor plants in field. Results indicated that if microspores were sampled in bud stage instead of blooming stage to ...

Genetic stability among date palm plantlets regenerated from petiole ...

African Journals Online (AJOL)

The regeneration capacity between the shoot tip and petiole explants that excised from two date palm (Phoenix dactylifera L.) cutivar namely Unknown and Ferhi was compared. It was noted that the shoot tip explants started to swill after several subculture when placed on Murashige and Skoog (MS) medium supplemented ...

Enhancement of Diosgenin Production in Plantlet and Cell Cultures ...

African Journals Online (AJOL)

HSCCC),. Sephadex LH-20 chromatography and preparative .... phase of acetonitrile-water (90:10, v/v) at a flow rate of 1 mL/min at 30 ºC, and an LC ..... Spirobisnaphthalenes from fungi and their biological activities. Mini-Rev Med Chem 2010; 10: ...

Identification of phytochemical components of aloe plantlets by gas ...

African Journals Online (AJOL)

MRT Pack 20 DVDs

2013-12-04

Dec 4, 2013 ... mented with 0.5 mgl-1 benzyl adenine + 0.5 mgl-1 α-naphthalene acetic acids into ... 5973) coupled to H.P. gas chromatograph (model 6890) equipped .... effects of Aloe vera leaf gel extract on oxidative stress in rats treated.

7 CFR 319.59-1 - Definitions.

Science.gov (United States)

2010-01-01

..., DEPARTMENT OF AGRICULTURE FOREIGN QUARANTINE NOTICES Wheat Diseases § 319.59-1 Definitions. Administrator... disease caused by the fungus Tilletia indica (Mitra) Mundkur. Plant. Any plant (including any plant part) for or capable of propagation, including a tree, a tissue culture, a plantlet culture, pollen, a shrub...

African Journal of Biotechnology - Vol 10, No 40 (2011)

African Journals Online (AJOL)

Effect of yeast extract and chitosan on shoot proliferation, morphology and antioxidant activity of Curcuma mangga in vitro plantlets · EMAIL FREE FULL TEXT EMAIL FREE FULL ... Characteristics and community diversity of a wheat straw-colonizing microbial community · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT

Rice is the seed of the monocot plants Oryza sativa (Asian rice) or ...

African Journals Online (AJOL)

BIOTECH

2013-10-16

Oct 16, 2013 ... of culture. The regenerated plantlets were transferred to pots for acclimatization. About 80% of plants were survived in the greenhouse condition. Key words: Somatic embryogenesis, immature zygotic embryos, Indica rice, plant regeneration. INTRODUCTION. Rice (Oryza sativa L.) is one of the most ...

Iridoid and caffeoyl phenylethanoid glycosides of the endangered carnivorous plant Pinguicula lusitanica L. (Lentibulariaceae)

NARCIS (Netherlands)

Grevenstuk, T.; Hooft, van der J.J.J.; Vervoort, J.J.M.; Waard, de P.; Romano, A.

2009-01-01

This work reports for the first time the identification of the major compounds of Pinguicula lusitanica, an endangered carnivorous plant species, using minimal amounts of plant material. A methanol extract was prepared from in vitro cultured plantlets and analyzed by HPLC–SPE–NMR/HPLC–MS. Three

Cryopreservation of medicinal plants: role of melatonin

Science.gov (United States)

Many useful plant species found in Canada are of conservation concern. In vitro storage and cryopreservation techniques guarantees safety of these species and have potential applications which may result in sustainable agriculture. Shoot tips of in vitro-grown plantlets of American elm, St John’s Wo...

Arbuscular mycorrhizal fungi improve the growth of olive trees and ...

African Journals Online (AJOL)

Two native Algerian mycorrhizal fungi (Glomus mosseae and Glomus intraradices) were tested for their effect on the growth of micropropagated olive tree (Olea europaea L.). The effect of inoculation of plantlets with G. mosseae was also compared with chemical fertilization using osmocote. Specific molecular techniques ...

The development of an efficient cultivar-independent plant regeneration system from callus derived from both apical and non-apical root segments of garlic (Allium sativum L.)

NARCIS (Netherlands)

Zheng, S.J.; Henken, G.; Krens, F.A.; Kik, C.

2003-01-01

Callus induction and later plant regeneration were studied in four widely grown garlic (Allium sativum L.) cultivars from Europe. Root segments from in vitro plantlets were used as starting material. In addition to cultivar effects, the effects of auxin and cytokinin levels and the position of the

Shoot and root morphogenesis from Eucalyptus grandis x urophylla ...

African Journals Online (AJOL)

Eucalyptus grandis x urophylla plantlets were regenerated via indirect organogenesis. Histological assessment of their development focused on identifying the calli, the differentiation of shoots from the calli and the shoot-root junction from the nascent shoots. Vascular tissue formation within the callus preceded that of ...

Certain growth related attributes of bunchy top virus infected banana ...

African Journals Online (AJOL)

Effect of banana bunchy top virus (BBTV) on morpho-physiological characteristics of banana (Musa sp.) cv., Basrai plants was assessed. Healthy and BBTV infected samples of banana were collected from its open fields and micro-propagated aseptically. These plantlets were established in wire-house for three months.

Mutagenesis for ACMV resistance in a Ghanian cassava cultivar `Bosom nsia`

Energy Technology Data Exchange (ETDEWEB)

Ahiabu, R K; Lokko, Y; Danso, K; Klu, G Y.P. [Biotechnology and Nuclear Agricultural Research Inst., Ghana Atomic Energy Commission, Legon (Ghana)

1997-07-01

Breeding for resistance to the African Cassava Mosaic Virus (ACMV) disease in the Ghanian cassava cultivar `Bosom nsia` has been on-going for the past four years at the Biotechnology and Nuclear Agricultural Research Institute. Protocols for in vitro culture from shoot meristems and acclimation of plantlets were established. Radiosensitivity tests on the regenerated plantlets indicated LD{sub 50} of 40 Gy, and doses of 25, 30 and 35 Gy were suitable for mutagenesis. These doses were applied to in vivo and in vitro grown plants, and selection was carried out in three propagations. Four variants, selected under field conditions with high viral incidence, were analyzed for virus particles with three virus indexing techniques. Polymerase chain reaction (PCR) product analysis of DNA extracts from Nicotiana benthamiana test plants, inoculated with sap from the leaves of variants, confirmed the presence of virus particles in all variants. Inoculation and ELISA tests suggested ACMV tolerance in selected variants. (author). 24 refs, 3 figs, 1 tab.

Production of polyhydroxybutyrate in oil palm (Elaeis guineensis Jacq. mediated by microprojectile bombardment of PHB biosynthesis genes into embryogenic calli

Directory of Open Access Journals (Sweden)

Ahmad Parveez eGhulam Kadir

2015-08-01

Full Text Available Biodegradable plastics, mainly polyhydroxybutyrate (PHB, which are traditionally produced by bacterial cells, have been produced in the cells of more than 15 plant species. Since the production of biodegradable plastics and the synthesis of oil in plants share the same substrate, acetyl-coenzyme A (acetyl-CoA, producing PHB in oil bearing crops, such as oil palm, will be advantageous. In this study, three bacterial genes, bktB, phaB and phaC, which are required for the synthesis of PHB and selectable marker gene, bar, for herbicide Basta resistant, were transformed into embryogenic calli. A number of transformed embryogenic lines resistant to herbicide Basta were obtained and were later regenerated to produce few hundred plantlets. Molecular analyses, including PCR, Southern blot and real-time PCR have demonstrated stable integration and expression of the transgenes in the oil palm genome. HPLC and Nile Blue A staining analyses confirmed the synthesis of PHB in some of the plantlets.

The study of genotype, cold pretreatment, low-dose gamma irradiation and 2,4-D concentration effect on wheat doubled haploid production

International Nuclear Information System (INIS)

Naserian, B.; Vedadi, C.; Karbalaii, S.

2005-01-01

Full text: In this study, response of a cultivar (Atrak) and two lines (F3 2005 and F3 2104) of wheat , and effect of cold pretreatment, low dose gamma irradiation and 2,4-D (2,4 mg l -1 ) were investigated to anther culture response. Donar plants were grown under field conditions in early spring. Anthers were plated on modified CHB medium containing 2, 4-D (2, 4 mg l -1 ), 0.5 mg l -1 Kinetin and 90 g l -1 Sucrose. Percent of Callus formation in 100 anther and percent of plantlet in 100 calli were measured. Results indicated that genotype, cold pretreatment and 2,4-D concentrations had significant effect on anther culture response. F3 2005 showed highest callus induction and plantlet production and F3 2104 had lowest response. This experiment indicated that androgenic traits are controlled by genotype and environmental factors. Further more these traits are controlled independently. (author)

Efficient micropropagation and chlorocholine chloride induced stevioside production of Stevia rebaudiana Bertoni.

Science.gov (United States)

Dey, Avishek; Kundu, Sayanti; Bandyopadhyay, Abhijit; Bhattacharjee, Aloke

2013-01-01

A promising method of micropropagation of Stevia rebaudiana Bertoni has been developed with an aim to increase the biomass, survivability of the plantlets and stevioside production, using chlorocholine chloride (CCC). Microshoots transferred to the MS medium containing different combinations CCC and IBA were found to be most effective in terms of growth pattern, hardening ability of the plantlets and stevioside content, compared to MS medium containing either IBA or CCC. Among other combinations tested, MS medium supplemented with 3 mg/l CCC and 3 mg/l IBA was found most effective in inducing significant changes like reduced shoot length, increased number of roots, higher leaf size, increased biomass and chlorophyll retaining capacity, higher survival percentage and most importantly the elevated stevioside content. Collectively, the major observations of this research indicate that application of CCC in micropropagation of S. rebaudiana Bertoni is a promising approach and has commercial prospects. Copyright © 2012 Académie des sciences. Published by Elsevier SAS. All rights reserved.

Evaluation of genetic variability in potato cv. 'Parda-Pastusa' obtained through the physical mutagenic agents

International Nuclear Information System (INIS)

Angarita-Zerda, A.; Mosquera, T.; Nustez, C.

1997-01-01

'Parda-Pastusa' is probably the most important variety of potato in Colombia. Its susceptibility to frost and Phytophthora infestans can cause important losses in the crop. In order to induce genetic variability to select clones resistant to frost and late blight, disease-free micropropagated plantlets, obtained from meristem culture of indexed tubers were irradiated with gamma rays from a 60 Co source. Virus-free mericlones were initially irradiated with 0,25, 50, 75 and 100 Gy or with 0, 10, 20 and 30 Gy. The optimal radiation dose was found to be 20 Gy. A mass propagation was carried out, and plantlets were irradiated with the optimal radiation dose. Clones derived from irradiated material were propagated for selection under simulated frost conditions (-7 deg. C for 360 minutes) and co-cultured with extracts of fungus, Phytophthora infestans. The selected clones will be transferred to field conditions for evaluation of agronomic and genetic characteristics. (author). 1 ref

Certain growth related attributes of micropropagated banana under different salinity levels

International Nuclear Information System (INIS)

Haq, I.U.; Soomro, F.

2011-01-01

The effect of salinity (NaCl) was assessed on banana (Musa spp.) cv., Sindhri Banana (Basrai) propagating plantlets in aseptic condition. Four different NaCl levels [0 (control) 50, 100 and 150 mM] were maintained at shoot multiplication stage for 6-weeks. Salinity reduced the number of plantlets per explants and plant biomass significantly. A proportional relationship was observed for Na/sup +/ and Cl/sub -/ but K/sup +/, Ca/sup 2+/and NO/sub 3/ were observed to be inversely proportioned with NaCl stress. Similarly, total proteins as well as carbohydrate contents were decreased significantly. Increasing mode of secondary metabolites (proline, betaine contents and reducing sugars) were showing a negative relationship of saline stress with plant micro-propagation efficiency. Among photosynthetic pigments, total carotenoids were increased while chlorophyll contents (Chl a and b) decreased. Similarly, nitrate reductase activity also reduced. Overall, vegetative propagation of banana was affected significantly by NaCl stress under in-vitro conditions. (author)

Effect of low dose {gamma} radiation on the dormancy breaking and physiological activity of 'Dejima' seed potato

Energy Technology Data Exchange (ETDEWEB)

Kim, J. S.; Kim, D. H.; Baek, M. H.; Lee, Y. K. [KAERI, Taejon (Korea, Republic of); Lee, Y. B. [Chungnam National Univ., Taejon (Korea, Republic of)

2001-05-01

To observe the stimulating effects of low dose {gamma}-radiation on the dormancy breaking and physiological activity, potato (Solanum tuberosum L. cv. Dejima) were irradiated at the dose of 0.5 {approx} 30 Gy. Low dose {gamma}-radiation had promoting effects on the srouting rate the optimum dose of 1 Gy and 2 Gy and on the sprout length at the optimum dose of 2 Gy and 4 Gy. In case of number of sprouts, there were increased in all of the low dose irradiation groups and period storages, which showed very significant promoting effects at 4,8,16 Gy of 15 DAP. The field growth of 45 DAP were increased at all of the low dose irradiation group, which particularly were very significant promoting effect at 4 Gy irradiation group. Irradiation didn't have significant effects on the antioxidant enzyme activity of potato plantlet, but the peroxidase activity of 45 DAP plantlet increased at 4 Gy irradiation group.

Evaluation of genetic variability in potato cv. `Parda-Pastusa` obtained through the physical mutagenic agents

Energy Technology Data Exchange (ETDEWEB)

Angarita-Zerda, A; Mosquera, T; Nustez, C [Tissue Culture Lab., Agronomy School, National Univ. of Colombia, Santafe de Bogota (Colombia)

1997-07-01

`Parda-Pastusa` is probably the most important variety of potato in Colombia. Its susceptibility to frost and Phytophthora infestans can cause important losses in the crop. In order to induce genetic variability to select clones resistant to frost and late blight, disease-free micropropagated plantlets, obtained from meristem culture of indexed tubers were irradiated with gamma rays from a {sup 60}Co source. Virus-free mericlones were initially irradiated with 0,25, 50, 75 and 100 Gy or with 0, 10, 20 and 30 Gy. The optimal radiation dose was found to be 20 Gy. A mass propagation was carried out, and plantlets were irradiated with the optimal radiation dose. Clones derived from irradiated material were propagated for selection under simulated frost conditions (-7 deg. C for 360 minutes) and co-cultured with extracts of fungus, Phytophthora infestans. The selected clones will be transferred to field conditions for evaluation of agronomic and genetic characteristics. (author). 1 ref.

Production of polyhydroxybutyrate in oil palm (Elaeis guineensis Jacq.) mediated by microprojectile bombardment of PHB biosynthesis genes into embryogenic calli

Science.gov (United States)

Parveez, Ghulam Kadir Ahmad; Bahariah, Bohari; Ayub, Nor Hanin; Masani, Mat Yunus Abdul; Rasid, Omar Abdul; Tarmizi, Ahmad Hashim; Ishak, Zamzuri

2015-01-01

Biodegradable plastics, mainly polyhydroxybutyrate (PHB), which are traditionally produced by bacterial cells, have been produced in the cells of more than 15 plant species. Since the production of biodegradable plastics and the synthesis of oil in plants share the same substrate, acetyl-coenzyme A (acetyl-CoA), producing PHB in oil bearing crops, such as oil palm, will be advantageous. In this study, three bacterial genes, bktB, phaB, and phaC, which are required for the synthesis of PHB and selectable marker gene, bar, for herbicide Basta resistant, were transformed into embryogenic calli. A number of transformed embryogenic lines resistant to herbicide Basta were obtained and were later regenerated to produce few hundred plantlets. Molecular analyses, including polymerase chain reaction (PCR), Southern blot, and real-time PCR have demonstrated stable integration and expression of the transgenes in the oil palm genome. HPLC and Nile blue A staining analyses confirmed the synthesis of PHB in some of the plantlets. PMID:26322053

Mutagenesis for ACMV resistance in a Ghanian cassava cultivar 'Bosom nsia'

International Nuclear Information System (INIS)

Ahiabu, R.K.; Lokko, Y.; Danso, K.; Klu, G.Y.P.

1997-01-01

Breeding for resistance to the African Cassava Mosaic Virus (ACMV) disease in the Ghanian cassava cultivar 'Bosom nsia' has been on-going for the past four years at the Biotechnology and Nuclear Agricultural Research Institute. Protocols for in vitro culture from shoot meristems and acclimation of plantlets were established. Radiosensitivity tests on the regenerated plantlets indicated LD 50 of 40 Gy, and doses of 25, 30 and 35 Gy were suitable for mutagenesis. These doses were applied to in vivo and in vitro grown plants, and selection was carried out in three propagations. Four variants, selected under field conditions with high viral incidence, were analyzed for virus particles with three virus indexing techniques. Polymerase chain reaction (PCR) product analysis of DNA extracts from Nicotiana benthamiana test plants, inoculated with sap from the leaves of variants, confirmed the presence of virus particles in all variants. Inoculation and ELISA tests suggested ACMV tolerance in selected variants. (author). 24 refs, 3 figs, 1 tab

A comparative study of Curcuma zedoaria and Zingiber zerumbet ...

African Journals Online (AJOL)

Curcuma zedoaria and Zingiber zerumbet plantlets could be micropropagated via induction of multiple shoots from in vitro shoot explants using different culture systems such as the agar-gelled medium cultures, shake flask system and temporary immersion system (TIS). The immersion period in TIS did not influence shoot ...

Studies on seed germination and in vitro shoot multiplication of ...

African Journals Online (AJOL)

Yomi

2011-12-21

Dec 21, 2011 ... vitro seed germination and plantlet regeneration of this plant. ... medical books, a few varieties of herbs are described ... Seed viability was determined by tetrazolium (TZ) test. (Perry ..... Dictionary of Drugs and Difficult Terms of 'The Book of ... establishment of stem cuttings of oregano, pepermint and balm.

78 FR 24633 - Restructuring of Regulations on the Importation of Plants for Planting

Science.gov (United States)

2013-04-25

...) capable of propagation, including a tree, a tissue culture, a plantlet culture, pollen, a shrub, a vine, a... sterile cultures of orchid plants) from any country or locality except Canada may be imported into the... including Africa, Australia, Brazil, Bulgaria, Japan and adjacent islands, Korea, New Zealand, Oceania, the...

Development of technique on the induction and selection of in vitro mutant lines(Potato, Solanum tuberosum L.)

International Nuclear Information System (INIS)

Hong, Joo Bong; Lee, Young Il; Song, Hee Sup; Kim, Jae Sung; Byun, Myung Woo; Lee, Young Keun; Shin, In Chul; Lee, Sang Jae; Lee, Ki Woon; Lim, Yong Taek

1992-08-01

The radiosensitivity and salt resistance on the single cell and callus of potato, mass production method of plantlet and microtuber of potato by in vitro culture and microtuber formation from the stem irradiated with radiation were investigated to obtain a optimum condition for selection of mutant cell line. (Author)

In vitro storage of synthetic seeds: Effect of different storage ...

African Journals Online (AJOL)

GREGORY

2010-08-30

Aug 30, 2010 ... to grow on poor soils even in arid conditions, many countries are interested in ... efficient delivery of cloned plantlets and also to meet the interna-tional ... DHL Courier Service, under ambient temperature conditions within four days to ... by using analysis of variance (ANOVA) technique and differences.

Micropropagation of Dioscorea alata L. through nodal segments

African Journals Online (AJOL)

SUPRIYA

2013-11-20

Nov 20, 2013 ... Regenerated plants were transplanted in hardening medium containing Brick bats + Charcoal. + Dried moss + Leaf molds + Soil in 1: 1: 1:1:1 proportion. .... Then the plantlets were transferred to sterile small plastic cups containing vermiculite. (a sterile inert medium for planting transferred plants) and kept.

Phyto-agglutinin, total proteins and amino assimilating enzymatic ...

African Journals Online (AJOL)

Biochemical studies were carried out with in vitro micropropagated plantlets of two indigenous cultivars of chickpea (Cicer arietinum L.), KK-1 and Hassan-2K, where extract from shoots, leaves, roots, and reproductive organs were determined for human erythrocyte agglutination (by mixing the lectin containing extract 1:1 ...

concentrations, two light regimes of in vitro rooting and ...

African Journals Online (AJOL)

PRECIOUS

2009-11-16

Nov 16, 2009 ... Mahmood AW (1998). Viability of Planting Teak and Sentang in. Malaysia. Forest Research Institute Malaysia. pp. 23-27. Lee SK, Rao AN (1981). In vitro plantlets development in tropical trees-. Calophyllum inophyllum and Eugenia grandis. In. Tissue culture of economically important plants. Rao AN (ed.).

Genotype dependent callus induction and shoot regeneration in ...

African Journals Online (AJOL)

SERVER

2007-07-04

Jul 4, 2007 ... tems have been used for improvement of sunflower, but these techniques are mainly limited by the tissue culture response of commercial varieties (Nestares et al., 2002). In sunflower, reports on shoot regeneration from hypocotyls, cotyledons, leaves and meristematic tissues of young plantlets are available ...

The bean rhizosphere Pseudomonas aeruginosa strain RZ9 strongly reduces Fusarium culmorum growth and infectiveness of plant roots

Energy Technology Data Exchange (ETDEWEB)

Haddoudi, I.; Sendi, Y.; Batnini, M.; Romdhane, S.B.; Mhadhbi, H.; Mrabet, M.

2017-07-01

A faba bean rhizospheric Pseudomonas aeruginosa isolate RZ9 was used for studying its antifungal activity and protecting effects of faba bean and common bean against the root pathogen Fusarium culmorum strain MZB47. The dual culture tests showed that RZ9 inhibits MZB47 in vitro growth by 56%. When mixing RZ9 cell suspension with MZB47 macroconidia at equal proportion, the macroconidia viability was reduced with 70%. Pathogenicity tests conducted in sterile conditions showed that MZB47 caused an intense root rotting in faba bean ‘Aquadulce’ plantlets and a slight level in common bean ‘Coco blanc’. This was associated to significant decreases in plant growth only in ‘Aquadulce’, reducing shoot dry weight (DW) by 82% and root DW by 70%. In soil samples, MZB47 caused severe root rotting and induced significant decreases in shoot DW (up to 51%) and root DW (up to 60%) for both beans. It was associated to a decrease in nodule number by 73% and 52% for faba bean and common bean, respectively. Biocontrol assays revealed that the inoculation of RZ9 to MZB47-treated plantlets enhanced shoot DWs (25% and 110%) and root DWs (29% and 67%), in faba bean and common bean, respectively. Moreover, root rotting levels decreased and nodule number increased in treated compared to untreated plantlets. Collected data highlighted the disease severity of F. culmorum and demonstrated the potential of using RZ9 in controlling Fusaria root diseases in beans. Thereby, the current study represents the first report on the biocontrol effectiveness of P. aeruginosa against F. culmorum in beans.

In-vitro regeneration of sugarcane (saccharum officinarum L.)

International Nuclear Information System (INIS)

Azu, E.

2009-01-01

Sugarcane (Saccharum officinarum L.) cultivars Barbados 46 (B46), Natal Coimbatore 339 (NC0339), Ragna, local sugarcane cultivar (LSC) and Kenana (Kn) series sourced from Kyebi, Akorley, Subriso, Jejeti and the University of Ghana Agriculture Research Station, Kpong, were evaluated for sterilisation, multiplication and rooting. An efficient double sterilisation protocol was achieved by immersing axillary buds pre-treated with Goldazim (active ingredient carbendazim) in 0.2% mercuric chloride for 7 minutes followed by 0.1 % mercuric chloride for 3 minutes. At this optimal sterilisation regime, 70% of the cultured buds were decontaminated. However, post sterilisation survival required the incorporation of 0.002g/L and 0.001g/L of amphotericin Band cefotaxime respectively in the culture medium indicating that the contaminants were endophytic. Evaluation for multiple shoot induction was conducted using Murashige and Skoog (1962) basal salts (MS) medium supplemented with 25g/L sucrose, 2mg/L IAA, 2mg/L GA 3 , 3g/L activated charcoal and varying concentrations (mg/L) of BAP or kinetin. The response of the cultivars to shoot induction varied with Ragna producing the highest number of shoots (2.17) on 5mg/L BAP indicating genotypic differences. Naphthalene acetic acid (NAA) at a concentration of 5mg/L induced the highest frequency (65%) of roots in all the cultivars tested. Post-flask acclimatization and survival of plantlets was high and independent of the age at which regenerated plantlets were transferred to the plant barn for weaning. This procedure could therefore be useful for regenerating sugarcane plantlets as well as provide target tissues for genetic transformation studies (au).

Transcriptome analysis of Pseudostellaria heterophylla in response to the infection of pathogenic Fusarium oxysporum.

Science.gov (United States)

Qin, Xianjin; Wu, Hongmiao; Chen, Jun; Wu, Linkun; Lin, Sheng; Khan, Muhammad Umar; Boorboori, Mohammad Reza; Lin, Wenxiong

2017-09-18

Pseudostellaria heterophylla (P. heterophylla), a herbaceous perennial, belongs to Caryophyllaceae family and is one of the Chinese herbal medicine with high pharmacodynamic value. It can be used to treat the spleen deficiency, anorexia, weakness after illness and spontaneous perspiration symptoms. Our previous study found that consecutive monoculture of Pseudostellaria heterophylla could lead to the deterioration of the rhizosphere microenvironment. The specialized forms of pathogenic fungus Fusarium oxysporum f.Sp. heterophylla (F. oxysporum) in rhizosphere soils of P. heterophylla plays an important role in the consecutive monoculture of P. heterophylla. In this study, F. oxysporum was used to infect the tissue culture plantlets of P. heterophylla to study the responding process at three different infection stages by using RNA-sequencing. We obtained 127,725 transcripts and 47,655 distinct unigenes by de novo assembly and obtained annotated information in details for 25,882 unigenes. The Kyoto Encyclopedia of Genes and Genomes pathway analysis and the real-time quantitative PCR results suggest that the calcium signal system and WRKY transcription factor in the plant-pathogen interaction pathway may play an important role in the response process, and all of the WRKY transcription factor genes were divided into three different types. Moreover, we also found that the stimulation of F. oxysporum may result in the accumulation of some phenolics in the plantlets and the programmed cell death of the plantlets. This study has partly revealed the possible molecular mechanism of the population explosion of F. oxysporum in rhizosphere soils and signal response process, which can be helpful in unraveling the role of F. oxysporum in consecutive monoculture problems of P. heterophylla.

The bean rhizosphere Pseudomonas aeruginosa strain RZ9 strongly reduces Fusarium culmorum growth and infectiveness of plant roots

Directory of Open Access Journals (Sweden)

Imen Haddoudi

2017-07-01

Full Text Available A faba bean rhizospheric Pseudomonas aeruginosa isolate RZ9 was used for studying its antifungal activity and protecting effects of faba bean and common bean against the root pathogen Fusarium culmorum strain MZB47. The dual culture tests showed that RZ9 inhibits MZB47 in vitro growth by 56%. When mixing RZ9 cell suspension with MZB47 macroconidia at equal proportion, the macroconidia viability was reduced with 70%. Pathogenicity tests conducted in sterile conditions showed that MZB47 caused an intense root rotting in faba bean ‘Aquadulce’ plantlets and a slight level in common bean ‘Coco blanc’. This was associated to significant decreases in plant growth only in ‘Aquadulce’, reducing shoot dry weight (DW by 82% and root DW by 70%. In soil samples, MZB47 caused severe root rotting and induced significant decreases in shoot DW (up to 51% and root DW (up to 60% for both beans. It was associated to a decrease in nodule number by 73% and 52% for faba bean and common bean, respectively. Biocontrol assays revealed that the inoculation of RZ9 to MZB47-treated plantlets enhanced shoot DWs (25% and 110% and root DWs (29% and 67%, in faba bean and common bean, respectively. Moreover, root rotting levels decreased and nodule number increased in treated compared to untreated plantlets. Collected data highlighted the disease severity of F. culmorum and demonstrated the potential of using RZ9 in controlling Fusaria root diseases in beans. Thereby, the current study represents the first report on the biocontrol effectiveness of P. aeruginosa against F. culmorum in beans.

7 CFR 301.89-1 - Definitions.

Science.gov (United States)

2010-01-01

... 7 Agriculture 5 2010-01-01 2010-01-01 false Definitions. 301.89-1 Section 301.89-1 Agriculture..., DEPARTMENT OF AGRICULTURE DOMESTIC QUARANTINE NOTICES Karnal Bunt § 301.89-1 Definitions. Actual price... part) for or capable of propagation, including a tree, a tissue culture, a plantlet culture, pollen, a...

Plant regeneration from leaf-derived callus in Plectranthus barbatus ...

African Journals Online (AJOL)

RAMA

2013-05-01

May 1, 2013 ... produces an ancient root-derived drug traditionally used in the treatment of ... has opened up vistas for bio medical research. The fasciculated ... stored mainly in the cytoplasmic vesicles of cork cells in ... After screening of suitable ..... vitro of embryos and plantlets in stem tissue cultures of Tylophora indica.

Micropropagation of Origanum sipyleum L., an endemic medicinal ...

African Journals Online (AJOL)

PRECIOUS

2009-11-02

Nov 2, 2009 ... the shoots rooted in a culture medium with 0.5 mg/L indolebutyric acid (IBA) after 3 weeks. The plantlets were acclimatized into ... of tissue cultures for propagation and multiplication of. Origanum sipyleum. There are reports on .... Geographic variation of Greek oregano (Origanum vulgare ssp. hirtum) ...

Environmentally safe in vitro regeneration protocol for Curcuma ...

African Journals Online (AJOL)

not contain HgCl2, a toxic pollutant for Curcuma amada, Curcuma longa, Zingiber barbatum and Kaempferia galanga. Plantlets were regenerated from the buds without any intervention of the callus phase. The contamination free survival of the bud explants from Curcuma, Zingiber and Kaempferia was more than 75, 57 and ...

Abscisic acid effects on water and photosynthetic characteristics of ...

African Journals Online (AJOL)

The aim of this study is to compare the water and photosynthetic characteristics of two xerophilic ecotypes of Atriplex halimus (L.). Seeds collected from two different sites Djelfa and Oran are germinated in controlled greenhouse. After 6 months, the plantlets were treated 21 days with increasing concentrations of abscisic ...

Optimization of somatic embryogenesis procedure for commercial ...

African Journals Online (AJOL)

Tuoyo Aghomotsegin

2016-09-07

Sep 7, 2016 ... scalpel into small pieces (~30 pieces/cotyledon; each square piece measured ... sets of permanent leaves and a well-developed root system. Plantlets from SSE ... dish as an experimental unit), and the mean of the total SSEs produced .... environmental/seasonal fluctuation over the course of the extended ...

Efficient regeneration of the endangered banana cultivar 'Virupakshi ...

African Journals Online (AJOL)

Plantlets of the banana cultivar 'Virupakshi' (AAB) were regenerated from somatic embryos derived from embryogenic cells of calli from immature male flower explants. Induction of calli from explants was favored by a relatively moderate concentration of 2,4-dichlorophenoxyacetic acid (2,4-D) (4 mg/L), high concentrations of ...

Leaf anatomy of genotypes of banana plant grown under coloured ...

African Journals Online (AJOL)

This study aimed to evaluate the effect of spectral light quality on different anatomical features of banana tree plantlets grown under coloured shade nets. Banana plants of five genotypes obtained from micropropagation, were grown under white, blue, red and black nets, with shade of 50%, in a completely randomized ...

Agrobacterium-mediated transformation of two Serbian potato cultivars

African Journals Online (AJOL)

STORAGESEVER

2010-07-26

Jul 26, 2010 ... al., 2004) or to modify common metabolic pathways, such as starch and sucrose ... known for their high and regular yields, universal cooking features and good ... Shoot cultures were established from sprouts and propagated in vitro by monthly ... Plantlets with well developed roots were multiplied and used ...

Effect of seasonal collection on callus Induction, proliferation and ...

African Journals Online (AJOL)

Hevea brasiliensis that is grown in Thailand and Southeast Asia is a very valuable source of natural rubber. However, few studies have been conducted to evaluate the effects of seasonal effect on plantlet regenera-tion through somatic embryogenesis from anther culture. Thus, this study discussed the effect of season on ...

In vitro regeneration of lychee (Litchi chinensis Sonn.)

African Journals Online (AJOL)

Administrator

In vitro plantlet regeneration in the main commercial variety of lychee (Litchi chinensis Sonn. cv Tai So) in Mauritius was achieved from callus cultures derived from young, tender leaf explants on Murashige and Skoog (MS) medium. Callogenesis was obtained in all media supplemented with auxin, but was most prominent ...

Laboratory and field studies of guayule modified to overexpress HMGR

Science.gov (United States)

We report the genetic modification of guayule to overexpress the isoprenoid pathway enzyme HMGR. The rubber content of two-month old in vitro transformed plantlets showed a 65% increase in rubber over the control for one line (HMGR6), and lower resin for another (HMGR2). In field evaluations HMGR6...

In vitro regeneration of a common medicinal plant, Ocimum sanctum ...

African Journals Online (AJOL)

Embracing micro-propagation method for large scale production of plantlets and also for protection of appropriate germplasm is a prerequisite that needs to be undertaken in order to develop a rapid in vitro regeneration protocol for Ocimum sanctum L. Shoot tips as well as nodal segments were subjected to numerous ...

Responses of Calathea species in different growing media ...

African Journals Online (AJOL)

... substrate of top soil and poultry manure mixtures. Although C.nigerica produced higher number of leaves and taller plants than C. zebrina, the latter may be preferred because of its more attractive leaves and its many plantlets that quickly fill the growing container. Key words: growth , container, media, Calathea Species.

Establishment of an efficient protocol for micropropagation of stem ...

