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Sample records for fruit callus cultures

  1. callus culture

    African Journals Online (AJOL)

    steve

    and KH2PO4 -NH4NO3 depletion was fatal to grape cells and resulted in formation ... Individual phenolic compounds found in the grape cell cultures were ...... Characterization of in vitro anthocyanin-producing sour cherry ... Process Biochem.

  2. Compositional changes in cell wall polysaccharides from apple fruit callus cultures modulated by different plant growth regulators.

    Science.gov (United States)

    Alayón-Luaces, Paula; Ponce, Nora M A; Mroginski, Luis A; Stortz, Carlos A; Sozzi, Gabriel O

    2012-04-01

    The cell wall composition of apples callus cultures showed changes in the presence of 5 mg l(-1) of three different plant growth regulators (PGRs), namely picloram, abscisic acid and gibberellic acid. Although the structural functions of cell walls do not generally allow for pronounced variations of the total pectin and matrix glycan content, this work provides evidence that the addition of these plant growth regulators can rule, at least partly, cell wall metabolism in apple callus cultures. The chelator- and carbonate-extracts always had the analytical characteristics of pectins, with high proportions of uronic acids, arabinose and galactose as the main monosaccharides, and a significant proportion of rhamnose, but the cross-linking glycan fractions were still rich in RG-I-like material. The application of PGRs produced shifts of uronic acid and neutral sugars between fractions. Arabinose was the neutral sugar exhibiting more variations in apple callus cell wall. Picloram and abscisic acid produced an increase of the uronic acid contents of the cell walls. The AIRs obtained from calluses treated with different PGRs did not show large amounts of high molecular weight products, as determined by size-exclusion chromatography. For the carbonate-extract only the callus treated with picloram displayed two separated peaks for products of different molecular weights. The chromatographic profiles for the 4% KOH-extract displayed two peaks for all the treatments, one very sharp with high molecular weight, and another one wider of smaller molecular weight, whereas the difference between treatments can only be appraised through the areas of the peaks. This is the first report on cell wall composition from fruit calluses supplemented with different PGRs.

  3. Regeneration of Stevia Plant Through Callus Culture

    Science.gov (United States)

    Patel, R. M.; Shah, R. R.

    2009-01-01

    Stevia rebaudiana Bertoni that conventionally propagated by seed or by cuttings or clump division which has a limitation of quality and quantity seed material. In present study, callus culture technique was tried to achieve rapid plant multiplication for quality seed material. Callus induction and multiplication medium was standardized from nodal as well as leaf sagments. It is possible to maintain callus on Murashige and Skoog medium supplemented with 6-benzyl amino purine and naphthalene acetic acid. Maximum callus induction was obtained on Murashige and Skoog medium incorporated with 6-benzyl amino purine (2.0-3.0 mg/l) and naphthalene acetic acid (2.0 mg/l) treatments. However, Murashige and Skoog medium containing 2.0 mg/l 6-benzyl amino purine+2.0 mg/l naphthalene acetic acid was found to be the best for callus induction. Higher regeneration frequency was noticed with Murashige and Skoog medium supplemented with 2.0 mg/l 6-benzyl amino purine+0.2 mg/l naphthalene acetic acid. Regenerated plants were rooted better on ¼ Murashige and Skoog strength supplemented with 0.1 mg/l indole-3-butyric acid. The rooted plantlets were hardened successfully in tera care medium with 63 per cent survival rate. The developed protocol can be utilized for mass production of true to type planting material on large scale independent of season, i.e. external environmental conditions. PMID:20177455

  4. Callus and cell suspension cultures of carnation

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1972-01-01

    Callus cultures of carnation, Dianthus caryophyllus L. ev. G. J. Sim, were grown on a synthetic medium of half strength Murashige and Skoog salts, 3 % sucrose, 100 mg/l of myo-inositol, 0.5 mg/l each of thiamin, HCl, pyridoxin, HCl and nicotinic acid and 10 g/l agar. Optimal concentrations of gro......, but all attempts to induce formation of shoots or em-bryoids gave negative results....

  5. Callus cultures of tomato mutants: I. Nutritional requirements.

    Science.gov (United States)

    Ulrich, J M; Mackinney, G

    1969-01-01

    Callus from hypocotyl, stem, and fruit tissue of tomato mutants was grown on a complex pea extract medium. The genotypes responded differently to the levels of nutrients and stimulators or inhibitors in the medium. Hypocotyl callus of yellow (r) tomato required K(2) SO(4) for quick establishment and continued steady growth for several months; callus of this mutant could also grow with 0.5 % dimethyl sulfoxide in the medium, although growth was less than the control. The red ghost (r(+) gh) mutant is sensitive to a toxic component in the pea extract, and makes its best growth with the standard minerals and vitamins, but in 1/2 concentration pea extract plus 5 % coconut water. Tangerine (t), red lutescent stem (r(+) l(2) ), and r(+) gh are mutants which respond differently to thiourea: t grows about the same at all concentrations, r(+) gh grows best at low thiourea, and r(+) l(2) grows best at the specific level of 20 mg/l thiourea. The recent active t or r(+) l(1) and r(+) l(2) isolates require supplementary auxin to which the older, slow-growing isolates do not respond. However, there is variation in growth response of different isolates of the same mutant. The several red (r(+) ) cultures are similar in their slow growth, but somewhat different in responses to specific nutrients. The recent (+) isolate is one of the most active cultures, in comparison to the slow growth of t callus isolated in 1964. It is therefore concluded that growth is affected both by the specific requirements of the mutant and by the age and vigor of isolates.

  6. Bioproduction of neohesperidin and naringin in callus cultures of Citrus aurantium.

    Science.gov (United States)

    Del Río, J A; Ortuño, A; Marín, F R; Puig, D G; Sabater, F

    1992-10-01

    The accumulation of both neohesperidin and naringin as major flavonoids in callus cultures of bitter orange (Citrus aurantium) was demonstrated using high performance liquid chromatography with a diode-array detector. The identity of both compounds was confirmed by their corresponding nuclear magnetic resonance spectra. The levels of neohesperidin are higher than those of naringin in callus culture, as they are in immature fruit, and high concentrations of both are found in young tissues such as immature fruits and the outer zone of calli.

  7. Callus induction and biomass accumulation in vitro in explants from chokeberry (Aronia melanocarpa (Michx. Elliot fruit

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    Tatiana I. Calalb

    2014-09-01

    Full Text Available In this study, the following features were determined: biological (the optimal histogen as explant and the optimal age of donor fruit, biotechnological (type, dosage and combination of growth regulators supplements in culture medium Murashige and Skoog as well as sucrose dosage, and physical (light regime, to induce callusing and biomass accumulation in vitro from the succulent chokeberry (Aronia melanocarpa (Michx. Elliot fruit. It turned out that it was much easier to induce callus from explants composed of the epicarp and hypoderm cut from fruits at 50–60 days after flowering. The role of light regime and varied supplementation of the basic MS medium with different doses of growth regulators was established; they resulted in four pigmented carpomass: violet, cream-pink, cream-white and green. The best combinations for the proliferation of fruit callus were culture media with 0.2–2.5 mg × dm-3 2,4-D+0.5 mg × dm-3 KIN +60 g × dm-3sucrose, while for fruit biomass accumulation enriched with phenolic substances – 2.5–3.5 mg × dm-3 NAA+0.5 mg × dm-3 KIN+60 g × dm-3sucrose. The chemical study of phenolic compounds by HPLC coupled with the mass spectrometry method identified chlorogenic acid, hiperozide, quercetrin, isoquercitrin and rutozide quantitatively and qualitatively in all pigmented carpomass and fruits; an exception is p-coumaric present only qualitatively in green carpomass and absent in fruit and quercetol absent in green carpomass.

  8. Chrysophanol glycosides from callus cultures of monocotyledonous Kniphofia spp. (Asphodelaceae).

    Science.gov (United States)

    Ito, Hideyuki; Nishida, Yasuko; Yamazaki, Mie; Nakahara, Keisuke; Michalska-Hartwich, Malgorzata; Furmanowa, Miroslawa; Leistner, Eckhard; Yoshida, Takashi

    2004-10-01

    We established callus cultures of the monocotyledonous plants Kniphofia foliosa and K. tuckii (Asphodelaceae), which produce the anthraquinone derivatives chrysophanol and its glycosides. The minor product chrysophanol 8-O-beta-gentiobioside was fully characterized by spectroscopic analysis and synthesis.

  9. EFFECT OF PHYSIOLOGICAL AGE AND GROWTH REGULATORS ON CALLUS BROWNING OF COCONUT ENDOSPERM CULTURE IN VITRO

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    LAZARUS AGUS SUKAMTO

    2011-01-01

    Full Text Available The possibility of physiological age and growth regulators affecting callus browning ofcoconut endosperm was investigated. Solid endosperm explants of four coconut fruits fromsame brunches of two coconut cultivars “Samoan Dwarf ” were grown on modified Murashigeand Skoog (MS formula with addition of 10 mg l putresine, 2.50 g l activated charcoal (AC,1.70 g l phytagel, 0, 10 , 10 , 10 , 10 M 2,4-dichlorophenoxyacetic acid (2,4-D or 4-amino-3,5,6-trichloropicolinic acid (Picloram combined with 10 M 6-benzylaminopurine (BA.Callogenesis occurred on 98.83% of explants. Callus browning between different physiologicalages (antipodal and micropylar tissues of coconut endosperm at 9, 26 and 31 weeks of culture(WOC was significantly different, but not at 16 and 21 WOC. Auxins of 2,4-D and Picloramdid not affect significantly callus browning of endosperm cultures. Auxin doses at 10 , 10 , and10 M decreased significantly callus browning at 9 and 16 WOC, respectively, but at 10 Mbrowning was less significant compared to other doses at 21 WOC. Auxin dose at 10 M causedless significant browning compared to other doses at 31 WOC. The addition of BA decreasedsignificantly callus browning at 9 WOC, but did not affect callus browning thereafter.

  10. The durative use of suspension cells and callus for volatile oil by comparative with seeds and fruits in Capparis spinosa L.

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    Yongtai Yin

    Full Text Available Capparis spinosa is one of the most important eremophytes among the medicinal plants, and continued destruction of these plants poses a major threat to species survival. The development of methods to extract compounds, especially those of medicinal value, without harvesting the whole plant is an issue of considerable socioeconomic importance. On the basis of an established system for culture of suspension cells and callus in vitro, Gas Chromatograph-Mass Spectrometer (GC-MS was used for the volatile oil composition analyzing in seed, fruit, suspension cells and callus. Fatty acids were the major component, and the highest content of alkanes was detected in seed, with <1.0% in suspension cells and callus. Esters, olefins and heterocyclic compounds were significantly higher in fruit than in the other materials. The content of acid esters in the suspension cells and callus was significantly higher than in seed and fruit. This indicated that the suspension cells and callus could be helpful for increasing the value of volatile oil and replacing seeds and fruit partially as a source of some compounds of the volatile oil and may also produce some new medical compounds. The above results give valuable information for sustainable use of C. spinosa and provide a foundation for use of the C. spinosa suspension cells and callus as an ongoing medical resource.

  11. Shoot Differentiation in Callus Cultures of Datura Innoxia

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1973-01-01

    promoted shoot differentiation. Gibberellic acid inhibited shoot formation weakly, but inhibited proper leaf blade formation. Root differentiation was rare. The callus cultures of Datura innoxia grew rapidly (100-fold in 4 weeks) on a slightly modified Murashige and Skoog medium (0.5 mg/l thiamin · HCl, p...

  12. Effects of Culture Conditions, Carbon Source and Regulators on Saffron Callus Growth and Crocin Accumulation in the Callus

    Institute of Scientific and Technical Information of China (English)

    刘雪; 郭志刚; 刘瑞芝

    2002-01-01

    There are many factors influencing the growth and secondary metabolites of callus and saffron callus. In this paper, the effects of culture conditions, including culture temperatures, light levels, the carbon source and its concentration, and the preserve of regulators (mainly hormones), are studied for callus cultures. All the experiments used Murashige and Skoog (MS) solid medium as the basic medium with 10 g/L agar, pH 5.75.8. Saffron callus was cultured at 20℃ in the dark, with a sucrose concentration of 45 g/L (or starchy hydrolysate concentration of 40 g/L), but 30 g/L sucrose was best for the synthesis of crocin (for starchy hydrolysate the concentration can range from 20 to 40 g/L). To promote callus growth, the best auxin was α-naphthaleneacetic acid (NAA) and the optimum ratio of NAA (mg/L) to benzylaminopurine (BA) (mg/L) was and uniconazole (S-07) (1.25 mg/L) increased the crocin content remarkably as analyzed by high performance liquid chromatography (HPLC). NAA (2 mg/L) promoted the growth of saffron callus but had no benefit and may inhibit crocin synthesis while S-07 (1.25 mg/L) had the opposite effect. GA3 promoted both growth and synthesis.

  13. Acylated flavonoids in callus cultures of Citrus aurantifolia.

    Science.gov (United States)

    Berhow, M A; Bennett, R D; Poling, S M; Vannier, S; Hidaka, T; Omura, M

    1994-07-01

    Two new acylated flavonol glycosides were isolated along with kaempferol 3-O-beta-rutinoside from 10-year-old callus cultures of Mexican lime. The structures of these new compounds are kaempferol 3-O-beta-D-glucopyranoside-6"-(3-hydroxy-3-methyl glutarate) and kaempferol 3-O-beta-D-glucopyranoside-6"-(3-hydroxy-3-methyl glutarate)-7-O-beta-D-glucopyranoside.

  14. Plant regeneration from callus culture of vetiver (Vetiveria zizanioides Nash

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    Somporn Prasertsongskun

    2003-09-01

    Full Text Available The present research aimed to establish cell suspension culture of vetiver (Vetiveria zizanioides Nash from Surat Thani germplasm source and efficient plant regeneration from callus derived from such cultures. Cell suspension cultures were established from calli derived from inflorescence of vetiver. Optimum cell proliferation occurred in liquid N6 medium supplemented with 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D and 10 mM proline. The cell suspension formed the highest small colonies when plated on solid MS medium containing 0.45 μM 2,4-D. After subsequent transfer to regeneration medium (MS free medium 65% of plantlets were obtained.

  15. Production of chlorogenic acid in Varthemia persica DC (var. persica) callus cultures

    OpenAIRE

    2011-01-01

    Chlorogenic acid, a pharmacologically important compound, is a phenolic compound that occurs in certain commonly used medicinal herbs. We looked for the presence of this compound in the callus cultures of Varthemia persica DC (var. persica). We have evaluated the conditions for establishment of callus cultures of V. persica and the in vitro production of chlorogenic acid. Callus was initiated by culturing seedling of V. persica on MS basal medium supplemented with different concentrations of ...

  16. Callus production from photoautotrophic soybean cell culture protoplasts.

    Science.gov (United States)

    Chowhury, V K; Widholm, J M

    1985-10-01

    Protoplasts were prepared from a photoautotrophic (PA) cell line of Glycine max (soybean). A yield of 75 to 90% after two to three hours digestion in a mixture of 1% Cellulase R10, 0.2% Pectolyase Y23 and 2% Driselase was obtained. Cell division and colony formation occurred from approximately 18% of the plated protoplasts. The cultured protoplasts were as sensitive to the herbicide atrazine, a photosynthetic inhibitor, as the original PA cells under the same conditions. Protoplasts and cells of a heterotrophic (HT) soybean culture were not as sensitive to atrazine. The isolated protoplasts retained the PA characteristics of the parental culture in the callus and cell suspension cultures obtained from the protoplasts. The chromosome numbers in the parental cell line and in cells derived from the isolated protoplasts (both PA and HT) were found to be largely (99%) the normal diploid number of 40.

  17. Establishment of callus, cell suspension and shoot cultures of Leonurus cardiaca L. and diterpene analysis.

    Science.gov (United States)

    Knöss, W

    1995-10-01

    Callus cultures, cell suspension cultures and shoot cultures of Leonurus cardiaca L. (Motherwort) were established and growth conditions optimized. Shoot cultures showed constant growth whether in the dark or under continuous light, accumulating varying amounts of the furanic labdane diterpenes leosibiricin, preleosibirin, leosibirin and isoballotenol acetate, which are also present in the soil-grown plants. Only traces of leosibiricin were detected in callus cultures, while cell suspension cultures did not produce any furanic diterpenes. A small amount of furanic labdane diterpenes was found in the medium of shoot cultures. Callus and shoot culture induction of several other Lamiaceae species is also described.

  18. Formation flavonoid secondary metabolites in callus culture of Chrysanthemum cinerariefolium as alternative provision medicine

    Science.gov (United States)

    Purwianingsih, Widi; Febri, Santika; Kusdianti

    2016-02-01

    Increasing need of medicine ingredients require the discovery of other methods that can be used as an alternative. One method that can be used as an alternative is tissue culture. Quercetin is a flavonoid secondary metabolites that have been known to be useful as antiviral, anti-asthma and anti-cancer potential. The purpose of this study was to produce flavonoids, especially quercetin in callus culture Chrysanthemum cinerariefolium. Pieces of leaves of plantlets C. cinerariefolium used as explants for formation of callus tissue. To grow the callus, Murashige and Skoog (MS) medium used with addition of various concentrations of growth regulators 2.4-D, and kinetin. For multiplication, callus subcultured on similar medium. Callus that had formed, especially brown callus, further analyzed using Gas Chromatography Mass Spectrum (GCMS). Before analyzed callus was extracted in 95% ethanol. The result showed that callus potentially generate secondary metabolite are brown and friable. Based on these parameters, the best callus produced from leaf explants grown on MS medium with the addition of 4 mg / L 2,4-D and 0 mg / L kinetin. The callus contain secondary metabolites such as some of the flavonoid quercetin precursors such as acetic acid and tetrahydroxychalcone, and some other secondary metabolites.