African Journals Online (AJOL)

Microshoots thus formed, were cultured on indole 3-butyric acid (IBA) and half strength basal MS media for induction of roots. Regenerated plantlets with healthy shoots and roots were acclimatized in moist cotton followed by their hardening in soil : vermicompost potting mixture with 90% survival rate. Key words: Tylophora ...

Chickpea regeneration and genetic transformation

African Journals Online (AJOL)

STORAGESEVER

2010-02-08

Feb 8, 2010 ... Chick- pea is good as a source of carbohydrate (48.2 - 67.6%), protein (12.4 - 31.5%), fat (6%) and nutritionally important minerals. Among the legumes, chickpea is the best hypo- cholesteremic agent, followed by black gram and green gram. Direct shoot organogenesis and establishment of plantlets from ...

Influence of ventilation and media on potato (Solanum tuberosum L ...

African Journals Online (AJOL)

STORAGESEVER

2009-05-18

May 18, 2009 ... Considering the most suitable relationship between ventilation and media in this study, two mixtures of perlite and coir (3:1, light and 1:3, heavy v/v) were applied to determine the growth characteristics var. Sante of potato plantlets. As potato has high potential in minituber production, different levels of.

Isolation and sequencing of the cryIC-like delta endotoxin gene from ...

African Journals Online (AJOL)

use

2012-02-16

Feb 16, 2012 ... rooting from transfer of shoots on basal-MS medium. Potato plantlets adapted to ... amino acid residues and contained a potential repeat motif. The deduced ... containing MS media supplemented with 3 or 4% sucrose and MS + 1.5 mg/l .... investment in time before the expressed proteins can be analysed.

In vitro propagation of Alternanthera sessilis (sessile joyweed), a ...

African Journals Online (AJOL)

MS medium supplemented with 1 mg/l IAA and 1 mg/l BAP showed maximum shoot multiplication. For rooting, the optimal medium was half strength MS medium supplemented with 1 mg/l IBA. Rooted plantlets were then transferred to sunbag vessels which provided the high humidity environment. The hardened plants were ...

In vitro adventitious shoot regeneration and acclimatisation of ...

African Journals Online (AJOL)

Different potting media were assessed during plantlet acclimatization. The highest percentage of plant survival (83.33%) was on the medium that contained sand and soil (1:1), while maximum root length (4.37 cm) and plant height (7.87 cm) were attained in potting medium that consisted peat moss, perlite and vermiculite ...

Enhancing rooting consistency in Rosa damascena scions | Saffari ...

African Journals Online (AJOL)

The highest percentage of plantlet survival was observed in potting media mixture containing; sand, soil, organic materials and vermiculite. Several experiments were carried out to stimulate in vitro rooting of Damask rose. Application of different media such as MS, 1/2 MS and 1/4 MS with different concentrations of 3-indole ...

EFFECT OF PERMEABLE VESSEL CLOSURE AND GELLING AGENT ON REDUCTION OF HYPERHYDRICITY IN IN VITRO CULTURE OF CARNATION

Directory of Open Access Journals (Sweden)

B. Winarto

2016-10-01

Full Text Available Hyperhydricity, an abnormal morphological appearance and physiologicalfunction, is an important problem in carnation tissue culture. The problem causes premature flowering, high occurrence of abnormal shoots, difficulty in transferring hyperhydric plantlets to soil, and low survival rate of plantlets. High relative humidity and the water potential are considered as the key factors involved in the abnormality. Furthermore, permeable culture vessel and gelling agent were assured to be high potential treatment to eliminate it. Objective of this research was to reduce  hyperhydricity in regenerants of carnation using different permeable vessel closures and gelling agents and to assess the multiplication and  acclimatization abilities of recovered shoots. Experiment was arranged in randomized complete block design with four replications. First factor was different types of closure, i.e. cotton wool, plastic wrap, parafilm and aluminium foil, while second one was gelling agents, i.e. bacto agar, phytagel, swallow agar, and Type 900 agar. The recovered shoots were then multiplied, rooted, and acclimatized. The results showed that hyperhydricity was successfully reduced by applying permeable closure (cotton wool and plastic wrap in combination with Type 900 agar. The combination of plastic wrap and Type 900 agar was the most appropriate treatment in reducing hyperhydricity and producing good quality shoots. The treatment reduced the problem down to 23% of total condition of hyperhydricity (100% and increased leaf chlorophyll content from 0.0883 to 0.1288 mg mg-1. The plastic wrap was easily applied and cheaper material compared to cotton wool. The recovered shoots were able to produce 1-3 healthy axillary shoots and easily rooted on half-strength MS. The recovered plantlets were simply acclimatized with survival rate up to 100% on kossas peat + soil (1:1, v/v and flowered 4-5 months after acclimatization with decreasing in number and size of flower.

Cryopreservation of peach palm zygotic embryos.

Science.gov (United States)

Steinmacher, Douglas A; Saldanha, Cleber W; Clement, Charles R; Guerra, Miguel P

2007-01-01

Cryopreservation is a safe and cost-effective option for long-term germplasm conservation of non-orthodox seed species, such as peach palm (Bactris gasipaes). The objective of the present study was to establish a cryopreservation protocol for peach palm zygotic embryos based on the encapsulation-dehydration technique. After excision, zygotic embryos were encapsulated with 3 percent sodium alginate plus 2 M glycerol and 0.4 M sucrose, and pre-treated or not with 1 M sucrose during 24 h, followed by air-drying. Fresh weight water contents of beads decreased from 83 percent and 87 percent to 18 percent and 20 percent for pre-treated or non-pretreated beads, respectively, after 4 h of dehydration. Sucrose pre-treatment at 1 M caused lower zygotic embryo germination and plantlet height in contrast to non-treated beads. All the variables were statistically influenced by dehydration time. Optimal conditions for recovery of cryopreserved zygotic embryos include encapsulation and dehydration for 4 h in a forced air cabinet to 20 percent water content, followed by rapid freezing in liquid nitrogen (-196 degree C) and rapid thawing at 45 degree C. In these conditions 29 percent of the zygotic embryos germinated in vitro. However, plantlets obtained from dehydrated zygotic embryos had stunted haustoria and lower heights. Histological analysis showed that haustorium cells were large, vacuolated, with few protein bodies. In contrast, small cells with high nucleus:cytoplasm ratio formed the shoot apical meristem of the embryos, which were the cell types with favorable characteristics for survival after exposure to liquid nitrogen. Plantlets were successfully acclimatized and showed 41+/-9 percent and 88+/-4 percent survival levels after 12 weeks of acclimatization from cryopreserved and non-cryopreserved treatments, respectively.

Effect of Dicamba and 2,4 dichlorophenoxiacetic acid on sugarcane somatic embryogenesis

Directory of Open Access Journals (Sweden)

Beatriz Alvez

2015-07-01

Full Text Available In order to conserve sugarcane germplasm, produce microbial disease-free material, detect resistance to diseases and pests, etc., in vitro propagation of sugarcane has been established in many commercial varieties. In this sense, the aim of this work was to analyze the efficiency of 2,4-dichlorophenoxyacetic acid (2,4D and 3,6-dichloro -2- methoxybenzoic acid (Dicamba to induce somatic embryogenesis and regeneration of plantlets from sugarcane varieties C26670, RB855546, V99245, V756, V781, V0050, CC8592, CC8475. For induction of embryogenic callus, leaf discs in tillering stage of 1 cm diameter and 2 mm thick, were inoculated on Murashige-Skoog, 1962 medium (MS, supplemented with 50 ml.l-1 coconut water, 30 g.l-1sucrose and two different treatments: 3 mg.l-1 2,4D or 6.63 mg.l-1 Dicamba, both of them in total darkness at 25 °C, during 1 month. For plant regeneration, embryogenic calli were transferred to ½ MS salts supplemented with coconut water 200 ml.l-1 and sucrose 60 g.l-1 and incubated under continuous light, 25 °C, for 2 months. The highest percent of embryogenic callus induction was obtained in media supplemented with Dicamba, an average of 70.83 % of embryogenic callus by variety, while in media with 2,4-D, 62.08 % of embryogenic callus was obtained by variety. An average of 89,00 % of plantlets was obtained from calli induced on media with Dicamba and an average of 66.12% of plantlets was obtained from calli induced on media supplemented with 2,4D. Using Dicamba it was possible to establish an efficient somatic embryogenesis protocol for these sugarcane varieties.

Cytogenetic studies on stevia rebaudiana produced by tissue culture and affected by gamma rays and drought

International Nuclear Information System (INIS)

Awad, A.S.A

2009-01-01

The present investigation was under taken to carry out in the laboratories of the Natural Products Department, National Center for Radiation Research and Technology, Atomic Energy authority, Nasr city, Cairo, Egypt, to study the effect of gamma radiation doses, osmostress and the combined effects between them on tissue culture, some biochemical analysis and molecular genetic marker in stevia rebaudiana bertoni. The results obtained were: Tissue culture 1- micropropagation media: stevia rebaudiana plantlets cultured on MS medium hormones free for micropropagation.Hormones such as BAP and NAA with different concentrations induced callus formation and give slight growth.Study the effect of gamma radiation, osmostress and the combined effects between them : 1)The effect of gamma radiation on buds survival: Gamma radiation doses (10, 20 and 30 Gy) induced decreasing in bud survival percentage with increasing radiation dose in stevia rebaudiana. The dose 30 Gy was induced 60% mortality.2) Study the effect of gamma radiation on some biochemical analysis: Gamma radiation doses induced increase in the total carbohydrate with doses (20 and 30 Gy) but decreased with dose 10 Gy. Proline contents increased in plantlets with increasing doses . The total protein was increased with doses (10 and 20 Gy), but the dose 30 Gy induced decrease in total protein. Gamma radiation doses induced decreasing in total DNA while, the nucleic acid RNA increased.3) The effect of osmostress on buds survival: The concentrations (40000,50000,60000,70000 and 80000 ppm) from sucrose or sorbitol decreased the bud survival and shoot length in stevia plantlets with increasing sucrose or sorbitol levels. 4) The effect of osmostress on some biochemical analysis: Sucrose and sorbitol concentrations (40000,50000,60000,70000 and 80000 ppm) caused decrease in total carbohydrate.

Cisgenic apple trees; development, characterization and performance

Directory of Open Access Journals (Sweden)

Frans A. Krens

2015-04-01

Full Text Available Two methods were developed for the generation of cisgenic apples. Both have been successfully applied producing trees. The first method avoids the use of any foreign selectable marker genes; only the gene-of-interest is integrated between the T-DNA border sequences. The second method makes use of recombinase-based marker excision. For the first method we used the MdMYB10 gene from a red-fleshed apple coding for a transcription factor involved in regulating anthocyanin biosynthesis. Red plantlets were obtained and presence of the cisgene was confirmed. Plantlets were grafted and grown in a greenhouse. After three years, the first flowers appeared, showing red petals. Pollination led to production of red-fleshed cisgenic apples. The second method used the pM(arkerF(ree vector system, introducing the scab resistance gene Rvi6, derived from apple. Agrobacterium-mediated transformation, followed by selection on kanamycin, produced genetically modified apple lines. Next, leaves from in vitro material were treated to activate the recombinase leading to excision of selection genes. Subsequently, the leaf explants were subjected to negative selection for marker-free plantlets by inducing regeneration on medium containing 5-fluorocytosine. After verification of the marker-free nature, the obtained plants were grafted onto rootstocks. Young trees from four cisgenic lines and one intragenic line, all containing Rvi6, were planted in an orchard. Appropriate controls were incorporated in this trial. We scored scab incidence for three consecutive years on leaves after inoculations with Rvi6-avirulent strains. One cisgenic line and the intragenic line performed as well as the resistant control. In 2014 trees started to overcome their juvenile character and formed flowers and fruits. The first results of scoring scab symptoms on apple fruits were obtained. Apple fruits from susceptible controls showed scab symptoms, while fruits from cisgenic and intragenic

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.

Chromosome Doubling of Microspore-Derived Plants from Cabbage (Brassica oleracea var. capitata L.) and Broccoli (Brassica oleracea var. italica L.).

Science.gov (United States)

Yuan, Suxia; Su, Yanbin; Liu, Yumei; Li, Zhansheng; Fang, Zhiyuan; Yang, Limei; Zhuang, Mu; Zhang, Yangyong; Lv, Honghao; Sun, Peitian

2015-01-01

Chromosome doubling of microspore-derived plants is an important factor in the practical application of microspore culture technology because breeding programs require a large number of genetically stable, homozygous doubled haploid plants with a high level of fertility. In the present paper, 29 populations of microspore-derived plantlets from cabbage (Brassica oleracea var. capitata) and broccoli (Brassica oleracea var. italica) were used to study the ploidy level and spontaneous chromosome doubling of these populations, the artificial chromosome doubling induced by colchicine, and the influence of tissue culture duration on the chromosomal ploidy of the microspore-derived regenerants. Spontaneous chromosome doubling occurred randomly and was genotype dependent. In the plant populations derived from microspores, there were haploids, diploids, and even a low frequency of polyploids and mixed-ploidy plantlets. The total spontaneous doubling in the 14 cabbage populations ranged from 0 to 76.9%, compared with 52.2 to 100% in the 15 broccoli populations. To improve the rate of chromosome doubling, an efficient and reliable artificial chromosome doubling protocol (i.e., the immersion of haploid plantlet roots in a colchicine solution) was developed for cabbage and broccoli microspore-derived haploids. The optimal chromosome doubling of the haploids was obtained with a solution of 0.2% colchicine for 9-12 h or 0.4% colchicine for 3-9 h for cabbage and 0.05% colchicine for 6-12 h for broccoli. This protocol produced chromosome doubling in over 50% of the haploid genotypes for most of the populations derived from cabbage and broccoli. Notably, after 1 or more years in tissue culture, the chromosomes of the haploids were doubled, and most of the haploids turned into doubled haploid or mixed-ploidy plants. This is the first report indicating that tissue culture duration can change the chromosomal ploidy of microspore-derived regenerants.

Perbanyakan In Vitro Dendrobium Indonesia Raya ‘Ina’ melalui Embriogenesis Somatik Berbasis Sistem Bioreaktor

Directory of Open Access Journals (Sweden)

Fitri Rachmawati

2017-01-01

Full Text Available ABSTRACTAn effective and efficient in vitro propagation system has important roles in preparing and producing high quality-seedlings of Dendrobium for commercial scale. The objective of this research was to establish an effective and efficient embryogenic callus (EC proliferation method using bioreactor system and regeneration EC into plantlet for producing high quality seedlings of Dendrobium Indonesia Raya ‘Ina’. Differences in callus densities (5, 10, 15, and 20 g callus in 250 mL medium, aeration levels (2.5, 5.0, and 10.0 O2 volume  per  medium volume per minute; vvm, and regeneration media half-strength MS and 2 g L-1 NPK (32:10:10 combinated by 0.00, 0.05 mg L-1 BA, 150  mL L-1 coconut water and their combinations were tested in this experiment. The experiments were arranged using randomized completely block design (RCBD with three replications for EC proliferation and randomized completely desaign (RBD for EC regeneration. The results showed that combination of  aeration at 2.5 vvm and 10 g of EC was the most suitable aeration level and callus density for proliferation of EC in the 500 ml airlift bioreactor with 6.85 multiplication rate, 92.5% EC formation, and malformed callus morphology as low as 6.1%. The highest somatic embryos (SEs formation was 87.7% with 44.5 SEs per clump and 92.1% SEs germination with 41.0 germinated-SEs per clump, 85.1% normal germinated-SEs, and whereas the best performance of plantlet was obtained from 1/2 MS + 0.05 mg L-1 BA semi solid medium. Plantlets were successfully acclimatized using Cycas rumphii medium with high survival rate (91.6%. Keywords: aerations, callus densities, germination, media, somatic embryos

Growth, flowering and fruiting in vitro pineapple ( Ananas comosus L ...

African Journals Online (AJOL)

The experiment was carried out in a greenhouse, located in Tehran city, Iran. The objectives of this study was to evaluate the effects of fertilizer and acidic soil on the foliar and radicular growth of micropropagated plantlets of the pineapple cv. Merr (Ananas comosus L.). We evaluated the growth of that genotype in five ...

Efficient micropropagation of Citrus sinensis (L.) Osbeck from ...

African Journals Online (AJOL)

... shoots/explant) in comparison to control (0.00±0.00 shoots/explant). Microshoots were rooted best (75.00±14.43%) under the treatment 100μM NAA for 48 hrs. and rooted plantlets were transferred to soil, following acclimatization were taken to maturity in the polyhouse. Keywords: Malta, Himalaya, benzyl adenine (BA), ...

A rapid two step protocol of in vitro propagation of an important ...

African Journals Online (AJOL)

Attempts were made to evolve a rapid in vitro technology to conserve, as well as, mass propagate this valuable medicinal herb in very short duration. The combinations of ... MS media supplemented with BAP with IBA (4:0.5 and 5:0.5) resulted in shooting, as well as, rooting simultaneously. Micro propagated plantlets were ...

Application of tissue culture to cashew ( Anacardium occidentale L ...

African Journals Online (AJOL)

Summary of the previous works on the in vitro culture of cashew is highlighted with emphasis on the critical factors that influence the explants response and plantlet regeneration. The recalcitrant nature of cashew has been attributed to the limited success recorded so far in the in vitro culture of the crop and abnormal ...

Responses of potatoes plants inoculated with arbuscular ...

African Journals Online (AJOL)

A pot experiment was set to examine the impact of the foliar litter (Hardwickia binata and Azadirachta indica) and an arbuscular mycorrhizal (AM) fungus on the development of two varieties of potato plants (Aida, Atlas). Three litter doses (0, 25 and 50 g) were applied to the pots after bedding plantlets. The plants were ...

Evaluation of Selected Pre-Emergence Herbicides for Weed Control ...

African Journals Online (AJOL)

Pendimentalin had no significant effect on the plantlets across the rates tested. The best weed control was achieved with diuron applied at 25% and above, while the least weed control occurred on the control plot and these treated with 10% of atrazine and primextra Gold. However, there were no significant variations in the ...

Tobacco clones derived from tissue culture with supersensitivity to ozone

International Nuclear Information System (INIS)

Sun, E.J.; Kang, H.W.

2003-01-01

New tobacco clones supersensitive to ozone were obtained from tissue culture. - At least two supersensitive tobacco somaclones were obtained from tissue culture (TC) , when this approach was used to asexually propagate Bel-W3 tobacco indicator plants. These somaclones can detect as low as 30 ppb ozone for a 4-h exposure duration both within CSTR exposure chambers and in ambient air. Comparison of the injury index and their coefficient of variance showed that the TC plantlets usually have more uniform performance in response to ozone in addition to their higher sensitivity. A quick regeneration procedure was established to preserve the supersensitive germplasm immediately when it was found. The TC plantlets will flower and produce seed similar to seed-grown tobacco. The TC approach proved to be a better propagation system for valuable indicator plant species. The mechanism that causes the variation and the possible difference in their genome from seed-grown tobacco is still unknown. Further studies are needed in the future to determine if factors in the TC system may be responsible for the sensitivity difference

Effect of γ-ray irradiation on in vitro culture and plant regeneration of alfalfa

International Nuclear Information System (INIS)

Zhang Xiaodong; Lin Tingan

1992-01-01

60 Co γ-ray irradiation ranged 0-16 kR was used to treat the cotyledons and hypocotyls of 5 cultivars of a alfalfa (Medicago sativa L.). The effects of irradiation on the frequency of callus, fresh weight of callus, the frequency of somatic embryo induction and plantlet regeneration were studied. The results showed as follows: the radiosensitivities of cotyledon was significantly higher than that of hypocotyl. Exposure of 2 kR could improve the growth of callus, embryogenesis and plantlet regeneration at various levels. Exposure of 12 kR completely inhibited the callus formation from explants of cotyledon. Callus from two cultivars, England 648 and Jining alfalfa, were irradiated with 0-8 kR. The results showed that exposure below 2 kR could promote the growth of callus and the formation of somatic embryo at different levels. The exposure of 8 kR had the effect of lethal. The optimum exposure for genetic improvement of alfalfa in vitro by inducing mutation was considered to be 4-6 kR

Establishment of an efficient plant regeneration culture protocol and achievement of successful genetic transformation in Jatropha curcas L.

Science.gov (United States)

Liu, Ying; Liu, Guoxuan; Yang, Yali; Niu, Sufang; Yang, Fuguang; Yang, Shaoxia; Tang, Jianian; Chen, Jianping

2017-12-01

An efficient and reproducible protocol is described for shoot-bud regeneration and Agrobacterium tumefaciens-mediated genetic transformation of J. curcas. Treating the explants with high concentrations (5-120 mg/L) of TDZ for short durations (5-80 min) before inoculation culture increased significantly the regeneration frequency and improved the quality of the regenerated buds. The highest shoot-buds induction rate (87.35%) was achieved when petiole explants were treated with 20 mg/L TDZ solution for 20 min and inoculated on hormone-free MS medium for 30 days. Regenerated shoots of 0.5 cm or a little longer were isolated and grafted to seedling stocks of the same species, and then the grafted plantlets were planted on half-strength MS medium containing 0.1 mg/L IBA and 2 mg/L sodium nitroprusside (SNP). This grafting strategy was found to be very effective, to obtain that healthy grafted plantlets ready for acclimatization within 20 days. By the above mentioned protocol and with general Agrobacterium - mediated genetic transformation methods only 65 days were needed to obtain intact transgenic plants.

Plant regeneration of bananas Ambon kuning and Barangan mutant lines were carried out by using female organ and shoot-tip as explants source

International Nuclear Information System (INIS)

Dewi, Azri K; Ishak

1998-01-01

Plant regeneration of bananas Ambon Kuning and Barangan mutant lines were carried out by using female organ and shoot-tip as explants source. Female organ was taken from heart of banana stem, while shoot-tip taken from sucker in banana plantation at Pasar Jumat, Jakarta. Those explants were cultured on MS medium containing 3 mg/l BAP, 0.5 mg/l IAA and supplemented by 100 tyrosin and 80 mg/l adenin hemisulphate. Observation showed that 180 and 42 buds were obtained from JBR 02 mutant lines respectively, while 84 and 79 buds for JAK 01 and JAK 02 respectively. The highest shoot formation was 1.013 shoots were obtained from BRC variety and lowest one was JBR 01 mutant line. statistical data analysis indicated that shoot formation between BRC variety and another mutant lines were significant difference using LSD test at level 0.05. Plantlet formation derived from female organ as well as shoot-tip showed that BRC variety produced number of plantlets per bottle was higher that another one. (author)

In vitro regeneration of Drosera burmannii Vahl.: a carnivorous plant of north-east India.

Science.gov (United States)

Yanthan, J Sureni; Kehie, Mechuselie; Kumaria, Suman; Tandon, Pramod

2017-06-01

An efficient in vitro regeneration protocol has been developed from shoot tips of Drosera burmannii Vahl., a carnivorous plant of north-east India. Various plant growth regulators were used to study their efficacy in the induction of multiple shoots and roots. Of the various treatments, the maximum number of shoots (28.8 ± 1.5) and roots (9.7 ± 0.6) was observed in one-fourth strength standard medium (MS with 50 mg/l citric acid and 10 mg/l ascorbic acid) supplemented with 4 mg/l 6-benzylaminopurine (BAP) and 4 mg/l α-naphthalene acetic acid (NAA) followed by 26.8 ± 1.4 shoots in one-fourth strength SM fortified with 4 mg/l kinetin (KN) and 4 mg/l NAA. The well-developed plantlets with shoots and roots were potted in small plastic glasses filled with a mixture of sand and farmyard manure (3:1); these plantlets when transferred to a glasshouse for hardening and acclimatization showed 90% survival.

Germination in vitro embryo of Walnut (Juglans boliviana

Directory of Open Access Journals (Sweden)

Pérez-Guzmán Jheanete

2015-05-01

Full Text Available Bolivian Juglans is an important forest species found in the rain forests of Bolivia. The seed of this species is recalcitrant with hardened cover, which hinders germination and propagation of the species. The aim of this study was to determine the culture medium for in vitro germination of mature embryos of Bolivian Juglans. Technique initially scarification and disinfection process was determined. Subsequently in vitro culture was performed using the culture medium Woody Plant Medium (WPM with the addition of plant growth regulators (indole butyric acid and 6-benzyl aminopurine in different concentrations. As control WPM, culture medium was used 100%. Response variables evaluated were percentage of contamination and germination; vitroplant length, number of leaves, number of shoots, number of roots per vitroplant, root length and percentage of survival. The plantlets in vitro germination in treatments and the control in the middle l culture WPM supplemented with 0.15 mg / l of IBA and 1.5 mg / l BAP was 90%; other treatments inhibit the growth of the stem and roots of plantlets.

one step method of plantlet regeneration in trichosanthes dioica roxb

African Journals Online (AJOL)

User

2011-01-03

Jan 3, 2011 ... Tissue Culture Laboratory, Department of Botany, Patna University, Patna, India. E-mail: rashmi0911@gmail.com. Accepted 23 December, 2010. In order to reduce the cost and time of in vitro raised plants of Trichosanthes dioica Roxb., a minimal .... 1951), tomato juice (Nitsch, 1951; Straus and La Rue,.

Effect of plant growth regulators, explants type and efficient plantlet ...

African Journals Online (AJOL)

use

2011-12-05

Dec 5, 2011 ... Plant Pathology, Tissue Culture and Biotechnology Laboratory, Department of Botany,. University of ... variability in response to growth regulators. In vitro rooting ..... an adult tree Wrightia tomentosa through enhanced axillary.

Rapid Screening of In-Vitro Regenerated Plantlets of Four Nigerian ...

African Journals Online (AJOL)

Prof. Ogunji

Tissue culture technique provides a rapid means of studying plant-pathogen interaction in a controlled ... Keywords: cowpea, in-vitro regeneration, hormones, Fusarium wilt, protocols. .... solanacearum and evaluation of tomato in Nepal.

Characteristics of micro-propagated banana (Musa spp.) cultures ...

African Journals Online (AJOL)

Administrator

2011-05-23

May 23, 2011 ... was conducted to assess the effect of NaCl and PEG separately as well as in combination on plant micro- propagation efficiency of banana (Musa spp.) cv., Basrai. In this experiment, 4-weeks old plantlets of the 3rd sub- culture with well propagation on MS2b nutrient were sub- cultured on three differentially ...

In vitro regeneration of solanum aethiopicum L. (scarlet eggplant), an african vegetable crop with potential ornamental value

Science.gov (United States)

Successful in vitro regeneration of plantlets was obtained from shoot tips of five Solanum aethiopicum (African eggplants) accessions evaluated in two media, M1 and M2. The M1 medium consisted of Murashige and Skoog (MS) basal salt mixture supplemented with 20 g/L sucrose, 0.75 g/L MgCl2, and 2 g/L ...

Contrôle des germes endogènes par usage d'agents antimicrobiens ...

African Journals Online (AJOL)

SARAH

31 janv. 2018 ... lisse (Hilo et Saint Michael) et Pain de sucre ont été ensemencés sur le milieu de Murashige et Skoog additionné de 5 mg/l ... In this study, the effects of ...... Impact of Thermotherapy And Chlorothalonil On. Plantlets Production Of Some Genotypes Of. Cassava (Manihot Esculenta Crantz) Produce In. Benin.

Concentração de bap e a eficiência de micropropagação de bananeira tetraplóide (grupo AAAB Bap concentration and tetraploid banana micropropagation efficiency (AAAB group

Directory of Open Access Journals (Sweden)

Roberto Pedroso de Oliveira

2001-03-01

Full Text Available Com o objetivo de desenvolver um procedimento eficiente para a produção de mudas de bananeiras tetraplóides (Musa sp. cv. FHIA-01, grupo AAAB estudou-se o efeito do clone no desenvolvimento in vitro dos explantes e as taxas de contaminação, multiplicação e nível de oxidação em meios de cultura contendo cinco concentrações de BAP. Na fase de introdução in vitro dos explantes, foi observada elevada taxa de contaminação (22,32%, causada, principalmente, por bactérias. No entanto, os níveis de contaminação decresceram na seqüência dos subcultivos, tendo sido de 1,07% no oitavo subcultivo. As maiores taxas de multiplicação foram obtidas no meio de cultura MS com 4,0 mg L-1 de BAP, em média de 2,65 plântulas por subcultivo, possibilitando a produção estimada de 584 plântulas de FHIA-01 por explante inicial no sexto e 3451 no oitavo subcultivo. Em função do clone, houve a produção estimada de 305 a 4497 plântulas por explante inicial, após oito subculturas. A eficiência de aclimatização das plântulas foi de 94%, não sendo encontrados variantes somaclonais em condições de casa-de-vegetação.The objective of the present work was to establish an efficient procedure for the in vitro production of tetraploid banana plantlets (Musa sp. cv. FHIA-01, AAAB group. The cloning effect on the in vitro development of the explants, the rates of contamination and multiplication, and the level of oxidation in culture media with five benzylaminopurine (BAP concentrations were studied. In the phase of in vitro introduction of explants, a high level of contamination (22.32%, caused mainly by bacteria, was observed. However, the degree of contamination decreased along the subcultures, reaching 1.07% in the eighth subculture. Higher multiplication rates were obtained, averaging 2.65 per subculture, on the MS media supplemented with 4.0 mg L-1 BAP, leading to an estimated production of 584 and 3451 plantlets/initial explant after

Cultivo in vitro de embriões zigóticos e aclimatação de plântulas de coqueiro-anão In vitro culture of zygotic embryos and acclimatization of green dwarf coconut palm

Directory of Open Access Journals (Sweden)

Ana da Silva Lédo

2007-02-01

Full Text Available O objetivo deste trabalho foi ajustar os protocolos de cultivo in vitro de embriões zigóticos e de aclimatação de plântulas de coqueiro-anão-verde do Brasil de Jiqui (Cocos nucifera L.. Os embriões zigóticos maduros, colocados assepticamente em meio de cultura líquido Y3 suplementado com 27,8 mg L-1 de Fe2SO4.7H2O e 60 g L-1 de sacarose, apresentaram maior germinação e formação de plântulas normais. As plântulas transferidas para meio indutor de enraizamento Y3, gelificado e suplementado com 1 mg L-1 de ANA, 0,5 mg L-1 de BAP e 2,5 g L-1 de carvão ativado, apresentaram incremento do desenvolvimento radicular e da parte aérea. Na fase de aclimatação, o substrato composto por areia lavada e pó de casca de coco seco, na proporção de 1:1, proporcionou maior sobrevivência das plântulas (58,33%, maior crescimento da parte aérea (39,08 cm e maior número de folhas (6,33. Os protocolos estabelecidos para o cultivo in vitro de embriões zigóticos e a aclimatação de coqueiro-anão-verde de Jiqui podem ser utilizados no intercâmbio e conservação de germoplasma.The objective of this work was to improve the protocols for culture and acclimatization of green dwarf coconut palm (Cocos nucifera L.. Mature zygotic embryos, cultivated in vitro aseptically in Y3 liquid medium culture, supplemented with 27.8 mg L-1 of Fe2SO4.7H2O and 60 g L-1 of sucrose, presented higher germination and normal plantlets formation. The plantlets transferred to Y3 semi-solid medium culture supplemented with 1 mg L-1 of ANA, 0.5 mg L-1 of BAP and 2.5 g L-1 of activated charcoal, promoted better root and shoot development. In the acclimatization phase, the 1:1 substrate composed of washed sand and coconut coir dust provided higher survival of plantlets (58.33%, larger growth of the aerial part (39.08 cm and larger number of leaves (6.33. The established protocols for in vitro culture of zygotic embryos and acclimatization of green dwarf coconut palm

Anatomical, germination and in vitro studies on shea tree (Vitellaria paradoxa.) seed

International Nuclear Information System (INIS)

Iddrisu, A.

2013-07-01

(1962) basal salts modified with 6-benzylaminopurine (BAP) produced no plantlets although 80% of the deshelled seeds developed long pseudoradicles on a medium supplemented with lower concerntration of BAP (1.0 or 2.0 mg/1). Similarly, the culture of TTC identified embrayonic axes did not produced plantlets, but rather significantly long pseudoradicles development. Contrastingly, in vitro culture of excised rudimentary shoots on the same MS medium modified with BAP and naphthaleneacetic acid (NAA) produced plantlets with distinct shoots and leaves. Significant reduction days to emergence of seedlings from deshelled seeds and successful in vitro plantlet development using rudimentary shoot explants will enhence nursery establishment of this economically important tree species for domestication and reafforestation programmes in sub-Saharan Africa. (au)

Effect of multiple subcultures on Musa shoots derived from cassava ...

African Journals Online (AJOL)

Shoot tip explants excised from in vitro plantlets of two Musa genotypes (TM3X 15108-6 and TMBX 612-74) were seeded singly into test tubes containing twenty milliliters each of Musa multiplication medium gelled differently in 60 and 70 gL-l cassava starch as well as 5 gL-l agar and placed on shelves under 14 h photo ...

In vitro mutagenesis and production of agronomically useful potato variants

International Nuclear Information System (INIS)

Das, A.; Gosal, S.S.; Sidhu, J.S.; Dhaliwal, H.S.

2001-01-01

In vitro grown shoot cultures of two Indian potato varieties 'Kufri jyoti' and 'Kufri Chandramukhi' were subjected to gamma irradiation at 20 and 40 Gy. The irradiated shoot cultures were subcultured to yield a generation of plantlets. After 4-6 weeks of incubation, these shoots were transferred onto MS medium supplemented with benzylaminopurine, BAP (10mg/1) and sucrose (8% w/v) and incubated at 20 deg. C. The M 1 V 3 plants were screened in vitro for late blight resistance by detached leaf method. The resistant plants were screened in M 1 V 4 generation by artificial inoculation of sporangial inoculum on the pot sown plants. Chlorophyll persistence is a simple screening method for heat tolerance. Chlorophyll persistence of different plantlets showed that the percentage of injury was less in the case of plants, which had been obtained from irradiated material. In the case of control plants, there was one hundred-percent damage to the plants. The mutation frequency was calculated for characters like late blight resistance and heat tolerance (in vitro microtuberisation and chlorophyll persistence). The gamma ray dose of 40 Gy was observed to produce a higher mutation frequency

High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

Directory of Open Access Journals (Sweden)

Dikash Singh THINGBAIJAM

2014-03-01

Full Text Available An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%. Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25. Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%. Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

Directory of Open Access Journals (Sweden)

Dikash Singh THINGBAIJAM

2014-03-01

Full Text Available An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%. Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25. Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%. Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

Establishment of an in vitro micropropagation protocol for Boscia senegalensis (Pers.) Lam. ex Poir.