  19. Effective salt criteria in callus-cultured tomato genotypes.

    Science.gov (United States)

    Dogan, Mahmut; Tipirdamaz, Rukiye; Demir, Yavuz

    2010-01-01

    Na+, Cl-, K+, Ca2+, and proline contents, the rate of lipid peroxidation level in terms of malondialdehyde (MDA) and chlorophyll content, and the changes in the activity of antioxidant enzymes, such as superoxide dismutase (SOD: EC 1.15.1.1), catalase (CAT: EC 1.11.1.6), ascorbate peroxidase (APX: EC 1.11.1.11), and glutathione reductase (GR: EC 1.6.4.2), in tissues of five tomato cultivars in salt tolerance were investigated in a callus culture. The selection of effective parameters used in these tomato genotypes and to find out the use of in vitro tests in place of in vivo salt tolerance tests were investigated. As a material, five different tomato genotypes during a 10-day time period were used, and 150 mM NaCl was applied at callus plant tissue. The exposure to NaCl induced a significant increase in MDA content in both salt-resistant and salt-sensitive cultivars. But the MDA content was higher in salt-sensitive cultivars. The chlorophyll content was more decreased in salt-sensitive than in salt-resistant ones. The proline amount was more increased in salt-sensitive than in salt-resistant ones. It has been reported that salt-tolerant plants, besides being able to regulate the ion and water movements, also exhibit a strong antioxidative enzyme system for effective removal of ROS. The degree of damage depends on the balance between the formation of ROS and its removal by the antioxidative scavenging system that protects against them. Exclusion or inclusion of Na+, Cl-, K+, and Ca2+, antioxidant enzymes and MDA concentration play a key protective role against stress, and this feature at the callus plant tissue used as an identifier for tolerance to salt proved to be an effective criterion.

  20. Homoeologous gene silencing in tissue cultured wheat callus

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    Chapman Natalie H

    2008-10-01

    Full Text Available Abstract Background In contrast to diploids, most polyploid plant species, which include the hexaploid bread wheat, possess an additional layer of epigenetic complexity. Several studies have demonstrated that polyploids are affected by homoeologous gene silencing, a process in which sub-genomic genomic copies are selectively transcriptionally inactivated. This form of silencing can be tissue specific and may be linked to developmental or stress responses. Results Evidence was sought as to whether the frequency of homoeologous silencing in in vitro cultured wheat callus differ from that in differentiated organs, given that disorganized cells are associated with a globally lower level of DNA methylation. Using a reverse transcription PCR (RT-PCR single strand conformation polymorphism (SSCP platform to detect the pattern of expression of 20 homoeologous sets of single-copy genes known to be affected by this form of silencing in the root and/or leaf, we observed no silencing in any of the wheat callus tissue tested. Conclusion Our results suggest that much of the homoeologous silencing observed in differentiated tissues is probably under epigenetic control, rather than being linked to genomic instability arising from allopolyploidization. This study reinforces the notion of plasticity in the wheat epi-genome.

  1. Growth and accumulation of flavan-3-ol in Camellia sinensis through callus culture and suspension culture method

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    Sutini Sutini

    2017-02-01

    Full Text Available This study was aimed to assess flavan-3-ol biomass in C. sinensis through callus cultures and suspension cultures derived from leaf explants. Callus initiation of both cultures were using Murashige and Skoog medium were enriched with plant growth regulators Naphtha-lene Acetic Acid 3.0 mg/L and kinetin 2.0 mg/L. The procedures in this study were: (1 callus initiation by cutting the leaves of C. sinen-sis shoots then planted on Murashige and Skoog medium that were enriched with plant growth regulators, (2 sub callus culture on fresh medium that enriched with the same growth regulators, (3 suspension culture initiation of liquid callus, (4 growth examination of callus and suspension cultures in week 12, (5 examination of qualitative-quantitative content of flavan-3-olin suspension cultures at week 4. The results show that suspension cultures contain biomass flavan-3-ol that increase in the same manner of the increase of callus age and weight

  2. Somaclonal variation in Asparagus officinalis plants regenerated by organogenesis from long-term callus cultures

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    Pontaroli Ana Clara

    2005-01-01

    Full Text Available Somaclonal variation in plants regenerated by organogenesis from long-term cultured calluses of two diploid staminate genotypes of Asparagus officinalis cv. Argenteuil was characterized by plant phenotype, ploidy, meiotic behavior, pollen viability, fruit and seed set, and AFLP profiles. Phenotypic deviations from the donors were detected in foliage color, flower size, and cladode and flower morphology. Ploidy changes were observed in 37.8% of the 37 regenerants studied. Meiotic alterations in 12 out of 21 regenerants included laggards, dicentric bridges, micronuclei, restitution nuclei and polyads. Of the 408 AFLP markers screened in 43 regenerants and the donors, 2.94% showed polymorphism. High pollen viability was observed in the 22 regenerants analyzed. All crosses between one pistillate plant and 35 regenerants, as well as the controls, produced fruits and seeds; however, no plump seeds resulted in 35.3% of the crosses with regenerants, and no seeds germinated in 12.5% of those with apparently normal seeds. Fruit and seed set was similar in crosses with diploid regenerants with normal meiosis and the controls but was lower in crosses with diploid and polyploid regenerants with abnormal meiosis. Our results show that the regenerated plants exhibited conspicuous somaclonal variation that could be eventually exploited for in vitro selection systems.

  3. Embryogenic callus formation, growth and regeneration in callus and suspension cultures of Miscanthus x ogiformis Honda 'Giganteus' as affected by proline

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Krogstrup, Peter; Hansen, Jürgen

    1997-01-01

    The effects of proline additions to culture systems of Miscanthus x ogiformis Honda Giganteus' were investigated. Proline was added in concentrations of 0, 12.5, 25, 50, 100 or 300 mM to the callus induction and suspension culture media containing either Murashige and Skoog or N6 basal salts and 22...... with Murashige and Skoog salts increased embryogenic callus formation on shoot apices and leaf explants while proline had no effect on embryogenic callus formation in medium with N6 salts. Increased growth with increasing proline concentration was obtained in suspension aggregates grown in medium with N6 salts......, whereas proline only increased growth of suspension aggregates grown in medium with Murashige and Skoog salts at concentrations of 12.5 or 25 mM. A stimulating effect of proline on plant regeneration was observed in short-term cultures of callus as well as in long-term cultures of suspension aggregates...

  4. Search for alkaloids on callus culture of Passiflora alata

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    Michelli Wesz Machado

    2010-08-01

    Full Text Available Preliminary work on Passiflora alata leaves failed to detect harmane alkaloids using LC. The aim of this work was to investigate the production of harmane alkaloids through the cell culture of P. alata, inducing its precursor (L-tryptophan. The leaf explants presented satisfactory results after disinfection, and the callus formation was initiated in MS media with adequate quantities of phytohormones. Sixty days after inoculation, calli were inoculated in the optimized semi-solid MS media, with and without the addition of L-tryptophan (50, 100, 200 mg/L and kept in standard conditions for 90 days. Calli were collected on days 6, 16, 26, 36, and 90, followed by acid-base extraction, and analysed by LC. The results showed an absence of harmane, harmin, harmol, harmalol, and harmaline. With L-tryptophan feeding, two peaks were detected, collected and analysed through positive mode electrospray [ESI(+-MS] and sequential analysis in tandem ESI(+-MS/MS. The spectra obtained were very similar, with a repetition of the more intense ions, and consecutive loss of 68 Da units, attributed to the heterocycle pyrazole. It appeared that this transformation was not related to any enzymatic pathway previously described for the plant from L-tryptophan, and the biosynthesis of β-carboline alkaloids in callus culture of P. alata were not observed in this work.As folhas de varias espécies de Passiflora são utilizadas como ansioliticas e sedativas. Passiflora alata Curtis, Passifloraceae consta em três edições da farmacopéia brasileira, porem não há muitos estudos sobre sua composição química. No passado, enfatizava-se a ação conjunta de alcalóides e flavonóides. Em trabalho anterior, não foi detectada a presença de alcalóides harmanicos através de CLAE. Assim, decidiu-se investigar a produção dos mesmos através de cultivo celular, introduzindo seu precursor metabólico L-triptofano. Os explantes foliares apresentaram resultados satisfatorios

  5. Induction of flavonoid production by UV-B radiation in Passiflora quadrangularis callus cultures.

    Science.gov (United States)

    Antognoni, Fabiana; Zheng, Suiping; Pagnucco, Cristina; Baraldi, Rita; Poli, Ferruccio; Biondi, Stefania

    2007-07-01

    Callus cultures from several species of Passiflora were initiated in vitro, and their capacity to produce four glycosyl flavonoids (orientin, isoorientin, vitexin, isovitexin) was analysed. The aim of the present work was to examine the possible role of UV-B irradiation and elicitation with methyl jasmonate (MJ) on the production of these compounds in callus cultures. All the species tested (P. incarnata, P. quadrangularis, P. edulis) formed friable callus from leaf explants after 4 weeks on medium supplemented with kinetin and 2,4-dichlorophenoxyacetic acid. Among them, P. quadrangularis turned out to have a faster growth rate and a more friable texture, and was therefore chosen for experiments with elicitors. In callus cultures only small amounts of isoorientin were found, while the concentration of the other flavonoids was below the detection limit. UV-B irradiation of calluses was able to increase the production of all four glycosyl flavonoids. After a 7-day exposure of cultures to UV-B light, the production of isoorientin reached concentrations similar to those found in fresh leaves from glasshouse-grown plants. Elicitation with methyl jasmonate also enhanced orientin, vitexin and isovitexin concentrations, even though the stimulation was about 6-fold weaker for orientin and vitexin and about 40-fold for isovitexin, than that exerted by UV-B treatment. Callus cultures treated with the UV-B dose which most enhanced flavonoid production showed a higher antioxidant activity compared to untreated calluses, with an increase ranging from 28% to 76%. Results show that the secondary metabolite biosynthetic capacity of Passiflora tissue cultures can be enhanced by appropriate forms of elicitation.

  6. Screening for disease resistance in barley cultivars against Bipolaris sorokiniana using callus culture method.

    Science.gov (United States)

    Chand, Ramesh; Sen, Devyani; Prasad, K D; Singh, A K; Bashyal, B M; Prasad, L C; Joshi, A K

    2008-04-01

    Screening for resistant barley genotypes in response to fungal toxin of Bipolaris sorokiniana was assessed on standing barley plants as well as in selected callus lines of the same. For the standing lines tested, those manifesting chlorosis in response to toxin infiltration showed a significantly slower disease progress as compared to the necrotic lines. Also, necrosis in the callus tissues of the susceptible cultivar in MS medium supplemented with different concentrations of the crude toxin was significantly higher than in the callus tissues of the chlorotic lines studied. Similar host response to the toxin in in vitro and field situations open up the possibility of screening barley cultivars for resistance to spot blotch using callus culture as against classical methods of screening in order to increase accuracy and save time and space.

  7. Metabolic changes associated with shoot formation in tobacco callus cultures

    Energy Technology Data Exchange (ETDEWEB)

    Grady, K.L.

    1982-08-01

    Callus tissue derived from Nicotiana tabacum L. stem pith parenchyma cells was grown either on medium which maintains the callus in an undifferentiated state, or on medium which induces the formation of shoots. Two complementary types of studies were performed with the goal of establishing metabolic markers for the initiation of shoot formation: one designed to characterize the flow of radioactive sucrose into various metabolic pools, and one which allowed measurement of intermediary metabolite concentrations. In the former, callus tissue was incubated in (U-/sup 14/C)sucrose for periods up to one hour, and patterns of metabolite labelling in tissue grown on shoot-forming and non-shoot-forming media were compared. In the latter studies, tissue was grown for an entire subculture period on non-shoot-forming medium labelled with (U-/sup 14/C)sucrose, then subcultured to labelled non-shoot-forming or shoot-forming media, and sampled at intervals during the first week of growth. 189 references.

  8. The early ontogeny of embryoids and callus from pollen and subsequent organogenesis in anther cultures of Datura metel and rice.

    Science.gov (United States)

    Iyer, R D; Raina, S K

    1972-06-01

    Haploidy induction through anther culture has been examined in Datura metel and rice with a view to tracing the precise sequence of development of the pollen, either directly or through an intervening callus, into an embryo and seedling. In D. metel, the vegetative cell of the young pollen grain assumes the major role in formation of embryos whereas the generative cell and its few derivatives degenerate. Embryos and seedlings arising directly from pollen without an intervening callus phase always proved to be haploids, whereas those differentiating from pollen-derived callus gave haploid, diploid and even triploid plants. Cytological analysis of callus tissue showed cells of various ploidy levels ranging from haploid to triploid, and in rare instances even with higher chromosome numbers.In rice anther cultures the embryoids arose from an initial callus phase. Of 15 different rice cultivars tried, only four produced a callus, and in only one, was there differentiation of plants, both haploid and diploid ones. Among other species tried, egg plant has also yielded plantlets through a callus phase whereas only callus production has been achieved in jute, tea and petunia. No response has been obtained in wheat, maize, cotton and coconut.Coconut milk (CM) appears to be the most important component of the medium for the initial induction of embryoids and callus in anther cultures of most of the species tried. However, further growth and differentiation of plants may require a simpler medium; in D. metel, continued culture on CM led to dedifferntiation.

  9. Factors Affecting Embryogenic Callus Production and Plant Regeneration in Anther Culture of Bupleurum chinense

    Institute of Scientific and Technical Information of China (English)

    YANG Cheng-min; ZHAO Yu-kai; WEI Jian-he; ZHAO Li-zi; SUI Chun; ZHANG Zheng; CUI Lu-ying

    2011-01-01

    Objective To evaluate the influences of the genotypes,anther developmental stages,and cultural conditions on the efficiency of embryogenic callus induction and plant regeneration in the anthers culture of Bupleurum chinense.Methods The different effects such as four genotypes,plant growth regulators,and temperature condition were compared in the experiments.The histological study was performed with the process of the anther culture.Results The highest inducing rate of embryogenic calli were achieved for the genotypes Zhongcaiyihao(ZCYH),Z4,and Z5 at the early-to middle-uninucleate stages,except for genotype ZPM1 at the tetrad stage.Cold pretreatment increased the production of the embryogenic callus,in which 4-day cold pretreatment improved the production of embryogenic callus from 0% to 2.2% and 5.0% for genotypes ZPM1 and ZCYH,respectively.No embryogenic callus was induced in the medium containing less than 0.75 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D).The highest regeneration rate (34.6%)was obtained in 1/2 MS salts regeneration medium supplemented with 0.1 mg/L 6-benzylmaminopurine (BA).The low concentration of BA was able to promote the embryogenic callus formation and subsequent plantlet regeneration via somatic embryogenesis.Chromosome counting of regenerated plantlets showed mostly diploid plant (2n = 12)with only one haploid plant(n = 6).Because of the low rate of microspore embryo formation,we only tracked the process of embryogenesis from the connective tissue,instead of microspore by histological observations.Conclusion This study establishes an efficient system for embryogenic callus induction and plant regeneration system.This is the first report on the haploid plantlet through the anther culture orB.chinense.

  10. Factors affecting the accumulation of 9-methoxycanthin-6-one in callus cultures of Eurycoma longifolia

    Institute of Scientific and Technical Information of China (English)

    N. Rosli; M. Maziah; K. L. Chan; S. Sreeramanan

    2009-01-01

    A study was conducted to improve 9-methoxycanthin-6-one productivity (potential anti-tumour compound) from callus cultures of Eurycoma longifolia (Tongkat Ali). Several factors affecting 9-methoxycanthin-6-one production in callus cultures such as different medium compositions and physical factors were investigated and analyzed. Results show that a higher production of 9-methoxycanthin-6-one (3.84 mg(g-1 DW (Dry Weight)) is obtained from callus cultured in 1/4 MS basal media. At fructose of 2% (w/v), the production of 9-methoxycanthin-6-one (4.59 mg(g-1 DW) is promoted to gain the highest yield, compared to other carbon sources tested. The addition of 2.0-mg(L-1 dicamba also increases 9-methoxycanthin-6-one production (12.3 mg(g-1 DW). Higher production of 9-methoxycanthin-6-one was obtained at pH 5.5 (1.53 mg(g-1 DW). Production of 9-methoxycanthin-6-one (2.34 mg(g-1 DW) in callus cultures is also increased when the medium is added with 1(10-1 μM phenylalanine. This study suggests that the successful production of 9-methoxycanthin-6-one in vitro cultures has a potential in large-scale production using bioreactor technology.

  11. Extraction and quantification of gymnemic acids through gymnemagenin from callus cultures of Gymnema sylvestre.

    Science.gov (United States)

    Kanetkar, P V; Singhal, R S; Laddha, K S; Kamat, M Y

    2006-01-01

    The phyto-constituents of Gymnema sylvestre are used in the treatment of diabetes and obesity. The present work reports on the extraction of gymnemic acid through gymnemagenin from callus cultures of G. sylvestre. Components were separated on pre-coated silica gel 60 GF254 plates with chloroform:methanol (8:2) and scanned using a densitometric scanner at 205 nm in the near-UV region. Linearity of determination of gymnemagenin was observed in the range 2-10 microg. The average percentage recovery of gymnemagenin from leaf callus extracts was 98.9+/-0.3.

  12. High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species

    NARCIS (Netherlands)

    Valk, P. van der; Scholten, O.E.; Verstappen, F.; Jansen, R.C.; Dons, J.J.M.

    1992-01-01

    The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum × A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS)

  13. High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species

    NARCIS (Netherlands)

    Valk, P. van der; Scholten, O.E.; Verstappen, F.; Jansen, R.C.; Dons, J.J.M.