Science.gov (United States)

Khalafalla, Mutasim M; Daffalla, Hussien M; Abdellatef, Eltayb; Agabna, Elsadig; El-Shemy, Hany A

2011-04-01

This report describes in vitro micropropagation of Boscia senegalensis, so-called famine foods, that helped the people in Darfur and Kordofan, Sudan survive during the 1984-1985 famine. Four types of explants prepared from green mature zygotic embryos were cultured on Murashige and Skoog (MS) medium augmented with 1-5 mg/L 6-benzyladenine (BA). The highest number of shoots per explant (14.3±0.9) was achieved on MS medium supplemented with 3 mg/L BA, while the highest shoot length [(3.5±0.4) cm] was obtained with 1 mg/L BA. The shoot cluster, when subcultured to its same medium, significantly increased the rate of shoot multiplication by the end of the third subculture. The maximum mean number of shoots per explant (86.5±3.6) was produced after three multiplication cycles on 3 mg/L BA-supplemented medium. In vitro induced shoots were excised and rooted on half strength MS medium fortified with 0.25 mg/L indole-3-butyric acid (IBA) to obtain complete plantlets. B. senegalensis-regenerated plantlets obtained in vitro for the first time, were hardened and 95% survived under greenhouse conditions.

Bioactive compounds of fourth generation gamma-irradiated Typhoniumflagelliforme Lodd. mutants based on gas chromatography-mass spectrometry

Science.gov (United States)

Sianipar, N. F.; Purnamaningsih, R.; Rosaria

2016-08-01

Rodent tuber (Typhonium flagelliforme Lodd.) is an Indonesian anticancer medicinal plant. The natural genetic diversity of rodent tuber is low due to vegetative propagation. Plant's genetic diversity has to be increased for obtaining clones which contain a high amount of anticancer compounds. In vitro calli were irradiated with 6 Gy of gamma ray to produce in vitro mutant plantlets. Mutant plantlets were acclimated and propagated in a greenhouse. This research was aimed to identify the chemical compounds in the leaves and tubers ofthe fourth generation of rodent tuber's vegetative mutant clones (MV4) and control plantsby using GC- MS method. Leaves and tubers of MV4 each contained 2 and 5 anticancer compounds which quantities were higher compared to control plants. MV4 leaves contained 5 new anticancer compounds while its tubers contained 3 new anticancer compounds which were not found in control. The new anticancer compounds in leaves were hexadecanoic acid, stigmast-5-en-3-ol, ergost-5-en-3-ol, farnesol isomer a, and oleic acid while the new anticancer compounds in tubers were alpha tocopherol, ergost-5-en-3-ol, and beta-elemene. Rodent tuber mutant clones are very potential to be developed into anticancer drugs.

Role of extrinsic arbuscular mycorrhizal fungi in heavy metal-contaminated wetlands with various soil moisture levels.

Science.gov (United States)

Zheng, S; Wang, C; Shen, Z; Quan, Y; Liu, X

2015-01-01

This study presents an efficient heavy metal (HM) control method in HM-contaminated wetlands with varied soil moisture levels through the introduction of extrinsic arbuscular mycorrhizal fungi (AMF) into natural wetland soil containing indigenous AMF species. A pot culture experiment was designed to determine the effect of two soil water contents (5-8% and 25-30%), five extrinsic AMF inoculants (Glomus mosseae, G. clarum, G. claroideum, G. etunicatum, and G. intraradices), and HM contamination on root colonization, plant growth, and element uptake of common reed (Phragmites australis (Cav.) Trin. ex Steudel) plantlets in wetland soils. This study showed the prevalence of mycorrhizae in the roots of all P. australis plantlets, regardless of extrinsic AMF inoculations, varied soil moisture or HM levels. It seems that different extrinsic AMF inoculations effectively lowered HM concentrations in the aboveground tissues of P. australis at two soil moisture levels. However, metal species, metal concentrations, and soil moisture should also be very important factors influencing the elemental uptake performance of plants in wetland ecosystems. Besides, the soil moisture level significantly influenced plant growth (including height, and shoot and root dry weight (DW)), and extrinsic AMF inoculations differently affected shoot DW.

In Vitro Regeneration of Foxtail Millet (Setaria italica (L. Beauv. cv. Buru Hotong

Directory of Open Access Journals (Sweden)

Iriawati

2017-09-01

Full Text Available In vitro regeneration of foxtail millet (Setaria italica (L. Beauv. was done using basal shoot explants of 10-day old seedlings. Explants were cultured in MS basal medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D, kinetin, 6-benzylaminopurine (BAP and 1.5 ppm NiSO4. Shoot multiplication and root induction were done in Murashige and Skoog (MS basal media. Plantlets were then acclimatized in rice husk charcoal, cocopeat, or mixed media. Results showed that MS basal medium containing 0.5 ppm kinetin, 2 ppm BAP, and 0.1 ppm 2,4-D was the optimal medium for shoot induction, in which 60% of explants developed direct shoot organogenesis. In addition, callus was optimally formed in MS medium supplemented by 1 ppm kinetin, 1 ppm BAP, and 0.5 ppm 2,4-D. Regenerated shoots spontaneously developed roots after being transferred into MS basal media without growth regulator. During acclimatization, the highest survival rate of plantlets (47% was obtained in rice husk charcoal. The developed method could be useful towards improvement of this species using in vitro tissue culture techniques.

Mutation breeding in Philippine fruits

International Nuclear Information System (INIS)

Espino, R.R.C.

1987-09-01

Studies were made to establish standard conditions for mutation induction by gamma-irradiation to be performed in combination with in-vitro culture for banana and citrus spp. Besides this, radio-sensitivity of seeds and/or plantlets of mango, sugar apple, soursop, lanzones and Jack fruit was investigated and primary observation on the occurrence of mutation was made. For the mutagenesis of banana shoot tip cultures, radio-sensitivity of plantlets derived from the culture as well as fresh-cultured shoots was examined and phenotypes indicative of mutation, such as chlorophyl streaking, slow growth, pigmentation and varied bunch orientation were recorded. Isozyme analysis for mutated protein structure was not conclusive. In the in-vitro culture of Citrus spp., seeds placed on fresh media as well as germinating seeds and two-leaf stage seedlings in test tubes were examined for their radio-sensitivity. Irradiated materials were propagated for further observation. In these two crops, basic methodology for mutation induction with combined use of in-vitro culture and gamma-irradiation was established. In mango, sugar apple, soursop, lanzones and Jack fruit, basic data on radiosensitivity were obtained. In mango, leaf abnormalities were observed after the treatment of scions

Effect of pollen irradiation on hybridization of wheat and Eltrigia intermedia and their embryo development

International Nuclear Information System (INIS)

Li Guiying; Wang Linqing; Shi Jinguo

2005-01-01

Spikes of Eltrigia intermedia were radiated with 5-100 Gy γ-ray during anthesis, and then their pollens were collected to pollinate to the common wheat 'J-11' and 'Chinese Spring'. The effects of pollen irradiation on the seed setting, embryo development, embryo culture and plantlet rate were studied. The results showed that low dose (5-9 Gy) of irradiation enhanced the seed setting for Chinese spring x E. intermedia, but no such effect for J-11xE. intermedia. Irradiation with all doses damaged embryo development, percentage of seeds with embryos; rate of immature hybrid embryos developing into plantlets decreases with the increased doses. Percentage of seeds with abnormal embryos increased significantly with the doses. 12.9%-14.5% of embryos could develop into plants in 30 Gy treatment, which seldom occur to wheat. Embryos in 50 Gy-100 Gy treatment were affected so serious that even none of them could develop into plants in vitro culture. It may be an effective approach to obtaining more mutants and enhancing induced mutation breeding by using combination of pollen irradiation, immature embryo rescue and anther culture of the resultant progenies. (authors)

Use of temporal immersion systems for the in vitro multiplication of Anthurium andraeanum Lind. var. Lambada shoots

Directory of Open Access Journals (Sweden)

Nydia del Rivero Bautista

2004-04-01

Full Text Available Two methods to improve the efficiency in the multiplication of shoots of Anthurium andraeanum Lind., they were used to optimize the frequencies of immersion of the explants using a System of Temporary Immersion. The relative efficiency of the method conventional means of culture semisolid for the multiplication of shoots was compared with the method of System of Temporary Immersion. The highest rate of multiplication of shoots was reached using six immersions four times a day during two minutes, controlling the immersion cycles achieves plants better developed and bigger multiplication coefficient. The plantlets multiplied through this method were harvested with 2 to 3 cm of longitude containing from two to five leaves after a period of culture of 50 days. The formation of callus, hiperhidricity and other abnormalities were not observed during the process. The plants obtained by this method were taken to rooting in medium of culture semisólid. This system avoids risks of contamination, it increases yields, it reduces manpower costs and of containers. This technique for its relationship benefit-cost has significant implications for the commercial propagation of the Anthurium. Key words: Culture medium, multiplication rate, plantlets.

Screenhouse and field persistence of nonpathogenic endophytic Fusarium oxysporum in Musa tissue culture plants.

Science.gov (United States)

Paparu, Pamela; Dubois, Thomas; Gold, Clifford S; Niere, Björn; Adipala, Ekwamu; Coyne, Daniel

2008-04-01

Two major biotic constraints to highland cooking banana (Musa spp., genome group AAA-EA) production in Uganda are the banana weevil Cosmopolites sordidus and the burrowing nematode Radopholus similis. Endophytic Fusarium oxysporum strains inoculated into tissue culture banana plantlets have shown control of the banana weevil and the nematode. We conducted screenhouse and field experiments to investigate persistence in the roots and rhizome of two endophytic Fusarium oxysporum strains, V2w2 and III4w1, inoculated into tissue-culture banana plantlets of highland cooking banana cultivars Kibuzi and Nabusa. Re-isolation of F. oxysporum showed that endophyte colonization decreased faster from the rhizomes than from the roots of inoculated plants, both in the screenhouse and in the field. Whereas rhizome colonization by F. oxysporum decreased in the screenhouse (4-16 weeks after inoculation), root colonization did not. However, in the field (17-33 weeks after inoculation), a decrease was observed in both rhizome and root colonization. The results show a better persistence in the roots than rhizomes of endophytic F. oxysporum strains V2w2 and III4w1.

Prolific plant regeneration through organogenesis from scalps of ...

African Journals Online (AJOL)

Four types of potting media comprising of sand, peat, sand + top soil + goat dung (3:2:1 v/v) and top soil + sand (1:1 v/v) were evaluated during acclimatization of the plantlets. Prolific shoot regeneration from scalps was obtained on MS medium containing 2.5 mM BAP, at 9.61 and 40.6 shoots per explant after 4 and 8 weeks ...

Continuous high and low temperature induced a decrease of photosynthetic activity and changes in the diurnal fluctuations of organic acids in Opuntia streptacantha.

Directory of Open Access Journals (Sweden)

Zaida Zarely Ojeda-Pérez

Full Text Available Opuntia plants grow naturally in areas where temperatures are extreme and highly variable in the day during the entire year. These plants survive through different adaptations to respond to adverse environmental conditions. Despite this capability, it is unknown how CAM photosynthetic activity and growth in Opuntia plantlets is affected by constant heat or cold. Therefore, the main objective of this research was to evaluate the short-term effect of high (40°C and low (4°C continuous temperatures on the photosynthetic efficiency, the organic acid content (malic acid and the relative growth rate (RGR in seven-month-old Opuntia streptacantha plantlets during 5, 10, and 15 days. Chlorophyll fluorescence analysis allowed us to determine that high temperatures negatively impact the photosynthetic efficiency of O. streptacantha plantlets, which exhibited the lowest values of maximum quantum efficiency of the photosystem II (Fv/Fm = 52%, Fv/F0 = 85%, operational quantum yield of PS (ΦPSII = 65% and relative electron transport rate (rETR = 65%, as well as highest values of basal fluorescence (F0 = 226% during 15 days of treatment. Similarly, low temperatures decreased Fv/Fm (16%, Fv/F0 (50%, ΦPSII and rETR (16%. High temperatures also decreased nocturnal acidification in approximately 34-50%, whereas low temperatures increased it by 30-36%. Additionally, both continuous temperatures affected drastically diurnal consumption of malic acid, which was related to a significant RGR inhibition, where the specific photosynthetic structure area component was the most affected. Our results allowed determining that, despite the high tolerance to extreme temperatures described for Opuntia plants, young individuals of O. streptacantha suffered photosynthetic impairment that led to the inhibition of their growth. Thus, the main findings reported in this study can help to predict the potential impact of climatic change on the establishment and survival of succulent

EFFECT OF SEED XYLOGLUCANS AND DERIVATES ON THE GROWTH OF Arabidopsis thaliana

Directory of Open Access Journals (Sweden)

Adriana Tourinho Salamoni

2009-10-01

Full Text Available Studies on xyloglucan (XG extracted from Hymenaea courbaril L. (jatoba seeds showed that this biopolymer has biological activity that enhanced wheat coleoptiles growth. In apple tree micropropagation, the culture medium containing XG combined with agar induced a higher multiplication rate, rooting rate and root length than medium solidified with agar only. The purpose of this study was to determine the effect of XG from jatobá seeds extracted from jatoba seeds collected in Sinope/MT (XGS and Cuiabá/MT (XGC, and from XGC hydrolysed with a cellulase (XGCH, as well from Tamarindus indica seeds (XGT collected in Bahia/BA, on the growth of in vitro cultured Arabidopsis thaliana plantlets. In the first experiment, XGCH (0.25, 25 and 250 nM or XGC (0.5, 50 and 500 nM were added to a liquid half-strength MS medium. In the second experiment, XGs from several origins were compared: XGC (500 nM, XGS (1200 nM and XGT (800 nM, using culture medium solidified with 6 g.L-1agar. Arabidopsis thaliana L. seeds germinated in Petri plates for 4 to 5 days were transferred to culture media containing the different concentrations of XGs and cultured in a growing room. When the plantlets were cultured in a liquid medium, their growth was very slow in the presence of XGC and XGCH at the highest concentration tested, and it was faster at the lowest concentration. In the semi-solid culture medium, XGs also reduced growth. It was concluded that XGs can play a biological role in Arabidopsis thaliana (L. Heynh. plantlets, stimulating or inhibiting the root system growth and the lateral root formation. These opposite effects varied according to the plant specie that furnished the seeds containing XG, as well as the place where the seeds were collected, to the XG form used (hydrolyzed or not and to its concentration in the culture media. 

Parameters necessary for in vitro hydroponic pea plantlet flowering and fruiting

Science.gov (United States)

To date, the development of culture systems to stimulate the reproductive and fruiting processes in vitro has not been earnestly exploited. Nonetheless, the ability to promote sterile flowering and fruiting would have commercial applications in the agricultural, food and medical industries. Non-ir...

Effect of gamma irradiation and ethyl methane sulphonate on flavour formation in garlic (Allium sativum L. ) cultures

International Nuclear Information System (INIS)

Malpathak, N.P.; David, S.B.

1990-01-01

In callus cultures both gamma irradiation and EMS treatments produced stimul ation of flavour formation at lower as well as at higher dosages. Different results were obtained when the plantlets differentiated from both the treatments were analysed. There was a gradual increase in flavour formation with increasing dosages, which declined at a higher dosages. Cytological observations showed an increase in polyploidy with increasing dosages. (author). 5 refs., 5 tabs

A novel growth-promoting microbe, Methylobacterium funariae sp. nov., isolated from the leaf surface of a common moss

OpenAIRE

Schauer, S; Kutschera, U

2011-01-01

Land plants (embryophytes) evolved in the presence of prokaryotic microbes. As a result, numerous mutually beneficial associations (symbioses) developed that can be analyzed using a variety of methods. Here we describe the isolation and characterization of a new pink-pigmented facultatively methylotrophic symbiotic bacterium of the genus Methylobacterium (laboratory strain F3.2) that was isolated from the gametophytic phylloids of the common cord moss Funaria hygrometrica Hedw. Plantlets were...

Monitoring Genetic Stability in Quercus serrata Thunb. Somatic Embryogenesis Using RAPD Markers

OpenAIRE

Ramesh C., Thakur; Susumu, Goto; Katsuaki, Ishii; S. Mohan, Jain; Forestry and Forest Products Research Institute; Fukuoka Prefecture Forest Research and Extension Center; Forestry and Forest Products Research Institute; University of Helsinki

1999-01-01

Genetic stability of propagules regenerated via somatic embryogenesis is of paramount importance for its application to clonal forestry. Random amplified polymorphic DNA (RAPD) markers were used to determine the genetic stability in somatic embryogenesis of Quercus serrata Thunb. (Japanese white oak). Forty samples from an embryogenic line, consisting of regenerated plantlets, somatic embryos, and embryogenic calli, were examined using 54 decanucleotide primers. A total of 6520 clear reproduc...

Ação do estresse térmico na sobrevivência de mudas e produção de camomila originadas de sementes importadas e nacionais Action of termic stress on the survival and yield of chamomile plantlets from imported and national seeds

Directory of Open Access Journals (Sweden)

José Roberto P de Souza

2006-06-01

Full Text Available Avaliou-se o efeito de baixas temperaturas sobre a sobrevivência de mudas de camomila (originadas de sementes importadas e coletadas localmente e sobre a produtividade da matéria-prima vegetal. Quarenta mudas com 7 a 10 cm de altura foram transferidas para câmara de germinação, tipo BOD, de duas procedências, onde permaneceram por 24 h nas condições: 10 h, com luz e 15ºC; 8 h, sem luz e 10ºC; 3 h, sem luz e 5ºC; 3 h; sem luz e temperaturas de 0ºC (testemunha, -2ºC, -4ºC e -6ºC. As mudas de sementes local apresentaram maior número de plantas mortas quando aplicadas às temperaturas de -4 e -6ºC, do que as importadas. O número de flores por planta e o acúmulo de matéria seca de flores por planta foram 462% e 226% superior, respectivamente, nas plantas originadas de sementes local comparadas com as importadas. O tratamento térmico de -6ºC proporcionou retomada do crescimento e aumento da produção das plantas de camomila, contrastando com aquelas submetidas às temperaturas de -2ºC e -4ºC. Os extratos das flores das plantas originadas de sementes local e importada mostraram igual quantidade de quercetina (substância marcadora.In the present trial the effects of low temperatures on the surviving of chamomile seedlings originated from two different origins (local and imported seeds was evaluated on the development, yield and concentration of quercetin. Forty to 10 cm high seedlings from the two origins were transferred to a germination chamber (BOD during 24 hours and submitted to the conditions: 10 h with light and 15ºC; 8 h without light and 10ºC; 3 h without light and 5ºC; 3 h without light and temperatures ranging from 0 (check, -2, -4, and -6ºC. The seedlings from national seeds were more susceptible to the temperatures of -4 and -6ºC. There were 462% more flowers per plant and 226% more dry matter accumulation per plant in the plants originated from national seeds compared to the plantlets from imported seeds

Exogenous salicylic acid protects phospholipids against cadmium stress in flax (Linum usitatissimum L.).

Science.gov (United States)

Belkadhi, Aïcha; De Haro, Antonio; Obregon, Sara; Chaïbi, Wided; Djebali, Wahbi

2015-10-01

Salicylic acid (SA) promotes plant defense responses against toxic metal stresses. The present study addressed the hypothesis that 8-h SA pretreatment, would alter membrane lipids in a way that would protect against Cd toxicity. Flax seeds were pre-soaked for 8h in SA (0, 250 and 1000µM) and then subjected, at seedling stage, to cadmium (Cd) stress. At 100µM CdCl2, significant decreases in the percentages of phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylethanolamine (PE) and monogalactosyldiacylglycerol (MGDG) and changes in their relative fatty acid composition were observed in Cd-treated roots in comparison with controls. However, in roots of 8-h SA pretreated plantlets, results showed that the amounts of PC and PE were significantly higher as compared to non-pretreated plantlets. Additionally, in both lipid classes, the proportion of linolenic acid (18:3) increased upon the pretreatment with SA. This resulted in a significant increase in the fatty acid unsaturation ratio of the root PC and PE classes. As the exogenous application of SA was found to be protective of flax lipid metabolism, the possible mechanisms of protection against Cd stress in flax roots were discussed. Copyright © 2015 Elsevier Inc. All rights reserved.

Micropropagation of Phalaenopsis orchids via protocorms and protocorm-like bodies.

Science.gov (United States)

Paek, Kee Yoeup; Hahn, Eun Joo; Park, So Young

2011-01-01

Phalaenopsis orchids have high economic value in the floriculture industry. Hybridization or cross-pollination in the breeding program have proven to be very reliable techniques for the production of a wide range of successful cultivars with attractive combinations of spray length, bud number, flower color and type, fragrance, seasonality, and compactness. In vitro propagation makes it possible to clonally mass propagate hybrids of commercial value and conserved species. However, in vitro culture technologies are still a challenge because of the slow growth of plantlets, low multiplication rate, poor rooting, and somaclonal variation. Although seed-raised plants can be used for conservation and breeding for the selection of superior features, genetic characteristics including seasonality, inflorescence, flower color, and type are not uniform. In this regard, micropropagation through protocorm-like bodies obtained from germinating embryos and somatic tissues is an important strategy in obtaining genetically stable plants and the improvement of quality. However, not all genotypes of Phalaenopsis respond to the same protocol under the same culture conditions and often result in the development of undesirable characteristics. In this chapter, plantlet production in Phalaenopsis orchids via the culture of protocorms from seeds and protocorm-like bodies from leaf sections and root tips are detailed.

Isozyme modifications and plant regeneration through somatic embryogenesis in sweet potato (Ipomoea batatas (L.) Lam.).

Science.gov (United States)

Cavalcante Alves, J M; Sihachakr, D; Allot, M; Tizroutine, S; Mussio, I; Servaes, A; Ducreux, G

1994-05-01

The potential of somatic embryogenesis was evaluated for 10 cultivars of sweet potato through extensive embryogenic response and isozyme analysis. Embryogenic callus was induced by incubating lateral buds on Murashige and Skoog medium containing 10 μM 2,4-dichlorophenoxyacetic acid for 6-8 weeks. The frequency of embryogenic response was low, and varied with genotypes, ranging from 0 to 17%. Embryo to plantlet formation could be enhanced by the use of the combination of 2,4-dichlorophenoxyacetic acid with kinetin, both used at 0.01 μM. Embryogenic callus with its potential of plantlet formation has constantly been maintained for over two years. However, after several subcultures, 0.5 to 12% of embryogenic callus reverted irreversibly into friable fast-growing non-embryogenic callus whose ability to regenerate shoots was then definitively lost. The isozymes of esterase, peroxidase, glutamate oxaloacetate transaminase and acid phosphatase investigated in this study were found appropriate to distinguish compact embryogenic from friable non-embryogenic callus in sweet potato. In fact, the callus reversion was associated with a loss of bands or a decline in isozyme activity. On the contrary, very small changes in isozyme activity or no specific changes at all were observed during the differentiation of embryogenic callus into globular embryos.

Mutation induction of roses through in vitro propagation

International Nuclear Information System (INIS)

Amran Abd Halim; Azhar Mohamad; Rusli Ibrahim

2002-01-01

The genus Rosa has been exploited for a variety used extensively in commercial production for many cultivars. Mutation induction is an alternative way of creating more variations in a single variety. Radiosensitivity test was carried out to identify optimum doses in mutation induction. Shoot tips of excises from in vitro plantlets were irradiated with doses of 0, 20, 40, 60, 80 and 100 Gy using a gamma cell with a 60 Co source at dose rate of 0.25 Gy s -1 . Irradiated shoot tips were cultured on semi solid modified Murashige and Skoog (MS) containing cytokinin (BAP) hormone and incubated at 24 with a photoperiod of 16 hours (3500 lux). Radiosensitivity was assessed by the rate of shoot proliferation four weeks after treatment. Increasing gamma ray doses caused a reduction of survival rate as well as the average shoots produced per plantlet. Through linear estimation of buds and shoots proliferation, radiation dose that reduced the growth to 50% of the control treatment (LD 50 ) was 53 Gy for miniature roses and 60 Gy for cut roses. Based on the radiosensitivity test 20 and 40 Gy were selected for irradiation to get new mutant varieties of roses. Changes in flower characters were found in three treated plants after three months in the green house. (Author)

Better Rooting Procedure to Enhance Survival Rate of Field Grown Malaysian Eksotika Papaya Transformed with 1-Aminocyclopropane-1-Carboxylic Acid Oxidase Gene

Science.gov (United States)

Sekeli, Rogayah; Abdullah, Janna Ong; Namasivayam, Parameswari; Muda, Pauziah; Abu Bakar, Umi Kalsom

2013-01-01

A high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. We report in this paper an improved rooting method for the production of transformed Malaysian Eksotika papaya with high survival rate when transferred to the field. Shoots were regenerated from embryogenic calli transformed with antisense and RNAi constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes using the Agrobacterium tumefaciens-mediated transformation method. Regenerated transformed shoots, each measuring approximately 3-4 cm in height, were cultured in liquid half-strength Murashige and Skoog (MS) medium or sterile distilled water, and with either perlite or vermiculite supplementation. All the culturing processes were conducted either under sterile or nonsterile condition. The results showed that rooting under sterile condition was better. Shoots cultured in half-strength MS medium supplemented with vermiculite exhibited a 92.5% rooting efficiency while perlite showed 77.5%. The survival rate of the vermiculite-grown transformed papaya plantlets after transfer into soil, contained in polybags, was 94%, and the rate after transfer into the ground was 92%. Morpho-histological analyses revealed that the tap roots were more compact, which might have contributed to the high survival rates of the plantlets. PMID:25969786

An effective protocol for micropropagation of edible bamboo species (Bambusa tulda and Melocanna baccifera) through nodal culture.

Science.gov (United States)

Waikhom, Sayanika Devi; Louis, Bengyella

2014-01-01

High demand for edible bamboo shoots of Bambusa tulda and Melocanna baccifera in many Asian ethnic groups has led to the need for developing intensive bamboo farming. To achieve this, in vitro regeneration of bamboo plantlets is needed due to the long and irregular bamboo flowering cycle and scarcity of bamboo seeds. An effective protocol for plantlets regeneration in B. tulda and M. baccifera from nodal explants following validation of the species using the sequence of trnL-F intergenic spacer region is described. Effective axillary bud breaking was achieved at 3 mg/L of 6-benzylaminopurine (BAP) in MS medium. Importantly, combining 2 mg/L of kinetin (Kn) with 3 mg/L of BAP produced a synergistic effect for shoot multiplication in B. tulda and M. baccifera. Under optimized conditions in half-strength MS medium supplemented with 3 mg/L of indole-3-butyric acid (IBA), 10 mg/L of coumarin, and 3% sucrose, profuse production of dark-brown rhizome in B. tulda and abundant rooting (81.67%, P < 0.05, F = 15.46) for M. baccifera within 30 days were achieved. The established protocol and the validation of the reported species at the molecular level will be of help to stakeholders in edible bamboo trade to conserve gene-pool and increase productivity.

Plant regeneration of Erigeron breviscapus (vant.) Hand. Mazz. and its chromatographic fingerprint analysis for quality control.

Science.gov (United States)

Liu, Chun-Zhao; Gao, Min; Guo, Bin

2008-01-01

An efficient micropropagation system for Erigeron breviscapus (vant.) Hand. Mazz., an important medicinal plant for heart disease, has been developed. Shoot organogenesis occurred from E. breviscapus leaf explants inoculated on a medium supplemented with a combination of plant growth regulators. On average, 17 shoots per leaf explant were produced after 30 days when they were cultured on MS basal salts and vitamin medium containing 5 microM 6-benzylaminopurine (BAP) and 5 microM 1-naphthaleneacetic acid (NAA). All the regenerated shoots formed complete plantlets on a medium containing 2.5-10 microM indole-3-butyric acid (IBA) within 30 days, and 80.2% of the regenerated plantlets survived and grew vigorously in field conditions. Based on the variation in common peaks and the produced amount of the most important bioactive component, scutellarin, a high performance liquid chromatography (HPLC) fingerprinting system was developed for quality control of these micropropagated plants. Chemical constituents in E. breviscapus micropropagated plants varied during plant development from regeneration to maturation, the latter of which showed the most similar phytochemical profile in comparison with mother plants. The regeneration protocol and HPLC fingerprint analysis developed here provided a new approach to quality control of micropropagated plants producing secondary metabolites with significant implications for germplasm conservation.

Mutation breeding for tomato bacterial with disease resistance through in vitro techniques

International Nuclear Information System (INIS)

Keoboonrueng, S.; Charaensatapon, R.; Panichsukpatana, C.; Jatisation, J.

1994-01-01

Cotyledons of half month and one month-old seedlings of tomato varieties Sida, SVRDC 4, Sidatip 2 and VF 137 irradiated with gamma rays at the dose of 10 Gy were cultured on most suitable medium found, MS supplemented with 0.01 mg/1 NAA and 2.0 mg/1 BAP. Sometimes calli and multiple shoots derived from normal seedlings were irradiated with gamma rays at the doses of 5 and 8 Gy, respectively. Plantlets from in vitro culture were screened in the greenhouse by soaking roots in bacterial suspension at transplanting time or by pouring bacterial suspension on injured roots at 2-3 wk. after transplant. The concentration of bacterial suspension was 10 5 -10 7 cells/ml. Total of 2541 tomato plantlets were screened and only 11 plants survived. They were, 5 plants from non-irradiated Sida, 1 and 2 plants from 5 and 10 Gy Sida, respectively, 2 plants from 5 Gy Sidatip 2 and single plant from non-irradiated VF 137. Most of the surviving plants were susceptible to tomato yellow leaf curl virus and only fruits from one Sida plant irradiated with 10 Gy could be harvested. Plants from these seeds will be further selected

Preliminary Assessment about Genetic Diversity, the Stability of Potential Mutants from Two Varieties of Chrysanthemum morifolium Ramat. (Bronze Doa and Purple Farm) via Gamma Irradiation

International Nuclear Information System (INIS)

Nguyen Tuong Mien; Le Ngoc Trieu; Le Tien Thanh; Pham Van Nhi; Huynh Thi Trung

2013-01-01

The aim of this study was to evidence the efficiency of irradiation by gamma ray from 60 Co source on chrysanthemum artificial seeds, through that select a number of potential mutants from two varieties of Chrysanthemum morifolium Ramat artificial seeds (Bronze and purple chrysanthemum). The experimental result showed that LD 50 for the Bronze Doa variety was 50 Gy and Purple Farm variety was 100 Gy. Irradiated in vitro artificial seeds were transferred into fresh MS medium and placed in the growth room with three replications at each of dose. The completed in vitro plants were transplanted into ex vitro condition in green house. After 30 days in the greenhouse, survival rate of plantlets were and growth in 20 Gy and 40 Gy for Purple chrysanthemum, while the number of survival Bronze chrysanthemum plantlets were reduced gradually toward the increasing of gamma doses. In this study, on farm, through screening 18 phenotypic mutants of both chrysanthemums were recorded and collected including 6 potential mutants that selected for next research based on their phenotypic differences to the originals, their aesthetic and low mosaic. These 6 potential mutants together with their original varieties were micro-propagated to induce the potential mutant lines for estimation on farm of mutant characteristic segregation rates. (author)

Demonstration of the economic feasibility of plant tissue culture for jojoba (Simmondsia chinensis) and Euphorbia spp

Energy Technology Data Exchange (ETDEWEB)

Sluis, C.

1980-09-01

The economic feasibility of plant tissue culture was demonstrated as applied to two plants: jojoba (Simmondsia chinensis) and Euphorbia spp. The gopher weed (Euphorbia lathyris) was selected as the species of Euphorbia to research due to the interest in this plant as a potential source of hydrocarbon-like compounds. High yield female selections of jojoba were chosen from native stands and were researched to determine the economic feasibility of mass producing these plants via a tissue culture micropropagation program. The female jojoba selection was successfully mass produced through tissue culture. Modifications in initiation techniques, as well as in multiplication media and rooting parameters, were necessary to apply the tissue culture system, which had been developed for juvenile seedling tissue, to mature jojobas. Since prior attempts at transfer of tissue cultured plantlets were unsuccessful, transfer research was a major part of the project and has resulted in a system for transfer of rooted jojoba plantlets to soil. Euphorbia lathyris was successfully cultured using shoot tip cultures. Media and procedures were established for culture initiation, multiplication of shoots, callus induction and growth, and root initiation. Well-developed root systems were not attained and root initiation percentages should be increased if the system is to become commercially feasible.

Efficient transformation and regeneration of transgenic cassava using the neomycin phosphotransferase gene as aminoglycoside resistance marker gene.