    1992-01-01

    The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum × A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS)

  14. EFFICIENT CALLUS INDUCTION AND PLANT REGENERATION FROM IMMATURE EMBRYO CULTURE OF TRITICUM AESTIVUM L.- THINOPYRUM INTERMEDIUM ALIEN DISOMIC ADDITION LINES

    Institute of Scientific and Technical Information of China (English)

    LI Hong-mei; LI Xing-feng; GAO Ju-rong; WANG Hong-gang

    2005-01-01

    An efficient plant regeneration system was developed from the immature embryos of Triticum aestivum L. - Thinopyrum intermedium alien disomic addition lines, which resistant to powdery mildew. The protocol was based on a series of experiments involving the callus induction and differentiation. The experiment studied the effects of embryo size on callus induction and differentiation of the immature embryos. We found that the embryo size is critical for the establishment of embryogenic callus. Immature embryos (0.8~1.5 mm) showed high ability to produce embryogenic callus capable of regenerating green plants. The medium Murashige and Skoog's (MS) added with 2mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) gave the best embryogenic callus induction, maintenance and regeneration. The embryogenic callus maintained high regeneration during six subcultures in the callus induction medium. Suitable time of partial desiccation could effectively improve the regeneration capacity of the callus cultured for 3~4 month.Bud green spot and root green spot were observed during the differentiation of callus and the difference between them was described. Regenerated shoots were rooted on half-strength MS medium containing 0.2 mg/L Naphthalene acetic acid (NAA). Plants were successfully transferred to soil and grew well. This efficient plant regeneration system provides a foundation for the study of somaclonal variation of Triticum aestivum L. - Thinopyrum intermedium alien disomic addition lines.

  15. Production of paeoniflorin and albiflorin by callus tissue culture of Paeonia lactif lora Pall☆

    Institute of Scientific and Technical Information of China (English)

    Sheng Hu; Yingying Ma; Hailong Jiang; Dingjun Feng; Wei Yu; Dongmei Dai; Lehe Mei

    2015-01-01

    In order to facilitate the preparation of paeoniflorin (PF) and albiflorin (AF), two chief bioactive constituents in Paeonia lactiflora Pal (PL), induction and culture of callus from PL were studied. With a modified woody plant medium supplemented with 0.5 mg·L−1 6-benzylaminopurine, 1.0 mg·L−1 naphthylacetic acid, 0.1 mg·L−1 thidiazuron and 30 g·L−1 sucrose, callus was induced from four kinds of explants:leaf, stems, petiole, and root. The potency to form callus varies between different explants and leaf explants exhibits the highest capacity (100%). On the other hand, root-derived cal us (R-callus) produces the highest level of total amount of PF and AF, 31.8 mg·g−1 dry mass, which is higher than the corresponding level in the root of field cultivated PL. Further-more, the time needed is only 40 days, remarkably shorter than the cultivation time of PL, about 4–5 years. Higher accumulation levels of PF and AF with shorter production time indicate that cal us culture of PL is a promising powerful tool for production of PF and AF in the future.

  16. Organogenesis and plant formation from cotyledon and callus culture of rape

    Directory of Open Access Journals (Sweden)

    Janina H. Rogozińska

    2014-01-01

    Full Text Available Cotyledon explants of rape were excised from aseptically germinated seedlings and cultured during 2 weeks on M u r a s h i g e and S k o o g medium supplemented with auxins, cytokinins, auxin-cytokinin combinations and abscisic acid. Callus formation occurred on medium with 2,4-dichlorophenoxyacetic acid (2,4-D, naphthalene-l-acetic acid (NAA, indole-3-acetic acid (IAA and on their combinations with kinetin (K or 6-benzylaminopurine (BAP. Regeneration of roots was achieved on media with NAA, IAA and indole-3-butyric acid (IBA and on combinations of these auxins with cytokinins. The presence of 2,4-D in the medium, though it promoted compact callus growth, had an inhibitory effect on root formation. Callus derived from the cotyledons had somewhat different requirements for growth in subculture and the root formation ability diminished in the course of the culture. Lower ABA concentrations stimulated callus growth whereas higher concentrations inhibited it similary as in the case of cotyledons. Shoot buds regenerated from the cotyledons after ca. 3 weeks on media supplemented with NAA + BAP. The 9-week-old plantlets transferred to the soil developed into complete plants. The plants which underwent vernalization formed flowers and normal seeds.

  17. Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L.

    Institute of Scientific and Technical Information of China (English)

    Nurul Izzati Osman; Norrizah Jaafar Sidik; Asmah Awal

    2016-01-01

    Objective: To induce callus from the medicinally valuable species, Barringtonia racemosa L.(B. racemosa) whereby the formation of callus is essential for micropropagation studies and in vitro plant secondary metabolites production.Methods: The callus induction potential in B. racemosa was assessed from endosperm explant cultured on different culture media and plant hormonal treatments. Lloyd and Mc Cown’s woody plant medium and Murashige and Skoog’s medium were used in the study as culture media. On the other hand, various concentrations and combinations of2,4-dichlorophenoxyacetic acid(1.0–2.0 mg/L) and kinetin(0.5–2.5 mg/L) had been incorporated in the culture media to exert the effects of auxin and cytokinin on callus induction.Results: From the present study, it was found that the profuse [(1.681 ± 0.770) g fresh weight,(0.239 ± 0.239) g dry weight] and friable callus formation was optimally produced with desirable morphology and considerable percentage of callus induction(56.70%) in endosperm explants cultured on 1.0 mg/L 2,4-dichlorophenoxyacetic acid and 1.5 mg/L kinetin in Murashige and Skoog’s medium.Conclusions: A reliable protocol for inducing callus formation of profuse and friable morphology in endosperm explant of B. racemosa had therefore been successfully established.

  18. Additions of precursors and elicitors improve geranylgeraniol production in Croton stellatopilosus callus cultures

    Directory of Open Access Journals (Sweden)

    Juraithip Wungsintaweekul

    2015-02-01

    Full Text Available Strategies for enhancing GGOH production in Croton stellatopilosus callus culture included additions of precursors (sodium acetate-NA, sodium pyruvate-NP, mevalonic acid lactone-MVA and elicitors (methyl jasmonate-MJ, acetylsalicylic acid-ASA, yeast extract-YE. Treated cells were evaluated for their GGOH contents by GC-FID and compared with the nontreated cells as controls. Additions of NA (25 mg/L, NP (50 mg/L and MVA (100 mg/L resulted in an enhancement of GGOH productivity to 0.61 mg/g DW, 0.52 mg/g DW and 0.70 mg/g DW, respectively, compared to the control culture (0.29 mg/g DW. Callus cultures elicited with MJ at 30 mg/L for 24 h stimulated GGOH production to 0.35 mg/g DW compared to the control culture (0.07 mg/g DW. Cells also responded to ASA (20 mg/L, 2 days and YE (0.25 g/L, 4 days and produced GGOH contents of 0.46 mg/g DW and 1.37 mg/g DW, respectively. This study has shown that isoprenoid precursors and conventional elicitors enhanced GGOH production in the C. stellatopilosus callus culture.

  19. Production of isoflavones, daidzein and genistein in callus cultures of Pueraria candollei Wall. ex Benth. var. mirifica

    OpenAIRE

    Sanha Panichajakul; Sudarat Thanonkeo

    2006-01-01

    Callus cultures of Pueraria candollei var. mirifica were first established from various parts of explants with the objective of isoflavones, daidzein and genistein production. The cultures were studied on their growth and isoflavone production by various combinations of growth regulators, auxins and cytokinins at 25±2ºC. Daidzein and genistein accumulated in the cells were determined. The results revealed that callus of P. candollei var. mirifica was capable of producing high level of both is...

  20. The Effect of Abscisic Acid on the Freezing Tolerance of Callus Cultures of Lotus corniculatus L.

    Science.gov (United States)

    Keith, C N; McKersie, B D

    1986-03-01

    The effects of growth temperature (2 degrees C and 24 degrees C), abscisic acid (ABA) concentration, duration of exposure to ABA, and light were assessed for their ability to induce acclimation to freezing temperatures in callus cultures of Lotus corniculatus L. cv Leo, a perennial forage legume. The maximal expression of freezing tolerance was achieved on B(5) media containing 10(-5) molar ABA, at 24 degrees C for 7 or 14 days. Under these culture conditions, the freezing tolerance of the callus approximated that observed in field grown plants. In contrast, low temperatures (2 degrees C) induced only a limited degree of freezing tolerance in these cultures. Viability was assessed by tetrazolium reduction and by regrowth of the callus. The two assays often differed in their estimates of absolute freezing tolerance. Regression analysis of the temperature profile suggested that there may be two or more distinct populations of cells differing in freezing tolerance, which may have contributed to the variability between viability assays.

  1. Micropropagation and Callus Culture of Saussurea laniceps, an Alpine Medicinal Plant

    Institute of Scientific and Technical Information of China (English)

    Chen Yu-zhen; Li Feng-lan

    2005-01-01

    Cottonhead windhairdaisy (Saussurea laniceps Hand.-Mazz.) is one of the most famous and important medicinal herbs in China. Illegal collection from wild populations is increasingly threatening the present environment of S. laniceps. Establishment of an efficient method for micropropagation is the best way to change its endangered situation. When mature seeds of S.laniceps were cultured on hormone-free MS medium, plantlets were formed from germinated seeds in 7-10 d. Then 0.5 cm × 0.5 cm leaf explants were transplanted to MS medium supplemented with 1-naphthalene-acetic acid (NAA)/2,4-D and benzyladenine(BA)/KT and callus was achieved 10 d after transfer. Shoot bud regeneration occurred from callus cultured on MS medium supplemented with different growth regulators 20 d after culturing. The regeneration percentages varied with the different components of plant growth regulators. The percent regeneration from callus pretreated at low temperature of 5℃ increased significantly compared with those incubated at 23/20℃ directly. Optimal regeneration was observed with explants on media supplemented with 1.5 mg·L-1 BA plus 0.2 mg·L-1 NAA. In the presence of 0.2 mg·L-1 NAA in half-strength MS, 78% of the shoots formed roots. Plantlets from explants showed 63% survival after acclimatization.

  2. The Extraction Process of Trimethyl Xanthina in Vitro Culture of Callus Camellia Sinensis with ethyl Acetate Solvent

    Directory of Open Access Journals (Sweden)

    Sutini

    2016-01-01

    Full Text Available Trimethyl xanthina is one of the compounds contained bioactive culture in vitro Cammelia sinensis callus which is widely used in the field of food, beverage, agriculture and health industries. The presence of trimethyl xanthina on food, beverages and health is needed in a certain amount depending on the use which is achieved by the user. To get a certain amount of trimethyl xanthina from callus culture of Cammelia sinensis, the extraction process is performed on the water solvent, as well as non-solvent water / organic solvent such as ethyl acetate. The purpose of this study was to obtain profile of trimethyl xanthina in the extraction of Cammelia sinensis callus. The experimental methods used consisted of dissolution, filtration, extraction with water solvent and ethyl acetate, then followed by identification of trimethyl xanthina using HPLC. The results shows the profile form of trimethyl xanthina of Cammelia sinensis callus have similarities with the standard form of trimethyl xanthina.

  3. Production of isoflavonoids in callus cultures of Pueraria candollei var. mirifica.

    Science.gov (United States)

    Udomsuk, Latiporn; Jarukamjorn, Kanokwan; Tanaka, Hiroyuki; Putalun, Waraporn

    2009-01-01

    Pueraria candollei Wall. ex Benth. var. mirifica (Airy Shaw & Suvat.) Niyomdham was investigated for callus induction using Murashige and Skoog (MS) medium containing different plant growth regulators. After 8 weeks of culture, 66-100% of leaf or stem explants formed calli. Calli from stem explants cultured on MS medium supplemented with 0.5 mg/l thidiazuron (TDZ) gave the maximum of shoot induction (16%) and the highest level of total isoflavonoids [(50.39 +/- 7.06) mg/g dry wt], which was 7-fold higher than that of the native tuber [(7.04 +/- 0.29) mg/g dry wt]. These results suggest that addition of TDZ to the culture medium markedly enhances the production of isoflavonoids in calli induced from stem explants of P. candollei var. mirifica.

  4. Inhibition of aconitase in citrus fruit callus results in a metabolic shift towards amino acid biosynthesis

    NARCIS (Netherlands)

    Degu, A.; Hatew, B.; Nunes-Nesi, A.; Shlizerman, L.; Zur, N.; Fernie, A.R.; Blumwald, E.; Sadka, A.

    2011-01-01

    Citrate, a major determinant of citrus fruit quality, accumulates early in fruit development and declines towards maturation. The isomerization of citrate to isocitrate, catalyzed by aconitase is a key step in acid metabolism. Inhibition of mitochondrial aconitase activity early in fruit development

  5. Primary characterization and evaluation of anti ulcerogenic activity of an aqueous extract from callus culture of Cereus peruvianus Mill. (Cactaceae).

    Science.gov (United States)

    Jayme, Milena O; Ames, Franciele Q; Bersani-Amado, Ciomar A; Machado, Maria de Fatima P S; Mangolin, Claudete A; Goncalves, Regina A C; de Oliveira, Arildo J B

    2015-01-01

    In the current study we reported cultivation, extraction procedure, analysis and preliminary characterization of the aqueous extract from Cereus peruvianus callus culture and evaluated its anti ulcerogenic activity in vivo models of experimental ulcers in Wistar rats. The obtained aqueous extract from callus (AC) was dialyzed and subjected to freeze-thaw process, providing a possible polysaccharide. The carbohydrate and protein contents of the aqueous extract were estimated at 53.4% and 0.66%, respectively, composed primarily of galactose, arabinose and galacturonic acid, with minor amounts of glucose. This appeared heterogeneous when analyzed by high-performance size-exclusion chromatography and a multiangle laser light scattering detector (HPSEC-MALLS). The AC was found to be significantly effective against ethanol-induced lesions but was ineffective against indomethacin-induced lesions. The callus culture of C. peruvianus is an alternative source for the synthesis of substances originally produced by plants. The calluses grown indefinitely in vitro under controlled conditions are stable tissues, and the aqueous extract from calluses may be used instead of fully developed plants using the protocols described in this study.

  6. Effect of plant growth regulators on fatty acids composition in Jatropha curcas L. callus culture.

    Science.gov (United States)

    Hernandez, Ludwi Rodríguez; Mendiola, Martha A Rodríguez; Castro, Carlos Arias; Gutiérrez-Miceli, Federico A

    2015-01-01

    The influence of Naphtaleneacetic acid (NAA) and 6-Benzylaminopurine (BAP) on callus formation, its morphology and fatty acids profile were examined from Jatropha curcas L. Embryo from seeds of J. curcas L. were sown in Murashige and skoog (MS) medium with NAA and BAP. All treatments induced callus formation, however callus morphology was different in most of the treatments. Higher callus biomass was presented with 1.0 NAA + 0.5 BAP mg/L. Plant growth regulators modifies the fatty acids profile in callus of J. curcas L. BAP was induced linoleic and linolenic acids.

  7. Cold Response of Dedifferentiated Barley Cells at the Gene Expression, Hormone Composition, and Freezing Tolerance Levels: Studies on Callus Cultures

    OpenAIRE

    Vashegyi, I.; Marozsan-Toth, Z.; Galiba, G.; Dobrev, P. (Petre); Vaňková, R. (Radomíra); Toth, B.

    2013-01-01

    In this study, data is presented how dark-grown, embryogenic barley callus cells respond to cold without any light-dependent, chloroplast-related mechanism, independently of the systemic signals. The expression of HvCBF9, HvCBF14, and HvCOR14b genes, members of one of the most important cold-inducible regulatory system, was measured by real-time PCR. Characteristic of the cold response was similar in the crowns of seedlings and in dark-grown callus cultures, however, gene expression levels we...

  8. Responses to NaCl stress of cultivated and wild tomato species and their hybrids in callus cultures.

    Science.gov (United States)

    Cano, E A; Pérez-Alfocea, F; Moreno, V; Bolarin, M C

    1996-06-01

    If in vitro culture is to be used for evaluating the salt tolerance of tomato hybrids and segregant populations in a breeding programme, it is previously necessary to get quick and reliable traits. In this work, growth and physiological responses to salinity of two interspecific hybrids between the cultivated tomato (Lycopersicon esculentum Mill) and its wild salt-tolerant species L pennellii are compared to those of their parents. The leaf callus of the first subculture was grown on media amended with 0, 35, 70, 105, 140, 175 and 210 mM NaCl for 40 days. Relative fresh weight growth of callus in response to increased salinity in the culture medium was much greater in L pennellii than in the tomato cultivars, and greater in the hybrids than in the wild species. Moreover, the different salt tolerance degree of hybrids was related to that of female parents. At high salt levels, only Cl(-) accumulation was higher in L pennellii than in tomato cultivars, whereas in the hybrids both Cl(-), and Na(+) accumulation were higher than in their parents. Proline increased with salinity in the callus of all genotypes; these increases were much higher in the tomato cultivars than in L pennellii, and the hybrids showed a similar response to that of the wild species. Salt-treated callus of the tomato cultivars showed significant increases in valine, isoleucine and leucine contents compared to control callus tissue. In contrast, these amino acids in callus tissues of the wild species and hybrids showed a tendency to decrease with increasing salinity.