Science.gov (United States)

Niklaus, Michael; Gruissem, Wilhelm; Vanderschuren, Hervé

2011-01-01

Cassava is one of the most important crops in the tropics. Its industrial use for starch and biofuel production is also increasing its importance for agricultural production in tropical countries. In the last decade cassava biotechnology has emerged as a valuable alternative to the breeding constraints of this highly heterozygous crop for improved trait development of cassava germplasm. Cassava transformation remains difficult and time-consuming because of limitations in selecting transgenic tissues and regeneration of transgenic plantlets. We have recently reported an efficient and robust cassava transformation protocol using the hygromycin phosphotransferase II (hptII) gene as selection marker and the aminoglycoside hygromycin at optimal concentrations to maximize the regeneration of transgenic plantlets. In the present work, we expanded the transformation protocol to the use of the neomycin phosphotransferase II (nptII) gene as selection marker. Several aminoglycosides compatible with the use of nptII were tested and optimal concentrations for cassava transformation were determined. Given its efficiency equivalent to hptII as selection marker with the described protocol, the use of nptII opens new possibilities to engineer transgenic cassava lines with multiple T-DNA insertions and to produce transgenic cassava with a resistance marker gene that is already deregulated in several commercial transgenic crops.

Better rooting procedure to enhance survival rate of field grown malaysian eksotika papaya transformed with 1-aminocyclopropane-1-carboxylic Acid oxidase gene.

Science.gov (United States)

Sekeli, Rogayah; Abdullah, Janna Ong; Namasivayam, Parameswari; Muda, Pauziah; Abu Bakar, Umi Kalsom

2013-01-01

A high survival rate for transformed papaya plants when transferred to the field is useful in the quest for improving the commercial quality traits. We report in this paper an improved rooting method for the production of transformed Malaysian Eksotika papaya with high survival rate when transferred to the field. Shoots were regenerated from embryogenic calli transformed with antisense and RNAi constructs of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) genes using the Agrobacterium tumefaciens-mediated transformation method. Regenerated transformed shoots, each measuring approximately 3-4 cm in height, were cultured in liquid half-strength Murashige and Skoog (MS) medium or sterile distilled water, and with either perlite or vermiculite supplementation. All the culturing processes were conducted either under sterile or nonsterile condition. The results showed that rooting under sterile condition was better. Shoots cultured in half-strength MS medium supplemented with vermiculite exhibited a 92.5% rooting efficiency while perlite showed 77.5%. The survival rate of the vermiculite-grown transformed papaya plantlets after transfer into soil, contained in polybags, was 94%, and the rate after transfer into the ground was 92%. Morpho-histological analyses revealed that the tap roots were more compact, which might have contributed to the high survival rates of the plantlets.

Sinorhizobium meliloti can protect Medicago truncatula against Phoma medicaginis attack

Directory of Open Access Journals (Sweden)

Moncef MRABET

2011-09-01

Full Text Available The Sinorhizobium meliloti microsymbiont of Medicago spp. was used in an antibiosis test against Phoma medicaginis and in bioprotection assays of Medicago truncatula JA17 from the pathogen. Among 17 S. meliloti strains isolated from root nodules of M. truncatula and Medicago laciniata grown in Tunisian soils, six showed up to 60% growth inhibition of five P. medicaginis strains isolated from infected field-grown M. truncatula. Two S. meliloti strains with differing in vitro effects on P. medicaginis, 10.16/R6 antagonist and 5M6 non antagonist, were used in a bioprotection assay of M. truncatula JA17 from the pathogen. The inoculation of P. medicaginis caused complete root and stem rotting, and the mortality of all treated plantlets. Inoculation of the antagonist S. meliloti strain 10.16/R6 to M. truncatula JA17 infected with P. medicaginis was associated with a significant 65% decrease of vegetative rotting length, an 80% decrease of plant mortality, an increase of root length, and enhancement of root and shoot biomass comparatively to control plantlets treated with P. medicaginis. The inoculation of the non antagonistic S. meliloti strain 5M6 slightly decreased disease and slightly increased plant growth parameters.

High-Throughput Agrobacterium-mediated Transformation of Medicago Truncatula in Comparison to Two Expression Vectors

International Nuclear Information System (INIS)

Sultana, T.; Deeba, F.; Naqvi, S. M. S.

2016-01-01

Legumes have been turbulent to efficient Agrobacterium-mediated transformation for a long time. The selection of Medicago truncatula as a model legume plant for molecular analysis resulted in the development of efficient Agrobacterium-mediated transformation protocols. In current study, M. truncatula transformed plants expressing OsRGLP1 were obtained through GATEWAY technology using pGOsRGLP1 (pH7WG2.0=OsRGLP1). The transformation efficiency of this vector was compared with expression vector from pCAMBIA series over-expressing same gene (pCOsRGLP1). A lower number of explants generated hygromycin resistant plantlet for instance, 18.3 with pGOsRGLP1 vector as compared to 35.5 percent with pCOsRGLP1 vector. Transformation efficiency of PCR positive plants generated was 9.4 percent for pGOsRGLP1 while 21.6 percent for pCOsRGLP1. Furthermore 24.4 percent of explants generated antibiotic resistant plantlet on 20 mgl/sup -1/ of hygromycin which was higher than on 15 mgl/sup -1/ of hygromycin such as 12.2 percent. T/sub 1/ progeny analysis indicated that the transgene was inherited in Mendelian manner. The functionally active status of transgene was monitored by high level of Superoxide dismutase (SOD) activity in transformed progeny. (author)

IN VITRO PROPAGATION OF DENDROBIUM AND PHALAENOPSIS THROUGH TISSUE CULTURE FOR CONSERVATION

Directory of Open Access Journals (Sweden)

Lita Soetopo

2012-06-01

Full Text Available The studies were focused on developing an efficient and effective propagation protocol for orchid species from genera Dendrobioum and Phalaenopsis through tissue culture. The Materials used were explants from adventive shoot tip, floral stalk buds and PLBs derived from seeds. The results indicated growth and development of adventive shoot tip explants of Dendrobium: a high survival percentage for explant with green color was shown by D. racianum, followed by D. laxiflorum, D. pseudo-conantum, D. strebloceras, D. lineale, and D. veratrifolium. However, plantlets regeneration occurred only on D. pseudoconantum, and D. strebloceras. Explant regeneration from seed derived protocorm-like bodies on D. spectabile occurred 40 days after inoculation transfer and subculture. High survival percentage of explant from floral stalk shoot was shown by P. amabilis. There were several plantlets surviving in acclimatisation. Explant regeneration from seed derived from protocorm-like bodies on P. hieroglypha occurred 40 days after inoculation and subculture. It was suggested that for ex situ conservation on certain species of Dendrobium and Phalaenopsis in the category of rare germplasms, tissue culture could be applied effectively and efficiently by using explant from adventive shoot tip, floral stalk buds and seed derived protocorm-like body explant for vegetative seed multiplication.

A new species of Burkholderia isolated from sugarcane roots promotes plant growth

Science.gov (United States)

Paungfoo-Lonhienne, Chanyarat; Lonhienne, Thierry G A; Yeoh, Yun Kit; Webb, Richard I; Lakshmanan, Prakash; Chan, Cheong Xin; Lim, Phaik-Eem; Ragan, Mark A; Schmidt, Susanne; Hugenholtz, Philip

2014-01-01

Sugarcane is a globally important food, biofuel and biomaterials crop. High nitrogen (N) fertilizer rates aimed at increasing yield often result in environmental damage because of excess and inefficient application. Inoculation with diazotrophic bacteria is an attractive option for reducing N fertilizer needs. However, the efficacy of bacterial inoculants is variable, and their effective formulation remains a knowledge frontier. Here, we take a new approach to investigating diazotrophic bacteria associated with roots using culture-independent microbial community profiling of a commercial sugarcane variety (Q208A) in a field setting. We first identified bacteria that were markedly enriched in the rhizosphere to guide isolation and then tested putative diazotrophs for the ability to colonize axenic sugarcane plantlets (Q208A) and promote growth in suboptimal N supply. One isolate readily colonized roots, fixed N2 and stimulated growth of plantlets, and was classified as a new species, Burkholderia australis sp. nov. Draft genome sequencing of the isolate confirmed the presence of nitrogen fixation. We propose that culture-independent identification and isolation of bacteria that are enriched in rhizosphere and roots, followed by systematic testing and confirming their growth-promoting capacity, is a necessary step towards designing effective microbial inoculants. PMID:24350979

Rice improvement involving altered flower structure more suitable to cross-pollination, using in vitro culture in combination with mutagenesis

International Nuclear Information System (INIS)

Min, S.K.

1998-01-01

Anther and somatic tissue culture in combination with mutagenesis were carried out to evaluate the efficiency of different mutagenic treatments of various in vitro culture materials, and to obtain some promising variants for rice improvement. Results indicated that in japonica rice radiation treatment of dry seeds and young panicles influenced the percentage of green plantlets regeneration from anther culture. Both treatments increased significantly the percentage of regenerated green plant lets in comparison with the control

Draft Genome Sequence of Bacillus velezensis 3A-25B, a Strain with Biocontrol Activity against Fungal and Oomycete Root Plant Phytopathogens, Isolated from Grassland Soil.

Science.gov (United States)

Martínez-Raudales, Inés; De La Cruz-Rodríguez, Yumiko; Vega-Arreguín, Julio; Alvarado-Gutiérrez, Alejandro; Fraire-Mayorga, Atzin; Alvarado-Rodríguez, Miguel; Balderas-Hernández, Victor; Gómez-Soto, José Manuel; Fraire-Velázquez, Saúl

2017-09-28

Here, we present the draft genome of Bacillus velezensis 3A-25B, which totaled 4.01 Mb with 36 contigs, 3,948 genes, and a GC content of 46.34%. This strain, which demonstrates biocontrol activity against root rot causal phytopathogens in horticultural crops and friendly interactions in roots of pepper plantlets, was obtained from grassland soil in Zacatecas Province, Mexico. Copyright © 2017 Martínez-Raudales et al.

Leaf-specific pathogenesis-related 10 homolog, PgPR-10.3, shows in silico binding affinity with several biologically important molecules

Directory of Open Access Journals (Sweden)

Jin Haeng Han

2015-10-01

Conclusion: Although ginseng PR-10.3 gene is expressed in all organs of 3-wk-old plantlets, its expression is restricted to leaves in mature 2-yr-old ginseng plants. The putative binding property of PgPR-10.3 with Re is intriguing. Further verification of binding affinity with other biologically important molecules in the large hydrophobic cavity of PgPR-10.3 may provide an insight into the biological features of PR-10 proteins.

Wheat biotechnology: A minireview

OpenAIRE

Patnaik, Debasis; Khurana, Paramjit

2001-01-01

Due to the inherent difficulties associated with gene delivery into regenerable explants and recovery of plantlets with the introduced transgene, wheat was the last among cereals to be genetically transformed. This review attempts to summarize different efforts in the direction of achieving genetic transformation of wheat by various methods. Particle bombardment is the most widely employed procedure for the introduction of marker genes and also for the generation of transformed wheat with int...

Cryopreservation of chayote (Sechium edule JACQ.SW.) zygotic embryos and shoot-tips from in vitroplantlets

OpenAIRE

Abdelnour-Esquivel, Ana; Engelmann, Florent

2002-01-01

This paper presents the development of cryopreservation protocols for zygotic embryos and apices of chayote (Sechium edule Jacq. Sw.), a tropical plant species with recalcitrant seeds. Zygotic embryos of two cultivars, Ccocro negro (CN) and Claudio (Cl) could withstand cryopreservation, with survival percentages of 10 and 30 %, after desiccation to 23 and 19 % moisture content (fresh weight basis), respectively. Apices sampled on in vitro plantlets of cultivars Cl, 13 and JM we...

Gelling agents and culture vessels affect in vitro multiplication of banana plantlets.

Science.gov (United States)

Kaçar, Y A; Biçen, B; Varol, I; Mendi, Y Y; Serçe, S; Cetiner, S

2010-03-09

Agar is the most commonly used gelling agent in media for plant tissue culture. Because of the high price of tissue-culture-grade agar, attempts have been made to identify suitable alternatives. The type of culture vessel and lid also affects the gaseous composition inside the vessel as well as light penetration. In turn, the vessel affects growth parameters, such as shoot elongation, proliferation and fresh weight, as well as hyperhydric degradation processes. We examined the effects of different culture vessels, including commercial glass jars, magenta boxes, and disposable containers, as well as different gelling agents (agar-agar, Agargel, Phytagel, and plant agar) on the micropropagation of Dwarf Cavendish bananas in an effort to find a combination that yields large numbers of high-quality seedlings. The different culture vessels did not significantly affect seedling culture success. The medium significantly affected shoot weight. Phytagel resulted in the highest shoot weight (overall mean = 2.4 g), while agar, Agargel and plant agar resulted in 1.7, 2.2 and 2.2 g, respectively. Disposable container/Phytagel and Magenta/Agargel combinations yielded the highest shoot weights (2.9 and 3.0 g, respectively). Mean shoot length increased progressively with subculture (four subcultures were made). The highest mean shoot length was obtained with Phytagel and Agargel media (6.4 and 6.3 cm, respectively). Shoot number was significantly affected by medium only at subculture 4. Overall, the highest mean shoot length was obtained with the Magenta/Agargel combination (8.5 cm). Phytagel and plant agar gave higher mean shoot number than agar and Agargel (2.1, 2.1 and 1.7 and 1.9, respectively). The costs of the media and of the culture vessels need to be taken into account for final choice of the banana shoot culture system.

Regeneration of plantlets under NaCl stress from NaN3 treated ...

African Journals Online (AJOL)

use

2011-11-16

Nov 16, 2011 ... plant regeneration under NaCl stressed conditions was assessed in some sugarcane ( ... cell is a useful work for the establishment of pure form of ... balance. The relative water contents (RWC) were calculated .... J. Life Sci.

An assessment of genetic fidelity of in vitro grown plantlets of rose ...

African Journals Online (AJOL)

A simple and routine method for the analysis of somaclonal variation among tissue culture derived rose plants is a prerequisite for precise monitoring of quality control during rapid mass micropropagation. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) molecular marker techniques ...

Manipulating the physiological quality of in vitro plantlets and transplants of potato

NARCIS (Netherlands)

Mehari, T.

2000-01-01

In vitro techniques have been introduced in potato seed production systems in recent years. This research project aimed at studying the morphological and physiological changes in plants and crops in the last three phases of a seed production system that included an

Agrobacterium-mediated transformation of Citrus sinensis and Citrus limonia epicotyl segments

OpenAIRE

Almeida,Weliton Antonio Bastos de; Mourão Filho,Francisco de Assis Alves; Mendes,Beatriz Madalena Januzzi; Pavan,Alexandra; Rodriguez,Adriana Pinheiro Martinelli

2003-01-01

Genetic transformation allows the release of improved cultivars with desirable characteristics in a shorter period of time and therefore may be useful in citrus breeding programs. The objective of this research was to establish a protocol for genetic transformation of Valencia and Natal sweet oranges (Citrus sinensis L. Osbeck) and Rangpur lime (Citrus limonia L. Osbeck). Epicotyl segments of germinated in vitro plantlets (three weeks in darkness and two weeks in a 16-h photoperiod) were used...

CRESCIMENTO E DESEMPENHO DE BERTALHA (Basella alba L.) EM FUNÇÃO DO TIPO DE PROPAGAÇÃO

OpenAIRE

RENÊ ARNOUX DA SILVA CAMPOS; GIUSEPPINA PACE PEREIRA LIMA; SANTINO SEABRA JÚNIOR; WILLIAM HIROSHI SUEKANE TAKATA; EWERTON GASPARETTO DA SILVA

2012-01-01

Malabar spinach is a non-conventional vegetable propagated mainly by seeds. This research evaluated the propagation by stem cuttings as an alternative method of propagation and verified its effect on growth of the vegetable. Plantlets propagated by seeds and stem cuttings were grown in greenhouse and their growth were evaluated in five successive intervals of 21 days each, with the first evaluation carried out 60 days after the transplanting. The results showed that the propagation methods ha...

Cytokinin induced shoot regeneration and flowering of Scoparia dulcis L. (Scrophulariaceae)-an ethnomedicinal herb.

Science.gov (United States)

Premkumar, G; Sankaranarayanan, R; Jeeva, S; Rajarathinam, K

2011-06-01

To develop an improved protocol for micropropagation of ethnomedicinally important Scoparia dulcis (S. dulcis) L. Explants were inoculated on MS basal medium supplemented with kinetin and 6-benzylaminopurine for shoot bud induction. To enhance the shoot induction, various auxins like 3-indoleacetic acid or 3-indolebutyric acid or α-naphthylacetic acid were tested along with 2.32 M KI and 4.44 µM BAP. The regenerated shoots were rooted in half strength MS medium supplemented with various concentrations of IAA, IBA or NAA. After roots were developed, the plantlets were transplanted to pots filled with vermiculate and sand and kept in growth chamber with 70%-80% humidity under 16 h photoperiod. After acclimatization, the plantlets were transferred to the garden and survival percentage was calculated. Data were statistically analyzed and means were compared using Duncan's multiple range test (Pdulcis. Shoot induction on young leaf explants was most successful in MS medium supplemented with combination of two cytokinins (2.32 µM KI and 4.44 µM BAP) 2.85 µM IAA, 10% CM and 1 483.79 µM adenine sulfate. A single young leaf explant was capable of producing 59 shoots after 13 days of culture. Flower was induced in medium supplemented with combination of KI and BAP. Cytokinins are the key factor to induce the direct shoot regeneration and flowering of S. dulcis.

Gerbera micropropagation.

Science.gov (United States)

Cardoso, Jean C; Teixeira da Silva, Jaime A

2013-12-01

Gerbera jamesonii (gerbera) is an important cut-flower in the global floricultural industry. Micropropagation is the main system used to clonally propagate gerbera in vitro resulting in the production of millions of plantlets each year. Numerous types of explants and protocols for micropropagation have been established and used for gerbera. Shoot tips are the commonly used explant while adventitious shoot induction from the capitulum is also a popular method. Most papers in the literature have focused on testing the influence of different types and combinations of plant growth regulators with the aim of improving the regeneration and multiplication stage of one or few cultivars. Genotype is one of the most influential factors on the response of gerbera in vitro. Despite this, no successful universal protocol has yet been developed for multiple cultivars, limiting the usefulness of current protocols for commercial biotechnology labs. Slow-growing endogenous bacteria are one of the most important problems in gerbera micropropagation but require more studies on control and prevention. Individual shoots are normally easy to root, usually in excess of 90% of plantlets, but the acclimatization stage requires improvements and new technologies to increase the survival of plants. Epigenetic variations in micropropagated gerbera are frequently observed only with high concentrations of cytokinins in the culture medium but somaclonal variation is rare. Copyright © 2013 Elsevier Inc. All rights reserved.

In vitro propagation of silybum marianum (l.) gaertn. and genetic fidelity assessment of micropropagated plants

International Nuclear Information System (INIS)

Lv, Y.W.; Wamg, R.J.; Yang, Z.S.; Wang, Y.J.

2017-01-01

Silybum marianum (milk thistle), an annual and biennial herbaceous plant, has been used to treat liver disease for 2000 years. An efficient system for micropropagation from leaf explants of wild-grown S. marianum was successfully established and the genetic fidelity of micropropagated plants was assessed using PCR-based random amplified polymorphic DNA (RAPD). The highest frequency (98.9%) of callus induction was obtained from explants cultivated for 4 weeks on Murashige and Skoog (MS) medium supplemented with 1.5 mg l/sup -1/N6-Benzylaminopurine (BA) and 2.0 mg l/sup -1/ alpha-Naphthaleneacetic acid (NAA). The optimal medium for shoots organogenesis was MS supplemented with 3.0 mg l/sup -1/ Gibberellic acid (GA/sub 3/) and 1.0 mg l/sup -1/NAA, while shoot proliferation was accomplished in MS supplemented with 1.0 mg l/sup -1/BA and 0.3 mg l/sup -1/NAA. Rooting (94.5%) of these shoots was achieved after 4 weeks on 1/2 MS medium supplemented with 1.0 mg l/sup -1/ IBA. A total of 92% of the plants were surviving 6 weeks after transplantation of plantlets to soil. RAPD analysis revealed that the genetic fidelity between the parent and the In vitro-raised plantlets exhibited 100% similarity. This confirmed the true-to-type nature of the In vitro-raised clones. |(author)

An Improved Micropropagation Protocol by Ex Vitro Rooting of Passiflora edulis Sims. f. flavicarpa Deg. through Nodal Segment Culture.

Science.gov (United States)

Shekhawat, Mahipal S; Manokari, M; Ravindran, C P

2015-01-01

A procedure for rapid clonal propagation of Passiflora edulis Sims. f. flavicarpa Deg. (Passifloraceae) has been developed in this study. Nodal explants were sterilized with 0.1% HgCl2 and inoculated on Murashige and Skoog (MS) basal medium. The addition of 2.0 mgL(-1) 6-benzylaminopurine (BAP) to MS medium caused an extensive proliferation of multiple shoots (8.21 ± 1.13) primordial from the nodal meristems. Subculturing of these multiple shoots on the MS medium augmented with 1.0 mgL(-1) of each BAP and Kinetin (Kin) was successful for the multiplication of the shoots in vitro with maximum numbers of shoots (25.73 ± 0.06) within four weeks of incubation. Shoots were rooted best (7.13 ± 0.56 roots/shoots) on half strength MS medium supplemented with 2.0 mgL(-1) indole-3 butyric acid (IBA). All in vitro regenerated shoots were rooted by ex vitro method, and this has achieved 6-7 roots per shoot by pulsing of cut ends of the shoots using 200 as well as 300 mgL(-1) IBA. The plantlets were hardened in the greenhouse for 4-5 weeks. The hardened plantlets were shifted to manure containing nursery polybags after five weeks and then transferred to a sand bed for another four weeks for acclimatization before field planting with 88% survival rate.

Production of virus-free orchid Cymbidium aloifolium (L.) Sw. by various tissue culture techniques.

Science.gov (United States)

Pradhan, Shreeti; Regmi, Tripti; Ranjit, Mukunda; Pant, Bijaya

2016-10-01

Orchids are affected by many viruses resulting in poor growth, yield and quality, and an overall decline in population. Cymbidium mosaic virus (CymMV) is one of the common orchid viruses found in Cymbidium species but it infects different orchid genera. In this study Cymbidium aloifolium was propagated in vitro using MS medium at different strength (1.0, ½, and ¼) with or without 0.5 mg/l BAP (6-benzylaminopurine) and 0.5 mg/l NAA (Naphthalene acetic acid). To provide disease-free planting material, source plant for in vitro propagation needs to be screened for pathogenic viruses. In the present study, in vivo -grown source (mother) plants and tissue culture-derived plants of C. aloifolium were tested for CymMV virus using Double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA). All the tissue cultured plants were found to be 100% virus-free whereas the in vivo grown source plants were highly affected by CymMV virus (83.33%). The virus-free in vitro plantlets were multiplied in large scale and then acclimatized on earthen pot containing a mixture of cocopeat, litter and clay in the ratio of 3:2:1. Eighty five percent of acclimatized plantlets survived making this method an efficient mass production system for high quality virus-free C. aloifolium for commercial floriculture and germplasm preservation.

Production of virus-free orchid Cymbidium aloifolium (L. Sw. by various tissue culture techniques

Directory of Open Access Journals (Sweden)

Shreeti Pradhan

2016-10-01

Full Text Available Orchids are affected by many viruses resulting in poor growth, yield and quality, and an overall decline in population. Cymbidium mosaic virus (CymMV is one of the common orchid viruses found in Cymbidium species but it infects different orchid genera. In this study Cymbidium aloifolium was propagated in vitro using MS medium at different strength (1.0, ½, and ¼ with or without 0.5 mg/l BAP (6-benzylaminopurine and 0.5 mg/l NAA (Naphthalene acetic acid. To provide disease-free planting material, source plant for in vitro propagation needs to be screened for pathogenic viruses. In the present study, in vivo-grown source (mother plants and tissue culture-derived plants of C. aloifolium were tested for CymMV virus using Double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA. All the tissue cultured plants were found to be 100% virus-free whereas the in vivo grown source plants were highly affected by CymMV virus (83.33%. The virus-free in vitro plantlets were multiplied in large scale and then acclimatized on earthen pot containing a mixture of cocopeat, litter and clay in the ratio of 3:2:1. Eighty five percent of acclimatized plantlets survived making this method an efficient mass production system for high quality virus-free C. aloifolium for commercial floriculture and germplasm preservation. Keywords: Biological sciences, Plant biology

Development of seedless fruits mutants in citrus including tangerine (C. reticulata) and pummelo (C. grandis) through induced mutations and biotechnology

Energy Technology Data Exchange (ETDEWEB)

Somsri, S. [Horticulture Research Institute, Department of Agriculture (DOA), Chatuchak, Bangkok (Thailand); Jompook, P. [Department of Applied Radiation and Isotopes, Faculty of Science, Kasetsart University, Chatuchak, Bangkok (Thailand); Kanhom, P. [Phare Horticultural Research Center, Muang, Phare (Thailand); Thayamanont, P.; Meecharoen, S. [Pichit Horticultural Research Center, Muang, Pichit (Thailand); Kumcha, U. [Srisaket Horticultural Research Center, Muang, Srisaket (Thailand)

2009-05-15

The development of seedless fruit mutants in citrus, including Tangerine (C. reticulata) and Pummelo (C. grandis), through induced mutation and biotechnology was studied at the Gamma Irradiation Service and Nuclear Technology Center, Pichit and Phare Horticultural Research Center for 4 years (August 2000 to September 2004). The results showed successful induction of mutants with gamma irradiation using both chronic and acute procedures for pot plants, scions and in vitro plantlets of tangerine (Citrus reticulata var. Shogun and Sai Nam Puaeng) and pummelo (Citrus grandis viz. Kao Thong Dee). MS medium with 2 mgL{sup -1} of BA was found to be the most suitable medium for shoot proliferation. The seedlings were sub-cultured at least 4 times, and then they were treated with acute and chronic irradiation. Shoot induction from M{sub 1}V{sub 0} to M{sub 1}V{sub 4} generation was performed in basic MS medium with 2 mgL{sup -1} added BA. Rooting was induced in the M{sub 1}V{sub 4} in halfstrength MS enriched with BA 2 mgL{sup -1}. Later, the shoots were excised and grafted on mature plants or the plantlets directly transferred in the field and later the fruits from mature trees were evaluated for seedlessness in M{sub 1}V{sub 4} at Pichit and Phare Horticultural Research Center. (author)

Phenotypic aberrations during micropropagation of Soymida febrifuga (Roxb. Adr. Juss

Directory of Open Access Journals (Sweden)

Kishore Kumar CHIRUVELLA

2014-03-01

Full Text Available Like most of the medicinal plants Soymida febrifuga (Meliaceae possess significance for its valuable secondary metabolites. Multiplication of this endemic plant is limited by difficulty in rooting of stem cuttings, high seedling mortality rates and low seed viability period. Hence efficient protocols for in vitro mass propagation has been established from field grown and aseptic seedlings explants. Strikingly, we observed aberrant structures such as vitrified shoots, faciated shoots, albino shoots as well shoot necrosis during its micropropagation. These phenotypic maladies were observed during organogenesis and rooting. Compared to other abnormalities, shoot necrosis nonetheless was frequent and pronounced leading to plant death. Shoots when subjected to rooting also displayed necrosis which was controlled by transferring to MS medium containing various concentrations and combinations of calcium levels, activated charcoal, glucose, fructose and auxins. Microshoots initiated roots on half strength MS medium with IBA and IAA individually or in combination within two weeks. MS half strength solid medium supplemented with CAN (556 mg l–1, CAP (1.0 mg l–1, IAA (2.0 mg l–1 and IBA (2.0 mg l–1 in combination was found to be more efficient in showing high frequency (95% of root regeneration. Rooted plantlets were successfully hardened and 70-85% of regenerated plants were successfully acclimatized to natural environment. In vitro derived plantlets were morphologically similar to in vivo plants.

Evaluation of genetic variability in micropropagated propagules of ornamental pineapple [Ananas comosus var. bracteatus (Lindley) Coppens and Leal] using RAPD markers.

Science.gov (United States)

Santos, M D M; Buso, G C S; Torres, A C

2008-10-21

The objective of the present study was to evaluate the genetic variability in micropropagated plantlets of ornamental pineapple, after the fourth period of subculture. The basal culture medium consisted of MS salts, vitamins, 3% sucrose, liquid formulation, supplemented with 6-benzylaminopurine (BAP) at concentrations of 0.125, 0.25, 0.5, 1.0, and 2.0 mg/L. The addition of BAP influenced the occurrence of genetic variation revealed using random amplified polymorphic DNA (RAPD) markers. Of a total of 520 primers tested, 44 were selected and amplified; 402 monomorphic bands (97.2%) and 18 polymorphic bands (2.8%) resulted among regenerated plantlets. The polymorphic fragments were produced by 12 primers (OPA-01, OPA-20, OPB-01, OPB-19, OPC-19, OPF-13, OPL-17, OPM-13, OPP-16, OPT-07, OPV-19, and OPX-03). Among the primers that identified polymorphism, OPA-01, OPA-20, OPB-19, OPC-19, OPL-17, OPP-16, and OPX-3 each showed, one polymorphic band and OPF-13 amplified a maximum of three bands. In this study, the RAPD technique was effective in showing the occurrence of somaclonal variations that occur during the micropropagation process of ornamental pineapple cultivation in BAP-supplemented medium, and it is possible to detect the presence of genetic variation in early stages of plant development.

Alginate Encapsulation of Begonia Microshoots for Short-Term Storage and Distribution

Directory of Open Access Journals (Sweden)

Hamidou F. Sakhanokho

2013-01-01

Full Text Available Synthetic seeds were formed from shoot tips of two in vitro grown Begonia cultivars using 3% sodium alginate in Murashige and Skoog medium (MS salt solution as the gel matrix and 100 mM calcium chloride for complexation. Synthetic seed formation was achieved by releasing the sodium alginate/explant combination into 100 mM calcium chloride (CaCl2·H2O solution for 30 or 45 min. Both control and encapsulated shoots were transferred into sterile Petri dishes and stored at 4°C or 22°C for 0, 2, 4, 6, or 8 weeks. Conversion of synthetic seeds into plantlets for both storage environments was assessed in MS medium or peat-based substrate. No significant difference was found between the 30 and 45 min CaCl2·H2O treatments or the two cultivars. Encapsulation of explants improved survival rate over time irrespective of the medium type or storage environment. Survival rates of 88, 53, 28, and 11% for encapsulated microshoots versus 73, 13, 0, and 0% for control explants were achieved in microshoots stored for 2, 4, 6, and 8 weeks, respectively. The best results were obtained when synthetic seeds were stored at 4°C and germinated on MS medium. Regenerated plantlets were successfully established in potting soil.

Cryopreservation of in vitro grown shoot tips and apical meristems of the forage legume Arachis pintoi.

Science.gov (United States)

Rey, Hebe Y; Faloci, Mirta; Medina, Ricardo; Dolce, Natalia; Mroginski, Luis; Engelmann, Florent

2009-01-01

A cryopreservation protocol using the encapsulation-dehydration procedure was established for shoot tips (2-3 mm in length) and meristems (0.3-0.5 mm) sampled from in vitro plantlets of diploid and triploid cytotypes of Arachis pintoi. The optimal protocol was the following: after dissection, explants were precultured for 24 h on establishment medium (EM), encapsulated in calcium alginate beads and pretreated in liquid EM medium with daily increasing sucrose concentration (0.5, 0.75, 1.0 M) and desiccated to 22-23 percent moisture content (fresh weight basis). Explants were frozen using slow cooling (1 C per min from 25C to -30C followed by direct immersion in liquid nitrogen), thawed rapidly and post-cultured in liquid EM medium enriched with daily decreasing sucrose concentrations (0.75, 0.50, 0.1 M). Explants were then transferred to solid EM medium in order to achieve shoot regeneration, then on Murashige and Skoog medium supplemented with 0.05 microM naphthalene acetic acid to induce rooting of shoots. With this procedure, 53 percent and 56 percent of cryopreserved shoot tips of the diploid and triploid cytotypes, respectively, survived and formed plants. However, only 16 percent of cryopreserved meristems of both cytotypes regenerated plants. Using ten isozyme systems and seven RAPD profiles, no modification induced by cryopreservation could be detected in plantlets regenerated from cryopreserved material.

Propagation of Drosera rotundifolia and Drosera capensis in an in vitro Culture System

OpenAIRE

Ileana MICLEA; Marius ZĂHAN

2017-01-01

Drosera rotundifolia and Drosera capensis (Droseraceae) are carnivorous plants grown as ornamentals and sources for homeopathic medicine. The aim of this study was to optimize nutrient and growth regulator concentrations for the in vitro propagation of these species. Half strength MS medium (1/2MS) was supplemented with kinetin (0.5, 2, 5 mg/l) or 6-benzyladenine (3, 5 mg/l) and plantlets were transferred to 1/2MS with or without cytokinins. After 8 weeks rosette diameter, plant height, numbe...

Effect of Silver Nitrate DuringEx vitro Acclimatization of Micropropagated Ginger Cultivars

Directory of Open Access Journals (Sweden)

Dikash Singh THINGBAIJAM

2014-03-01

Full Text Available Silver nitrate (AgNO3 was used under in vitro conditions to study the response of ginger cultivars ‘Nadia’ and ‘Baishey’ under ex vitro. Micropropagated plants treated with AgNO3 showed significant difference (p<0.05 compared to those plantlets without AgNO3 and control type in almost all the different quantitative traits analyzed. Significant difference in number of finger per plant and minirhizome yield indicated the repercussion of AgNO3 during acclimatization.