  9. Production and optimisation of rosmarinic acid by Satureja hortensis L. callus cultures.

    Science.gov (United States)

    Tepe, Bektas; Sokmen, Atalay

    2007-11-01

    In this study, production and optimisation of rosmarinic acid, a phenolic acid and an economically important metabolite, was investigated in the callus cultures established from the mature seeds of Satureja hortensis L. (summer savory) plant. Gamborg's B5 basal medium, supplemented with indol butyric acid (IBA) (1.00 mg L(-1)), N6-benzyl aminopurine (6-BA) (1.00 mg L(-1)) and sucrose (2.5%, w/v), was employed for the establishment and maintenance of the callus cultures. Applications were individually prepared by preparing the media containing different IBA/6-BA combinations and sucrose concentrations. All of the applications were carried out in the continuous dark. In the applications, where the effects of IBA/6-BA combinations on the growth and rosmarinic acid accumulation were assayed (1-15 applications), the highest biomass yield was obtained from the medium supplemented with 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA. In the case of the rosmarinic acid accumulation, an opposite relationship was determined between the growth and rosmarinic acid production. While the highest biomass yield was obtained from the medium containing 1.00 mg L(-1) IBA and 5.00 mg L(-1) 6-BA, the highest rosmarinic acid accumulation was obtained from the medium supported with 1.00 mg L(-1) IBA and 1.00 mg L(-1) 6-BA. In the applications where the effects of sucrose concentrations on the growth and rosmarinic acid accumulation were examined, the highest biomass yield was obtained from the medium which is supplemented with 5.0% (w/v) sucrose. In this category, the highest rosmarinic acid accumulation was obtained from the medium which is supported with 3.0% (w/v) sucrose. According to the experiments carried out with the wild S. hortensis, it is found to have 25.02+/-1.21 mg g(-1) rosmarinic acid. No differentiation was observed in any callus during the course of this study.

  10. Impact of UV-B radiation on some biochemical changes and growth parameters in Echinacea purpurea callus and suspension culture

    Directory of Open Access Journals (Sweden)

    Hossam H. Manaf

    2016-12-01

    Full Text Available The effects of UV-B light force, exposure time and incubation period on producing caffeic acid derivatives and growth parameters in Echinacea purpurea callus and suspension culture were assessed. UV-B led to an increment of all growth parameters and antioxidant activity in callus and cell suspension and caffeic acid derivatives in cell suspension by increasing incubation period. The reverse was true for G-POD activity in cell suspension and PAL activity in both types of cultures. Incubation period 2 weeks was more effective in caffeic acid, total phenols and G-POD activity in callus cells and incubation period one week only for total phenols in cell suspension. The two exposure times 2 and 4 h increased antioxidant activity in the two types of cultures. Exposure time 2 h led to increase caffeic acid and total phenols in callus cells. The maximum increase in caffeic acid, total phenols and PAL activity in cell suspension was achieved by 4 h exposure time. Likewise, using 2 UV-B lamps for 2 h was the most effective in creating more biochemical components than the other treatments.

  11. Embryogenic Callus Induction of Pencil Orchid (Papilionanthe hookeriana Rchb.f. Through in Vitro Culture

    Directory of Open Access Journals (Sweden)

    Atra Romeida

    2016-03-01

    Full Text Available Papilionthe hookeriana  Rchb.f.  (Orchidaceae,  popularly known as ‘Anggrek Pinsil’  in Indonesia, is a perennial ephiphyte orchid, found only at Dendam Tak Sudah Lake in Bengkulu. The aims of this research were to find the best sterilization technique of P.  hookeriana explant and to induce maximum formation of embryogenic calli. Rapid multiplication of this orchid was achieved through culture of shoot tips and young leaf segments of mature plants by in vitro cultured on Murashige and Skoog (MS basal medium  enriched with 50 g L-1 banana pulp.    The experiment used Completely Randomized Design (CRD with three replications. In the first stage, the explants were sterilized using three compositions of sterilant. In the second stage, the explants were planted on the MS basal medium with addition of five levels of 2,4-Diclhorophenoxy Acetis Acid concentrations, namely 0, 0.25,  0.50, 0.75, and 1.00 mg L-1.  The result showed that the best sterilization method to reduce explant contaminant  was method 3, in which the explant  was washed with detergent, rinsed with flowing water, soaked in 0.1%  (v/v HgCl2 solution for 30 minutes, soaked again in the 10% (v/v Natrium hypocloride solution for 20 minutes,  rinsed three times with sterile water before planted and then soaked in  sterile  water + 10% (v/v  betadhine before planted on treated medium. This method was able to reduce contamination levels up to 70% from explants cultured for 5 months on MS medium.  MS medium added with 1.00 mg L-1 2,4-D produced the highest number of embryogenic calli, and the biggest callus diameter ( 3.5 cm , characterized by  transparent green color and  friable callus structure

  12. Optimization of callus and cell suspension cultures of Barringtonia racemosa (Lecythidaceae family) for lycopene production

    OpenAIRE

    Behbahani, Mandana; Shanehsazzadeh, Mehrnaz; Hessami,Mohamad Javad

    2011-01-01

    Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC) and dark. Leaf explants of Barringtonia racemosa we...

  13. Biomass Yield and Steviol Glycoside Production in Callus and Suspension Culture of Stevia rebaudiana Treated with Proline and Polyethylene Glycol.

    Science.gov (United States)

    Gupta, Pratibha; Sharma, Satyawati; Saxena, Sanjay

    2015-06-01

    Enhanced production of steviol glycosides (SGs) was observed in callus and suspension culture of Stevia rebaudiana treated with proline and polyethylene glycol (PEG). To study their effect, yellow-green and compact calli obtained from in vitro raised Stevia leaves were sub-cultured on MS medium supplemented with 2.0 mg l(-1) NAA and different concentrations of proline (2.5-10 mM) and PEG (2.5-10 %) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16 h. Callus and suspension culture biomass (i.e. both fresh and dry weight content) was increased with 5 mM proline and 5 % PEG, while at further higher concentrations, they got reduced. Further, quantification of SGs content in callus (collected at 15th day) and suspension culture (collected at 10th and 15th day) treated with and without elicitors was analysed by HPLC. It was observed that chemical stress enhanced the production of SGs significantly. In callus, the content of SGs increased from 0.27 (control) to 1.09 and 1.83 % with 7.5 mM proline and 5 % PEG, respectively, which was about 4.0 and 7.0 times higher than control. However, in the case of suspension culture, the same concentrations of proline and polyethylene glycol enhanced the SG content from 1.36 (control) to 5.03 and 6.38 %, respectively, on 10th day which were 3.7 times and 4.7 times higher than control.

  14. Light-induced biochemical variations in secondary metabolite production and antioxidant activity in callus cultures of Stevia rebaudiana (Bert).

    Science.gov (United States)

    Ahmad, Naveed; Rab, Abdur; Ahmad, Nisar

    2016-01-01

    Stevia rebaudiana (S. rebaudiana) is a very important species with worldwide medicinal and commercial uses. Light is one of the major elicitors that fluctuate morphogenic potential and biochemical responses. In the present study, we investigated the effect of various spectral lights on biomass accumulation and secondary metabolite production in callus cultures of S. rebaudiana. Leaf explants were placed on Murashige and Skoog (MS) medium and exposed to various spectral lights. 6-Benzyle adenine (BA) and 2, 4-dichlorophenoxy acetic acid (2, 4-D; 2.0 mgl(-1)) were used for callus induction. The control light (16/8h) produced optimum callogenic response (92.73%) than other colored lights. Compared to other colored lights, control grown cultures displayed maximum biomass accumulation (5.78 gl(-1)) during a prolonged log phase at the 18th day of growth kinetics. Cultures grown under blue light enhanced total phenolic content (TPC; 102.32 μg/g DW), total flavonoid content (TFC; 22.07 μg/g DW) and total antioxidant capacity (TAC; 11.63 μg/g DW). On the contrary, green and red lights improved reducing power assay (RPA; 0.71Fe(II)g(-1) DW) and DPPH-radical scavenging activity (DRSA; 80%). Herein, we concluded that the utilization of colored lights is a promising strategy for enhanced production of antioxidant secondary metabolites in callus cultures of S. rebaudiana.

  15. Synthesis of deuterium-labelled substrates for the study of oleuropein biosynthesis in Olea europaea callus cultures.

    Science.gov (United States)

    Serrilli, Anna Maria; Maggi, Agnese; Casagrande, Valentina; Bianco, Armandodoriano

    2016-01-01

    We propose the cell culture approach to investigate oleuropein (1) biogenesis in Olea europaea L. We suggest employing olive callus cultures to identify the iridoidic precursor of oleuropein. In fact, we confirmed that callus cells from olive shoot explants are able to produce key secoiridoid as 1. To enable this approach, we synthesised and characterised deuterium-labelled iridoidic precursors belonging both to the loganin and the 8-epiloganin series. These iridoids are [7,8-(2)H2]-7-deoxy-8-epi-loganin (2(D)), [8,10-(2)H2]-8-epi-loganin (4(D)) and [7,8-(2)H2]-7-deoxy-loganin (3(D)).

  16. Callus culture development of two varieties of Tagetes erecta and carotenoid production

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    Israel Benítez-García

    2014-05-01

    Conclusions: WF callus appeared to be a suitable candidate as a source of different carotenoids, and tested varieties could represent an alternative for further studies about in vitro pigment production.

  17. Production of isoflavones, daidzein and genistein in callus cultures of Pueraria candollei Wall. ex Benth. var. mirifica

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    Sanha Panichajakul

    2006-03-01

    Full Text Available Callus cultures of Pueraria candollei var. mirifica were first established from various parts of explants with the objective of isoflavones, daidzein and genistein production. The cultures were studied on their growth and isoflavone production by various combinations of growth regulators, auxins and cytokinins at 25±2ºC. Daidzein and genistein accumulated in the cells were determined. The results revealed that callus of P. candollei var. mirifica was capable of producing high level of both isoflavones consistently. The culture temperature played an important role in the growth and isoflavone production. Over twofold of growth and threefold of isoflavone production were demonstrated at 32±2ºC. The callus established from the stems in MS medium supplemented with 4.5 µM 2,4-D and 0.46 µM kinetin produced the highest yield of daidzein (5.12 mg/g, DW and genistein (2.77 mg/g, DW, which was remarkably higher than the intact plants.

  18. Agrobacterium-mediated genetic transformation of Coffea arabica (L. is greatly enhanced by using established embryogenic callus cultures

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    Lashermes Philippe

    2011-05-01

    Full Text Available Abstract Background Following genome sequencing of crop plants, one of the main challenges today is determining the function of all the predicted genes. When gene validation approaches are used for woody species, the main obstacle is the low recovery rate of transgenic plants from elite or commercial cultivars. Embryogenic calli have frequently been the target tissue for transformation, but the difficulty in producing or maintaining embryogenic tissues is one of the main problems encountered in genetic transformation of many woody plants, including Coffea arabica. Results We identified the conditions required for successful long-term proliferation of embryogenic cultures in C. arabica and designed a highly efficient and reliable Agrobacterium tumefaciens-mediated transformation method based on these conditions. The transformation protocol with LBA1119 harboring pBin 35S GFP was established by evaluating the effect of different parameters on transformation efficiency by GFP detection. Using embryogenic callus cultures, co-cultivation with LBA1119 OD600 = 0.6 for five days at 20 °C enabled reproducible transformation. The maintenance conditions for the embryogenic callus cultures, particularly a high auxin to cytokinin ratio, the age of the culture (optimum for 7-10 months of proliferation and the use of a yellow callus phenotype, were the most important factors for achieving highly efficient transformation (> 90%. At the histological level, successful transformation was related to the number of proembryogenic masses present. All the selected plants were proved to be transformed by PCR and Southern blot hybridization. Conclusion Most progress in increasing transformation efficiency in coffee has been achieved by optimizing the production conditions of embryogenic cultures used as target tissues for transformation. This is the first time that a strong positive effect of the age of the culture on transformation efficiency was demonstrated. Our

  19. Production of rosmarinic acid and salvianolic acid B from callus culture of Salvia miltiorrhiza with cytotoxicity towards acute lymphoblastic leukemia cells.

    Science.gov (United States)

    Wu, Ching-Fen; Karioti, Anastasia; Rohr, Doris; Bilia, Anna Rita; Efferth, Thomas

    2016-06-15

    Salvia miltiorrhiza (SM) Bunge is one of the widely-used Chinese medicinal herbs. In this study, the chemical constituents and anticancer potential of SM stems and leaves were examined with those of respective callus cultures. The callus culture for stem and leaf explants was initiated in modified Murashige and Skoog (MS) medium. Active constituents of respective extracts were analyzed by high performance liquid chromatography coupled with DAD and MS (HPLC-DAD-MS). Rosmarinic acid (RA) and salvianolic acid B (Sal B) were determined to be the main phenolic compounds. Quantitative analyses revealed that callus stem extracts produced higher amount of RA and Sal B (stem RA: 1.27±0.38%; stem Sal B: 0.87±0.20%) than callus leaf did (leaf RA: 0.28±0.02%; leaf Sal B: 0.07±0.03%). Stem and leaf callus extracts exerted cytotoxic effects towards CCRF-CEM cells (stem: 13.1±0.90 μg/ml; leaf: 18.1±0.33 μg/ml). As expected, stem extract with higher amount of RA and Sal B showed lower IC50 value than leaf extract. These findings suggest the possibility to isolate bioactive constituents with anticancer properties from in vitro callus cultures of stems and leaves of SM.

  20. Callus cell proliferation from broccoli leaf slice using IBA and BAP in vitro culture: Its biochemical and antioxidant properties

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    A.B.M. Sharif Hossain

    2016-03-01

    Full Text Available Plant tissue or cell culture keeps a significant role in micro-propagation in the plant production industry. Combination of 6-Benzylaminopurine (BAP and other plant growth regulators like 1-Naphthaleneacetic acid (NAA or Indole-3-acetic acid (IAA or indole-3-butyric acid (IBA was used in the most of the research in tissue culture. The study was carried out to investigate the optimization of the concentration of IBA and BAP combination (0, 0.25, 0.50, 1.0, 1.50, 2.0, 2.5, 3.0 and 3.5 mg/l for the root, callus and leaf proliferation from the leaf cutting slice. The highest number (6.75 of root proliferation was observed in the concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination. The callus initiation was found in the concentration of IBA 1.0–3.5 mg/l+BAP 1.0–2.0 mg/l. However, the highest callus weight was observed at the concentration of IBA 1.5 mg/l+BAP 1.0 mg/l combination than other combination of concentrations. Positively leaf initiation and formation was better in the concentration of IBA 1–3.5 mg/l+BAP 1.0–2.0 mg/l combination. In addition, the 2,2-diphenyl-2-picrylhydarzyl (DPPH free radical scavenging potential was higher (70.1% in leaves extract than in callus extracts (46.3% at the concentration of 10 mg/ml though both extracts had lower DPPH free radical scavenging activity compared to the positive control, vitamin C and BHT. Theresults conclude that the optimum concentration was IBA 1.5 mg/l+BAP 1.0 mg/l combination to produce callus cell proliferation and concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination was the optimum for root proliferation of broccoli in vitro.

  1. Callus cell proliferation from broccoli leaf slice using IBA and BAP in vitro culture: Its biochemical and antioxidant properties.

    Science.gov (United States)

    Sharif Hossain, A B M; Haq, Imdadul; Ibrahim, Nasir A; Aleissa, Mohammed Saad

    2016-03-01

    Plant tissue or cell culture keeps a significant role in micro-propagation in the plant production industry. Combination of 6-Benzylaminopurine (BAP) and other plant growth regulators like 1-Naphthaleneacetic acid (NAA) or Indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) was used in the most of the research in tissue culture. The study was carried out to investigate the optimization of the concentration of IBA and BAP combination (0, 0.25, 0.50, 1.0, 1.50, 2.0, 2.5, 3.0 and 3.5 mg/l) for the root, callus and leaf proliferation from the leaf cutting slice. The highest number (6.75) of root proliferation was observed in the concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination. The callus initiation was found in the concentration of IBA 1.0-3.5 mg/l+BAP 1.0-2.0 mg/l. However, the highest callus weight was observed at the concentration of IBA 1.5 mg/l+BAP 1.0 mg/l combination than other combination of concentrations. Positively leaf initiation and formation was better in the concentration of IBA 1-3.5 mg/l+BAP 1.0-2.0 mg/l combination. In addition, the 2,2-diphenyl-2-picrylhydarzyl (DPPH) free radical scavenging potential was higher (70.1%) in leaves extract than in callus extracts (46.3%) at the concentration of 10 mg/ml though both extracts had lower DPPH free radical scavenging activity compared to the positive control, vitamin C and BHT. Theresults conclude that the optimum concentration was IBA 1.5 mg/l+BAP 1.0 mg/l combination to produce callus cell proliferation and concentration of 2.0 mg/l IBA+0.25 mg/l BAP combination was the optimum for root proliferation of broccoli in vitro.

  2. Comparative analysis of lycorine in wild plant and callus culture samples of Hymenocallis littoralis by HPLC-UV method.

    Science.gov (United States)

    Subramaniam, Sreeramanan; Sundarasekar, Jeevandran; Sahgal, Geethaa; Murugaiyah, Vikneswaran

    2014-01-01

    The Hymenocallis littoralis, an ornamental and medicinal plant, had been traditionally used for wound healing. In the present study, an analytical method using HPLC with ultraviolet detection was developed for the quantification of lycorine in the extracts of different parts of wild plant and tissue culture samples of H. littoralis. The separation was achieved using a reversed-phase column. The method was found to be accurate, repeatable, and sensitive for the quantification of minute amount of lycorine present in the samples. The highest lycorine content was found in the bulb extract (2.54 ± 0.02 μg/mg) whereas the least was in the root extract (0.71 ± 0.02 μg/mg) of the wild plants. Few callus culture samples had high content of lycorine, comparable to that of wild plants. The results showed that plant growth regulators, 2,4-dichlorophenoxyacetic acid (2,4-D) alone at 4.5 μM (2.58 ± 0.38 μg/mg) or a combination of 2,4-D at 9.00 μM with 4.5 μM of 6-benzylaminopurine (BAP), were the optimum concentrations for the production of high lycorine (2.45 ± 0.15 μg/mg) content in callus culture. The present analytical method could be of value for routine quantification of lycorine in the tissue culture production and standardization of the raw material or extracts of H. littoralis.

  3. Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae: proteomic and metabolic responses at early stage

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    Payet Bertrand

    2010-05-01

    Full Text Available Abstract Background Vanilla planifolia is an important Orchid commercially cultivated for the production of natural vanilla flavour. Vanilla plants are conventionally propagated by stem cuttings and thus causing injury to the mother plants. Regeneration and in vitro mass multiplication are proposed as an alternative to minimize damage to mother plants. Because mass production of V. planifolia through indirect shoot differentiation from callus culture is rare and may be a successful use of in vitro techniques for producing somaclonal variants, we have established a novel protocol for the regeneration of vanilla plants and investigated the initial biochemical and molecular mechanisms that trigger shoot organogenesis from embryogenic/organogenic callus. Results For embryogenic callus induction, seeds obtained from 7-month-old green pods of V. planifolia were inoculated on MS basal medium (BM containing TDZ (0.5 mg l-1. Germination of unorganized mass callus such as protocorm -like structure (PLS arising from each seed has been observed. The primary embryogenic calli have been formed after transferring on BM containing IAA (0.5 mg l-1 and TDZ (0.5 mg l-1. These calli were maintained by subculturing on BM containing IAA (0.5 mg l-1 and TDZ (0.3 mg l-1 during 6 months and formed embryogenic/organogenic calli. Histological analysis showed that shoot organogenesis was induced between 15 and 20 days after embryogenic/organogenic calli were transferred onto MS basal medium with NAA (0.5 mg l-1. By associating proteomics and metabolomics analyses, the biochemical and molecular markers responsible for shoot induction have been studied in 15-day-old calli at the stage where no differentiating part was visible on organogenic calli. Two-dimensional electrophoresis followed by matrix-assisted laser desorption ionization time-of-flight-tandem mass spectrometry (MALDI-TOF-TOF-MS analysis revealed that 15 protein spots are significantly expressed (P Conclusion The

  4. Induction of secondary metabolite production by UV-C radiation in Vitis vinifera L. Öküzgözü callus cultures

    OpenAIRE

    Cetin, Emine Sema

    2014-01-01

    Background The aim of the present work was to examine the role of UV-C irradiation on the production of secondary metabolites (total phenolic, total flavanols, total flavonols, catechin, ferulic acid and trans-resveratrol in phenolic compounds and α-, β-, γ- δ-tocopherols) in callus cultures. Studies on the effects of UV-C treatment on callus culture are seldom and generally focused on UV-B. However UV-C radiation play an important role in accumule secondary metabolites. Results In this study...

  5. 匙羹藤愈伤组织培养研究%Study on Callus Culture from Gymnema sylvestre

    Institute of Scientific and Technical Information of China (English)

    苏钛; 黄宁珍

    2015-01-01

    Using Gymnema sylvestre as explants to study how stems, stem tips, leaves and hormone affect callus. Using 4 factors and 4 levels orthogonal to design, using SPSS to analyze results. Results displayed that all kinds of explants have different callus induction abilities, followed by stems and leaves. Leaves inducedi non-embryogenic callus. Stems and stem tips induced embryogenic callus and embryoid. The best tissue culture of callus induction is MS+2,4-D 2. 0 mg/L+6-BA 0. 5 mg/L, the induction rate is 84. 5%, callus grown more vigorous, splinter grown faster, the color is faint yellow. The optimal medi-um for differentiation was MS+IBA 0. 1 mg/L+NAA 1. 0 mg/L. The inducing rate was 90. 6%, The optimal rooted medium was 1/2 MS+NAA 1. 6 mg/L. The rooting rate was 100%. This provides refer-ence for dedifferentiation, differentiation, embryoid formation mechanism of Gymnema sylvestre organiza-tion.%以匙羹藤组培苗为外植体,研究茎段、茎尖和叶片以及不同植物生长调节剂组合对愈伤组织形成的影响;采用4因素4水平正交设计,用SPSS软件其结果进行分析。结果显示:各种外植体诱导愈伤的能力不同,依次是茎段、叶片。叶片诱导出非胚性愈伤组织,茎段、茎尖利于诱导胚性愈伤组织和胚状体;愈伤诱导的最佳培养基为MS+2,4-D 2.0 mg/L+6-BA 0.5 mg/L,诱导率为84.5%,愈伤生长旺盛,分裂快,颜色为淡黄色。愈伤组织分化成苗效果最好的条件是MS+NAA 1.0 mg/L+IBA 0.1 mg/L,分化率达到90.6%,最佳的生根条件是1/2 MS+NAA 1.6 mg/L,生根率为100%。这为研究匙羹藤的组织的脱分化、分化以及胚状体的形成机制提供了参考。

  6. [Plants regeneration from genetically transformed root and callus cultures of periwinkle Vinca minor L. and foxglove purple Digitalis purpurea L].

    Science.gov (United States)

    Leshina, L G; Bulko, O V

    2014-01-01

    Plants regenerated from hairy roots and calluses of foxglove purple and periwinkle have been obtained. It was found that organogenesis in hairy root culture occurs spontaneously on hormone-free medium but with different efficiencies. The frequency of direct shoot formation from root cultures was up to 60% in Digitalis and 3.7% in Vinca. Addition of 1 mg/l BA, 0.1 mg/l NAA and 5% sucrose to B5 medium increased regenerative capacity of Vinca roots up to 19.1%. Regenerated plants showed morphological features typically seen in Ri-transgenic plants. They include growth and plagiotropism of the root system, increased shoot formation, changed leaf morphology and short internodes.

  7. 中药白芍愈伤组织的培养及其中芍药苷的测定%Study on Callus Culture of Paeonia lactiflora pall and Determination of Paeoniflorin in Callus

    Institute of Scientific and Technical Information of China (English)

    凌庆枝; 董丽辉; 章聪敏; 高莉莉; 江海龙; 冯定军; 周海滨

    2013-01-01

    [Objective] To establish the methods of callus culture,and determine the content of paeoniflorin in callus cells of Paeonia lactiflora pall.[Method] Leaves and stem sterilized were cut into tissue pieces and inoculated on different induction medium and regeneration medium.Paeoniflorin was extracted from callus and the content of paeoniflorin was determined by TLC and HPLC.[Result] The results showed that medium of 1/2MS + 0.2 mg/L 2,4-D + 2.0 mg/L 6-BA + 0.2 mg/L NAA were fit for stem callus induction,and medium of 1/2MS + 1.0 mg/L 2,4-D + 1.5 mg/L 6-BA were fit for leaves callus induction.Regeneration medium of callus is 1/2MS + 0.1 mg/L 2,4-D + 1.0 mg/L ZT + 0.5 mg/L NAA.The content of paeoniflorin of callus is 0.068 9 mg/g by HPLC.[Conclusion] Leaf and stem of Paeonia lactiflora pall could be induced to form callus.There are paeoniflorin in callus cells of Paeonia lactiflora pall.%[目的]建立中药白芍愈伤组织培养的方法,并测定其中芍药苷的含量.[方法]以白芍叶、茎为外植体,接种于不同的愈伤诱导培养基和继代培养基上培养愈伤组织,并提取愈伤组织中的芍药苷,用TLC和HPLC法测定愈伤组织中芍药苷的含量.[结果]适合白芍茎的诱导愈伤培养基是1/2MS +0.2 mg/L2,4-D +2.0 mg/L 6-BA +0.2 mg/L NAA,叶的最适诱导培养基是1/2MS+ 1.0 mg/L 2,4-D+ 1.5 mg/L 6-BA,白芍愈伤继代培养适合培养基为1/2MS +0.1 mg/L2,4-D+ 1.0 mg/L ZT +0.5 mg/L NAA.经HPLC测定每克干白芍愈伤组织中的芍药苷含量为0.068 9 mg.[结论]中药白芍的叶和茎可诱导形成愈伤组织,且形成的愈伤组织中含有芍药苷.

  8. Innovative use of Mucuna monosperma (Wight) callus cultures for continuous production of melanin by using statistically optimized biotransformation medium.

    Science.gov (United States)

    Inamdar, Shrirang; Joshi, Swati; Bapat, Vishwas; Jadhav, Jyoti

    2014-01-20

    Melanins are predominantly indolic polymers which are having extensive applications in cosmetics, agriculture and medicine. In the present study, optimization of nutritional parameters influencing melanin production by Mucuna monosperma callus cultures was attempted using the response surface methodology (RSM). Standardization of four factors was carried out using the Box-Behnken design. The optimized levels of factors predicted by the model include tyrosine 0.978gL(-1), pH 5.85, SDS 34.55mgL(-1)and copper sulphate 21.14mgL(-1) tyrosine, which resulted in highest melanin yield of 0.887gL(-1). The optimization of medium using RSM resulted in a 3.06-fold increase in the yield of melanin. The ANOVA analysis showed a significant R(2)-value (0.9995), model F-value (1917.72) and probability (0.0001), with insignificant lack of fit. Optimized medium was used in the laboratory scale column reactor for the continuous production of melanin. Uninterrupted flow column exhibited maximum melanin production rate of 250mgL(-1)h(-1) which is the highest value ever reported using plant as a biotransformation source. Melanin production was confirmed by spectrophotometric and chemical analysis. Thus, this study demonstrates the production of melanin by M. monosperma callus, using a laboratory scale column reactor.

  9. Malate dehydrogenase isozymes (MDH; EC 1.1.1.37) in long-term callus culture of Cereus peruvianus (Cactaceae) exposed to sugar and temperature stress.

    Science.gov (United States)

    Jorge, I C; Mangolin, C A; Machado, M F

    1997-06-01

    Malate dehydrogenase (MDH; EC 1.1.1.37) isozymes in long-term callus tissue culture of Cereus peruvianus were studied in starch gel electrophoresis to investigate the control of differential Mdh gene expression under sugar and temperature stress. While two cytosol MDH isozymes showed an unchanged phenotype when the callus tissues were transferred to medium maintained at 22 or 37 degrees C and containing different concentrations of sucrose, glucose, and fructose, the different combinations of five mitochondrial MDH (mtMDH) and two micro-body MDH (mbMDH) showed different MDH isozyme patterns in the callus populations. Differential expression of mtMDH isozymes seems to be modulated at the posttranslational level in callus tissues exposed to different concentrations and types of sugar and to high-temperature and low-temperature stress. An inductor effect on the expression of mbMDH isozymes was observed under stress conditions and in long-term callus tissue, and they may also present different responses.

  10. Changes in the effective gravitational field strength affect the state of phosphorylation of stress-related proteins in callus cultures of Arabidopsis thaliana

    OpenAIRE

    Barjaktarović, Žarko; Schütz, Wolfgang; Madlung, Johannes; Fladerer, Claudia; Nordheim, Alfred; Hampp, Rüdiger

    2009-01-01

    In a recent study it was shown that callus cell cultures of Arabidopsis thaliana respond to changes in gravitational field strengths by changes in protein expression. Using ESI-MS/MS for proteins with differential abundance after separation by 2D-PAGE, 28 spots which changed reproducibly and significantly in amount (P

  11. ANTIOXIDANT ACTIVITY IN CALLUS CULTURES AND IN VITRO REGENERATED PLANTS OF ARTEMISIA NILAGIRICA (CLARKE PAMP.- AN IMPORTANT MEDICINAL PLANT SPECIES

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    A. K. BALIARSINGH 1* , L. SAMANTA 2 AND S. S. MOHANTY 3

    2015-01-01

    dichlorophenoxyacetic  acid  (2,4-D  with  1.0  mg/L  Kinetin  (Kin  yielded maximum  callogenic  response.  Shoot  organogenesis  in  callus  cultures  was most favoured in MS containing 2.0 mg/L 6-benzylaminopurine (BAP and 0.5 mg/L indole-3-acetic acid (IAA. In vitro regenerated plantlets, emerged from culture medium, were acclimatized and the survival rate of ex vitro plants after soil transplantation  was 80-83% with  no apparent  phenotypic variations. The antioxidant  potential  of  natural  (in  vivo  plants,  callus  tissues  and  in  vitro regenerated plants before and after field transplantation (ex vitro plants were compared. DPPH scavenging activity was the highest in aqueous extracts of 10 week-old ex vitro plants than  other  sources.  Superoxide  anion  and  nitric  oxide  radical  scavenging  activity  was  the  highest  in ethanolic  extracts  of  10 week-old  ex  vitro  plants  where  as  the  hydroxyl  radical  was  the  maximum  in  6 week-old in vivo plants. Lipid peroxidation was neither observed in calli nor in regenerated plants of A. nilagirica.

  12. Production of the Anti-Inflammatory Compound 6-O-Palmitoyl-3-O-β-D-glucopyranosylcampesterol by Callus Cultures of Lopezia racemosa Cav. (Onagraceae

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    Roberta Salinas

    2014-06-01

    Full Text Available Lopezia racemosa Cav. is a plant used in Mexican traditional medicine to heal inflammatory diseases. From this plant we isolated the novel compound 6-O-palmitoyl- 3-O-β-D-glucopyranosylcampesterol (1 and 6-O-palmitoyl-3-O-β-D-glucopyranosyl-β-sitosterol (2, previously reported to have cytotoxic activity on several cancer cell lines. We evaluated the anti-inflammatory activity of 1 in vivo by mouse ear edema induced with 12-O-tetradecanoylphorbol-13-acetate (TPA and 57.14% inhibition was observed. The aim of our study was to obtain callus cultures derived from this plant species with the ability to produce the compounds of interest. Callus cultures were initiated on MS basal medium amended with variable amounts of naphthaleneacetic acid (NAA, or 2,4-dichlorophenoxyacetic acid (2,4-D, combined or not with 6-benzylaminopurine (BAP. Ten treatments with these growth regulators were carried out, using in vitro germinated seedlings as source of three different explants: hypocotyl, stem node, and leaf. Highest yield of 1 was observed on callus derived from leaf explants growing in medium containing 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Selected callus lines produced less 1 than wild plants but the in vitro cultured seedlings showed higher production. So we conclude that it could be attractive to further investigate their metabolic potential.

  13. In Vitro Production of Echioidinin, 7-O-Methywogonin from Callus Cultures of Andrographis lineata and Their Cytotoxicity on Cancer Cells.

    Directory of Open Access Journals (Sweden)

    Arifullah Mohammed

    Full Text Available Andrographis lineata is an herbal medicinal plant used in traditional medicine as a substitute for Andrographis paniculata. Here, using mature leaf explants of A. lineata we demonstrate for the first time the callus induction established on MS medium containing 1.0 mg l-1 IAA. Dried callus was subjected to solvent extraction with acetone. Further the acetone residue was separated by silica gel column chromatography, crystallized and characterized on the basis of nuclear magnetic resonance (proton and c13 and liquid chromatographic mass spectroscopy. This analysis revealed the occurrence of two known flavones namely, 7-O-methylwogonin (MW and Echioidinin (ED. Furthermore, these compounds were tested for their cytotoxicity against leukemic cell line, CEM. We identify that ED and MW induced cytotoxicity in a time- and concentration-dependent manner. Further increase in the LDH release upon treatment with ED and MW further confirmed our cytotoxicity results against leukemic cell line. Strikingly, MW was more potent than ED when compared by trypan blue and MTT assays. Our results recapitulate the utility of callus cultures for the production of plant specific bioactive secondary metabolites instead of using wild plants. Together, our in vitro studies provide new insights of A. lineata callus cultures serving as a source for cancer chemotherapeutic agents.

  14. Badanie wzrostu kultur korzeniowych oraz tworzenia i wzrostu in vitro kalusów z różnych organów marchmi odmiany Terfekcja [Growth and callus formation of tissue cultures derived from various carrot organs

    Directory of Open Access Journals (Sweden)

    Anna M. Domańska

    2015-06-01

    Full Text Available The clones of excised roots, leaves, petioles, cotylenods, hypocotyls and root calluses derived from the respective carrot fragments (cv. 'Perfekcja' commonly cultivated in Poland were cultured in vitro. An influence of thiamine concentrations on the growth of root tissue was examined. Several various media were tested for callus cultures. Bee bread extract was also applied. The growth of isolated clones during early and later culture periods was compared.

  15. Effect of polyamine biosynthetic inhibitors on alkaloids and organogenesis in tobacco callus cultures.

    Science.gov (United States)

    Tiburcio, A F; Kaur-Sawhney, R; Galston, A W

    1987-01-01

    We studied the effects of inhibitors of ornithine decarboxylase (ODC), arginine decarboxylase (ADC) and spermidine synthase (Spd synthase) on organogenesis and the titers of polyamines (PA) and alkaloids in tobacco calli. DL-alpha-diffluromethylarginine (DFMA) and D-arginine (D-Arg), both inhibitors of ADC activity, were more effective than DL-alpha-difluromethylorinithine (DFMO), an inhibitor of ODC, in reducing titers of PA and the putrescine (Put)-derived alkaloids (nornicotine and nicotine). Dicyclohexylammonium sulfate (DCHA), an inhibitor of Spd synthase, was also more efficient than DFMO in reducing PA and alkaloid levels. Root organogenesis is inversely related to the titers of Put and alkaloids. Thus, DFMA and D-Arg, which strongly inhibit Put and alkaloid biosynthesis, markedly promote root organogenesis, while control callus with high Put and alkaloid content showed poor root organization. These results suggest that morphological differentiation is not required for activation of secondary metabolic pathways and support the view that ADC has a major role in the generation of Put going to the pyrrolidine ring of tobacco alkaloids.

  16. Corns and calluses

    Science.gov (United States)

    ... rowers get calluses on their hands that prevent blisters from forming. People with bunions often develop a callus over the bunion because it rubs against the shoe. Corns and calluses are not serious problems. Symptoms ...