In vitro organogenesis optimization and plantlet regeneration in Citrus sinensis and C. limonia

Directory of Open Access Journals (Sweden)

Almeida Weliton Antonio Bastos de

2002-01-01

Full Text Available Exogenous genes can be introduced in plants by genetic transformation techniques. However, an efficient tissue culture system with high rates of plant recovery is necessary for gene introduction. This work aimed to define organogenesis and plant regeneration protocols for sweet orange varieties Natal, Valencia and Hamlin (Citrus sinensis L. Osbeck and Rangpur lime (Citrus limonia L. Osbeck which can be used in plant transformation experiments. Seeds of which teguments were removed, were germinated in vitro and maintained in the dark for three weeks, followed by one week at 16-h photoperiod (40 µmol m-2 s-1 and 27 ± 2°C. Organogenesis induction was done by introducing epicotyl segments in MT medium with 25 g L-1 sucrose and different BAP concentrations. After adventitious bud growth, the shoots were transferred to MT medium with either NAA or IBA (1 mg L-1, or absence of auxin, for rooting. The best results were obtained with 1 mg L-1 BAP for bud induction and 1 mg L-1 IBA for rooting for all three sweet orange cultivars. The use of 0.5-2.5 mg L-1 BAP, followed by 1 mg L-1 IBA were the best growth regulator combinations for bud induction and rooting, respectively, for 'Rangpur' lime. The protocols presented in this work are suitable for associations with genetic transformation experiments for these cultivars.

Selection of suitable propagation method for consistent plantlets production in Stevia rebaudiana (Bertoni)

OpenAIRE

Khalil, Shahid Akbar; Zamir, Roshan; Ahmad, Nisar

2014-01-01

Stevia rebaudiana (Bert.) is an emerging sugar alternative and anti-diabetic plant in Pakistan. That is why people did not know the exact time of propagation. The main objective of the present study was to establish feasible propagation methods for healthy biomass production. In the present study, seed germination, stem cuttings and micropropagation were investigated for higher productivity. Fresh seeds showed better germination (25.51–40%) but lost viability after a few days of storage. In o...

Protein as a sole source of nitrogen for in vitro grown tobacco plantlets

Czech Academy of Sciences Publication Activity Database

Synková, Helena; Hýsková, V.; Garčeková, K.; Křížová, S.; Ryšlavá, H.

2016-01-01

Roč. 60, č. 4 (2016), s. 635-644 ISSN 0006-3134 Institutional support: RVO:61389030 Keywords : glutamate-dehydrogenase * inorganic nitrogen * nitrate transport * organic nitrogen * amino-acids * metabolism * seedlings * leaves * roots * assimilation * ammonium * casein * chlorophyll fluorescence * nitrate * photosynthesis Subject RIV: EF - Botanics Impact factor: 1.551, year: 2016

The mechanism of ethylene signaling induced by endophytic fungus Gilmaniella sp. AL12 mediating sesquiterpenoids biosynthesis in Atractylodes lancea

Directory of Open Access Journals (Sweden)

Jie eYuan

2016-03-01

Full Text Available Ethylene, the first known gaseous phytohormone, is involved in plant growth, development as well as responses to environmental signals. However, limited information is available on the role of ethylene in endophytic fungi induced secondary metabolites biosynthesis. Atractylodes lancea is a traditional Chinese herb, and its quality depends on the main active compounds sesquiterpenoids. This work showed that the endophytic fungus Gilmaniella sp. AL12 induced ethylene production in Atractylodes lancea. Pre-treatment of plantlets with ethylene inhibiter aminooxyacetic acid (AOA suppressed endophytic fungi induced accumulation of ethylene and sesquiterpenoids. Plantlets were further treated with AOA, salicylic acid (SA biosynthesis inhibitor paclobutrazol (PAC, jasmonic acid inhibitor ibuprofen (IBU, hydrogen peroxide (H2O2 scavenger catalase (CAT, nitric oxide (NO-specific scavenger 2-(4-Carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO. With endophytic fungi inoculation, IBU or PAC did not inhibit ethylene production, and JA and SA generation were suppressed by AOA, showing that ethylene may act as an upstream signal of JA and SA pathway. With endophytic fungi inoculation, CAT or cPTIO suppressed ethylene production, and H2O2 or NO generation was not affected by 1-aminocyclopropane-1-carboxylic acid (ACC, showing that ethylene may act as a downstream signal of H2O2 and NO pathway. Then, plantlets were treated with ethylene donor ACC, JA, SA, H2O2, NO donor sodium nitroprusside (SNP. Exogenous ACC could trigger JA and SA generation, whereas exogenous JA or SA did not affect ethylene production, and the induced sesquiterpenoids accumulation triggered by ACC was partly suppressed by IBU and PAC, showing that ethylene acted as an upstream signal of JA and SA pathway. Exogenous ACC did not affect H2O2 or NO generation, whereas exogenous H2O2 and SNP induced ethylene production, and the induced sesquiterpenoids

Early selection of elite clones of an ornamental bromeliad in vitro Seleção precoce in vitro de clones elite de uma bromélia ornamental

Directory of Open Access Journals (Sweden)

Candida Elisa Manfio

2010-07-01

Full Text Available Orthophytum grossiorum is a typical bromeliad from Atlantic forestry threatened of extinction. The objectives of this research were to select O. grossiorum clones with ornamental values easy to propagate in vitro, and establish in vitro propagation protocols for these clones. The project was developed in three steps: germination and in vitro selection of seedlings responsive to BAP (6-benzylaminopurine, selection of clones with ornamental values, and establishment of protocol for in vitro propagation of the selected clones. In the first step only 18.33% of plantlets germinated in vitro were responsive to BAP. These plantlets were selected and replicated in vitro several times, each replicated plantlet constituting a clone. In the second step these clones were established ex vitro and surveyed for ornamental attributes. Five out of 11 clones were selected in this step. These clones presented distinct phenotypic traits and were considered of high ornamental quality. In the third step a protocol for in vitro propagation was developed for each selected clone.Orthophytum grossiorum é uma bromélia ameaçada de extinção típica de Mata Atlântica. Os objetivos deste trabalho foram selecionar clones de O. grossiorum com potencial ornamental e de fácil propagação in vitro e estabelecer protocolo de propagação in vitro para esses clones. O trabalho foi desenvolvido em três etapas: germinação e em seleção in vitro de plântulas responsivas a BAP (6-benzylaminopurine, seleção de clones com valores ornamentais e estabelecimento de protocolo para propagação in vitro dos clones selecionados. Na primeira etapa, foi observado que apenas 18.33% das plântulas germinadas in vitro eram responsivas a BAP. Essas plântulas foram selecionadas e reproduzidas em in vitro, e cada plântula selecionada e reproduzida constituiu um clone. Na segunda etapa, esses clones foram estabelecidos ex vitro e selecionados em relação aos atributos ornamentais

Regeneration of three sweet potato (Ipomea batatas (L.)) accessions via meristem, Nodal and callus induction

International Nuclear Information System (INIS)

Addae-Frimpomaah, F.

2012-11-01

In vitro regeneration of three sweet potato accessions UE007, UK-BNARI and SA-BNARI using meristem, nodal cuttings or callus induction was studied. Meristematic explants cultured on Murashige and Skoog (1962) basal medium supplemented with low concentration of benzylaminopurine (BAP) or kinetin resulted in callus with or without shoot development which delayed shoot emergence. The degree of callus development increased as the concentration of the cytokinin in the culture medium increased. Although, callus development was comparatively lower on kinetin amended medium than BAP amended medium, Murashige and Skoog medium supplemented with 0.25mg/1BAP had the highest shoot induction (80%). For further differentiation of callus or shoots into distinct stem and leaves, the culture were transferred into fresh MS medium supplemented with 0.25mg/1 BAP, 0.1 mg/1 NAA and 0.1 mg/1 Gibberellic acid (GA 3 . To overcome the delay in shoot initiation using meristem culture, nodal cuttings of sweet potato were used as explants and cultured on MS medium amended with 0.3 - 0.9mg/1 BAP. All explants cultured on 0.3 or 0.6mg/1 BAP developed shoots. Furthermore, liquid MS medium amended with 0.25mg/1 BAP, 0.1mg/I NAA, and 0.1mg/1 GA 3 also enhanced early shoot development from nodal cutting explants compared to solid culture. Post flask acclimatisation of meristem or nodal cutting-derived plantlets showed that meristem derived plantlets were better acclimatised than nodal cutting plants due to vigorous root development leading to higher percentage survival in pots and subsequent tuber production. Callusogenesis was achieved when leaf lobe explants were cultured on CLC/ Ipomoea medium supplemented with 1.0 - 4.0mg/1 2,4-D with 4.0mg/1 2,4-D being the optimal concentration. However, the calli were non-embryogenic and therefore could not produce embryos when transferred to 0.1mg/1 BAP amended medium but rather produced either single or multiple shoots. The highest percentage shoot (83

Utilization of gamma radiation for the induction of phenotypical variability in hybrid petunia cultivated in vitro

International Nuclear Information System (INIS)

Miranda Jimenez, Javier I.

2006-01-01

Conditions of in vitro culture of two lines of yellow petunia (hybrid petunia) were studied. The effect of two controllers of growth in the establishment was evaluated. It was observed that the lines improved their growth in the absence of BAP (6-benzil amino purine) and with addition of AIA (3-indoleacetic acid) in doses of 0,5 to 2 mg 1 - 1 1 (milligrams by litre). The BAP in the used concentrations (0, 1, 0,2, 0,5 mg 1 -1 ), produced toxic effects to the plantlets growth, presented as hyperhydricity and backwardness. Every line answered better to a different combination of regulators. Therefore, it was defined for the meristems sowing the use of a MS means (Murashige and Skoog, 1962) without regulators for the AMW3A line and a MS means with addition of 2 mg 1 -1 of AIA for the YELLOWAAS line. A radio sensibility curve for the AMW3A line was established. The irradiation process was continued only with the AMW3A line because this is the principal cause of the commercial hybrids that are proving in the country. The mortality increase followed to a lineal model with the increase of the gamma radiation doses was observed. It was established that lethal media doses was of 3 krad. Criteria of surviving and growth of irradiated material was used and it was selected a doses of 2 krad for treatment of a more quantity of plantlets. The growth, with respect to witness, was reduced to half to the four weeks of the irradiation and did not make root. Plantlets with deformation of leafs and chlorophyllous chimeras were observed. The plants that survived to the irradiation process were multiplied and, later, they were acclimatized in greenhouse conditions. An visual analyses of the morphological variations was performed, and the possible phenotypical variation of these plants were determined. Modifications in the form of the plant, the leaf and the flower were found. Nevertheless, the presence of some of these variations in the witness plants suggests that much of the variations

Rooting of west indian cherry cuttings under indo-butiric acid concentrations / Enraizamento de estacas de acerola sob concentrações de ácido indol-butírico

Directory of Open Access Journals (Sweden)

Thiago Luiz Ragugnetti Furlaneto

2003-05-01

Full Text Available Oncidium varicosum is a native Brazilian orchid popularly known as ‘Golden Shower´ because of its very ramified inflorescence and many yellow flowers. The carboydrate type and concentration are important in promoting plantlet development of in vitro orchids. The present study was carried out to asses the effect of different carbohydrate sources and concentrations on the in vitro growth of O. varicosum plantlets. Murashige e Skoog culture medium was used modified with half concentration of the macronutrients. The plantlets, derived from seeds that were already established in vitro and 0.8 + 0.2 cm in height, were inoculated in the culture media containing the following carbohydrate sources: saccharine, maltose and glucose, at concentrations of 0, 10, 20, 30, 60 and 90 g.L-1. The following variables were analyzed 8 months later: canopy height, number of roots, greatest root length, pseudobulb diameter and fresh weight. A completely randomized block experimental design was used with five replications per treatment. Analysis of variance and the Tukey test (5% were performed to compare the means. It was concluded that 60 g.L-1 saccharose was the best treatment for all the parameters assessed. The sugars 30 g.L-1 glucose and 60 g.L-1 maltose were also suitable, but presented lower pseudobulb diameter and lower fresh weight when compared to 60 g.L-1saccharose.O trabalho teve o objetivo de avaliar a influência do ácido indol-butírico sobre o enraizamento de estacas de acerola (Malpighia emarginata D. C. na Universidade Estadual de Londrina. As plantas matrizes utilizadas pertencem a seleção Camb-6 do IAPAR, tiveram suas estacas colhidas com 10 a 12 cm de comprimento, com três a quatro pares de folhas e foram submetidas a uma imersão rápida (10 segundos em soluções de ácido indol-butírico (AIB com cinco concentrações (0, 500, 1.000, 1.500, 2.000 mg L-1. O delineamento estatístico adotado foi o de blocos ao acaso com cinco repeti

GROWTH AND ROOTING SYSTEM OF ACACIA MANGIUM OBTAINED BY TISSUE CULTURE

Directory of Open Access Journals (Sweden)

SUPRIYANTO

1991-01-01

Full Text Available Since 1980/1981, the government of Indonesia through the Ministry of Forestry has started to reforest logged-over, alang-alang, unproductive areas and to convert them to Forest Industry Plantation. The target is 300 000 ha per year. It means, 750 million seedlings should be provided per year (planting distance 2 m x 2 m. The tree species to be planted in forest industry plantation should have shorter life cycle (8 - 10 years, good stem-form, good rooting system, and should be fast growing. Acacia mangium has been selected as one of the important tree species for forest industry plantation due to its growth, quality of fiber wood (pulp and paper industry and rooting system (produce a lot of secondary root and nitrogen fixater (Soebardjo 1986. The reforestation of logged-over Dipterocarp forests in Malaysia with A. mangium has also been considered (Appanah and Weinland 1989. Generally, reforestation with A. mangium is done with seedlings obtained by seed germination. A. mangium produce a lot of seeds but its production is still limited by the season, while the conventional method of vegetative propagation through cuttings gave very low percentage of rooted-cuttings (1% (Umboh and Syamsul Yani 1989. The micropropagation of A. mangium through tissue culture is a promising method. The production of A. mangium plantlets through that method has been done at the Forest Genetic Laboratory, Tropical Forest Biology, SEAMEO BIOTROP (Situmorang 1988, Umboh 1988, Umboh et al. 1989, 1990. These rooted-plantlets (plantlings were first put in the green house (acclimatization before planting in the field. Field tests of some agricultural plants have been done but information on forest trees species is still lacking because the production of plantlings through tissue culture is still limited as there are still problems of their rooting. In fact, the progress of reproducing woody plants by tissue culture has been much slower than with herbaceous plants. The major

Development of a vinasse culture medium for plant tissue culture

International Nuclear Information System (INIS)

Silva, A.L.L.D.; Gollo, L.

2014-01-01

Vinasse is the main pollutant (effluent) obtained from the distillation of sugarcane in the production of fuel alcohol. However, this residue is rich in nutrients that are required by plants. We developed a new culture medium using vinasse for the In vitro propagation of an orchid. The vinasse was treated (decanted and filtered), and the nutrients were determined and quantified. Different formulations using vinasse were tested for an In vitro culture. The vinasse dilutions demonstrated a good buffering effect. The ideal vinasse dilution for media formulation was 2.5%. The best KC formulations with vinasse were KCV1 and KCV5. Compared to KC medium, these formulations demonstrated similar results for In vitro multiplication, with the exception of protocorm-like body number, which was inferior in the vinasse formulations. Conversely, for In vitro elongation and rooting, these vinasse media were superior to KC medium. KC medium promotes a low rooting rate (8%) compared to 68 and 100% obtained by KCV1 and KCV5, respectively. Moreover, plantlets cultured on KC medium become protocorm-like body clusters, which impeded the acclimatization of these explants. Plantlets elongated and rooted on KCV1 and KCV5 were successfully acclimatized with a 91% survival rate for both KC vinasse formulations. This study shows the great potential of this technology as a rational alternative to vinasse disposal and adds value to what is currently considered a waste product. (author)

Identification and evaluation of two diagnostic markers linked to Fusarium wilt resistance (race 4) in banana (Musa spp.).

Science.gov (United States)

Wang, Wei; Hu, Yulin; Sun, Dequan; Staehelin, Christian; Xin, Dawei; Xie, Jianghui

2012-01-01

Fusarium wilt caused by the fungus Fusarium oxysporum f. sp. cubense race 4 (FOC4) results in vascular tissue damage and ultimately death of banana (Musa spp.) plants. Somaclonal variants of in vitro micropropagated banana can hamper success in propagation of genotypes resistant to FOC4. Early identification of FOC4 resistance in micropropagated banana plantlets is difficult, however. In this study, we identified sequence-characterized amplified region (SCAR) markers of banana associated with resistance to FOC4. Using pooled DNA from resistant or susceptible genotypes and 500 arbitrary 10-mer oligonucleotide primers, 24 random amplified polymorphic DNA (RAPD) products were identified. Two of these RAPD markers were successfully converted to SCAR markers, called ScaU1001 (GenBank accession number HQ613949) and ScaS0901 (GenBank accession number HQ613950). ScaS0901 and ScaU1001 could be amplified in FOC4-resistant banana genotypes ("Williams 8818-1" and Goldfinger), but not in five tested banana cultivars susceptible to FOC4. The two SCAR markers were then used to identify a somaclonal variant of the genotype "Williams 8818-1", which lost resistance to FOC4. Hence, the identified SCAR markers can be applied for a rapid quality control of FOC4-resistant banana plantlets immediately after the in vitro micropropagation stage. Furthermore, ScaU1001 and ScaS0901 will facilitate marker-assisted selection of new banana cultivars resistant to FOC4.

Cadmium-induced genomic instability in Arabidopsis: Molecular toxicological biomarkers for early diagnosis of cadmium stress.

Science.gov (United States)

Wang, Hetong; He, Lei; Song, Jie; Cui, Weina; Zhang, Yanzhao; Jia, Chunyun; Francis, Dennis; Rogers, Hilary J; Sun, Lizong; Tai, Peidong; Hui, Xiujuan; Yang, Yuesuo; Liu, Wan

2016-05-01

Microsatellite instability (MSI) analysis, random-amplified polymorphic DNA (RAPD), and methylation-sensitive arbitrarily primed PCR (MSAP-PCR) are methods to evaluate the toxicity of environmental pollutants in stress-treated plants and human cancer cells. Here, we evaluate these techniques to screen for genetic and epigenetic alterations of Arabidopsis plantlets exposed to 0-5.0 mg L(-1) cadmium (Cd) for 15 d. There was a substantial increase in RAPD polymorphism of 24.5, and in genomic methylation polymorphism of 30.5-34.5 at CpG and of 14.5-20 at CHG sites under Cd stress of 5.0 mg L(-1) by RAPD and of 0.25-5.0 mg L(-1) by MSAP-PCR, respectively. However, only a tiny increase of 1.5 loci by RAPD occurred under Cd stress of 4.0 mg L(-1), and an additional high dose (8.0 mg L(-1)) resulted in one repeat by MSI analysis. MSAP-PCR detected the most significant epigenetic modifications in plantlets exposed to Cd stress, and the patterns of hypermethylation and polymorphisms were consistent with inverted U-shaped dose responses. The presence of genomic methylation polymorphism in Cd-treated seedlings, prior to the onset of RAPD polymorphism, MSI and obvious growth effects, suggests that these altered DNA methylation loci are the most sensitive biomarkers for early diagnosis and risk assessment of genotoxic effects of Cd pollution in ecotoxicology. Copyright © 2016 Elsevier Ltd. All rights reserved.

The effect of Piriformospora indica inoculation on salt and drought stress tolerance in Stevia rebaudiana under in vitro conditions

Directory of Open Access Journals (Sweden)

Fahimeh Seraj

2016-09-01

Full Text Available In order to investigate the effect of Piriformospora indica under salt and drought stresses on some vegetative characteristics and physiological parameters of stevia (Stevia rebaudiana Bertoni medicinal plant, an experiment was conducted in factorial arrangement based on completely randomized design with three replicates at Genetics and Agricultural Biotechnology Institute in Sari Agricultural Sciences and Natural Resources University. Factors include three levels of osmatic potential (0, -5, and -10 bar and with three osmotic sources including NaCl (Na, Mannitol (M and NaCl+Mannitol (N+M and inoculation of mycorrhizae like fungi at two levels (non-inoculated and inoculation with fungi. The plantlets were treated for 30 days and then some morphological and physiological parameters were measured. Results of ANOVA showed that there was a significant interaction between osmatic source and levels with fungi inoculation for the most determined parameters. Inoculation of stevia plantlets with P. indica at osmatic level of -5 bar caused either by M or M+Na markedly improved dry weight of leaf (112 and 156%, respectively and aerial parts (49 and 144%, respectively as compared to the uninoculated control. Fungi inoculation positively improved vegetative parameters of stevia plant under most osmatic levels and sources. The most ameliorate effect, however, was observed where M as drought stress or M+Na were adjusted to -5 bar. Therefore, the results of this study represented a positive effect of P. indica inoculation in inproving osmotic tolerance of stevia medicinal plant.

Morphological and transcript changes in the biosynthesis of lignin in oil palm (Elaeis guineensis) during Ganoderma boninense infections in vitro.

Science.gov (United States)

Goh, Kar Mun; Dickinson, Matthew; Supramaniam, Christina V

2018-03-01

Lignification of the plant cell wall could serve as the first line of defense against pathogen attack, but the molecular mechanisms of virulence and disease between oil palm and Ganoderma boninense are poorly understood. This study presents the biochemical, histochemical, enzymology and gene expression evidences of enhanced lignin biosynthesis in young oil palm as a response to G. boninense (GBLS strain). Comparative studies with control (T1), wounded (T2) and infected (T3) oil palm plantlets showed significant accumulation of total lignin content and monolignol derivatives (syringaldehyde and vanillin). These derivatives were deposited on the epidermal cell wall of infected plants. Moreover, substantial differences were detected in the activities of enzyme and relative expressions of genes encoding phenylalanine ammonia lyase (EC 4.3.1.24), cinnamate 4-hydroxylase (EC 1.14.13.11), caffeic acid O-methyltransferase (EC 2.1.1.68) and cinnamyl alcohol dehydrogenase (CAD, EC 1.1.1.195). These enzymes are key intermediates dedicated to the biosynthesis of lignin monomers, the guaicyl (G), syringyl (S) and ρ-hydroxyphenyl (H) subunits. Results confirmed an early, biphasic and transient positive induction of all gene intermediates, except for CAD enzyme activities. These differences were visualized by anatomical and metabolic changes in the profile of lignin in the oil palm plantlets such as low G lignin, indicating a potential mechanism for enhanced susceptibility toward G. boninense infection. © 2017 Scandinavian Plant Physiology Society.

Combining zygotic embryo culture and mutation induction to improve salinity tolerance in avocado (Persea americana Mill)

Energy Technology Data Exchange (ETDEWEB)

Fuentes, J. L.; Santiago, L.; Alvarez, A.; Valdés, Y.; Vernhe, M.; Guerra, M.; Altanez, S.; Prieto, E. F. [Centro de Aplicaciones Tecnológicas y Desarrollo Nuclear (CEADEN), Miramar, Playa, C. Habana (Cuba); Rodríguez, N. N.; Arbelo, O. Coto; Velázquez, B.; Rodríguez, J. A.; Sourd, D. G.; Fuentes, V. R. [Instituto de Investigaciones de Fruticultura Tropical (IICF), Miramar, Playa, C. Habana (Cuba); Leal, M. R. [Departamento de Microbiología, Facultad de Biología, Universidad de la Habana, Vedado, C. Habana (Cuba)

2009-05-15

Mutation induction and biotechnological techniques are some of the current approaches used in plant breeding. In the present work radiation-induced mutation followed by in vitro culture of zygotic embryos and high osmotic pressure selection methods to improve salt tolerance in avocado are investigated. The in vitro germination, rooting, bud multiplication and plantlet acclimatization of Cuban avocado varieties were recorded. The germination rates of whole embryos in vitro were found to be higher when using mature rather than immature embryos. Almost 80% of the whole embryos derived plantlets produced were successfully acclimatized under greenhouse conditions. An in vitro propagation method for avocado breeding purposes was optimized and documented. However, in vitro multiplication results indicated the need to improve bud multiplication methods in avocado. The survival rates of gamma rays irradiated and salt pressured avocado embryos were also investigated. Both mutagenic (LD{sub 50} = 27-28 Gy) and selective (LD{sub 20} = 157 mM of NaCl) doses were established. A procedure combining zygotic embryo culture and mutation induction was used to obtain. Putative mutant lines derived from salt tolerant rootstocks were developed. Putative M{sub 1}V{sub 3} progenies were planted in the field for segregation analysis. An avocado gene bank was established under the same study. Therefore this methodology appears as an alternative to traditional breeding methods, particularly for improving agronomic characteristics such as salt tolerance in avocado. (author)

A Novel Protocol for Stevia rebaudiana (Bert. Regeneration

Directory of Open Access Journals (Sweden)

Parto Roshandel

2013-01-01

Full Text Available Stevia rebaudiana Bertoni has sweet substances (stevioside in its leaves that are free of calories and their consumption is beneficial for diabetic patients and is also helpful in high blood pressure also. Because of low capability in seed germination, tissue culture is an appropriate method for propagation of this plant. In the current study, optimization of stevia in vitro cultivation via direct organogenesis with different explants, light intensities and plant hormones has been examined. These treatments included BAP (at 0.5, 1, 1.5 and 2mg/l in combination with 2, 4-D, IBA and NAA (each with concentrations of 0.1, 0.2 and 0.5mg/l and different light intensities (Dark, 2000, 4000 and 6000 lux. MS was utilized as a basal medium. Results indicated the highest rate of organogenesis (85% occurred on the auxiliary buds explants with a medium containing 1.5mg/l BAP + 0.1mg/l NAA under 6000 lux light intensity. Also, the highest range of primary organ per explant (42 with 0.3cm length was achieved at this condition. The most efficient medium for rhizogenesis i.e. 100% root production along with the highest root number (11 with approximately 7.13cm length was obtained in presence of activated charcoal and 1mg/l of IBA. At the end of rhizogenesis experiments, the plantlet length and node multiplicity were 12.8cm and 7 respectively. Greenhouse cultivation of these plantlets was successful.

LHC II system sensitivity to magnetic fluids

CERN Document Server

Cotae, Vlad

2005-01-01

Experiments have been designed to reveal the influences of ferrofluid treatment and static magnetic field exposure on the photosynthetic system II, where the light harvesting complex (LHC II) controls the ratio chlorophyll a/ chlorophyll b (revealing, indirectly, the photosynthesis rate). Spectrophotometric measurement of chlorophyll content revealed different influences for relatively low ferrofluid concentrations (10-30 mul/l) in comparison to higher concentrations (70-100 mul/l). The overlapped effect of the static magnetic field shaped better the stimulatory ferrofluid action on LHC II system in young poppy plantlets.

Somatic seeds of Plantago asiatica L.

Directory of Open Access Journals (Sweden)

Emilia Andrzejewska-Golec

2011-01-01

Full Text Available Somatic seeds of Plantago asiatica L. were produced for the first time. Shoot-tips isolated from in vitro obtained 4-week shoots were encapsulated using sodium alginate and calcium chloride. Capsules with or without sucrose and with and without cytokinin - indole-3-butyric acid (IBA were used. Sucrose presence in capsules very distinctly influences somatic seeds of Plantago asiatica germination and their conversion into plants. However, addition of IBA to capsules has not clear influence on the ability of plant regrowth. Plantlets transplanted to soil grew to phenotypically normal plants.

Study on in-vitro propagation for preparation of mutatio materials of Vietnamese rare of Vietnam

International Nuclear Information System (INIS)

Le Quang Luan; Vo Thi Thu Ha; Nguyen Thi Nu; Nguyen Thi Vang; Phan Dinh Thai Son; Hoang Thi My Linh; Le Xuan Tham; Nguyen Quang Khanh

2007-01-01

The shoot tips, flower-stalk and leaves of Cymbidium sayonara Raritan and Paphiopedilum delenatii were used as initial samples. All samples were steriled by Ca(OCl) 2 solution. Protocorm like bodies (PLBs), shoot buds and leaves from 30 days samples were used for callus formation, PLB generation, shoot proliferation and plantlet formation. To investigate the effect of gamma Co-60 onto the in vitro grow and development of two mentioned orchids, the PLB segments were derived and then irradiated in various dose with the dose rate of 0.2 Gy/s. (NHA)

Micropropagation of Lavandula spp.

Science.gov (United States)

Gonçalves, Sandra; Romano, Anabela

2013-01-01

Lavandula species are some of the most popular ornamental and medicinal plants with great economic values. These species are vegetative propagated by stem cuttings. However, the poor rooting ability and vulnerability of plantlets to contamination are major limiting factors for propagation. In vitro culture methods are suitable to overcome these limitations. This chapter describes protocols for in vitro propagation of Lavandula viridis L'Hér and Lavandula vera DC. Nodal shoot proliferation of L. viridis and plant regeneration from leaf-derived callus of L. vera by an "open culture system" are highlighted.

Micropropagation of Plantago asiatica L. through culture of shoot-tips

Directory of Open Access Journals (Sweden)

Joanna Makowczyńska

2011-01-01

Full Text Available Shoot-tip multiplication of the medicinal species - Plantago asiatica was carried on MS medium with IAA and BAP or kinetin. Best results in micropropagation were achieved by adding 0.1 mg/dm3 IAA and 1 mg/dm3 BAP. After 6 weeks shoots were transferred to MS medium for rooting. The resulting plantlets were transferred after 8 weeks into pots and after a period of adaptation into the ground (field culture. The species Plantago asiatica was propagated in vitro by shoot-tip multiplication for the first time.

An efficient protocol for regeneration and transformation of Symphyotrichum novi-belgii

DEFF Research Database (Denmark)

Mørk, Eline Kirk; Henriksen, Karin; Brinch-Pedersen, Henrik

2012-01-01

) fusion gene, and an enhanced green fluorescent protein gene. Pre-cultured hypocotyl explants were transformed in the presence of 100 μM acetosyringone using 90 s sonication plus 10 min vacuum-infiltration. Kanamycin at 20 mg l−1 was used for selecting transformed cells. Adventitious shoots regenerated......II genes in GUS-positive shoots were confirmed by PCR and copy number of the nptII gene in PCR-positive shoots was determined by Southern blotting. Three transgenic plantlets were acclimatized to the greenhouse. This transformation and regeneration system using hypocotyls provides a foundation...

COMPORTAMENTO FISIOLÓGICO IN VITRO DE BANANEIRA (Musa sp., AAA e AAB cvs. NANICA E PRATA ANÃ: INFLUÊNCIA DE DIFERENTES NÍVEIS DE POTÁSSIO PYSIOLOGICAL BEHAVIOR OF BANANA (Musa sp., AAA and AAB cvs. 'NANICA' AND 'PRATA ANÃ' IN VITRO CONDITION: INFLUENCE OF DIFFERENT LEVELS OF POTASSIUM

Directory of Open Access Journals (Sweden)

Humberto Actis Zaidan

1999-01-01

cultivated in asseptic conditions in modified MS media for 90 days in the presence of six different concentrations of K: 5, 10, 15, 20, 25 and 30 mM. The concentration of 20 mM is found in the basic MS medium. The cultures were then transferred to rooting media with the same concentrations of K. Dry matter weight and k concentration in shoots, roots and the whole plantlet, and number of plantlets produced were determined. Visual symptoms of K deficiency, such as chlorosis and necrosis in older leaves, were observed in all plantlets under low concentration of K. Quantitative differences were observed between the two cultivars in relation to dry matter weight and number of in vitro regenerated banana plantlets; K concentration was optimum around 15-20 mM.

In vitro rescue of interspecific embryos from Elaeis guineensis x E. oleifera (Arecaceae

Directory of Open Access Journals (Sweden)

Paula Cristina da Silva Angelo

2011-09-01

Full Text Available The African oil palm (Elaeis guineensis is the most effective oil producer in tons per hectare. Nevertheless, its increasing cultivation in Latin America is harmed by the “lethal yellowing”. Genetic resistance to this anomaly can be found in the germplasm of American oil palm or caiaué (E. oleifera, a native species from the Amazon rainforest. However, the procedures adopted to induce seeds of E. guineensis to germination frequently result mild for interespecific hybrids. Embryo in vitro cultivation can be a viable option. This work was aimed initially to test liquid MS medium supplemented with different glucose or sucrose concentrations for the in vitro cultivation of zygotic embryos from E. guineensis x E. oleifera controlled pollinations. Additionally we investigated different compost mixtures to acclimatize the regenerated hybrid plantlets. Concentrations of 10, 20 and 30g/L of both sugars were tested on flasks containing five mature zygotic embryos, with 15 repetitions per treatment in a total of 450 explants. The number of embryos displaying shoots and radicles at least 2mm in length per experimental unit was evaluated during phase one of in vitro cultivation. Plantlets displaying shoots and radicles were transferred to phase two of in vitro cultivation and subsequently to acclimatization, under 70% shading with manual water supply. The experiments of acclimatization were conducted with 130 plantlets randomly distributed in pure horticultural compost, 3:1 or 1:1 compost:sand mixtures and each plantlet was defined as an experimental unit. Data were submitted to ANOVA, t test and analyzes of correlation (p≤0.05. Highest emergence rates were 97% for shoots and 73% for radicles, observed in MS medium supplemented with 20g/L (110mM of glucose. This sugar in concentrations of 20 or 30g/L provided balanced shoot/root development, and this was considered one of the reasons for the higher frequency of plantlet establishment. The survival

A Gaijin-like miniature inverted repeat transposable element is mobilized in rice during cell differentiation

Directory of Open Access Journals (Sweden)

Dong Hai-Tao

2012-04-01

Full Text Available Abstract Background Miniature inverted repeat transposable element (MITE is one type of transposable element (TE, which is largely found in eukaryotic genomes and involved in a wide variety of biological events. However, only few MITEs were proved to be currently active and their physiological function remains largely unknown. Results We found that the amplicon discrepancy of a gene locus LOC_Os01g0420 in different rice cultivar genomes was resulted from the existence of a member of Gaijin-like MITEs (mGing. This result indicated that mGing transposition was occurred at this gene locus. By using a modified transposon display (TD analysis, the active transpositions of mGing were detected in rice Jiahua No. 1 genome under three conditions: in seedlings germinated from the seeds received a high dose γ-ray irradiation, in plantlets regenerated from anther-derived calli and from scutellum-derived calli, and were confirmed by PCR validation and sequencing. Sequence analysis revealed that single nucleotide polymorphisms (SNPs or short additional DNA sequences at transposition sites post mGing transposition. It suggested that sequence modification was possibly taken place during mGing transposition. Furthermore, cell re-differentiation experiment showed that active transpositions of both mGing and mPing (another well studied MITE were identified only in regenerated plantlets. Conclusions It is for the first time that mGing active transposition was demonstrated under γ-ray irradiation or in cell re-differentiation process in rice. This newly identified active MITE will provide a foundation for further analysis of the roles of MITEs in biological process.