  17. Distribution of Trans-Anethole and Estragole in Fennel (Foeniculum vulgare Mill of Callus Induced from Different Seedling Parts and Fruits

    Directory of Open Access Journals (Sweden)

    Abd El-Moneim Mohamed Radwan AFIFY

    2011-03-01

    Full Text Available In the present study, seeds from local cultivar of fennel were germinated on Murashige and Skoog medium (MS without plant growth regulators. Different types of explants from the growing seedling such as cotyledonal leaves, hypocotyls, epicotyls and roots were cultured on MS medium, contained different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D either alone or with kinetin. Differential responses in the essential oil constituents were observed in the induction and development of callus. The major components of essential oils includes estragole, trans-anethole, limonene and fenchone were studied under different conditions to find out the best methods which could be used to reduce the amount of estragole (not favorite for human consumption and increase the amount of trans-anethole.

  18. Effect of salts (NaCl and Na2CO3) on callus and suspension culture of Stevia rebaudiana for Steviol glycoside production.

    Science.gov (United States)

    Gupta, Pratibha; Sharma, Satyawati; Saxena, Sanjay

    2014-03-01

    Steviol glycosides are natural non-caloric sweeteners which are extracted from the leaves of Stevia rebaudiana plant. Present study deals the effect of salts (NaCl and Na2CO3) on callus and suspension culture of Stevia plant for steviol glycoside (SGs) production. Yellow-green and compact calli obtained from in vitro raised Stevia leaves sub-cultured on MS medium supplemented with 2.0 mg l(-1) NAA and different concentrations of NaCl (0.05-0.20%) and Na2CO3 (0.0125-0.10%) for 2 weeks, and incubated at 24 ± 1 °C and 22.4 μmol m(-2) s(-1) light intensity provided by white fluorescent tubes for 16 h. Callus and suspension biomass cultured on salts showed less growth as well as browning of medium when compared with control. Quantification of SGs content in callus culture (collected on 15th day) and suspension cultures (collected at 10th and 15th days) treated with and without salts were analyzed by HPLC. It was found that abiotic stress induced by the salts increased the concentration of SGs significantly. In callus, the quantity of SGs got increased from 0.27 (control) to 1.43 and 1.57% with 0.10% NaCl, and 0.025% Na2CO3, respectively. However, in case of suspension culture, the same concentrations of NaCl and Na2CO3 enhanced the SGs content from 1.36 (control) to 2.61 and 5.14%, respectively, on the 10th day.

  19. Callus and azadirachtin related limonoids production through in vitro ...

    African Journals Online (AJOL)

    ONOS

    2010-01-25

    Jan 25, 2010 ... Full Length Research Paper. Callus and ... compounds of medical importance have been reported ... alternative supply of important compounds used in ..... in compact callus clusters cultures of Catharanthus roseus. App.

  20. Effect of low doses γ-irradiation on oxidative stress and secondary metabolites production of rosemary ( Rosmarinus officinalis L.) callus culture

    Science.gov (United States)

    El-Beltagi, Hossam S.; Ahmed, Osama K.; El-Desouky, Wael

    2011-09-01

    Effect of various γ-irradiation doses (0, 5, 10, 15 and 20 G) on the enhancement of secondary metabolites production and antioxidant properties of rosemary callus culture was investigated. The obtained data showed a highly metabolic modification of chemical constituents and various antioxidant defense enzymes (APX, CAT, SOD and GR), which gradually increased in response to radiation doses, while reduced (GSH), ascorbic acid (AsA) contents, total soluble protein, total soluble amino acids, total soluble sugars and PAL activity positively correlated with the increased doses. On the other hands the high irradiation levels significantly increased the accumulation of various oxidative burst (MDA, H 2O 2 and O 2-). Meanwhile, higher doses of gamma irradiation positively enhanced secondary products accumulation of total phenols and total flavonoids in rosemary callus culture.

  1. Antioxidant activity and high-performance liquid chromatographic analysis of phenolic compounds during in vitro callus culture of Plantago ovata Forsk. and effect of exogenous additives on accumulation of phenolic compounds.

    Science.gov (United States)

    Talukder, Pratik; Talapatra, Shonima; Ghoshal, Nirmalya; Sen Raychaudhuri, Sarmistha

    2016-01-15

    Plantago ovata, commonly called psyllium, is known to be a rich source of polyphenolic compounds. The present study was aimed at determining polyphenol content and studying their antioxidant activities in P. ovata during in vitro callus culture. An attempt was also made to enhance polyphenol content using external additives. The role of PAL gene in polyphenol accumulation was also studied. The study indicated the presence of significant amounts of polyphenols, including flavonoids, in P. ovata callus. A gradual increase in polyphenol and flavonoid content was observed up to the third passage (63 days) of callus culture, which declined at the next passage. The third-passage callus showed highest antioxidant activity. High-performance liquid chromatographic results indicated the presence of high amounts of gallic acid and rutin in P. ovata calli; however, other polyphenols were also present but to a lesser extent. Additive supplementation was effective in enhancing polyphenol production and in increasing antioxidant activity in P. ovata callus. The present research reported accumulation of polyphenols in callus culture of P. ovata, which could be applied to isolation of polyphenols for various beneficial purposes. It also indicated enhancement in the production of several polyphenols and also an increase in antioxidant activity in the additive-treated callus. © 2015 Society of Chemical Industry.

  2. 芦笋花药愈伤组织诱导技术研究%Preliminary Study on Anther Culture Callus Inducement of Asparagus

    Institute of Scientific and Technical Information of China (English)

    张元国; 李芳; 包艳存

    2012-01-01

    [目的]对芦笋花药愈伤组织诱导培养技术进行详细研究.[方法]以MS为基本培养基,附加外源激素对二倍体花药愈伤组织诱导技术进行研究.[结果]二倍体芦笋花蕾长度为1.5~2.0 mm时,其诱导效果最好;采用5%高糖浓度可显著抑制花药体细胞分裂和提高花粉愈伤组织诱导率;同一品种不同植株花药愈伤组织诱导率差异显著;适合于二倍体花药愈伤组织诱导的激素配比是1.0 mg/LNAA+ 1.0 mg/L BA.[结论]为芦笋的品种改良与选育奠定了基础.%To study anther culture callus inducement of asparagus in detail. [Method] The diploid anther callus inducement was studied by MS as basic medium with exogenous hormone. [ Result] The length of the diploid bud was 1. 5 - 2. 0 mm which induced result best; The high sugar density of 5% can repress the anther somatic cell division remarkably and improved the inducement rate of pollens callus; The callus inducement rate of the same plant with different species differed greatly; The suitable hormone proportion of diploid anther callus was 1.0 mg/L NAA + 1.0 mg/L BA. [ Conclusion] The study will lay a foundation for improvement and breeding of asparagus.

  3. Callus and cell culture of Tagetes species in relation to production of thiophenes.

    NARCIS (Netherlands)

    Ketel, D.H.

    1987-01-01

    The production of thiophene-biocides by cell cultures invitro was simultaneously investigated with Tageteserecta , T.patula and T.minuta . The calli from which the liquid cultures had to be derived differed between species in the appearance of

  4. Effect of low doses {gamma}-irradiation on oxidative stress and secondary metabolites production of rosemary (Rosmarinus officinalis L.) callus culture

    Energy Technology Data Exchange (ETDEWEB)

    El-Beltagi, Hossam S., E-mail: lbltg@yahoo.com [Biochemistry Department, Faculty of Agriculture, Cairo University, P.O. Box 12613, Gamma st, Giza, Cairo (Egypt); Ahmed, Osama K. [Biochemistry Department, Faculty of Agriculture, Cairo University, P.O. Box 12613, Gamma st, Giza, Cairo (Egypt); El-Desouky, Wael [Biochemistry Hot Laboratories Center, Atomic Energy Authority, Cairo (Egypt)

    2011-09-15

    Effect of various {gamma}-irradiation doses (0, 5, 10, 15 and 20 G) on the enhancement of secondary metabolites production and antioxidant properties of rosemary callus culture was investigated. The obtained data showed a highly metabolic modification of chemical constituents and various antioxidant defense enzymes (APX, CAT, SOD and GR), which gradually increased in response to radiation doses, while reduced (GSH), ascorbic acid (AsA) contents, total soluble protein, total soluble amino acids, total soluble sugars and PAL activity positively correlated with the increased doses. On the other hands the high irradiation levels significantly increased the accumulation of various oxidative burst (MDA, H{sub 2}O{sub 2} and O{sub 2}{sup -}). Meanwhile, higher doses of gamma irradiation positively enhanced secondary products accumulation of total phenols and total flavonoids in rosemary callus culture. - Highlights: > We model effects of {gamma}-irradiation on rosemary callus. > Highly changes of chemical contents, oxidative burst and antioxidant enzymes. > Treatments positively enhanced secondary products.

  5. Formation of Mycorrhizs-like Structures in Cultured Root/Callus of Cathays argyrophylla Chun et Kuang Infected with the Ectomycorrhizal Fungus Cenococcum geophilum Fr.

    Institute of Scientific and Technical Information of China (English)

    Xue Sun; Yu-Hua Li; Lu-Min Vaario

    2006-01-01

    An in vitro system was used for ectomycorrhizal synthesis of Cenococcum geophilum Fr. with Cathaya argyrophylla Chun et Kuang, an endangered species. Calli initiated from stem segments and adventitious roots differentiated from young seedlings were removed and cocultured with Cenococcum geophilum on a modified Murashige-Skoog medium. Fungal hyphae were visible within intercellular spaces of the callus 4 weeks after inoculation, but definite and well-developed Hartig net structures did not form in the calli 8 weeks after inoculation. The typical ectomycorrhizal structures (i.e. hyphal mantle and intracortical Hartig net) were observed in root segments 8 weeks after inoculation. This is the first report of aseptic ectomycorrhizal-like formation/infection between root organ/callus of Cathaya argyrophylla and the ectomycorrhizal fungus Cenococcum geophilum. This culture system is useful for further investigation of mycorrhizal synthesis in Cathaya trees.

  6. [Influence of genotype, explant type and component of culture medium on in vitro callus induction and shoot organogenesis of tomato (Solanum lycopersicum L.)].

    Science.gov (United States)

    Khaliluev, M R; Bogoutdinova, L R; Baranova, G B; Baranova, E N; Kharchenko, P N; Dolgov, S V

    2014-01-01

    The influence of explant type as well as of the type of growth regulators and concentration on callus induction processes and somatic organogenesis of shoots was studied in vitro on four tomato genotypes of Russian breeding. Cytological study of callus tissue was conducted. It was established that tomato varieties possess a substantially greater ability to indirect shoot organogenesis compared with the F1 hybrid. The highest frequency of somatic organogenesis of shoots, as well as their number per explant, was observed for most of the genotypes studied during the cultivation of cotyledons on Murashige-Skoog culture medium containing 2 mg/l of zeatin in combination with 0.1 mg/l of 3-indoleacetic acid. An effective protocol of indirect somatic organogenesis of shoots from different explants of tomato varieties with a frequency of more than 80% was developed.

  7. Isolation of protoplasts and regeneration of callus from suspension cultures of cultivated beets.

    Science.gov (United States)

    Bhat, S R; Ford-Lloyd, B V; Callow, J A

    1985-12-01

    Conditions necessary for the isolation and culture of protoplasts from suspension cultures of sugar, fodder and garden beets were investigated. Good yields of protoplasts were obtained by treating cells with a mixture of cellulase, Macerozyme and Driselase enzymes. Nutritional requirements of beet protoplasts were found to be quite simple: protoplasts could be cultured in MS, B5 or PGo based media with 0.4 M glucose with the optimum result being produced on KM8p medium. Plating efficiency (P.E) was genotype-dependent with the sugar beet giving better P.E. than the fodder or garden beets used, and higher values being achieved with the use of desalted Driselase for isolation followed by culture on KMBp medium.

  8. Palm fruit in traditional African food culture.

    Science.gov (United States)

    Atinmo, Tola; Bakre, Aishat Taiwo

    2003-01-01

    The centre of origin of the oil palm is the tropical rain forest region of West Africa. It is considered to be the 200-300 kilometre wide coastal belt between Liberia and Mayumbe. The oil palm tree has remained the 'tree of life' of Yoruba land as well as of other parts of southern West Africa to which it is indigenous. The Yoruba are adept at spinning philosophical and poetical proverbs around such ordinary things as hills, rivers, birds, animals and domestic tools. Hundreds of the traditional proverbs are still with us, and through them one can see the picture of the environment that contributed to the moulding of the thoughts of the people. Yoruba riddles or puzzles were also couched in terms of the environment and the solutions to them were also environmental items. They have a popular saying: A je eran je eran a kan egungun, a je egungun je egungun a tun kan eran: 'A piece of meat has an outer layer of flesh, an intermediate layer of bone and an inner layer of flesh'. What is it? A palm fruit: it has an outer edible layer, the mesocarp; then a layer of shell, inedible, and the kernel inside, edible. The solution to this puzzle summarises the botanical and cultural characteristics of the palm fruit.

  9. 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.

  10. The Callus Induction of Asparagus“Champion” Anther Culture%芦笋“冠军”花药培养愈伤组织的诱导

    Institute of Scientific and Technical Information of China (English)

    包艳存; 李书华; 李霞; 李芳

    2016-01-01

    Objective] The aim was to study asparagus“champion” anther culture technique. [Method] Using the F1 hybrid asparagus variety“champion” as the test material, callus induction in tissue culture process and effects of growth regulator on callus differentiation were studied. [Result] The results showed that using 0. 1% mercuric chloride sterilization 5 min, inoculated on the MS solid medium by adding NAA0. 8 mg/L + 6-BA 2. 0 mg/L of darkness first 10 days, and then light culture, anther callus induction rate reached about 16. 5%. [Conclusion] The asparagus“champion” anther culture technical system is established, which will lay a foundation for carrying out asparagus all male breeding.%[目的]研究芦笋“冠军”花药培养技术。[方法]以杂交一代品种“冠军”为试验材料,研究花药组培过程中影响愈伤组织诱导的因素及生长调节剂对愈伤分化成苗的影响。[结果]采用0.1%升汞灭菌5 min,再接种在添加NAA 0.8 mg/L+6-BA 2.0 mg/L的 MS固体培养基上先暗培养10 d,再光照培养,花药愈伤组织诱导率较高,达16.5%左右。[结论]建立了芦笋“冠军”花药培养技术体系,为开展芦笋全雄育种奠定基础。

  11. Physiological and biochemical effects of morphactin IT 3233 on callus and tumour tissues of Nicotiana tabacum L. cultured in vitro III. Transamination processes catalysed by aminotransferase L-alanine: 2-oxoglutarate

    Directory of Open Access Journals (Sweden)

    Z. Chirek

    2015-01-01

    Full Text Available An active alanine transaminase was found both in callus and tumour tissues of tobacco. The enzyme is more active in the latter tissue, and the reaction balance is strongly shifted towards alanine production, while in callus tissue towards glutamic acid formation. Morphactin applied to the tissue cultures stimulates markedly the enzyme activity only in callus. A negative correlation was observed between the intensity of transamination processes and enhanced synthesis of proteins in the tissues studied. Morphactin disturbs nitrogen metabolism in the callus tissue. Tumour tissue is more resistant to the action of this substance. The different hormonal activities in these tissues may be the cause of the different effects of morphactin.

  12. Blisters, Calluses, and Corns

    Science.gov (United States)

    ... Too Tall or Too Short All About Puberty Blisters, Calluses, and Corns KidsHealth > For Kids > Blisters, Calluses, ... the surfaces is your tender skin! What's a Blister? A blister is an area of raised skin ...

  13. Characterization of biomass production, cytology and phenotypes of plants regenerated from embryogenic callus cultures of Pennisetum americanum x P. purpureum (hybrid triploid napiergrass).

    Science.gov (United States)

    Rajasekaran, K; Schank, S C; Vasil, I K

    1986-11-01

    Five hundred and twenty-four plants of a triploid, sexually sterile hybrid napiergrass (Pennisetum americanum x P. purpureum; 3x=21) were regenerated from embryogenic callus cultures obtained from segments of young inflorescences. Replicated field trials were conducted for two consecutive years to compare the biomass yield, phenotype and cytology of tissue culture regenerants (TC) and vegetatively propagated (V) plants. In the first year total biomass yield of TC plants was significantly greater than V plants but there was no significant difference in the second year. TC plants had more tillers compared to V plants. V plants did not show any morphological variability. The TC population also exhibited a high degree of phenotypic stability (96%). There were 23 phenotypic variants in the TC population of 524, most of them being more dwarf and late-flowering. Detailed morphological analysis of the TC-variant plants suggests that they very likely arose from only a few variant cell lines. Cytological analysis indicated stability of the triploid status in randomly selected regenerants. Two of the morphological variants were hexaploids (6x=42). It is concluded that embryogenic callus cultures can provide useful alternative for the rapid propagation of hybrid napier-grass which is commonly propagated by cuttings.

  14. The production of class III plant peroxidases in transgenic callus cultures transformed with the rolB gene of Agrobacterium rhizogenes.

    Science.gov (United States)

    Shkryl, Y N; Veremeichik, G N; Bulgakov, V P; Avramenko, T V; Günter, E A; Ovodov, Y S; Muzarok, T I; Zhuravlev, Y N

    2013-10-10

    The production of plant peroxidases by plant cell cultures is of great interest because of the potential for industrial applications. We used plant cell cultures overexpressing the rolB gene to produce increased amounts of plant class III peroxidases. The rolB gene ensured the stable and permanent activation of peroxidase activity in the transformed callus cultures of different plants. In particular, the total peroxidase activity in transformed Rubia cordifolia cells was increased 23-86-fold, and the abundance of the major peroxidase gene transcripts was increased 17-125-fold (depending on the level of rolB expression) compared with non-transformed control calli. The peroxidase-activating effect of rolB was greater than that of other peroxidase inducers, such as external stresses and methyl jasmonate. Copyright © 2013 Elsevier B.V. All rights reserved.

  15. Ultrastructure of Single Cells, Callus-like and Monosore-like Cells in Porphyra yezoensis Ueda on Semi solid Culture Medium

    Institute of Scientific and Technical Information of China (English)

    梅俊学; 沈颂东; 姜明; 费修绠

    2003-01-01

    It had been demonstrated that individual cells or protoplasts isolated from Porphyra thallus by enzyme could develop into normal leafy thalli in the same way as monospores, and that isolated cells develop in different way in liquid and on semi solid media. The authors observed the ultrastructure of isolated vegetative cells cultured on semi solid media and compared them with those of monospores and isolated cells cultured in liquid media. The results showed that subcellular structures were quite different among cells in different conditions. In their development, isolated cells on semi solid media did not show the characteristic subcellular feature of monospore formation, such as production of fibrous vesicles. Callus like cells formed on semi solid media underwent a distinctive modification in cellular organization. They developed characteristic cell inclusions and a special 2 layer cell covering. Golgi bodies, ER, starch grains, mitochondria. Vacuoles were not commonly found in them.

  16. Optimized growth and plant regeneration for callus of Lilium longiflorum cv. Nellie White

    Science.gov (United States)

    The rates of growth and regeneration were compared for compact callus, friable callus, and suspension cells of Lilium longiflorum cv. Nellie White to determine the optimal culture conditions. The fresh weight was higher for compact callus induced from bulb scales cultured on Murashige and Skoog’s m...

  17. Production of anti-cancer triterpene (betulinic acid) from callus cultures of different Ocimum species and its elicitation.

    Science.gov (United States)

    Pandey, Harshita; Pandey, Pallavi; Singh, Sailendra; Gupta, Ruby; Banerjee, Suchitra

    2015-03-01

    Betulinic acid (BA), a pentacyclic triterpenoid, is gaining unmatched attention owing to its unique anti-cancer activity with selective melanoma growth inhibition without damaging normal cells. It is also well-known for its multifaceted pharmacokinetics, entailing antibacterial, antimalarial, anti-HIV and antioxidant merits. Considering the escalating demand with diminishing bioresource of this molecule, the present study was undertaken that revealed the untapped potentials of Ocimum calli, contrasting to that in the in vitro derived leaves, as effective production alternative of BA in three out of four tested species (i.e. Ocimum basilicum, Ocimum kilimandscharicum, Ocimum sanctum excluding Ocimum grattisimum). Callus inductions were obtained in all the four species with different 2,4-dichlorophenoxyacetic acid (2,4-D)/α-naphthaleneacetic acid (NAA) concentrations with kinetin. Notably, 2,4-D favoured maximum callus growth in all whereas NAA proved beneficial for the highest metabolite yield in the calli of each BA-producing species. The O. basilicum calli demonstrated the maximum growth (growth index (GI) 678.7 ± 24.47) and BA yield (2.59 ± 0.55 % dry weight [DW]), whereas those in O. kilimandscharicum (GI 533.33 ± 15.87; BA 1.87 ± 0.6 % DW) and O. sanctum (GI 448 ± 16.07; BA 0.39 ± 0.12 % DW) followed a descending order. The O. gratissimum calli revealed minimum growth (GI 159 ± 13.25) with no BA accumulation. Elicitation with methyl jasmonate at 200-μM concentration after 48-h exposure doubled the BA yield (5.10 ± 0.18 % DW) in NAA-grown O. basilicum calli compared to that in the untreated counterpart (2.61 ± 0.19 % DW), which further enthused its future application.

  18. Early fruit setting from tissue culture-derived mangosteen tree

    Directory of Open Access Journals (Sweden)

    Sompong Te-chato

    2004-05-01

    Full Text Available Vitro-plantlets of mangosteen derived from culturing young leaves were acclimatized in 1993. Small and large polybag seedlings were carefully raised under controlled environmental conditions until 1994 when they were ready to be transferred to the field. During this stage, morphological abnormalities of the seedlings were recorded. After transferring to the field for 5-6 years (1994-1999 at Yi Ngo District, Narathiwat Province and Klong Hoi Khong District, Songkhla Province, morphological characters of the plants were again observed in comparison with seed-derived plants. The results showed that tissue culture-derived plants were more bushy and started blooming 5 years after planting while the seed-derived plants still had tall canopy (not bushy and were not bearing fruit in the same period of time. However, the blooming of cultured plants did not give the fruit setting in the first blooming year. All flowers dropped off completely. Heavy fruit setting was observed in the following year (2000. Tissue culture trees had smaller but healthier leaves whereas seed-derived trees had pale yellowish green leaves. Fruit qualities in terms of total soluble solids (TSS and total acids (TA were not much different between the two types of these mangosteen trees.

  19. Anti-Inflammatory Effects of Concentrated Ethanol Extracts of Edelweiss (Leontopodium alpinum Cass. Callus Cultures towards Human Keratinocytes and Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Lulli Daniela

    2012-01-01

    Full Text Available Edelweiss (Leontopodium alpinum Cass. is traditionally employed in folk medicine as an anti-inflammatory remedy. In nature, the plant is sparsely available and protected; therefore production of callus cultures was established. A concentrated ethanolic extract of culture homogenate, with leontopodic acid representing 55±2% of the total phenolic fraction (ECC55, was characterized for anti-inflammatory properties in primary human keratinocytes (PHKs and endotheliocytes (HUVECs. Inflammatory responses were induced by UVA+UVB, lipopolysaccharide (LPS, oxidized low-density lipoprotein (oxLDL, and a mixture of proinflammatory cytokines. Trichostatin A, a sirtuin inhibitor, was used to induce keratinocyte inflammatory senescence. ECC55 (10–50 μg/mL protected PHK from solar UV-driven damage, by enhancing early intracellular levels of nitric oxide, although not affecting UV-induced expression of inflammatory genes. Comparison of the dose-dependent inhibition of chemokine (IL-8, IP-10, MCP-1 and growth factor (GM-CSF release from PHK activated by TNFα + IFNγ showed that leontopodic acid was mainly responsible for the inhibitory effects of ECC55. Sirtuin-inhibited cell cycle, proliferation, and apoptosis markers were restored by ECC55. The extract inhibited LPS-induced IL-6 and VCAM1 genes in HUVEC, as well as oxLDL-induced selective VCAM1 overexpression. Conclusion. Edelweiss cell cultures could be a valuable source of anti-inflammatory substances potentially applicable for chronic inflammatory skin diseases and bacterial and atherogenic inflammation.

  20. Effect of antibiotic cefotaxime and kanamycin on callus formation and plantlet regeneration from leaves and callus of mangosteen

    Directory of Open Access Journals (Sweden)

    Rugpheug, R.

    2002-10-01

    Full Text Available In order to get rid of contamination from Agrobacterium tumefaciens, the bacterium employed in gene transformation, various kinds and concentrations of antibiotics were added singly or in combinations. In this investigation, concentrations of cefotaxime and kanamycin were examined for callus formation and regenerability from leaves and callus. The results showed that cefotaxime at the concentration of up to 300 mg/l gave a non-significant difference in callus formation. In the case of direct shoot bud formation, concentration over 100 mg/l drastically reduced percentage of leaf-forming shoot buds. The calli which were cultured continuously in 300 mg/l cefotaxime-containing medium for 6 passages gave callus forming shoot buds of 35%. Higher concentration of cefotaxime drastically decreased bud formation. In the case of kanamycin, callus could be induced and maintained in the medium supplemented with a lower concentration than cefotaxime. However, the callus could not be maintained after 3 subculturings.

  1. Optimization of embryogenic-callus induction and embryogenesis of ...

    African Journals Online (AJOL)

    ONOS

    2010-09-06

    Sep 6, 2010 ... With the protocol reported herein, some green embryo-like cultures were ... glabra L., callus induction, embryogenesis, cell culture, histological section. .... All experiments were arranged in a completely randomized design.

  2. Callus induction and plant regeneration from different explant types of Miscanthus x ogiformis Honda 'Giganteus'

    DEFF Research Database (Denmark)

    Holme, Inger Bæksted; Petersen, Karen Koefoed

    1996-01-01

    . The explants were cultured on urashige and Skoog medium supplemented with 4.5, 13.6, 22.6 or 31.7 μM 2,4-dichlorophenoxyacetic acid. Three types of callus were formed but only one was embryogenic and regenerated plants. Callus induction and formation of embryogenic callus depended on the type and developmental...

  3. [Principles of callus distraction].

    Science.gov (United States)

    Hankemeier, S; Bastian, L; Gosling, T; Krettek, C

    2004-10-01

    Callus distraction is based on the principle of regenerating bone by continuous distraction of proliferating callus tissue. It has become the standard treatment of significant leg shortening and large bone defects. Due to many problems and complications, exact preoperative planning, operative technique and careful postoperative follow-up are essential. External fixators can be used for all indications of callus distraction. However, due to pin tract infections, pain and loss of mobility caused by soft tissue transfixation, fixators are applied in patients with open growth plates, simultaneous lengthening with continuous deformity corrections, and increased risk of infection. Distraction over an intramedullary nail allows removal of the external fixator at the end of distraction before callus consolidation (monorail method). The intramedullary nail protects newly formed callus tissue and reduces the risk of axial deviation and refractures. Recently developed, fully intramedullary lengthening devices eliminate fixator-associated complications and accelerate return to normal daily activities. This review describes principles of callus distraction, potential complications and their management.

  4. Establishing Embryogenic Callus Suspension Culture System in Black Locust%刺槐胚性细胞悬浮体系的建立1)

    Institute of Scientific and Technical Information of China (English)

    冯玥; 习洋; 陈串; 喻娃亚雄; 王少明; 徐惠敏; 孙宇涵; 李云

    2014-01-01

    以刺槐( Robinia pseudoacacia L.)未成熟合子胚为外植体材料,通过对外植体取材时期、基本培养基营养物质质量浓度、细胞起始密度、2,4-二氯苯氧乙酸(2,4-D)质量浓度等影响悬浮培养的因素的研究,建立了良好的刺槐胚性细胞悬浮系。研究结果表明:授粉后55 d的未成熟合子胚是为刺槐细胞悬浮培养提供胚性愈伤组织的最佳起始材料;添加有2.5 mg· L-1 NAA和0.5 mg· L-16-BA的1/2MS培养基为刺槐细胞悬浮培养的最适培养基;建立悬浮系的最适pH值范围在5.0~5.7;起始质量为2.0 g的悬浮系统其生长量增速较快,且细胞保持持续增长;当起始质量为3.0 g时,培养16 d后细胞进入缓慢增长期,且生长量有回落的趋势,起始质量为1.0 g时,细胞增殖速度非常缓慢,且容易出现褐化死亡的现象。%We employed embryogenic cell suspension ( ECS) induced from immature seeds of black locust ( Robinia pseudoaca-cia L.) as materials to study the effecting factors as explants period , nutrient concentration of basic medium , initial cell density and 2,4-D concentration of suspension culture medium to establish a successful embryogenic callus suspension cul -ture system for black locust.The seeds collected at eight weeks (about 55 days) post-anthesis obtained the highest ability to initiate embryogenic culture for embryogenic callus cell suspension establishment .The best callus initiation medium was 1/2MS+2.5 mg· L-1 NAA+0.5 mg· L-1 6-BA and the suitable pH ranged in 5.0-5.7.The cells maintained a sustained growth as initial density of suspension system was 2.0 g, the cells proliferation was slowed down after 16 days when the ini-tial density was 3.0 g, and the growth rate was with down trend .The cells were brown and died in slow proliferation rate with the initial density of 1.0 g.

  5. Analysis of the Components in Callus and Cell Suspension Cultures of Ligusticum chuanxiong Hort. By Gas Chromatography-Mass Spectrometry%川芎愈伤组织及悬浮培养物化学成分气相色谱-质谱联用分析

    Institute of Scientific and Technical Information of China (English)

    詹玉莲; 马小军; 戴均贵; 刘德华

    2006-01-01

    Objective To analyze the components in callus and cell suspension cultures of Ligusticum chuanxiong Hort. by gas chromatography-mass spectrometry (GC-MS). Methods Initiate the callus and cell suspension cultures of Ligusticum chuanxiong Hort.. Collect the callus and three sequent generations cell suspension cultures, and the components in callus and cell suspension cultures were analyzed by gas chromatography-mass spectrometry. Results The callus and cell suspension cultures of Ligusticum chuanxiong Hort. contained similar constituenents, but the components in callus and cell suspension cultures were quite different from those of the plant, and the main bioactive constituentes of the plant were not found in callus and cell suspension cultures. Conclusion The characteristics of callus and cell suspension cultures were different from those of the plant.%目的用气相色谱-质谱联用法分析川芎愈伤组织及悬浮培养物中的化学成分.方法建立川芎愈伤组织及悬浮培养体系,收集愈伤组织及连续三代的悬浮培养物,采用气相色谱-质谱联用法进行川芎愈伤组织及悬浮培养物化学成分的分析.结果川芎愈伤组织及悬浮培养物含有相似的成分,但与原植物中的成分区别很大,原植物中的主要活性成分在愈伤组织及悬浮培养物中也未发现.结论愈伤组织及悬浮培养物的特性与原植物有很大的区别.

  6. A whole-genome microarray study of Arabidopsis thaliana semisolid callus cultures exposed to microgravity and nonmicrogravity related spaceflight conditions for 5 days on board of Shenzhou 8.

    Science.gov (United States)

    Fengler, Svenja; Spirer, Ina; Neef, Maren; Ecke, Margret; Nieselt, Kay; Hampp, Rüdiger

    2015-01-01

    The Simbox mission was the first joint space project between Germany and China in November 2011. Eleven-day-old Arabidopsis thaliana wild type semisolid callus cultures were integrated into fully automated plant cultivation containers and exposed to spaceflight conditions within the Simbox hardware on board of the spacecraft Shenzhou 8. The related ground experiment was conducted under similar conditions. The use of an in-flight centrifuge provided a 1 g gravitational field in space. The cells were metabolically quenched after 5 days via RNAlater injection. The impact on the Arabidopsis transcriptome was investigated by means of whole-genome gene expression analysis. The results show a major impact of nonmicrogravity related spaceflight conditions. Genes that were significantly altered in transcript abundance are mainly involved in protein phosphorylation and MAPK cascade-related signaling processes, as well as in the cellular defense and stress responses. In contrast to short-term effects of microgravity (seconds, minutes), this mission identified only minor changes after 5 days of microgravity. These concerned genes coding for proteins involved in the plastid-associated translation machinery, mitochondrial electron transport, and energy production.

  7. A Whole-Genome Microarray Study of Arabidopsis thaliana Semisolid Callus Cultures Exposed to Microgravity and Nonmicrogravity Related Spaceflight Conditions for 5 Days on Board of Shenzhou 8

    Directory of Open Access Journals (Sweden)

    Svenja Fengler

    2015-01-01

    Full Text Available The Simbox mission was the first joint space project between Germany and China in November 2011. Eleven-day-old Arabidopsis thaliana wild type semisolid callus cultures were integrated into fully automated plant cultivation containers and exposed to spaceflight conditions within the Simbox hardware on board of the spacecraft Shenzhou 8. The related ground experiment was conducted under similar conditions. The use of an in-flight centrifuge provided a 1 g gravitational field in space. The cells were metabolically quenched after 5 days via RNAlater injection. The impact on the Arabidopsis transcriptome was investigated by means of whole-genome gene expression analysis. The results show a major impact of nonmicrogravity related spaceflight conditions. Genes that were significantly altered in transcript abundance are mainly involved in protein phosphorylation and MAPK cascade-related signaling processes, as well as in the cellular defense and stress responses. In contrast to short-term effects of microgravity (seconds, minutes, this mission identified only minor changes after 5 days of microgravity. These concerned genes coding for proteins involved in the plastid-associated translation machinery, mitochondrial electron transport, and energy production.

  8. Cytochemical characterization and ultrastructural organization in calluses of the agarophyte Gracilariopsis tenuifrons (Gracilariales, Rhodophyta).

    Science.gov (United States)

    Bouzon, Zenilda Laurita; Schmidt, Eder Carlos; Almeida, Ana Carolina de; Yokoya, Nair S; Oliveira, Mariana Cabral de; Chow, Fungyi

    2011-01-01

    The culture and physiology of red macroalgae calluses are well documented. To date, however, no report has either performed a cytochemical analysis or characterized the ultrastructural organization of calluses at different stages of development and under the effect of plant growth regulators. Therefore, to undertake such analyses, this work studied the red seaweed Gracilariopsis tenuifrons (Bird et Oliveira) Fredericq et Hommersand. Morphology studies suggested three types of calluses: a) terminal callus having an irregular amorphous shape and filamentous projections originating from the cortical region of the thallus; b) apical callus growing on apical branches and having an elongated semispherical shape; and c) intercalary callus developing along the intermediary region of the thallus and having the appearance of small declivities with irregular edges. The abundance of intercalary calluses over terminal and apical calluses is most likely a result of a major cortical surface that would support the cellular growth required to generate calluses. Callus development was initially observed as a matrix of cellular disorganization with filamentous projections; then, the cellular mass seemed to become more compact with spherical uncolored aspect. The presence of starch grains in the inner part of the explant could be explained by absorption from the culture medium and by proper biosynthesis during callus development. Cell wall reaction to staining suggested cellulose and agar composition with acidic polysaccharides. Results suggest that none of the three morphological types of calluses showed any significant differences on the basis of either cytochemistry or ultrastructural organization.

  9. Effects of rare earth elements on callus growth, soluble protein ...

    African Journals Online (AJOL)

    USER

    2010-04-19

    Apr 19, 2010 ... The effects of La3+ on soluble protein content, peroxidase activity and shoot differentiation of callus were ... enzymes activity, nitrogen metabolism of in vitro culture .... ion might produce different effects on cell growth through.