Genetic stability and phytochemical analysis of the in vitro regenerated plants of Dendrobium nobile Lindl., an endangered medicinal orchid

Science.gov (United States)

Bhattacharyya, Paromik; Kumaria, Suman; Diengdoh, Reemavareen; Tandon, Pramod

2014-01-01

An efficient genetically stable regeneration protocol with increased phytochemical production has been established for Dendrobium nobile, a highly prized orchid for its economic and medicinal importance. Protocorm like bodies (PLBs) were induced from the pseudostem segments using thidiazuron (TDZ; 1.5 mg/l), by-passing the conventional auxin–cytokinin complement approach for plant regeneration. Although, PLB induction was observed at higher concentrations of TDZ, plantlet regeneration from those PLBs was affected adversely. The best rooting (5.41 roots/shoot) was achieved in MS medium with 1.5 mg/l TDZ and 0.25% activated charcoal. Plantlets were successfully transferred to a greenhouse with a survival rate of 84.3%, exhibiting normal development. Genetic stability of the regenerated plants was investigated using randomly amplified polymorphic DNA (RAPD) and start codon targeted (SCoT) polymorphism markers which detected 97% of genetic fidelity among the regenerants. The PIC values of RAPD and SCoT primers were recorded to be 0.92 and 0.76 and their Rp values ranged between 3.66 and 10, and 4 and 12 respectively. The amplification products of the regenerated plants showed similar banding patterns to that of the mother plant thus demonstrating the homogeneity of the micropropagated plants. A comparative phytochemical analysis among the mother and the micropropagated plants showed a higher yield of secondary metabolites. The regeneration protocol developed in this study provides a basis for ex-situ germplasm conservation and also harnesses the various secondary metabolite compounds of medicinal importance present in D. nobile. PMID:25606433

Micropropagation and non-steroidal anti-inflammatory and anti-arthritic agent boswellic acid production in callus cultures of Boswellia serrata Roxb.

Science.gov (United States)

Nikam, Tukaram D; Ghorpade, Ravi P; Nitnaware, Kirti M; Ahire, Mahendra L; Lokhande, Vinayak H; Chopra, Arvind

2013-01-01

Micropropagation through cotyledonary and leaf node and boswellic acid production in stem callus of a woody medicinal endangered tree species Boswellia serrata Roxb. is reported. The response for shoots, roots and callus formation were varied in cotyledonary and leafy nodal explants from in vitro germinated seeds, if inoculated on Murshige and Skoog's (MS) medium fortified with cytokinins and auxins alone or together. A maximum of 8.0 ± 0.1 shoots/cotyledonary node explant and 6.9 ± 0.1 shoots/leafy node explants were produced in 91 and 88 % cultures respectively on medium with 2.5 μM 6-benzyladenine (BA) and 200 mg l(-1) polyvinylpyrrolidone (PVP). Shoots treated with 2.5 μM IBA showed the highest average root number (4.5) and the highest percentage of rooting (89 %). Well rooted plantlets were acclimatized and 76.5 % of the plantlets showed survival upon transfer to field conditions. Randomly amplified polymorphic DNA (RAPD) analysis of the micropropagated plants compared with mother plant revealed true-to-type nature. The four major boswellic acid components in calluses raised from root, stem, cotyledon and leaf explants were analyzed using HPLC. The total content of four boswellic acid components was higher in stem callus obtained on MS with 15.0 μM IAA, 5.0 μM BA and 200 mg l(-1) PVP. The protocol reported can be used for conservation and exploitation of in vitro production of medicinally important non-steroidal anti-inflammatory metabolites of B. serrata.

Micropropagation of Cyclopia genistoides, an endemic South African plant of economic importance.

Science.gov (United States)

Kokotkiewicz, Adam; Luczkiewicz, Maria; Hering, Anna; Ochocka, Renata; Gorynski, Krzysztof; Bucinski, Adam; Sowinski, Pawel

2012-01-01

An efficient micropropagation protocol of Cyclopia genistoides (L.) Vent., an indigenous South African shrub of economic importance, was established. In vitro shoot cultures were obtained from shoot tip fragments of sterile seedlings cultured on solid Schenk and Hildebrandt (SH) medium supplemented with 9.84 microM 6-(gamma,gamma-dimethylallylamino)purine (2iP) and 1.0 microM thidiazuron (TDZ). Maximum shoot multiplication rate [(8.2 +/- 1.3) microshoots/explant)] was observed on this medium composition. Prior to rooting, the multiplied shoots were elongated for 60 days (two 30-days passages) on SH medium with one-half sucrose concentration, supplemented with 4.92 microM indole-3-butyric acid (IBA). The rooting of explants was only possible in the case of the elongated shoots. The highest root induction rate (54.8%) was achieved on solid SH medium with one-half sucrose and one-half potassium nitrate and ammonium nitrate concentration, respectively, supplemented with 28.54 microM indole-3-acetic acid (IAA) and 260.25 microM citric acid. The plantlets were acclimatized for 30 days in the glasshouse, with the use of peat/gravel/perlite substrate (1:1:1). The highest acclimatization rate (80%) was obtained for explants rooted with the use of IAA-supplemented medium. The phytochemical profile of the regenerated plants was similar to that of the reference intact plant material. HPLC analyses showed that C. genistoides plantlets obtained by the micropropagation procedure kept the ability to produce xanthones (mangiferin and isomangiferin) and the flavanone hesperidin, characteristic of wild-growing shrubs.

Preculturing effect of thidiazuron on in vitro shoot multiplication and micropropagation round in Capparis decidua (Forsk.) an important multipurpose plant.

Science.gov (United States)

Bukhari, Najat A W; Siddique, Iram; Perveen, Kahkashan

2016-09-01

An efficient protocol was developed for clonal multiplication of an important shrub: Capparis decidua (Forsk.) Edgew, through in vitro shoot induction and multiplication from nodal explants. Pretreatment of nodal explants in a liquid Murashige and Skoog (MS) medium augmented with various thidiazuron (TDZ) concentrations at relatively high levels (5-100 μM) for different time duration (4, 8, 12 and 16 d), proved a significant approach for in vitro shoot production. After an initial exposure time to TDZ, nodal explants were inoculated onto a MS basal medium devoid of TDZ for further induction and proliferation. The highest regeneration rate (85%), average number of shoots/explant (8.7 ± 0.22) and maximum shoot length (3.9 ± 0.33 cm) were obtained from the nodal explants exposed to 50 μM TDZ for 8 d. The nodal explants excised from the proliferated cultures of TDZ (50 μM) for 8 d were used as explants and showed an enhancement rate after next three round of in vitro propagation. Best results for rooting was obtained by ex vitro treatment of shoots with 200 μM indole-3-butyric acid (IBA) for 20 min. as it produced an average of 5.7 ± 0.41 roots per microshoot with 4.4 ± 0.39 cm root length in 84% shoots. Different planting substrates was tested for maximum survival of hardening off micropropagated plantlets and soilrite proved most effective than others as 97.1 ± 7.21 plantlets survived. All micropropagated plants grew well in natural conditions and showed similar morphology to the mother plant.

Effect of salinity on gene expression, morphological and biochemical characteristics of stevia rebaudiana Bertoni under in vitro conditions.

Science.gov (United States)

Fallah, F; Nokhasi, F; Ghaheri, M; Kahrizi, D; Beheshti Ale Agha, A; Ghorbani, T; Kazemi, E; Ansarypour, Z

2017-08-15

Stevia rebaudiana Bertoni is a famous medicinal plant for its low calorific value compounds which are named steviol glycosides (SGs) and they are 150-300 times sweeter than sugar. Among various SGs, stevioside and rebaudioside A considered to be the main sweetening compounds.  Soil salinity is one of the most essential stress in the world. Salinity affects the survival and yield of crops. In current study the effects of salinity and osmotic stress caused by different concentration of NaCl (0, 20, 40, 60 and 80 mM) on morphological traits, genes expressionand amount of both stevioside and rebaudioside Aunder in vitro conditions has been investigated. The morphological traits such as bud numbers, root numbers, shoot length (after 15 and 30 days) were evaluated. With increasing salinity, the values of all studied morphological traits decreased. To investigation of UGT74G1 and UGT76G1 genes expression that are involved in the synthesis of SGs, RT-PCR was done and there were significant differences between all media. The highest expression of both genes was observed in plantlets grown on MS media (with NaCl-free). Also, the lowest amounts of gene expression of the both genes were seen in MS+ 60 mM NaCl. Based on HPLC results, the highest amount of both stevioside and rebaudioside A were observed in plantlets grown in MS media (with NaCl-free). Finally, it can be concluded that stevia can survive under salt stress, but it has the best performance in the lower salinity.

Enhanced salt-induced antioxidative responses involve a contribution of polyamine biosynthesis in grapevine plants.

Science.gov (United States)

Ikbal, Fatima Ezzohra; Hernández, José Antonio; Barba-Espín, Gregorio; Koussa, Tayeb; Aziz, Aziz; Faize, Mohamed; Diaz-Vivancos, Pedro

2014-06-15

The possible involvement of polyamines in the salt stress adaptation was investigated in grapevine (Vitis vinifera L.) plantlets focusing on photosynthesis and oxidative metabolism. Salt stress resulted in the deterioration of plant growth and photosynthesis, and treatment of plantlets with methylglyoxal-bis(guanylhydrazone) (MGBG), a S-adenosylmethionine decarboxylase (SAMDC) inhibitor, enhanced the salt stress effect. A decrease in PSII quantum yield (Fv/Fm), effective PSII quantum yield (Y(II)) and coefficient of photochemical quenching (qP) as well as increases in non-photochemical quenching (NPQ) and its coefficient (qN) was observed by these treatments. Salt and/or MGBG treatments also triggered an increase in lipid peroxidation and reactive oxygen species (ROS) accumulation as well as an increase of superoxide dismutase (SOD) and peroxidase (POX) activities, but not ascorbate peroxidase (APX) activity. Salt stress also resulted in an accumulation of oxidized ascorbate (DHA) and a decrease in reduced glutathione. MGBG alone or in combination with salt stress increased monodehydroascorbate reductase (MDHAR), SOD and POX activities and surprisingly no accumulation of DHA was noticed following treatment with MGBG. These salt-induced responses correlated with the maintaining of high level of free and conjugated spermidine and spermine, whereas a reduction of agmatine and putrescine levels was observed, which seemed to be amplified by the MGBG treatment. These results suggest that maintaining polyamine biosynthesis through the enhanced SAMDC activity in grapevine leaf tissues under salt stress conditions could contribute to the enhanced ROS scavenging activity and a protection of photosynthetic apparatus from oxidative damages. Copyright © 2014 Elsevier GmbH. All rights reserved.

In vitro plant regeneration from leaf explants of Solanum pimpinellifolium L.

Directory of Open Access Journals (Sweden)

Shirley Valderrama-Alfaro

2011-01-01

Full Text Available Tomato is a species of agricultural importance. Besides, it is a source of vitamins, minerals and antioxidant compounds. Therefore it is essential to obtain varieties resistant to diseases. Solanum pimpinellifolium L. ‘Tomatillo wild’, one of the wild relatives of tomato, is considered multipurpose reservoir of genes. This characteristic must be exploited. In vitro plantlets from leaves of ‘wild tomatillo’ were regenerated with this objective. A system for in vitro germination of seeds obtained from ex vitro sources was created. The use of seed allowed obtaining adequate seedlings in a short time to start regeneration. Thein vitro response of explants was evaluated in four treatments. The Murashige and Skoog (MS basal culture medium supplemented with a-naphthaleneacetic acid (NAA and 6-benzyl aminopurine (BAP was used in different combinations. The higher percentage (30% of shoot induction was achieved with 0.1 mg l1 NAA / 1 mg l1 BAP. The presence of callus and roots was observed after seven days of culture in the combination treatment with 1 mg l1 NAA / 0.1 mg l1 BAP. Callus showed 1 or 2 adventitious shoots per explant after 30 days of culture and 3 to 6 shoots after seventy days. Furthermore, the presence of fully formed adventitious plantlets (shoot and root was observed after the fifth week of culture. Therefore, the best combination for in vitro regeneration is NAA 0.1 mg l1 / 1 mg l1 BAP. Keywords: auxin, cytokinins, indirect organogenesis

Optimization of a protocol for the micropropagation of pineapple

Directory of Open Access Journals (Sweden)

ALMEIDA WELITON ANTONIO BASTOS DE

2002-01-01

Full Text Available The present work aimed at maximizing the number of plantlets obtained by the micropropagation of pineapple (Ananas comosus (L. Merrill cv. Pérola. Changes in benzylaminopurine (BAP concentration, type of medium (liquid or solidified and the type of explant in the proliferation phase were evaluated. Slips were used as the explant source, which consisted of axillary buds obtained after careful excision of the leaves. A Sterilization was done in the hood with ethanol (70%, for three minutes, followed by calcium hypochlorite (2%, for fifteen minutes, and three washes in sterile water. The explants were introduced in MS medium supplemented with 2mg L-1 BAP and maintained in a growth room at a 16h photoperiod (40 mmol.m-2.s-1, 27 ± 2ºC. After eight weeks, cultures were subcultured for multiplication in MS medium. The following treatments were tested: liquid x solidified medium with different BAP concentrations (0.0, 1.5 or 3.0 mg L-1, and the longitudinal cut, or not, of the shoot bud used as explant. The results showed that liquid medium supplemented with BAP at 1.5 mg L-1, associated with the longitudinal sectioning of the shoot bud used as explant presented the best results, maximizing shoot proliferation. On average, the best treatment would allow for an estimated production of 161,080 plantlets by the micropropagation of the axillary buds of one plant with eight slips and ten buds/slips, within a period of eight months.

Comparison of cauliflower-insect-fungus interactions and pesticides for cabbage root fly control.

Science.gov (United States)

Razinger, Jaka; Žerjav, Metka; Zemljič-Urbančič, Meta; Modic, Špela; Lutz, Matthias; Schroers, Hans-Josef; Grunder, Jürg; Fellous, Simon; Urek, Gregor

2017-12-01

Cabbage root fly (Delia radicum L.) control represents a major challenge in brassica production, therefore different management strategies for its control were tested in conventionally managed open field cauliflower production. Strategies included treatments with low-risk methods such as nitrogen lime, the insecticide spinosad and the Beauveria bassiana ATCC 74040-based biopesticide Naturalis. Their effects were compared with treatments based on nonformulated fungal species Metarhizium brunneum, B. bassiana, Clonostachys solani, Trichoderma atroviride, T. koningiopsis, and T. gamsii and commercial insecticides λ-cyhalothrin and thiamethoxam. Spinosad and thiamethoxam were pipetted to individual plants before transplanting; λ-cyhalothrin was sprayed after transplanting; nitrogen lime was applied at first hoeing. Nonformulated fungi were delivered onto cauliflower plantlets' roots as a single pretransplantation inoculation. The cabbage root fly population dynamics exhibited a strong spatiotemporal variation. The lowest number of cabbage root fly pupae recovered from cauliflower roots in the field experiments was recorded in plants treated with spinosad (significant reduction), followed by Naturalis and one of the tested M. brunneum strains (nonsignificant reduction). Significantly more pupae were counted in the nitrogen lime treatment. The field experiments showed that a single drench of cauliflower plantlets with spinosad offered consistent and enduring cabbage root fly control. Naturalis and nonformulated fungal isolates did not decrease cabbage root fly pressure significantly, apparently due to lack of statistical power. The implications of the substantial intra- and inter-annual pest pressure variation and the benefits of using single plant treatments are discussed, and recommendations for improvement of rhizosphere-competence utilizing biological control strategies provided. © 2017 Institute of Zoology, Chinese Academy of Sciences.

Biofertilization with Azospirillum brasilense improves in vitro culture of Handroanthus ochraceus, a forestry, ornamental and medicinal plant.

Science.gov (United States)

Llorente, Berta E; Alasia, María A; Larraburu, Ezequiel E

2016-01-25

Biofertilization with plant growth-promoting rhizobacteria is a potential alternative to plant productivity. Here, in vitro propagation of Handroanthus ochraceus (yellow lapacho), a forest crop with high economic and environmental value, was developed using the Azospirillum brasilense strains Cd and Az39 during rhizogenesis. Epicotiles of in vitro plantlets were multiplied in Woody Plant Medium (WPM). For rooting, elongated shoots were transferred to auxin-free Murashige-Skoog medium with Gamborg's vitamins and WPM, both at half salt concentration (½MSG and ½WPM), and inoculated with Cd or Az39 at the base of each shoot. Anatomical studies were performed using leaves cleared and stained with safranin for optical microscopy and leaves and roots metalized with gold-palladium for scanning electron microscopy (SEM). In ½WPM auxin-free medium, A. brasilense Cd inoculation produced 55% of rooting, increased root fresh and dry weight (45% and 77%, respectively), and led to lower stomata size and density with similar proportion of open and closed stomata. Both strains selectively increased the size or density of glandular trichomes in ½MSG. Moreover, bacteria were detected on the root surface by SEM. In conclusion, the difference in H. ochraceus response to A. brasilense inoculation depends on the strain and the plant culture media. Cd strain enhanced rooting in auxin-free ½WPM and produced plantlets with features similar to those expected in ex vitro plants. This work presents an innovative in vitro approach using beneficial plant-microorganism interaction as an ecologically compatible strategy in plant biotechnology. Copyright © 2015 Elsevier B.V. All rights reserved.

Seeds and plantlets of Myrciaria dubia “camu-camu”: biometry, germination, and initial growth

Directory of Open Access Journals (Sweden)

Anderson E. Medina Bardales

2014-06-01

Full Text Available The objectives were to determine seed biometrics, recording cellular respiration in the imbibition process, evaluate the effect of inducing chemicals in germination and early growth analysis. Cellular respiration during imbibition was measured by reduction of methylene blue and Janus Green B. The inductor effect of five chemical compounds in germination was evaluated. Initial growth was recorded by stereoscopic. Results showed that the seeds have an average weight of 520 ± 172 mg, length of 13.50 ± 1.76 mm, width of 10.61 ± 1.14 mm, and thickness of 4.68 ± 0 62 mm. These variables showed significant correlations (r of 0.494 to 0.897 and p 90% and 65% respectively. Seedling initial growth consisted of six stages and occurred over a period of 22 days after radicle emergence. In conclusion, M. dubia seeds show wide variation in their biometrics (weight, length, width and thickness, but show significant correlations with each other. Likewise, seeds are metabolically active in imbibition process, and germination is induced by sodium nitrite and potassium cyanide. Furthermore, seedling initial growth to occur over a period of 22 days after emergence of the radicle and consists of six distinct stages.

Research work to obtain pepper (Piper negrum l.) mutants resistant to Fusarium disease

International Nuclear Information System (INIS)

Ando, A.; Menten, J.O.M.; Tulmann Neto, A.; Albuquerque, F.C. de

1984-01-01

Cuttings of black pepper (Piper nigrum L.) variety Singapore were irradiated at different doses (from 3.0-7.0 kR) of gamma radiation in the 60 Co source. After two successive prunings of surviving M 1 plants (from the irradiated cuttings), hundreds of vM 3 cuttings were planted separately in pots and artificially inoculated with Fusarium solani f. piperi. The surviving vM 3 plantlets were transplanted to an experimental field highly infested with this patogen. After two years in the infested field, only three UM 3 plants continue to grow. (M.A.C.) [pt

Somatic embryogenesis and plant regeneration in tissue cultures of sweet potato (Ipomea batatas Poir.).

Science.gov (United States)

Liu, J R; Cantliffe, D J

1984-06-01

Leaf, shoot-tip, stem, and root explants of sweet potato (Ipomea batatas Poir.) gave rise to two kinds of callus on nutrient agar medium containing 0.5 to 2.0 mg/l 2,4-D. One callus, bright- to pale-yellow, was compact and organized, while the other was dull-yellow and friable. The former callus gave rise to numerous globular and heart-shaped embryoids. When transferred onto hormone-free medium, the embryoids readily developed into a torpedo-shape before germination. The plantlets were transplanted to soil where they flowered and formed storage roots at maturity.

Embryogenesis in gamma-irradiated habituated ovular callus of the 'Shamouti' orange as affected by auxin and by tissue age

International Nuclear Information System (INIS)

Kochba, J.; Spiegel-Roy, P.

1977-01-01

The response of habituated embryogenic ovular callus of the 'Shamouti' orange, Citrus sinensis, to gamma irradiation was studied. Stimulation of embryogenesis was observed when only callus, but not when the medium was irradiated. Age of callus prior to subculture modified the response to irradiation as regards optimal dose for embryogenesis and radiosensitivity. Radiation intensities also modified callus response to radiation dose. Addition of unirradiated IAA to irradiated callus cultures increased embryo formation and furthered development of embryos into plantlets, with some combinations of radiation dose and IAA concentration. Addition of IAA also decreased radiosensitivity at high doses (32 kR). (author)

Clonal propagation of Stevia rebaudiana Bertoni by stem-tip culture.

Science.gov (United States)

Tamura, Y; Nakamura, S; Fukui, H; Tabata, M

1984-10-01

Clonal propagation of Stevia rebaudiana has been established by culturing stem-tips with a few leaf primordia on an agar medium supplemented with a high concentration (10 mg/l) of kinetin. Anatomical examination has suggested that these multiple shoots originate from a number of adventitious buds formed on the margin of the leaf. Innumerable shoots can be obtained by repeating the cycle of multiple-shoot formation from a single stem-tip of Stevia. These shoots produce roots when transferred to a medium containing NAA (0.1 mg/l) without kinetin. The regenerated plantlets can be transplanted to soil.

RESEARCH REGARDING THE POTENTIAL ACTIVITY OF SOME HETEROCYCLIC COMPOUNDS ON PLANTS GROWTH AND DEVELOPMENT

Directory of Open Access Journals (Sweden)

OANA-IRINA PATRICIU

2017-06-01

Full Text Available It is well known that growth and morphogenesis of plant tissue cultures can be improved by small amounts of some organic compounds. Heterocyclic compounds such as chromanones and thiazoles derivatives, valuable because of their potential biological activities, have also been reported as pesticides, herbicides and plant-growth regulators. In the present study, different concentrations of chromanones and thiazoles derivatives were employed to evaluate their effects on plantlets growth of Ocimum basilicum L. and Echinacea purpurea L. The studied compounds were proved to be growth inhibitors at high concentrations. A growth stimulation effect was registered at low concentration.

High frequency plant regeneration from leaf explants derived callus of evening primrose (oenothera biennis)

International Nuclear Information System (INIS)

Ghauri, E.G.; Shafi, N.; Ghani, S.; Fatima, A.

2008-01-01

The seeds of Evening primrose were aseptically grown and leaf explants were used for establishment of callus culture. The Excellent growth in callus biomass was achieved on MS medium supplemented with 2, 4, -D and TDZ. For optimal growth of bud and shoot regeneration, fortification of IAA along with TDZ, or BAP was found to be essential. Rooting (70%) could be inducted on hormone free MS-medium. This percentage improved to 98 when NAA was added to the medium. The plantlets thus obtained were transferred to the field successfully after passing through the process of hardening. (author)

In vitro techniques for mutation breeding of tropical root and tuber crops

International Nuclear Information System (INIS)

Novak, F.

1987-01-01

Full text: To assist IAEA Technical Co-operation projects, the Agricultural Section of the IAEA Laboratory in Seibersdorf is developing techniques for in vitro mutation breeding of cassava (Manihot esculenta) and yam (Dioscorea alata, D. rotundata). The first aim was to induce morphogenesis (plant regeneration) in tissue culture and establish techniques for in vitro propagation. Subsequently, the in vitro mutation breeding technology is being developed. (i) Cassava is one of the important staple food crops of tropical countries. Pest and disease resistance as well as low toxic cyanide content are among the objectives for genetic improvement. For in vitro mutation induction we use shoot-tip and node culture. Shoot apices (1 and 2 mm long) are aseptically dissected from cassava buds and cultured on MS medium with 1 mg/l thiamine, -naphthalene acetic acid, 6-benzyladenine and gibberellic acid. Elevated concentration of 6-benzyladenine is used for multiple shoot formation. The rapid multiplication was induced in liquid medium, when flasks were placed on a gyratory shaker with 60 rpm at 28 deg. C during 16/8 light/dark photoperiod. Nodes with axillary buds from in vitro growing plantlets were irradiated with gamma rays. Doses of 30 to 45 Gy allowed the survival of approx. 50 percent of explants and subsequent shoot proliferation from axillary buds. Radiosensitivity of cassava genotypes may be different and this will be investigated in future experiments. (ii) Yams are likewise important tuber crops, particularly in West Africa, South-East Asia and the Caribbean. The main breeding objectives are improved yield, shortened growth period, improved storability (resistance of tubers to fungal attack), shoot tip cultures have been utilized for clonal propagation, and germplasm preservation and exchange. At present, the IAEA Laboratory at Seibersdorf is trying this technique for mutation induction. Somatic embryogenesis in cell and tissue culture is worked on to develop a

Crescimento in vitro e aclimatização de Cattleya loddigesii Lindley (Orchidaceae com carvão ativado sob dois espectros luminosos In vitro growth and acclimatization of Cattleya loddigesii Lindley (Orchidaceae with actived charcoal in two light spectra

Directory of Open Access Journals (Sweden)

Renato Fernandes Galdiano Júnior

2012-05-01

Full Text Available O objetivo deste estudo foi avaliar a adição de concentrações de carvão ativado em meio de cultura ½ MS (com metade da concentração dos macronutrientes sob dois espectros luminosos para a obtenção de plântulas in vitro de Cattleya loddigesii. Plântulas com aproximadamente 90 dias foram subcultivadas em oito tratamentos, nos quais foi testada a adição ao meio de cultura ½ MS com carvão ativado (0; 0,5; 1,0 e 2,0g L-1 e combinados sob espectro de luz branca e luz vermelha. Após 180 dias da germinação, foram mensurados dados biométricos (raiz e parte aérea, massa fresca e teores de pigmentos fotossintéticos. Em plântulas aclimatizadas em casa de vegetação, foram avaliadas a taxa de sobrevivência após 120 dias. As concentrações de clorofila total, clorofila a e carotenoides foram maiores nos tratamentos sob luz branca, enquanto a luz vermelha influenciou significativamente maior clorofila b, plântulas com menos raízes e de menor comprimento e elevada mortalidade ex vitro. A adição de 2,0g L-1 de carvão ativado ao meio de cultura e o uso de luz branca proporcionaram maior eficiência de desenvolvimento tanto para as culturas in vitro quanto para a sobrevivência ex vitro das plantas.The aim of this study was to evaluate the effect of different concentrations of activated charcoal in ½ MS (half concentration of macronutrients culture medium under two light spectra on the in vitro growth of Cattleya loddigesii seedlings. Plantlets with approximately 90 days were subcultured under eight treatments, consisting of different active charcoal concentrations (0; 0.5; 1.0 and 2.0g L-1 in ½ MS medium combined with white and red light spectra. After 180 days of germination, biometric data, fresh weight, and the level of photosynthetic pigments were evaluated. Plantlets acclimatized in a greenhouse were evaluated for survival after 120 days. Total chlorophyll, chlorophyll a, and carotenoid concentrations were higher in

Growing halophytes floating at sea

Directory of Open Access Journals (Sweden)

Ricardo Radulovich

2017-11-01

Full Text Available Freshwater shortages are increasingly limiting both irrigated and rainfed agriculture. To expand possibilities for controlled plant production without using land nor freshwater, we cultivated potted halophytes floating at sea that were provided with rain- and seawater. Plantlets of two mangroves (Avicennia germinans and Rhizophora mangle and plants of two herbaceous species, sea purslane (Sesuvium portulacastrum and salt couch grass (Sporobolus virginicus were grown in near-coastal tropical Pacific waters of Costa Rica for 733 days. There were a total of 504 rainless days, including two dry periods of ca. 150 d long each, evidencing prolonged and exclusive reliance on seawater. Pots with a sandy soil mixture and the transplanted plants were placed on low-cost wooden floating rafts with their lower end perforated and immersed for capillary rise of water. Free seawater entry and exit through the bottom from bobbing with waves, which also occasionally added water from the top, effectively controlled soil salinity build-up even during the rainless seasons. Continuous leaching made necessary frequent fertilizer addition. No water deficit symptoms were observed and midday canopy temperature during rainless periods was not significantly different between species or from air temperature. With all-year-round growth, height increase of mangrove plantlets ranged from 208.1 to 401.5 mm yr−1. Fresh biomass production of sea purslane and the grass was 10.9 and 3.0 kg m−2 yr−1 respectively. High yield, edibility and protein content of 10.2% dry weight established sea purslane as a potential crop. While further research is needed, the method evidenced to be a viable plant production option of potentially far-reaching applications.

The effects of gamma irradiation on growth and expression of genes encoding DNA repair-related proteins in Lombardy poplar (Populus nigra var. italica).

Science.gov (United States)

Nishiguchi, Mitsuru; Nanjo, Tokihiko; Yoshida, Kazumasa

2012-07-01

In this study, to elucidate the mechanisms of adaptation and tolerance to ionizing radiation in woody plants, we investigated the various biological effects of γ-rays on the Lombardy poplar (Populus nigra L. var. italica Du Roi). We detected abnormal leaf shape and color, fusion, distorted venation, shortened internode, fasciation and increased axillary shoots in γ-irradiated poplar plants. Acute γ-irradiation with a dose of 100Gy greatly reduced the height, stem diameter and biomass of poplar plantlets. After receiving doses of 200 and 300Gy, all the plantlets stopped growing, and then most of them withered after 4-10 weeks of γ-irradiation. Comet assays showed that nuclear DNA in suspension-cultured poplar cells had been damaged by γ-rays. To determine whether DNA repair-related proteins are involved in the response to γ-rays in Lombardy poplars, we cloned the PnRAD51, PnLIG4, PnKU70, PnXRCC4, PnPCNA and PnOGG1 cDNAs and investigated their mRNA expression. The PnRAD51, PnLIG4, PnKU70, PnXRCC4 and PnPCNA mRNAs were increased by γ-rays, but the PnOGG1 mRNA was decreased. Moreover, the expression of PnLIG4, PnKU70 and PnRAD51 was also up-regulated by Zeocin known as a DNA cleavage agent. These observations suggest that the morphogenesis, growth and protective gene expression in Lombardy poplars are severely affected by the DNA damage and unknown cellular events caused by γ-irradiation. Copyright © 2012 Elsevier Ltd. All rights reserved.

Efficient plant regeneration through somatic embryogenesis from callus cultures of Oncidium (Orchidaceae).

Science.gov (United States)

Chen, J -T.; Chang, W -C.

2000-12-07

An efficient method was established for high frequency somatic embryogenesis and plant regeneration from callus cultures of a hybrid of sympodial orchid (Oncidium 'Gower Ramsey'). Compact and yellow-white embryogenic calli formed from root tips and cut ends of stem and leaf segments on 1/2 MS [11] basal medium supplemented with 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ, 0.1-3 mg/l), 2,4-dichlorophenoxyacetic acid (2,4-D, 3-10 mg/l) and peptone (1 g/l) for 4-7 weeks. Embryogenic callus was maintained by subculture on the same medium for callus induction and proliferated 2-4 times (fresh weight) in 1 month. Initiation of somatic embryogenesis and development up to the protocorm-like-bodies (PLBs) from callus cultures was achieved on hormone-free basal medium. Regenerants were recovered from somatic embryos (SEs) after transfer to the same medium and showed normal development. The optimized protocol required about 12-14 weeks from the initiation of callus to the plantlet formation. Generally, the frequency of embryo formation of root-derived callus was higher than stem- and leaf-derived calli. Combinations of naphthaleneacetic acid (NAA) and TDZ significantly promoted embryo formation from callus cultures. The high-frequency (93.8%) somatic embryogenesis and an average of 29.1 SEs per callus (3x3 mm(2)) was found in root-derived callus on a basal medium supplemented with 0.1 mg/l NAA and 3 mg/l TDZ. Almost all the SEs converted and the plantlets grew well with an almost 100% survival rate when potted in sphagnum moss and acclimatized in the greenhouse.

In vitro propagation and assessment of the genetic fidelity of Musa acuminata (AAA) cv. Vaibalhla derived from immature male flowers.

Science.gov (United States)

Hrahsel, Lalremsiami; Basu, Adreeja; Sahoo, Lingaraj; Thangjam, Robert

2014-02-01

An efficient in vitro propagation method has been developed for the first time for Musa acuminata (AAA) cv. Vaibalhla, an economically important banana cultivar of Mizoram, India. Immature male flowers were used as explants. Murashige and Skoog's (MS) medium supplemented with plant growth regulators (PGRs) were used for the regeneration process. Out of different PGR combinations, MS medium supplemented with 2 mg L(-1) 6-benzylaminopurine (BAP) + 0.5 mg L(-1) α-naphthalene acetic acid (NAA) was optimal for production of white bud-like structures (WBLS). On this medium, explants produced the highest number of buds per explant (4.30). The highest percentage (77.77) and number (3.51) of shoot formation from each explants was observed in MS medium supplemented with 2 mg L(-1) kinetin + 0.5 mg L(-1) NAA. While MS medium supplemented with a combination of 2 mg L(-1) BAP + 0.5 mg L(-1) NAA showed the maximum shoot length (14.44 cm). Rooting efficiency of the shoots was highest in the MS basal medium without any PGRs. The plantlets were hardened successfully in the greenhouse with 96% survival rate. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were employed to assess the genetic stability of in vitro regenerated plantlets of M. acuminata (AAA) cv. Vaibalhla. Eight RAPD and 8 ISSR primers were successfully used for the analysis from the 40 RAPD and 30 ISSR primers screened initially. The amplified products were monomorphic across all the regenerated plants and were similar to the mother plant. The present standardised protocol will find application in mass production, conservation and genetic transformation studies of this commercially important banana.

Propagation of goldenrod (Solidago canadensis L. from leaf and nodal explants

Directory of Open Access Journals (Sweden)

Jun Li

2012-02-01

Full Text Available Goldenrod (Solidago canadensis L. is an invasive plant species in many countries except North America but a cut-flower species worldwide. There is a need to generate and propagate goldenrod clones efficiently for research and commercial purposes. A callus induction and plantlet regeneration system was developed by studying the influence of explant type and different concentrations of plant growth regulators. The highest callus production from leaf segments was obtained on Murashige and Skoog’s medium (MS medium supplemented with 1.0 mg/L naphthalene acetic acid (NAA and 1.0 mg/L 6-benzylaminopurine (BA. Adventitious shoots could be regenerated directly from leaf explants without an intermediate callus phase with the highest shoot induction percentage of 87.2%. The largest number of adventitious shoots per leaf explant (3.2 was obtained on MS medium supplemented with 0.4 mg/L NAA and 2.0 mg/L BA. MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L BA was the best medium for axillary shoot regeneration from nodal segments. The highest root number and longest roots occurred on half-strength MS without the addition of any growth regulator. Rooted plantlets were then transferred to a soil-based growth medium, placed in a greenhouse, and acclimatized with 100% success. All surviving plants grew normally without showing any morphological varia­tion when compared to those grow from seed. This regeneration protocol may be used to produce certain biotypes of goldenrod suitable for genetic transformation rapid propagation of goldenrod for commercial purposes or for screening fungi and toxins as potential biocontrol agents against this weed.

Secondary Metabolite Content in Roots and Callus of Paeonia Anomala L.

Directory of Open Access Journals (Sweden)

A.A. ZARIPOVA

2014-06-01

Full Text Available Taking into account the fact that in the process of introduction in vitro culture the change of secondary metabolite content may take place we compared the chemical composition of plant material of wild-growing plants of Paeonia anomala introduced and produced using the methods of clonal micropropagation, callus tissue.The content of phenolic compounds, that is catechins and gallatos was compared. Paeoniflorin content was estimated by direct spectrophotometry of methanol extracts (λ = 231,7 nm, ε 1% 1 sm = 265.4. Integral characteristic of the obtained extracts was received by comparison of absorption spectra using spectrophotometer SP - 121 within wave diapason 300 - 460 nm.Light absorption curves of methanol extracts had two distinct peaks at λ = 232 nm and λ = 275 nm typical of paeoniflorin. Paeoniflorin content was 80 % higher in young peony roots than in control plant. This glycoside content in callus culture was 44 % higher than in wild-growing plant roots and 26 % lower than in plantlet roots.The use of ethanol as extragent showed a higher content of extracted substances in callus tissue. The comparison of the obtained spectra in the region corresponding to phenolic compound absorption shows the highest phenolic compound content in callus tissue and young plant roots. Judging by light absorption maximum it may be phenolic acids. The lowest phenolic compound content was determined in adult wild-growing plant roots, where a high phlobaphene content was visually observed.The conducted research confirms the fact that callus culture of Paeonia anomala L. is a perspective producer of monoterpene glycosides and phenolic compounds. Extracts from plantlets and callus culture exceed in biological active substance content rootstock extracts of open air plants.

Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine.

Science.gov (United States)

Manganaris, G A; Economou, A S; Boubourakas, I N; Katis, N I

2003-10-01

The plum pox virus (PPV) and prunus necrotic ringspot virus (PNRSV) cause serious disease problems in stone-fruit trees. In this work, the possibility of obtaining plant material free from these viruses through thermotherapy and meristem-tip culture from infected nectarine shoots (Prunus persica var. nectarina Max, cv. 'Arm King') was studied. In addition, the detection of these viruses in in vitro cultures and young acclimatized plantlets with double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) and multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) was studied. Meristem-tip explants (0.8-1.3 mm) derived from sprouted buds of winter wood and spring shoots from field grown plants had a 2-5% regeneration response. However, application of thermotherapy to potted nectarine trees (3 weeks at a maximum temperature of 35 degrees C) facilitated excision of longer meristem tips (1.3-2.0 mm) that resulted in a significantly higher regeneration response (38%) in woody plant medium (WPM) without plant growth regulators. Such explants formed multiple shoots with the addition of 8 microM benzylaminopurine and 0.8 microM indoleacetic acid. When they were tested for the presence of PPV and PNRSV, 86% and 81% were found to be virus-free as detected by DAS-ELISA and multiplex RT-PCR, respectively. Individual shoots excised from virus-free cultures readily rooted in vitro (half-strength WPM plus 2 microM indolebutyric acid) and grew to plantlets. The combination of an efficient protocol for virus elimination and the establishment of highly sensitive diagnostics resulted in the production of nectarine plants free from PPV and PNRSV.

Varietal performance of potato on induction and development of microtuber in response to sucrose

Directory of Open Access Journals (Sweden)

Md. Sadek Hossain

2017-06-01

Full Text Available Tissue culture techniques are the most momentous and extensively method to produce disease-free quality plantlets and microtubers in potato. In spite of this, there has been a lack of research information into in vitro microtuberization for seed tuber production program in Bangladesh. In this experiment, we assessed the appropriateness of different concentrations of sucrose with especial accent on their effects for induction and development of microtuber in potato varieties. In vitro plantlets of three potato varieties; Asterix, Granola, and Diamant were treated with eight level of sucrose as 0, 3, 4, 6, 8, 10, 12 and 14% for 70 days of incubation producing microtuber. Asterix induces microtuber after 10.69 days, it was statistically shorter duration than other two varieties. Tuberization did not occur without sucrose and were required a minimum (8.92 days with 8% sucrose, while it delayed with either increasing or decreasing rate of sucrose concentration. A single number of microtuber was not formed in absence of sucrose after 28 days of incubation. Microtuber plantlet−1 was the highest more or less in all varieties at harvest with 8% sucrose concentration. Hundred percent of microtuber below 250 mg induced in 3% sucrose concentration and from then the microtuber grade induction decreased with the increase of sucrose concentration. >500 mg grade microtuber produced by Granola (47.95% with 10% sucrose while Diamant produced 50.15% above 500 mg grade microtuber with 14% sucrose. It is also noticeable by Astrerix variety where >500 mg microtuber were produced about 46.95% with 8% sucrose.

Application of RNAi to confirm theobromine as the major intermediate for caffeine biosynthesis in coffee plants with potential for construction of decaffeinated varieties.

Science.gov (United States)

Ogita, Shinjiro; Uefuji, Hirotaka; Morimoto, Masayuki; Sano, Hiroshi

2004-04-01

The caffeine biosynthetic pathway in coffee plants has been proposed to involve three distinct N -methyltransferases, xanthosine methyltransferase (XMT), 7- N -methylxanthine methyltransferase (MXMT; theobromine synthase), and 3,7-dimethylxanthine methyltransferase (DXMT; caffeine synthase). We previously isolated all corresponding cDNAs designated as CaXMT1 , CaMXMT1 , CaMXMT2 and CaDXMT1 , respectively, and showed that caffeine was indeed synthesized in vitro by the combination of their gene products. In order to regulate caffeine biosynthesis in planta , we suppressed expression of CaMXMT1 by the double stranded RNA interference (RNAi) method. For this purpose, we first established a protocol for efficient somatic embryogenesis of Coffea arabica and C. canephora , and then Agrobacterium -mediated transformation techniques. The RNAi transgenic lines of embryogenic tissues derived from C. arabica and transgenic plantlets of C. canephora demonstrated a clear reduction in transcripts for CaMXMT1 in comparison with the control plants. Transcripts for CaXMT1 and CaDXMT1 were also reduced in the most cases. Both embryonic tissues and plantlets exhibited a concomitant reduction of theobromine and caffeine contents to a range between 30% and 50% of that of the control. These results suggest that the CaMXMT1 -RNAi sequence affected expression of not only CaMXMT1 itself, but also CaXMT1 and CaDXMT1 , and that, since the reduction in theobromine content was proportional to that for caffeine, it is involved in the major synthetic pathway in coffee plants. The results also indicate that the method can be practically applied to produce decaffeinated coffee plants.

A genetically stable rooting protocol for propagating a threatened medicinal plant—Celastrus paniculatus

Science.gov (United States)

Phulwaria, Mahendra; Rai, Manoj K.; Patel, Ashok Kumar; Kataria, Vinod; Shekhawat, N. S.

2012-01-01

Celastrus paniculatus, belonging to the family Celastraceae, is an important medicinal plant of India. Owing to the ever-increasing demand from the pharmaceutical industry, the species is being overexploited, thereby threatening its stock in the wild. Poor seed viability coupled with low germination restricts its propagation through sexual means. Thus, alternative approaches such as in vitro techniques are highly desirable for large-scale propagation of this medicinally important plant. Nodal segments, obtained from a 12-year-old mature plant, were used as explants for multiple shoot induction. Shoot multiplication was achieved by repeated transfer of mother explants and subculturing of in vitro produced shoot clumps on Murashige and Skoog's (MS) medium supplemented with various concentrations of 6-benzylaminopurine (BAP) alone or in combination with auxin (indole-3-acetic acid (IAA) or α-naphthalene acetic acid (NAA)). The maximum number of shoots (47.75 ± 2.58) was observed on MS medium supplemented with BAP (0.5 mg L−1) and IAA (0.1 mg L−1). In vitro raised shoots were rooted under ex vitro conditions after treating them with indole-3-butyric acid (300 mg L−1) for 3 min. Over 95 % of plantlets acclimatized successfully. The genetic fidelity of the regenerated plants was assessed using random amplified polymorphic DNA. No polymorphism was detected in regenerated plants and the mother plant, revealing the genetic fidelity of the in vitro raised plantlets. The protocol discussed could be effectively employed for large-scale multiplication of C. paniculatus. Its commercial application could be realized for the large-scale multiplication and supply to the State Forest Department.

Proline metabolism in the wild-type and in a salt-tolerant mutant of nicotiana plumbaginifolia studied by (13)C-nuclear magnetic resonance imaging

Science.gov (United States)

Roosens; Willem; Li; Verbruggen; Biesemans; Jacobs

1999-12-01

To obtain insight into the link between proline (Pro) accumulation and the increase in osmotolerance in higher plants, we investigated the biochemical basis for the NaCl tolerance of a Nicotiana plumbaginifolia mutant (RNa) that accumulates Pro. Pro biosynthesis and catabolism were investigated in both wild-type and mutant lines. (13)C-Nuclear magnetic resonance with [5-(13)C]glutamate (Glu) as the Pro precursor was used to provide insight into the mechanism of Pro accumulation via the Glu pathway. After 24 h under 200 mM NaCl stress in the presence of [5-(13)C]Glu, a significant enrichment in [5-(13)C]Pro was observed compared with non-stress conditions in both the wild type (P2) and the mutant (RNa). Moreover, under the same conditions, [5-(13)C]Pro was clearly synthesized in higher amounts in RNa than in P2. On the other hand, measurements of enzyme activities indicate that neither the biosynthesis via the ornithine pathway, nor the catabolism via the Pro oxidation pathway were affected in the RNa mutant. Finally, the regulatory effect exerted by Pro on its biosynthesis was evaluated. In P2 plantlets, exogenous Pro markedly reduced the conversion of [5-(13)C]Glu into [5-(13)C]Pro, whereas Pro feedback inhibition was not detected in the RNa plantlets. It is proposed that the origin of tolerance in the RNa mutant is due to a mutation leading to a substantial reduction of the feedback inhibition normally exerted in a wild-type (P2) plant by Pro at the level of the Delta-pyrroline-5-carboxylate synthetase enzyme.

Proline Metabolism in the Wild-Type and in a Salt-Tolerant Mutant of Nicotiana plumbaginifolia Studied by 13C-Nuclear Magnetic Resonance Imaging1

Science.gov (United States)

Roosens, Nancy H.; Willem, Rudolph; Li, Yan; Verbruggen, Ingrid; Biesemans, Monique; Jacobs, Michel

1999-01-01

To obtain insight into the link between proline (Pro) accumulation and the increase in osmotolerance in higher plants, we investigated the biochemical basis for the NaCl tolerance of a Nicotiana plumbaginifolia mutant (RNa) that accumulates Pro. Pro biosynthesis and catabolism were investigated in both wild-type and mutant lines. 13C-Nuclear magnetic resonance with [5-13C]glutamate (Glu) as the Pro precursor was used to provide insight into the mechanism of Pro accumulation via the Glu pathway. After 24 h under 200 mm NaCl stress in the presence of [5-13C]Glu, a significant enrichment in [5-13C]Pro was observed compared with non-stress conditions in both the wild type (P2) and the mutant (RNa). Moreover, under the same conditions, [5-13C]Pro was clearly synthesized in higher amounts in RNa than in P2. On the other hand, measurements of enzyme activities indicate that neither the biosynthesis via the ornithine pathway, nor the catabolism via the Pro oxidation pathway were affected in the RNa mutant. Finally, the regulatory effect exerted by Pro on its biosynthesis was evaluated. In P2 plantlets, exogenous Pro markedly reduced the conversion of [5-13C]Glu into [5-13C]Pro, whereas Pro feedback inhibition was not detected in the RNa plantlets. It is proposed that the origin of tolerance in the RNa mutant is due to a mutation leading to a substantial reduction of the feedback inhibition normally exerted in a wild-type (P2) plant by Pro at the level of the Δ-pyrroline-5-carboxylate synthetase enzyme. PMID:10594115

Recent advances in the cryopreservation of shoot-derived germplasm of economically important fruit trees of Actinidia, Diospyros, Malus, Olea, Prunus, Pyrus and Vitis.

Science.gov (United States)

Benelli, Carla; De Carlo, Anna; Engelmann, Florent

2013-01-01

This paper presents the advances made over the last decade in cryopreservation of economically important vegetatively propagated fruit trees. Cryopreservation protocols have been established using both dormant buds sampled on field-grown plants and shoot tips sampled on in vitro plantlets. In the case of dormant buds, scions are partially dehydrated by storage at -5 °C, and then cooled slowly to -30 °C using low cooling rates (c.a. 1 °C/h) before immersion in liquid nitrogen. After slow rewarming and rehydration of samples, regrowth takes place either through grafting of buds on rootstocks or excision of apices and inoculation in vitro. In the case of shoot tips of in vitro plantlets, the cryopreservation techniques employed are the following: controlled rate cooling procedures involving slow prefreezing followed by immersion in liquid nitrogen or vitrification-based procedures including encapsulation-dehydration, vitrification, encapsulation-vitrification and droplet-vitrification. The current status of cryopreservation for a series of fruit tree species including Actinidia, Diospyros, Malus, Olea, Prunus, Pyrus and Vitis is presented. Routine application of cryopreservation for long-term germplasm storage in genebanks is currently limited to apple and pear, for which large cryopreserved collections have been established at NCGRP, Fort Collins (USA), using dormant buds and in vitro shoot tips, respectively. However, there are a growing number of examples of pilot scale testing experiments under way for different species in various countries. Progress in the further development and application of cryopreservation techniques will be made through a better understanding of the mechanisms involved in the induction of tolerance to dehydration and cryopreservation in frozen explants. Copyright © 2012 Elsevier Inc. All rights reserved.

Melatonin-producing endophytic bacteria from grapevine roots promote the abiotic stress-induced production of endogenous melatonin in their hosts

Directory of Open Access Journals (Sweden)

Jian Jiao

2016-09-01

Full Text Available Endophytes form symbiotic relationships with plants and constitute an important source of phytohormones and bioactive secondary metabolites for their hosts. To date, most studies of endophytes have focused on the influence of these microorganisms on plant growth and physiology and their role in plant defenses against biotic and abiotic stressors; however, to the best of our knowledge, the ability of endophytes to produce melatonin has not been reported. In the present study, we isolated and identified root-dwelling bacteria from three grapevine varieties and found that, when cultured under laboratory conditions, some of the bacteria strains secreted melatonin and tryptophan-ethyl ester. The endophytic bacterium Bacillus amyloliquefaciens SB-9 exhibited the highest level of in vitro melatonin secretion and also produced three intermediates of the melatonin biosynthesis pathway: 5-hydroxytryptophan, serotonin, and N-acetylserotonin. After B. amyloliquefaciens SB-9 colonization, the plantlets exhibited increased plant growth. Additionally, we found that, in grapevine plantlets exposed to salt or drought stress, colonization by B. amyloliquefaciens SB-9 increased the upregulation of melatonin synthesis, as well as that of its intermediates, but reduced the upregulation of grapevine tryptophan decaboxylase genes (VvTDCs and a serotonin N-acetyltransferase gene (VvSNAT transcription, when compared to the un-inoculated control. Colonization by B. amyloliquefaciens SB-9 was also able to counteract the adverse effects of salt- and drought-induced stress by reducing the production of malondialdehyde and reactive oxygen species (H2O2 and O2− in roots. Therefore, our findings demonstrate the occurrence of melatonin biosynthesis in endophytic bacteria and provide evidence for a novel form of communication between beneficial endophytes and host plants via melatonin.

TDZ-Induced Plant Regeneration in Jatropha curcas: A Promising Biofuel Plant

KAUST Repository

Kumar, Nitish

2018-03-23

In recent years, Jatropha curcas has pronounced attention due to its capacity of production of biodiesel. Uniform large-scale propagation of J. curcas is one of the significant keys that will eventually decide victory. Direct regeneration is one of the methods which help in the production of uniform and homogenous plant, and TDZ plays an important role in the production of plantlets by direct organogenesis in several number of plant species including J. curcas. Measuring the economical importance of J. curcas and the role of TDZ in shoot regeneration, the present book chapter briefly reviews the impact of TDZ on shoot bud induction from various explants of J. curcas.

Tissue culture regeneration and radiation induced mutagenesis in banana

International Nuclear Information System (INIS)

Kulkarni, V.M.; Ganapathi, T.R.

2009-01-01

Radiation induced mutagenesis is an important tool for banana genetic improvement. At BARC, protocols for shoo-tip multiplication of commercial banana varieties have been developed and transferred to user agencies for commercial production. Excellent embryogenic cell suspensions were established in banana cvs. Rasthali and Rajeli, and were maintained at low temperatures for long-term storage. Normal plantlets were successfully regenerated from these cell suspensions. The cell suspensions and shoot-tip cultures were gamma-irradiated for mutagenesis. The mutagenized populations were field screened and a few interesting mutants have been isolated. The existence of genetic variation was confirmed using DNA markers. Further evaluation of these mutants is in progress. (author)

Development of technique on the induction and selection of in vitro mutant lines (Potato, Solanum tuberosum L.)

Energy Technology Data Exchange (ETDEWEB)

Yoo, Jang Ryoel; Lee, Yeong Il; Song, Hee Seop; Kim, Jae Seong; Sin, In Cheol; Lee, Sang Jae; Lee, Ki Un; Lim, Yong Taek [Korea Atomic Energy Res. Inst., Taejon (Korea, Republic of)

1993-09-01

For the development of the technique on the plant tissue culture and application of nuclear technique in the in vitro mutation breeding, present research laid emphasis on the development of techniques of potato tissue culture, and on the induction and selection of radiation mutation. Another culture for haploid induction, optimum radiation dosage for cybrid formation of potato and mutation induction from in vitro cultured microtuber and plantlets were investigated for modelling the technique on the induction and selection of in vitro mutant lines. Inheritance stability of the selected mutants were also studied in field condition. In vitro system of micropropagation and selection of mutation was summarized.

Reduced expression of AtNUP62 nucleoporin gene affects auxin response in Arabidopsis

DEFF Research Database (Denmark)

Boeglin, Martin; Fuglsang, Anja Thoe; Luu, Doan Trung

2016-01-01

seedlings and at the adult stage in stipules of cauline leaves. The atnup62-1 mutant, harbouring a T-DNA insertion in intron 5, was identified as a knock-down mutant. It displayed developmental phenotypes that suggested defects in auxin transport or responsiveness. Atnup62 mutant plantlets were found...... to be hypersensitive to auxin, at the cotyledon and root levels. The phenotype of the AtNUP62-GFP overexpressing line further supported the existence of a link between AtNUP62 and auxin signalling. Furthermore, the atnup62 mutation led to an increase in the activity of the DR5 auxin-responsive promoter, and suppressed...

The Influence of Explant Types and Orientation on in Vitro Culture

Directory of Open Access Journals (Sweden)

Kamnoon KANCHANAPOOM

2011-08-01

Full Text Available Inflorescence, apical and lateral buds of Musa balbisiana ‘Kluai Hin’ (BBB group were used to culture on MS medium supplemented with 22 μM BA and 15% (v/v coconut water. Comparison of bud orientation showed that the best response of in vitro shoot tip proliferation was obtained with abaxial surface of buds lying down i.e. one side touching the medium (tilt. Mass propagation of shoot tips was obtained when cultured buds on MS medium containing 44 μM BA. Rooting was achieved by transferring the isolated shoots to MS basal medium without growth regulators. Rooted plantlets were acclimatized and successfully established in soil.

TDZ-Induced Plant Regeneration in Jatropha curcas: A Promising Biofuel Plant

KAUST Repository

Kumar, Nitish; Bhatt, Vacha D.; Mastan, Shaik G.; Reddy, Muppala P.

2018-01-01

In recent years, Jatropha curcas has pronounced attention due to its capacity of production of biodiesel. Uniform large-scale propagation of J. curcas is one of the significant keys that will eventually decide victory. Direct regeneration is one of the methods which help in the production of uniform and homogenous plant, and TDZ plays an important role in the production of plantlets by direct organogenesis in several number of plant species including J. curcas. Measuring the economical importance of J. curcas and the role of TDZ in shoot regeneration, the present book chapter briefly reviews the impact of TDZ on shoot bud induction from various explants of J. curcas.

Development of technique on the induction and selection of in vitro mutant lines (Potato, Solanum tuberosum L.)

International Nuclear Information System (INIS)

Yoo, Jang Ryoel; Lee, Yeong Il; Song, Hee Seop; Kim, Jae Seong; Sin, In Cheol; Lee, Sang Jae; Lee, Ki Un; Lim, Yong Taek

1993-09-01

For the development of the technique on the plant tissue culture and application of nuclear technique in the in vitro mutation breeding, present research laid emphasis on the development of techniques of potato tissue culture, and on the induction and selection of radiation mutation. Another culture for haploid induction, optimum radiation dosage for cybrid formation of potato and mutation induction from in vitro cultured microtuber and plantlets were investigated for modelling the technique on the induction and selection of in vitro mutant lines. Inheritance stability of the selected mutants were also studied in field condition. In vitro system of micropropagation and selection of mutation was summarized

Effects of growth regulators and activated charcoal for the micropropagation improvement of Dendrobium friedericksianum Rchb.f.

Directory of Open Access Journals (Sweden)

Prasertsongskun, S. and

2004-09-01

Full Text Available The effects of different concentrations of growth regulators and activated charcoal on micropropagation were studied. Seedlings of orchid, Dendrobium friedericksianum Rchb.f, cultured on Vacin and Went medium containing 1.0 mg/l α-naphthaleneacetic acid (NAA and 1.0 mg/l kinetin (KN significantly yielded per cultured plant, the highest average number of leaves (4.40, roots (8.30, shoot length (1.67 centimeters and fresh weight (0.23 grams. Particularly when activated charcoal was added (0.3% w/v the average number of leaves, roots, shoot length and fresh weight per cultured plant increased. Percentage survival of the plantlet after 21 days of transplantation was 85%.

Growth promotion of pineapple 'vitória' by humic acids and burkholderia spp. during acclimatization Promoção do crescimento do abacaxizeiro 'vitória' por ácidos húmicos e Burkholderia spp. durante a aclimatização

Directory of Open Access Journals (Sweden)

Lílian Estrela Borges Baldotto

2010-10-01

Full Text Available In vitro propagation of pineapple produces uniform and disease-free plantlets, but requires a long period of acclimatization before transplanting to the field. Quicker adaptation to the ex vitro environment and growth acceleration of pineapple plantlets are prerequisites for the production of a greater amount of vigorous, well-rooted planting material. The combination of humic acids and endophytic bacteria could be a useful technological approach to reduce the critical period of acclimatization. The aim of this study was to evaluate the initial performance of tissue-cultured pineapple variety Vitória in response to application of humic acids isolated from vermicompost and plant growth-promoting bacteria (Burkholderia spp. during greenhouse acclimatization. The basal leaf axils were treated with humic acids while roots were immersed in bacterial medium. Humic acids and bacteria application improved shoot growth (14 and 102 %, respectively, compared with the control; the effect of the combined treatment was most pronounced (147 %. Likewise, humic acids increased root growth by 50 %, bacteria by 81 % and the combined treatment by 105 %. Inoculation was found to significantly increase the accumulation of N (115 %, P (112 % and K (69 % in pineapple leaves. Pineapple growth was influenced by inoculation with Burkholderia spp., and further improved in combination with humic acids, resulting in higher shoot and root biomass as well as nutrient contents (N 132 %, P 131 %, K 80 % than in uninoculated plantlets. The stability and increased consistency of the host plant response to bacterization in the presence of humic substances indicate a promising biotechnological tool to improve growth and adaptation of pineapple plantlets to the ex vitro environment.A propagação in vitro de abacaxizeiro produz mudas uniformes e sadias, mas exige longo período de aclimatização antes da transferência para o campo. A adaptação ao ambiente ex vitro seguida da

Effect of abscisic acid on photosynthetic parameters during ex vitro transfer of micropropagated tobacco plantlets

Czech Academy of Sciences Publication Activity Database

Pospíšilová, Jana; Synková, Helena; Haisel, Daniel; Baťková, Petra

2009-01-01

Roč. 53, č. 1 (2009), s. 11-20 ISSN 0006-3134 R&D Projects: GA ČR GA522/07/0227 Institutional research plan: CEZ:AV0Z50380511 Keywords : carotenoids * chlorophyll contents * chlorophyll fluorescence Subject RIV: ED - Physiology Impact factor: 1.656, year: 2009

Concentrações de ANA e BAP na micropropagação de abacaxizeiro L. Merrill (Ananas comosus e no cultivo hidropônico das plântulas obtidas in vitro The effect of ANA and BAP concentrations on the micropropagation and hydroponic cultures of pineapple

Directory of Open Access Journals (Sweden)

Cristiane Elizabeth Costa de Macêdo

2003-12-01

Full Text Available O efeito de diferentes concentrações de ANA e BAP na micropropagação do abacaxizeiro, bem como no cultivo em hidroponia das plântulas obtidas in vitro, foram estudados em brotos de abacaxizeiro da variedade Pérola, inoculados em meio de cultura básico MS, suplementado com os fitorreguladores BAP e ANA em diferentes concentrações. Caracteres morfológicos quantitativos relacionados ao crescimento dos brotos e das plântulas de abacaxizeiro foram avaliados respectivamente durante os cultivos in vitro e em hidroponia e mostraram que, o tratamento T1 (BAP = 1,0 mg L-1e ANA = 0,5 mg L-1 proporcionou a maior taxa média de regeneração de brotos e conseqüentemente uma maior produção de matéria fresca. Entretanto, a altura dos brotos e a formação de suas raízes foram maiores nos tratamentos T2 (BAP = 0,5 mg L-1 e ANA = 0,25 mg L-1 e T3 (BAP = 0,25 mg L-1 e ANA = 0,12 mg L-1 . Após sessenta dias de cultivo em hidroponia, todas as plântulas oriundas do tratamento T1 apresentaram um bom desenvolvimento, expresso pela maioria dos caracteres morfológicos avaliados. O sistema de micropropagação utilizado neste trabalho possibilitou a obtenção de brotos de abacaxizeiro Pérola, em quantidade suficiente e ao mesmo tempo de fácil individualização, seguida da regeneração de plântulas que foram cultivadas em hidroponia.The effect of different ANA and BAP concentrations on in vitro and hydroponic cultures were studied. Pineapple shoots derived from Pérola explants variety were inoculated in MS media containing BAP and ANA in different concentrations. Growth parameters of shoots and plantlets were measured for in vitro and hydroponic cultures. Showed a highest multiplication rates of shoots and consequently highest fresh matter production were obtained with BAP and ANA at the concentrations of 1,0 and 0,5 mg L-1 respectively. However, the shoot length as well as the root number formed were higher in the T2 (0,5 de BAP + 0,25 de ANA

Concentration of nutrient solution in the hydroponic production of potato minitubers Concentração da solução nutritiva na produção hidropônica de minitubérculos de batata

Directory of Open Access Journals (Sweden)

Manuel Benito Novella

2008-09-01

Full Text Available The effect of the nutrient solution concentration on potato plant growth and minituber yield were determined in a sand closed hydroponic system. Minitubers and micropropagated plantlets of the cv. 'Macaca' were used. Treatments were five nutrient solution concentrations at electrical conductivities (EC of 1.0 (T1, 2.2 (T2, 3.4 (T3, 4.7 (T4 and 5.8dS m-1 (T5. The split plot randomised experimental design was used with three replications. Plants from minitubers produced higher fresh and mean weight of minitubers, shoot dry mass and leaf area index than the micropropagated ones. However, higher dry mass of minitubers was found with micropropagated plantlets compared to minitubers. The concentration of the nutrient solution did not affect minituber number. Increasing the nutrient solution concentration decreased total and minituber dry mass production of micropropagated plantlets and plant growth and minituber production of minituber-originated plants. Low concentration of nutrient solution at an EC of about 1.0dS m-1 can be used in the hydroponic production of potato minitubers of both micropropagated and minituber-originated plants.Neste trabalho foi determinado o efeito da concentração da solução nutritiva no crescimento e na produtividade de minitubérculos de batata em um sistema hidropônico fechado empregando areia como substrato. Plântulas micropropagadas e minitubérculos foram plantados em 24 de março de 2004. Os tratamentos foram cinco soluções nutritivas com condutividades elétricas (CE de 1,0 (T1, 2,2 (T2, 3,4 (T3, 4,7 (T4 e 5,8dS m-1 (T5. O experimento foi conduzido em parcelas subdivididas no delineamento inteiramente casualizado com três repetições. Plantas originadas de minitubérculos produziram mais massa fresca total e média de minitubérculos, massa seca da parte aérea e maior índice de área foliar que plantas micropropagadas. Entretanto, maior massa seca dos minitubérculos foi obtida em plantas micropropagadas

Micropropagation, seed propagation and germplasm bank of Mandevilla velutina (Mart. Woodson Micropropagação, propagação por sementes e banco de germoplasma de Mandevilla velutina (Mart. Woodson

Directory of Open Access Journals (Sweden)

Ronaldo Biondo

2007-06-01

Full Text Available Mandevilla velutina (Mart. Woodson (Apocynaceae is a medicinal plant species with antivenom properties, native from Brazilian Savanna regions (Cerrado, which due to overexploitation and habitat deforestation is in danger of extinction. As an initiative for conserving this endangered but economically important plant species, a micropropagation protocol was developed and genotypes were stored in the Germplasm Bank "Cerrado In vitro". For the in vitro propagation of M. velutina, nodal segments were inoculated on Murashige and Skoog (MS medium supplemented with different concentrations of BA, Zeatin, 2ip, DTT and TDZ. Best multiplication ratio was achieved when to the medium 0.44 µM BA, ranging 1: 6.7, were added. Plantlets cultured on MS/2 medium supplemented with 26.85 µM NAA rooted successfully (50.5%. Although rooted and un-rooted plantlets acclimatized to soil conditions, great losses were observed within un-rooted plantlets, while the rooted presented 100 % survival. It was possible to maintain 43% of the M. velutina germplasm under healthy conditions for six months, with no subcultures, using the MS medium supplemented with 2% sucrose, 13.8 mM spermidine, 2% sorbitol and 2% dextrose.Mandevilla velutina (Mart. Woodson (Apocynaceae é uma planta medicinal de espécie nativa de regiões do Cerrado brasileiro com propriedades anti-ofídicas. Devido a alta exploração e devastação deste bioma, M. velutina está em perigo de extinção. Como uma iniciativa para a conservação desta espécie ameaçada e economicamente importante, um protocolo de micropropagação foi desenvolvido e genótipos foram colocados no Banco de Germoplasma "Cerrado In vitro". Para a propagação in vitro de M. velutina, segmentos nodais foram inoculados no meio Murashige and Skoog (MS suplementado com diferentes concentrações de BA, zeatina, 2ip, DTT e TDZ. A melhor resposta de multiplicação foi obtida em meio suplementado com 0.44 µM BA na razão 1: 6.7. Pl

Crescimento de mudas micropropagadas de bananeira aclimatizadas nas condições da Amazônia Sul Ocidental sob a influência de diferentes substratos e recipientes Growth of micropropagated banana plantlets acclimatized on conditions of South West Amazon under the influence of different substrates and recipients

Directory of Open Access Journals (Sweden)

Janiffe Peres de Oliveira

2008-06-01

. The aim of this work was to evaluate the growth of micropropagated banana plantlets during the acclimatization under the influence of different substrates and recipients on conditions of South West Amazon. Shoots of banana, cv. Grand Naine, were rooted in MS medium, being the rooting plants transferred to a nursery, in two types of plastic dibble tubes (115 cm³ and 180 cm³ and six different substrates formulated from different portions of soil, carbonized rice hulls and bovine manure. Evaluations of plant survival, height of the aerial part and pseudostem diameter were carried out each fifteen days, and at the end of 75 days, fresh and dry mass for the roots and aerial parts of the plants were also determined. It was verified that the plant survival was not influenced by the use of the 115 cm³ or 180 cm³ plastic dibble tubes. However, acclimatization accomplished in the 180 cm³ plastic dibble tubes provided larger growth in height and pseudostem diameter of the plants and, consequently, larger accumulation of fresh and dry mass of roots and aerial parts, when compared to the 115 cm³ plastic dibble tubes. Among the substrates, it was observed that bovine manure was fundamental as substrate component to obtain the best results.

Pengaruh penambahan auxin terhadap pertunasan dan perakaran kopi arabika perbanyakan Somatic Embryogenesis (The effects of shooting and rooting of arabica coffee propagation through Embryogenesis Somatic auxin uses.

Directory of Open Access Journals (Sweden)

Rina Arimarsetiowati

2012-08-01

Full Text Available Plantlet that has developed shoots and roots will have a high level adaptation in the field. The objective of this experiment was to improve the ability of planlet in shooting and rooting so that it is ready for acclimatization in the field. The increase ability in shooting and rooting of the planlet were conducted by adding various types of auxin in the media. The arabica coffee embryo of clone AS 2K which has entered the phase of the cotyledons was transfered into the treatment media containing half-strength of MS (Murashige & Skoog macro and micro nutrient, vitamin B5, 30 g/L glucose, 100 ml/L coconut water, 50 mg/L AgNO3 added with the combination of IAA, IBA and NAA. The research was conducted by using completely randomized design with seven combined treatment i.e. 0.1 mg/L IBA, 0.1 mg/L NAA, 0.1 mg/L IAA; 0 , 1 mg/L IBA + 0.1 mg/L NAA, 0.1 mg/L IBA + 0.1 mg/L IAA, 0.1 mg/L NAA + 0.1 mg/L IAA; without auxin. There were 12 replications in every treatment and each replication consisted of five cotyledonary embryos. The parameters of observation were the root length, leaf number, leaf area, stem diameter, and height of plantlets. The observations were conducted in eighth weeks after cotyledonary embryo had shoots. The results showed that in the number of leaves and height of planlet parameters, the treatment without auxin was the best result compared to planlet with auxin addition. The addition of auxin varians and their combination did not significantly influent leaf area, root length and stem diameter parameters. The medium tested was optimum for the growth of shoots and roots of AS 2K arabica coffee.

Generation of peanut drought tolerant plants by pingyangmycin-mediated in vitro mutagenesis and hydroxyproline-resistance screening.

Directory of Open Access Journals (Sweden)

Jiongming Sui

Full Text Available In order to enlarge the potential resources of drought-tolerant peanuts, we conducted in vitro mutagenesis with Pingyangmycin (PYM as the mutagen as well as directed screening on a medium supplemented with Hydroxyproline (HYP. After being extracted from mature seeds (cv. Huayu 20, the embryonic leaflets were cultured on somatic embryogenesis-induction medium with 4 mg/L PYM and the generated embryos were successively transferred to a germination medium with 4 and then 8 mmol/L HYP to screen HYP-tolerant plantlets. After that, these plantlets were grafted and transplanted to the experimental field. In the next generation, all seeds were sown in the field, and phenotype variation and trait segregation can be observed in most of the offspring (M2 generation. The M3 generation individuals were subjected to drought stress at the seedling stages. The activities of SOD and POD were substantially increased in eight offspring of 11 HYP-tolerant, regenerated plants than in their mutagenic parents. To determine the correlation between mutant phenotypes and genomic modification, we carried out a comparison of the DNA polymorphisms between the mutagenic parents and 13 M3 generation individuals from different HYP-tolerant, regenerated plants with SSR primers. Results showed that most mutants and parent plants had signs of polymorphisms. Under drought stress, some M3 generation individuals of 10 original HYP-tolerant, regenerated plants produced more pods than the mutagenic parent; twenty individuals among them produced >60 g pods/plant. M4-generation seeds were tested for quality characteristics by Near Infrared Spectroscopy (NIS and nine individuals with higher protein content (>30% and 21 individuals with higher oil content (>58% were screened. We concluded that the use of PYM-based in vitro mutagenesis in combination with directed screening with HYP is effective for the creation of potential drought-tolerant mutants of peanut.

Bacillus subtilis and Enterobacter cloacae endophytes from healthy Theobroma cacao L. trees can systemically colonize seedlings and promote growth.

Science.gov (United States)

Leite, Hianna Almeida Câmara; Silva, Anderson Barbosa; Gomes, Fábio Pinto; Gramacho, Karina Peres; Faria, José Cláudio; de Souza, Jorge Teodoro; Loguercio, Leandro Lopes

2013-03-01

Clonal genotypes resistant to fungal diseases are an important component of the cocoa production system in southeastern Bahia state (Brazil), so that technologies for faster production of stronger and healthier plantlets are highly desirable. In this study, the effects of inoculated bacterial endophytes isolated from healthy adult cacao plants on seedlings, and aspects related to inoculation methods, colonization patterns, and photosynthesis were investigated. Sequencing of 16S rRNA, hsp-60, and rpo-B genes placed the wild-type isolates within the species Enterobacter cloacae (isolates 341 and 344) and Bacillus subtilis (isolate 629). Spontaneous rifampicin-resistant (rif(R)) variants for 344 were also produced and tested. Endophytic application was either by immersion of surface sterilized seeds in bacterial suspensions or direct inoculation into soil, 20 days after planting non-inoculated seeds into pots. Results from in vitro recovery of inoculated isolates showed that the wild-type endophytes and rif(R) variants systemically colonized the entire cacao seedlings in 15-20 days, regardless of the inoculation method. Some endophytic treatments showed significant increases in seedlings' height, number of leaves, and dry matter. Inoculation methods affected the combined application of endophytes, which maintained the growth-promotion effects, but not in the same manner as in single applications. Interestingly, the 344-3.2 rif(R) variant showed improved performance in relation to both the wild type and another related variant. Photosynthetic rates and stomatal conductance increased significantly for some endophytic treatments, being partially associated with effects on growth and affected by the inoculation method. The results suggest that E. cloacae and B. subtilis endophytes from healthy adult plants (not transmitted by seeds) were able to promote vegetative growth on cacao seedlings. The development of products for large-scale use in seedlings/plantlets

ENRAIZAMENTO DE CRAVO (Dianthus caryophyllus L. IN VITRO E EX VITRO

Directory of Open Access Journals (Sweden)

G.R.F CUZZUOL

1996-01-01

Full Text Available Plântulas de cravo (Dianthus caryophyllus micropropagadas durante várias gerações pelo período de um ano, foram enraizadas in vitro com AIA, ANA e AIB nas concentrações de 0,0; 0,25; 0,5 e 1,0 mg/l, em fatorial do tipo 3 x 4, com todos os tratamentos promovendo a formação de raízes, mas não diferindo do controle. Foi confrontado em condição autotrófica, o desempenho entre plântulas enraizadas in vitro na presença e ausência do regulador AIA 0,5 mg/l e plântulas enraizadas ex vitro, sem nenhuma diferença quanto ao comprimento da parte aérea. Para a variável produção de massa de matéria seca os melhores resultados foram proporcionados pelas plantas que passaram pela fase de enraizamento in vitro, tendo o sistema radicular efeito sinergístico no crescimento da parte aérea.Plantlets of carnation (Dianthus caryophyllus L. micropropagated through several generations during one year, were observed with respect to rooting in vitro, in the presence of IAA , NAA and IBA, at the following concentrations: 0,0; 0,25; 0,5 and 1,0 mg/l. All treatments promoted root formation, however no differences were detected in comparison to control. As far as the lenght of the aerial part is concerned no difference was observed between in vitro rooting. in the presence or absence of IAA 0,5 mg/l, and ex vitro rooting. Plantlets which were rooted in vitro conditions showed higher production of fresh matter then those rooted ex vitro. The root system had a synergistic effect on the growth of the aerial part.

Establishment of an efficient protocol for micropropagation of some pakistani cultivars of date palm (phonex dactylifera l.) using novel inflorescence explants

International Nuclear Information System (INIS)

Jatoi, M. A.; Markhand, G. S.; Solangi, N.

2015-01-01

An efficient protocol for rapid and large scale In vitro propagation of some Pakistani cultivars of date palm has been established using inflorescence explants at Date Palm Research Institute (DPRI), Shah Abdul Latif University (SALU), Khairpur, Pakistan. Immature inflorescences of desired cultivars of date palm detached from mother palms followed by surface sterilization with low torrent of current tap water and then 30 percentage NaOCl/sub 2/ solution, the outer cover were removed in order to get spike explants and cut into the 2-3 cm small pieces and cultured on modified MS medium supplemented with 0.1 mg l-1 2, 4-D + 0.1 mg l-1 IAA + 5.0 mg l-1 NAA for initiation and establishment of cultures. The obtained somatic embryos were subjected to multiplication medium involved 0.1 mg l-1 NAA + 0.05 mg l-1BA. Rooting was achieved using quarter strength MS medium containing 0.1 mg l-1NAA without activated charcoal (AC) initially and then with 3 g l-1 AC. Strong rooted plantlets with 2-3 leaves were transferred to pots contained sand and peat moss mixture (1:1 v/v) with more than 95 percentage success in acclimatization. The acclimatized plants with at least one compound leaf were shifted to the open field conditions at SALU campus for further studying morphological and fruit characterization to ensure the true-to-type nature of tissue culture derived plantlets. High multiplication efficiency and survival percentage with no any somaclonal variation ensured the efficacy of the protocol developed for the production of elite cultivars of date palm of Pakistan and can be used to optimize production of other cultivars of date palm worldwide. (author)

Micropropagation and assessment of genetic fidelity of Dendrocalamus strictus (Roxb.) nees using RAPD and ISSR markers.

Science.gov (United States)

Goyal, Arvind Kumar; Pradhan, Sushen; Basistha, Bharat Chandra; Sen, Arnab

2015-08-01

Dendrocalamus strictus popularly known as 'Male bamboo' is a multipurpose bamboo which is extensively utilized in pharmaceutical, paper, agricultural and other industrial implements. In this study, in vitro regeneration of D. strictus through nodal culture has been attempted. Murashige and Skoog's medium supplemented with 4 mg/l BAP was found to be most effective in shoot regeneration with 3.68 ± 0.37 shoots per explant. The effect of Kn was found to be moderate. These hormones also had considerable effect on the shoot length. The highest shoot length after 6 weeks (3.11 ± 0.41 cm) was noted with 5 mg/l BAP followed by 3.07 ± 0.28 cm with 5 mg/l Kn, while decrease in the shoot length was noted with other treatments. The effect of IBA and NAA individually or in combination at different concentrations on rooting was evaluated. The highest number of root (1.36 ± 0.04) was regenerated on full-strength MS medium supplemented with 3 mg/l NAA, while maximum length of 1.64 ± 0.03 cm of roots was recorded with combination of 1 mg/l IBA and 3 mg/l NAA. Tissue-cultured plants thus obtained were successfully transferred to the soil. The clonal fidelity among the in vitro-regenerated plantlets was assessed by RAPD and ISSR markers. The ten RAPD decamers produced 58 amplicons, while nine ISSR primers generated a total of 66 bands. All the bands generated were monomorphic. These results confirmed the clonal fidelity of the tissue culture-raised D. strictus plantlets and corroborated the fact that nodal culture is perhaps the safest mode for multiplication of true to type plants.

Two-stage culture procedure using thidiazuron for efficient micropropagation of Stevia rebaudiana, an anti-diabetic medicinal herb.

Science.gov (United States)

Singh, Pallavi; Dwivedi, Padmanabh

2014-08-01

Stevia rebaudiana Bertoni, member of Asteraceae family, has bio-active compounds stevioside and rebaudioside which taste about 300 times sweeter than sucrose. It regulates blood sugar, prevents hypertension and tooth decay as well as used in treatment of skin disorders having high medicinal values, and hence there is a need for generating the plant on large scale. We have developed an efficient micropropagation protocol on half strength Murashige and Skoog (MS) media, using two-stage culture procedures. Varying concentrations of cytokinins, i.e., benzylaminopurine, kinetin and thidiazuron (TDZ) were supplemented in the nutrient media to observe their effects on shoot development. All the cytokinins promoted shoot formation, however, best response was observed in the TDZ (0.5 mg/l). The shoots from selected induction medium were sub-cultured on the multiplication media. The media containing 0.01 mg/l TDZ produced maximum number of shoot (11.00 ± 0.40) with longer shoots (7.17 ± 0.16) and highest number of leaves (61.00 ± 1.29). Rooting response was best observed in one-fourth strength on MS media supplemented with indole-3-butyric acid (1.0 mg/l) and activated charcoal (50 mg/l) with (11.00 ± 0.40) number of roots. The plantlets thus obtained were hardened and transferred to the pots with soil and sand mixture, where the survival rate was 80 % after 2 months. Quantitative analysis of stevioside content in leaves of in vivo mother plant and in vitro plantlets was carried out by high performance liquid chromatography. A remarkable increase in stevioside content was noticed in the in vitro-raised plants as compared to in vivo grown plants. The protocol reported here might be useful in genetic improvement and high stevioside production.

Micropropagation of Araucaria excelsa R. Br. var. glauca Carrière from orthotropic stem explants.

Science.gov (United States)

Sarmast, Mostafa Khoshhal; Salehi, Hassan; Khosh-Khui, Morteza

2012-07-01

The objectives of the present work were in vitro propagation of Araucaria excelsa R. Br. var. glauca Carrière (Norfolk Island pine) with focus on the evaluation of the mean number of shoots per explant (MNS/E) and mean length of shoots per explants (MLS/E) produced by different parts of the orthotropic stem of A. excelsa R. Br. var. glauca in response to plant growth regulators. Norfolk Island pine axillary meristems responded very well to the 2-iso-pentenyl adenine (2iP) and thidiazuron (TDZ) levels. Explants taken from stem upper segments in the media containing 2iP had a higher MNS/E (3.47) and MLS/E (6.27 mm) in comparison to those taken from stem lower segments, which were 0.71 and 0.51 mm, respectively. Using 0.045 μM TDZ in the MS medium not only resulted in 4.60 MNS/E with 7.08 mm MLS/E but proliferated shoots showed a good performance as well. Investigating the best position of stem explant on mother plant as well as the best concentrations of growth regulators were performed which were useful for efficient micropropagation of this plant. Thirty three percent of explants were rooted in the MS medium containing 3 % sucrose, supplemented with 7.5 μM of both NAA and IBA for 2 weeks before transferring to a half strength MS medium without any growth regulator. Plantlets obtained were acclimatized and transferred to the greenhouse with less than 20 % mortality. This procedure considered the first successful report for regeneration and acclimatization of A. excelsa R. Br. var. glauca plantlet through main stem explants.

The resistance response of sunflower genotypes to black stem disease under controlled conditions

Directory of Open Access Journals (Sweden)

Reza DARVISHZADEH

2010-09-01

Full Text Available Phoma black stem, caused by Phoma macdonaldii, is one of the most important diseases of sunflower in the world. The sources of resistance to Phoma black stem were investigated. A total of 184 genotypes, including some recombinant inbred lines (RILs, several M6 mutant lines obtained by gamma irradiation of seed of the genotype AS 613, and other genotypes from different countries, were evaluated against an aggressive French isolate (MP6 in controlled conditions. The study was carried out in a randomized complete block design with three replicates. Each replicate consisted of 10–12 seedlings. Twenty μL of spore suspension (106 pycnidiospores mL-1 were deposited on the intersection of the cotyledon petiole and the hypocotyl of sunflower plantlets at the two-leaf stage. The percentage of the area exhibiting disease symptoms was scored on the two cotyledon petioles of each of the plantlets three, five and seven days after inoculation. The disease progress rate (rd, as the slope of the regression line for disease severity against time, was also calculated. Analysis of variance detected significant differences among sunflower genotypes for disease severity 7 days after inoculation,as well as for the disease progress rate. A strong correlation (r=0.96, P<0.01 was found between disease severity 7 days after inoculation and the disease progress rate. The inbred lines F1250/03 (origin: Hungary, M5-54-1, M6-862-1 (mutant lines, SDR 18 (origin: USA and two wild Helianthus accessions, 1012 Nebraska and 211 Illinois, (wild type were highly resistant to Phoma black stem. These findings will assist breeders in choosing parent plants for breeding durable resistance to Phoma black stem.

A gene expression analysis of cell wall biosynthetic genes in Malus × domestica infected by ‘Candidatus Phytoplasma mali’

Science.gov (United States)

Guerriero, Gea; Giorno, Filomena; Ciccotti, Anna Maria; Schmidt, Silvia; Baric, Sanja

2016-01-01

Apple proliferation (AP) represents a serious threat to several fruit-growing areas and is responsible for great economic losses. Several studies have highlighted the key role played by the cell wall in response to pathogen attack. The existence of a cell wall integrity signaling pathway which senses perturbations in the cell wall architecture upon abiotic/biotic stresses and activates specific defence responses has been widely demonstrated in plants. More recently a role played by cell wall-related genes has also been reported in plants infected by phytoplasmas. With the aim of shedding light on the cell wall response to AP disease in the economically relevant fruit-tree Malus × domestica Borkh., we investigated the expression of the cellulose (CesA) and callose synthase (CalS) genes in different organs (i.e., leaves, roots and branch phloem) of healthy and infected symptomatic outdoor-grown trees, sampled over the course of two time points (i.e., spring and autumn 2011), as well as in in vitro micropropagated control and infected plantlets. A strong up-regulation in the expression of cell wall biosynthetic genes was recorded in roots from infected trees. Secondary cell wall CesAs showed up-regulation in the phloem tissue from branches of infected plants, while either a down-regulation of some genes or no major changes were observed in the leaves. Micropropagated plantlets also showed an increase in cell wall-related genes and constitute a useful system for a general assessment of gene expression analysis upon phytoplasma infection. Finally, we also report the presence of several ‘knot’-like structures along the roots of infected apple trees and discuss the occurrence of this interesting phenotype in relation to the gene expression results and the modalities of phytoplasma diffusion. PMID:23086810

Cell-specific expression of tryptophan decarboxylase and 10-hydroxygeraniol oxidoreductase, key genes involved in camptothecin biosynthesis in Camptotheca acuminata Decne (Nyssaceae

Directory of Open Access Journals (Sweden)

Santamaria Anna

2010-04-01

Full Text Available Abstract Background Camptotheca acuminata is a major natural source of the terpenoid indole alkaloid camptothecin (CPT. At present, little is known about the cellular distribution of the biosynthesis of CPT, which would be useful knowledge for developing new strategies and technologies for improving alkaloid production. Results The pattern of CPT accumulation was compared with the expression pattern of some genes involved in CPT biosynthesis in C. acuminata [i.e., Ca-TDC1 and Ca-TDC2 (encoding for tryptophan decarboxylase and Ca-HGO (encoding for 10-hydroxygeraniol oxidoreductase]. Both CPT accumulation and gene expression were investigated in plants at different degrees of development and in plantlets subjected to drought-stress. In all organs, CPT accumulation was detected in epidermal idioblasts, in some glandular trichomes, and in groups of idioblast cells localized in parenchyma tissues. Drought-stress caused an increase in CPT accumulation and in the number of glandular trichomes containing CPT, whereas no increase in epidermal or parenchymatous idioblasts was observed. In the leaf, Ca-TDC1 expression was detected in some epidermal cells and in groups of mesophyll cells but not in glandular trichomes; in the stem, it was observed in parenchyma cells of the vascular tissue; in the root, no expression was detected. Ca-TDC2 expression was observed exclusively in leaves of plantlets subjected to drought-stress, in the same sites described for Ca-TDC1. In the leaf, Ca-HGO was detected in all chlorenchyma cells; in the stem, it was observed in the same sites described for Ca-TDC1; in the root, no expression was detected. Conclusions The finding that the sites of CPT accumulation are not consistently the same as those in which the studied genes are expressed demonstrates an organ-to-organ and cell-to-cell translocation of CPT or its precursors.

Oxidative Stress in Horseradish (Armoracia lapathifolia Gilib. Tissues Grown in vitro

Directory of Open Access Journals (Sweden)

Petra Peharec

2011-01-01

Full Text Available In a previous study it was reported that transformed tissue of horseradish (Armoracia lapathifolia Gilib., obtained by infection of leaf explants with A. tumefaciens, developed two tumour lines with different morphology. One line grew as a completely unorganized tissue (TN – tumour tissue, while the other line grew as a partially organized teratogenous tumour with malformed hyperhydric shoots (TM – teratoma tissue, but did not regenerate the whole plant of normal morphology. The factor responsible for this problem could be the increased production of reactive oxygen species (ROS. Therefore, in this study a possible involvement of activated oxygen metabolism in dedifferentiation and hyperhydricity in TM and TN tissues is investigated. Elevated values of malondialdehyde and protein carbonyl contents found in TM and TN, in comparison with plantlet leaf, confirm the presence of oxidative stress. However, lower H2O2 content was measured in TM and TN. Lipoxygenase (LOX activity was more pronounced in TM and especially in TN compared to leaf, which suggests that the LOX-dependent peroxidation of fatty acids might be one of the causes of oxidative damage. Moreover, significantly higher peroxidase (PRX and ascorbate peroxidase (APX activity as well as the increased number of their isoforms was found in transformed TM and TN in comparison with leaf. On the other hand, significantly lower superoxide dismutase (SOD activity was found in TM and TN, which correlates with lower H2O2 content. High catalase (CAT activity measured in leaf and partially organized TM is consistent with the role of CAT in growth and differentiation. In conclusion, in horseradish transformed tissues that underwent dedifferentiation and hyperhydricity, prominent oxidative damage was found. This result suggests that oxidative stress could be associated with the inability of partially organized teratogenous TM to regenerate plantlets with normal morphology.

Significance of in vitro adventitious bud techniques for mutation breeding of vegetatively propagated crops

International Nuclear Information System (INIS)

Broertjes, C.

1982-01-01

It was investigated whether in vitro propagation techniques are of significance for the production of solid, non-chimeric mutants in mutation breeding programmes of vegetatively propagated crops. Irradiated explants of chrysanthemum, potato, begonia and carnation were used for the production of (adventitious) shoots and plantlets to determine the number and frequency of solid mutants and chimeras respectively. It was demonstrated that by the methods described high numbers of solid, non-chimeric mutants can be obtained and that the percentage of chimeras is comparable to the low figures reported after use of in vivo adventitious bud techniques. Consequently, the micro-propagation techniques seem very promising for the commercial plant breeder of vegetatively propagated crops. (author)

Induced mutations and in vitro culture techniques for improving crop plant resistance to diseases

International Nuclear Information System (INIS)

1993-12-01

This co-ordinated research program was undertaken in search of in vitro techniques to increase the resistance of plants to disease. The studies performed under the program ranged from the preparation of materials for mass screening to screening of mutagen-treated cells, tissues, organs or plantlets for resistance to viruses, fungi and other pathogens. The characteristics of the resulting mutants were evaluated to determine the relevance of these techniques for plant breeding. The present document contains the papers presented at the final Research Co-ordination Meeting of the program, as well as a summary of the conclusions and recommendations drawn from the work. The nine individual papers have been input separately to the database. Refs, figs and tabs

Results of Zea mays seeds β- irradiation in 0 - 5 Gy range

International Nuclear Information System (INIS)

Chicea, Dan; Racuciu, Mihaela

2008-01-01

In this study young plants, obtained from the control and beta irradiated seeds, were studied in laboratory experiments. Zea mays seeds with uniform genophond were irradiated with 90 Sr source in the 0 - 5 Gy range. Increased germination percentage was observed under the influence of different radiation doses. We found that small doses of β- radiation have a stimulating effect on the growth of the plantlets, the maximum simulation (among the doses we used) being induced by 0.615 Gy and is statistically significant. We investigated the chlorophyll a to b ratio and the average length variations with the irradiation dose and present the results. Key words: b- radiation, Zea mays, germination rate, plants growth, photoassimilatory pigments,

Growth and proliferation in vitro of Vaccinium corymbosum under different irradiance and radiation spectral composition

International Nuclear Information System (INIS)

Noe, N.; Eccher, T.; Signore, E. del; Montoldi, A.

1998-01-01

Plantlets of highbush blueberry (Vaccinium corymbosum) cvs. Atlantic, Barkeley and Elizabeth, were exposed in vitro to radiation of different spectral compositions obtained by filtering the cool-white light. Red colour of leaves was the first response to the light treatments. On average, cv. Atlantic yielded the highest number of shoots per explant (10.4), followed by cv. Elizabeth (9.1) and Berkeley (6.5). No-B-PMMA increased the proliferation rate in all the 3 genotypes, especially in cv. Atlantic. Cutting wavelengths between 650 and 760 nm (no-R-PMMA filter) generally depressed the proliferation rate. No-B-PMMA induced remarkable changes in the morphology of the shoots - more elongate leaves and longer internodes - especially in cv. Atlantic

Plant regeneration from organ culture in white Guinea Yam

Energy Technology Data Exchange (ETDEWEB)

Nwachukwu, E C; Mbanaso, E N.A. [National Root Crops Research Inst., Umudike, Umuahia, Abia State (Nigeria); Sonnino, A [Centro Recerche Energia, ENEA, Rome (Italy)

1997-07-01

Explants from leaves, leaf segments, petioles and internodal stem of in vitro grown seedlings of white guinea yam, Dioscorea rotundata Poir, cv. `Obiaoturugo` were cultured on defined media. NAA at concentrations of 0.5-1.0 mg/1 induced shoot regeneration from petiolar and inter-nodal stem pieces, and rooting occurred with little or no callusing from whole leaves or leaf segments. With concentration of 3.0-10.0 m/1 NAA, explants from petioles, inter-nodal stem, whole leaves and leaf segments formed callus which produced roots. These explants developed plantlets when subcultured on MS medium supplemented with 2.0 mg/1 BAP and 0.1 mg/1 NAA. (author). 11 refs, 1 tab.

Micropropagation and cytogenetic assessment of Zingiber species of Northeast India.

Science.gov (United States)

Das, Archana; Kesari, Vigya; Rangan, Latha

2013-12-01

An improved micropropagation protocol was developed for Zingiber moran and Z. zerumbet, two wild species of the genus Zingiber, found in Northeast India. The effects of growth regulators, sugar concentrations, and nutrients were tested on the rate of shoot initiation and multiplication. An increase in proliferation and multiplication occurred in modified Murashige and Skoog (MS) medium supplemented with benzyladenine and kinetin. About 2 % sucrose and 0.7 % agar were found to be the optimum for shoot multiplication and regeneration. Naphthalene acetic acid at 0.5 mg/L produced the best rooting response for both the species. Regenerated plantlets were acclimatized successfully and cytogenetic stability was confirmed by RAPD profiling and ploidy checks.

Micropropagation, Acclimatization, and Greenhouse Culture of Veratrum californicum.

Science.gov (United States)

White, Sarah A; Adelberg, Jeffrey; Naylor-Adelberg, Jacqueline; Mann, David A; Song, Ju Yeon; Sun, Youping

2016-01-01

Micropropagation and production of Veratrum californicum is most successful when using a premixed Murishage and Skoog basal medium with vitamins and a 5-week subculture cycle at 16 °C for multiplication. These culture conditions provide the best percent survival after acclimatization in the greenhouse. However, clone response to temperature and light quality within culture conditions varies. Micropropagated plants have mass and morphology similar to 2- or 3-year-old seedlings. Acclimatized plantlets can then be grown in the greenhouse using sub-irrigation (ebb and flood) to maintain substrate volumetric water content > 44 %. Growth cycle in the greenhouse must be about 100 days, followed by dormancy for 5 months at 5 °C.

Micropropagation of Rubus spp.

Directory of Open Access Journals (Sweden)

Åke Finne

1986-09-01

Full Text Available Rapid micropropagation of ‘Black Satin’, ‘Thornless Evergreen’ and ‘Bedford Giant’ was achieved by culturing shoot tips in a 1/1 MS. The best medium in the introduction stage contained 2.0 mg/l BAP. Shoot proliferation occurred with 3.0 mg/1BAP. Some problems with callus and phenolic compounds appeared in both introduction and shoot multiplication stages. The plantlets were rooted either in 1/10 MS without hormones or directly in peat. Two months later most of ‘Black Satin’ and ‘Bedford Giant’ and about half of ‘Thornless Evergreen’ were growing steadily. Theoretically it is possible to produce 60,000 plants within a half year by this method.

Micropropagation from cultured nodal explants of rose (Rosa hybrida L. cv. ‘Perfume Delight’

Directory of Open Access Journals (Sweden)

Kamnoon Kanchanapoom

2010-01-01

Full Text Available A method for the micropropagation of rose (Rosa hybrida L. cv. ‘Perfume Delight’ was developed. First to fifth nodal explants from young healthy shoots were excised and cultured on basal medium of Murashige and Skoog (1962, MS containing several concentrations of BA and NAA. Multiple shoot formation of up to 3 shoots was obtained on MS medium supplemented with 3 mg/l BA and 0.003 mg/l NAA. Shoot readily rooted on ¼MS medium devoid of growth regulators.Rooted plantlets were hardened and established in pots at 100% survival. In vitro flowering was observed on rose plantscultured on MS medium containing 3 mg/l BA and 0.003 mg/l NAA.

Plant regeneration from organ culture in white Guinea Yam

International Nuclear Information System (INIS)

Nwachukwu, E.C.; Mbanaso, E.N.A.; Sonnino, A.

1997-01-01

Explants from leaves, leaf segments, petioles and internodal stem of in vitro grown seedlings of white guinea yam, Dioscorea rotundata Poir, cv. 'Obiaoturugo' were cultured on defined media. NAA at concentrations of 0.5-1.0 mg/1 induced shoot regeneration from petiolar and inter-nodal stem pieces, and rooting occurred with little or no callusing from whole leaves or leaf segments. With concentration of 3.0-10.0 m/1 NAA, explants from petioles, inter-nodal stem, whole leaves and leaf segments formed callus which produced roots. These explants developed plantlets when subcultured on MS medium supplemented with 2.0 mg/1 BAP and 0.1 mg/1 NAA. (author). 11 refs, 1 tab

Induction of materials for mutation breeding of strawberry (Fragaria × Ananassa) by gamma irradiation

International Nuclear Information System (INIS)

Le Ngoc Trieu; Nguyen Tuong Mien; Le Tien Thanh; Huynh Thi Trung; Pham Van Nhi; Vu Thi Trac

2015-01-01

From collected New Zealand strawberry runners, micropropagation was executed to establish 500 shoot clusters for investigation effect of Gamma ray irradiation doses on survival rate. LD_5_0 at 52 Gy was recorded 45 days after re-injection and used as base for choosing 5 irradiation doses of 20, 40, 60, 80, 100 Gy for creation potentially existent mutant materials. 30 shoot clusters were irradiated at each chosen dose. Irradiated material was propagated by in vitro techniques to achieve 300 plantlets/chosen dose. There was no recorded alteration in survival rate and other morphological characteristics of irradiated materials compared to the control in nursery period. These materials were transplanted to plastic greenhouse to screen the mutant. (author)

Studies on the radiation application for development genetic resources

International Nuclear Information System (INIS)

Lee, Young Il; Song, Hi Sup; Kim, Jae Sung; Shin, In Chul; Lee, Sang Jae; Lee, Ki Woon; Lim, Young Tack; Lee, In Suk; Kang, Kwan Kyu.

1997-07-01

For the development of nuclear application technique on the induction and selection of radiation mutation in in vitro and in vivo mutagenesis, several crops plants such as rice, soybean, perilla potato and sweet potato were irradiated with gamma rays of various dose to the seeds. Variants of sweet potato plantlets were obtained from embryogenic callus irradiated with gamma rays and variants were analyzed by using DNA, RNA and protein band patterns. Various mutants were selected from rice, soybean and perilla for short plant, earliness, high yield, large seed size and disease resistance in the advanced mutant generations. Several promising mutants of rice, soybean and perilla will be released to farmers. (author). 198 refs., 26 tabs.,15 figs

Studies on the radiation application for development genetic resources

Energy Technology Data Exchange (ETDEWEB)

Lee, Young Il; Song, Hi Sup; Kim, Jae Sung; Shin, In Chul; Lee, Sang Jae; Lee, Ki Woon; Lim, Young Tack; Lee, In Suk; Kang, Kwan Kyu

1997-07-01

For the development of nuclear application technique on the induction and selection of radiation mutation in in vitro and in vivo mutagenesis, several crops plants such as rice, soybean, perilla potato and sweet potato were irradiated with gamma rays of various dose to the seeds. Variants of sweet potato plantlets were obtained from embryogenic callus irradiated with gamma rays and variants were analyzed by using DNA, RNA and protein band patterns. Various mutants were selected from rice, soybean and perilla for short plant, earliness, high yield, large seed size and disease resistance in the advanced mutant generations. Several promising mutants of rice, soybean and perilla will be released to farmers. (author). 198 refs., 26 tabs.,15 figs.

Germinação in vitro de embriões zigóticos de murmuru (Astrocaryum ulei In vitro germination of 'Murmuru' zygotic embryos (Astrocaryum ulei

Directory of Open Access Journals (Sweden)

Jonny Everson Scherwinski Pereira

2006-04-01

statistical design was a 2x3 factorial, with six repetitions and five embryos by experimental unit. After 30 days it was evaluated the germination percentage and plantlets height. It was verified that embryos from immature fruits presented higher germination percentage and the sucrose concentration of 30 g.L-1 was the one that provided the best results among the others tested. Embryos from mature fruits gave rise to plantlets with average height significantly greater than that of plantlets from immature fruits and 30 g.L-1 of sucrose was the concentration that provided the best results.