  10. Callus production and regeneration of the medicinal plant Papaver ...

    African Journals Online (AJOL)

    Administrator

    2011-09-19

    Sep 19, 2011 ... Papaver species produce a wide range of iso- quinolines ... thebaine in a green callus with relatively high levels of kinetin. .... software. RESULTS AND DISCUSSION. In this study, two different media for plant tissue culture.

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

    Science.gov (United States)

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

    2003-11-01

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

  12. Determination of abscisic acid and its glucosyl ester in embryogenic callus cultures of Vitis vinifera in relation to the maturation of somatic embryos using a new liquid chromatography-ELISA analysis method.

    Science.gov (United States)

    Prado, María Jesús; Largo, Asier; Domínguez, Cristina; González, María Victoria; Rey, Manuel; Centeno, María Luz

    2014-06-15

    The levels of abscisic acid (ABA), its conjugate ABA-GE, and IAA were determined in embryogenic calli of Vitis vinifera L. (cv. Mencía) cultured in DM1 differentiation medium, to relate them to the maturation process of somatic embryos. To achieve this goal, we developed an analytical method that included two steps of solid-phase extraction, chromatographic separation by HPLC, ABA-GE hydrolysis, and sensitive ELISA quantification. Because the ABA immunoassay was based on new polyclonal antibodies raised against a C4'-ABA conjugate, the assay was characterized (detection limit, midrange, measure range, and cross-reaction) and validated by a comparison of the ABA data obtained with this ELISA procedure and with a physicochemical method (LC-ESI-MS/MS). Radioactive-labeled internal standards were initially added to callus extracts to correct the losses of plant hormones, and thus assure the accuracy of the measurements. The endogenous concentration of ABA in the embryogenic callus cultured in DM1 medium was doubled at the fifth week of culture, concurring with the maturation process of somatic embryos, as indicated by the accumulation of carbohydrates observed through histological analysis. The ABA-GE content was higher than ABA, decreasing at 21 days of culture in DM1 medium but increasing thereafter. The data suggest the involvement of the synthesis and conjugation of ABA in the final stages of development in grapevine somatic embryos from embryogenic callus. IAA levels were low, suggesting that auxin plays no significant role during the maturation of somatic embryos. In addition, the lower ABA levels in calli cultured in DM differentiation medium with PGRs, a medium presenting high precocious germination and deficiencies in somatic embryo development indicate that an increase in ABA content during the development of somatic embryos in grapevine is necessary for their correct maturation. Copyright © 2014 Elsevier GmbH. All rights reserved.

  13. Callus induction and plant regeneration of Oncidium‘Gower Ramsey’ by means of protocorm-like body culture%文心兰类原球茎的愈伤组织诱导及其植株再生

    Institute of Scientific and Technical Information of China (English)

    黄霞; 卢禹

    2016-01-01

    the first time protocorm-like bodies of Oncidium‘Gower Ramsey’ were used as explants for callus induction and plant regeneration. The effects of different concentrations and combinations of TDZ and 2,4-D on callus proliferation were investigated. The results showed that 1/2MS medium supplemented with 1 mg.L-1 TDZ and 3 mg.L-1 2,4-D could promote the induction of whitish and friable callus, the induction rate was 100%. The proliferation rate of callus was mainly affected by the concentration of TDZ, while the callus subculture medium consisted of TDZ in combination with 2,4-D at the concentrations of 0.5 to 2.0 mg .L-1 . When the concentration of TDZ decreased from 1.0 to 0.5 mg.L-1 , pro-liferation rate of the callus fresh weigh increased significantly. And the optimal medium for callus proliferation contained 1/2MS basal medium, 0.5 mg.L-1 TDZ and 1.0 mg.L-1 2,4-D. After four weeks of subculture in the dark, the highest prolif-eration rate of the callus fresh weigh, which was 6.04 folds, appeared on the optimal medium, while the lowest proliferation rate of the callus fresh weigh, which was 4.50 folds, appeared on the medium containing 1/2MS basal medium, 1.0 mg.L-1 TDZ and 2.0 mg.L-1 2,4-D. The callus subcultured on the optimal proliferation medium for about one month were trans-ferred to T2 medium (3.5 g.L-1 Hyponex 1+20 g.L-1 sweet potato+25 g.L-1 banana+1 g.L-1 tryptone+20 g.L-1 sucrose + 3.5 g.L-1 phytagel) and cultured in the dark for PLBs induction. Approximately, 1 328.67 protocorm-like bodies could be generated in one month from an initial culture of 1 g callus fresh weight. Subsequently, the cultures transferred on-to the fresh T2 medium. The germination rate of protocorm-like bodies was as high as 90.12%after one month of culture un-der the light. When transplanted on 1/2MS medium supplemented with 1 g.L-1 active carbon, all shoots became healthy plants after two months. In this study, the callus induction and plant regeneration system of Oncidium

  14. Production of lignans in calluses of Schisandra chinensis.

    Science.gov (United States)

    Kohda, Hiroshi; Ozaki, Makoto; Namera, Akira

    2012-04-01

    Calluses were induced from leaves of Schisandra chinensis Baillon (Schisandraceae). Murashige-Skoog (MS) and Woody Plant (WP) media were used for the induction, in full and half strength (1/2 MS or 1/2 WP) salt formulations. Test media were solidified with 0.25% gelrite and supplemented with 2% sucrose and various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (Kin), 3-indolebutyric acid (IBA), and 6-benzylaminopurine (BAP). Optimal conditions for callus induction and growth were found to be 1/2 MS medium containing 0.02 mg/l Kin and 0.2 mg/l 2,4-D. Chloroform extracts of all induced calluses contained gomisin A and F as major components. Gomisin A and F contents of calluses that were cultured under the optimal conditions mentioned above were highest compared to the calluses incubated with other combinations of plant hormones and media. Subculture, by repeated transfer of cultured calluses to fresh medium, caused no decrease in the production of gomisin A and F. Optimal conditions for lignan production were found to be 1/2 MS medium supplemented with 0.05 mg/1 Kin and 0.2 mg/l 2,4-D. Under these conditions, gomisin A and gomisin F contents were 0.05 and 0.04% of callus dry weight, respectively.

  15. C/sub 4/ photosynthesis in Euphorbia degeneri and E. remyi: a comparison of photosynthetic carbon metabolism in leaves, callus cultures and regenerated plants

    Energy Technology Data Exchange (ETDEWEB)

    Ruzin, S.E.

    1984-04-01

    Based on analysis of /sup 14/CO/sub 2/ fixation kinetics and assays of enzymes related to C/sub 4/ metabolism (NAD-ME, NADP-ME, NAD-MDH, NADP-MDH, AST, ALT), leaves and regenerated plants of Euphorbia degeneri exhibit a modified NADP-ME-type photosynthesis. Apparently, both aspartate and malate are used for transport of CO/sub 2/ to bundle sheath cells. Callus grown on either non-shoot-forming or shoot-forming media fixes CO/sub 2/ into RPP-cycle intermediates and sucrose, as well as malate and aspartate. /sup 14/CO/sub 2/ pulse/chase kinetics show no significant loss of label from C/sub 4/ acids throughout a one minute chase. Analysis of PEPCase revealed the presence of 2 isoenzymes in both leaf and regenerated plant tissues (K/sub m/ (PEP) = 0.080 and 0.550) but only one isoenzyme in callus (K/sub m/ = 0.100). It appears that C/sub 4/ photosynthesis does not occur in callus derived from this C/sub 4/ dicot but is regenerated concomitant with shoot regeneration, and ..beta..-carboxylation of PEP in callus, mediated by the low K/sub m/ isoenzyme of PEPCase, produces C/sub 4/ acids that are not involved in the CO/sub 2/ shuttle mechanism characteristic of C/sub 4/ photosynthesis. 161 references, 19 figures, 12 tables.

  16. Thidiazuron-enhanced biosynthesis and antimicrobial efficacy of silver nanoparticles via improving phytochemical reducing potential in callus culture of Linum usitatissimum L.

    Directory of Open Access Journals (Sweden)

    Anjum S

    2016-02-01

    Full Text Available Sumaira Anjum, Bilal Haider AbbasiDepartment of Biotechnology, Quaid-i-Azam University, Islamabad, PakistanAbstract: Green synthesis of silver nanoparticles (AgNPs by using plants is an emerging class of nanobiotechnology. It revolutionizes all domains of medical sciences by synthesizing chemical-free AgNPs for various biomedical applications. In this report, AgNPs were successfully synthesized by using whole plant extract (WPE and thidiazuron-induced callus extract (CE of Linum usitatissimum. The phytochemical analysis revealed that the total phenolic and flavonoid contents were higher in CE than that in WPE. Ultraviolet-visible spectroscopy of synthesized AgNPs showed a characteristic surface plasmon band in the range of 410–426 nm. Bioreduction of CE-mediated AgNPs was completed in a shorter time than that of WPE-mediated AgNPs. Scanning electron microscopy showed that both types of synthesized AgNPs were spherical in shape, but CE-mediated AgNPs were smaller in size (19–24 nm and more scattered in distribution than that of WPE-mediated AgNPs (49–54 nm. X-ray diffraction analysis confirmed crystalline nature (face-centered cubic of both types of AgNPs. Fourier-transform infrared spectroscopy revealed that the polyphenols and flavonoids were mainly responsible for reduction and capping of synthesized AgNPs. Energy dispersive X-ray analysis further confirmed the successful synthesis of AgNPs. Moreover, the synthesized AgNPs were found to be stable over months with no change in the surface plasmon bands. More importantly, CE-mediated AgNPs displayed significantly higher bactericidal activity against multiple drug-resistant human pathogens than WPE-mediated AgNPs. The present work highlighted the potent role of thidiazuron in in vitro-derived cultures for enhanced biosynthesis of chemical-free AgNPs, which can be used as nanomedicines in many biomedical applications.Keywords: silver nanoparticles, Linum usitatissimum L., in vitro

  17. Gravitational and magnetic field variations synergize to cause subtle variations in the global transcriptional state of Arabidopsis in vitro callus cultures

    Directory of Open Access Journals (Sweden)

    Manzano Ana I

    2012-03-01

    Full Text Available Abstract Background Biological systems respond to changes in both the Earth's magnetic and gravitational fields, but as experiments in space are expensive and infrequent, Earth-based simulation techniques are required. A high gradient magnetic field can be used to levitate biological material, thereby simulating microgravity and can also create environments with a reduced or an enhanced level of gravity (g, although special attention should be paid to the possible effects of the magnetic field (B itself. Results Using diamagnetic levitation, we exposed Arabidopsis thaliana in vitro callus cultures to five environments with different levels of effective gravity and magnetic field strengths. The environments included levitation, i.e. simulated μg* (close to 0 g* at B = 10.1 T, intermediate g* (0.1 g* at B = 14.7 T and enhanced gravity levels (1.9 g* at B = 14.7 T and 2 g* at B = 10.1 T plus an internal 1 g* control (B = 16.5 T. The asterisk denotes the presence of the background magnetic field, as opposed to the effective gravity environments in the absence of an applied magnetic field, created using a Random Position Machine (simulated μg and a Large Diameter Centrifuge (2 g. Microarray analysis indicates that changes in the overall gene expression of cultured cells exposed to these unusual environments barely reach significance using an FDR algorithm. However, it was found that gravitational and magnetic fields produce synergistic variations in the steady state of the transcriptional profile of plants. Transcriptomic results confirm that high gradient magnetic fields (i.e. to create μg* and 2 g* conditions have a significant effect, mainly on structural, abiotic stress genes and secondary metabolism genes, but these subtle gravitational effects are only observable using clustering methodologies. Conclusions A detailed microarray dataset analysis, based on clustering of similarly expressed genes (GEDI software, can detect underlying global

  18. Systematic secretome analyses of rice leaf and seed callus suspension-cultured cells: workflow development and establishment of high-density two-dimensional gel reference maps.

    Science.gov (United States)

    Jung, Young-Ho; Jeong, Seung-Hee; Kim, So Hee; Singh, Raksha; Lee, Jae-Eun; Cho, Yoon-Seong; Agrawal, Ganesh Kumar; Rakwal, Randeep; Jwa, Nam-Soo

    2008-12-01

    Secreted proteins control a multitude of biological and physiological processes in multicellular organisms such as plants. Identification of secreted proteins in reference plants like Arabidopsis and rice under normal growth conditions and adverse environmental conditions will help better understand the secretory pathways. Here, we have performed a systematic in planta and in vitro analyses of proteins secreted by rice leaves (in planta) and seed callus suspension-cultured cells (SCCs; in vitro), respectively, using a combination of biochemical and two-dimensional gel electrophoresis (2-DGE) coupled with liquid chromatography mass spectrometry analyses. Secreted proteins prepared from either leaves or SCCs medium were essentially free from contamination of intracellular proteins as judged by biochemical and Western blot analyses. 2-DGE analyses of secreted proteins collectively identified 222 protein spots with only 6 protein spots common to both in planta and in vitro derived data sets. Data were used to establish high-resolution and high-density 2-D gel reference maps for both in planta and in vitro secreted proteins. Identified proteins belonged to 11 (in planta) and 6 (in vitro) functional classes. Proteins involved in carbon metabolism (33%) and cell wall metabolism having plant defense mechanism (18%) were highly represented in the in planta secreted proteins accounting for 51% of total identified proteins, whereas proteins of cell wall metabolism having plant defense mechanism (64%) were predominant in the in vitro secreted proteins. Interestingly, secreted proteins possessing signal peptides were significantly lower in an in planta (27%) prepared secreted protein population than in vitro (76%) as predicted by SignalP prediction tool, implying the notion that plant might possess yet unidentified secretory pathway(s) in addition to the classical endoplasmic reticulum/Golgi pathway. Taken together, this systematic study provides evidence for (i) significant

  19. Osmotic induced stimulation of the reduction of the viability dye 2,3,5-triphenyltetrazolium chloride by maize roots and callus cultures.

    Science.gov (United States)

    Duncan, David R; Widholm, Jack M

    2004-04-01

    Live cells can reduce colorless 2,3,5-triphenyltetrazolium chloride (TTC) to a red insoluble compound, formazan. Maize (Zea mays) callus, when osmotically stressed by 0.53 mol/L mannitol, produced 7-times or more formazan than untreated control callus. This result was seen with all osmotica tested and could not be attributed to differences in TTC uptake rate or accumulation, increased respiration rate as measured by O2 uptake, or to de novo protein synthesis. Increased formazan production could be detected after 2.5 h of exposure to osmotic stress and leveled off after 48 h of exposure. The increased formazan production was only detected when callus was moved from high osmotic medium to low osmotic, TTC-containing medium. Abscisic acid increased TTC reduction only when added in combination with 0.53 mol/L mannitol. Incubation of maize seedling roots with 0.53 mol/L mannitol also increased formazan production as seen visually. Further studies are needed to determine the cause of the increased formazan production. These results show that TTC viability measurements must be carefully evaluated with appropriate controls to confirm their validity.

  20. Somaclonal variation on in vitro callus culture potato cultivars Variação somaclonal in vitro em cultura de calos de cultivares de batata

    Directory of Open Access Journals (Sweden)

    Patricia N. Bordallo

    2004-06-01

    Full Text Available Synthetic seeds can be an alternative for those species in which botanical seeds are not viable. One of the major problems of in vitro plant cultivation is the high level of somaclonal variation. The most common factors affecting somaclonal variation are genotype, explant source, in vitro period and cultivation conditions in which the culture is established. In this work, calli were induced using leaf and stem explants of the commercial potato cultivars Achat, Baraka, Baronesa, Bintje, and Contenda in MS culture media supplemented with 1.65 mM of picloram and 11.5 mM of 2,4-D. Seventy and 90 days after induction, DNA samples of 40 calli were compared concerning the effects of the two explant (leaf and stem and two growth regulator sources on five potatoes cultivars. A total of 20 arbitrary sequence primers were evaluated. The RAPD pattern generated by these primers suggested a high percentage of polymorphic fragments among the five genotypes, indicating a high level of genetic variation among cultivars. Cultivar Baronesa showed the highest number of polymorphic fragments for all treatments. The cultivar Contenda showed the smallest somaclonal variation, for most of the treatments, except for the treatment which consisted of stem explants, picloram (1.65 mM application, and a 70-day period of callus formation. 'Contenda' is, therefore, the most suitable cultivar for synthetic seed production.A produção de sementes sintéticas pode ser uma altenativa para culturas, cuja produção de sementes botânicas não é viável. Um dos principais problemas do cultivo de planta in vitro é o alto nível de variação somaclonal resultante. Os fatores mais comuns que afetam a variação somaclonal são genótipo, fonte de explante e duração e condições de cultivo. Neste trabalho, calos foram induzidos utilizando explantes de folha e caule das cultivares comerciais de batata Achat, Baraka, Baronesa, Bintje e Contenda em meio de cultura MS, suplementado

  1. In Vitro Culture and Phytochemical Analysis of Passiflora tenuifila Killip and Passiflora setacea DC (Passifloraceae).

    Science.gov (United States)

    Sozo, Jenny Sumara; Cruz, Daniel Cuzziol; Pavei, Ana Flavia; Pereira, Isadora Medeiros da Costa; Wolfart, Marcia; Ramlov, Fernanda; Fiuza Montagner, Daiane; Maraschin, Marcelo; Viana, Ana Maria

    2016-01-01

    We have developed reproducible micropropagation, callus culture, phytochemical, and antioxidant analysis protocols for the wild passion fruit species P. tenuifila, and P. setacea, native to the Brazilian endangered biomes Atlantic Forest, Cerrado, and Caatinga, by using seeds and explants from seedlings and adult plants. Genotype and explant origin-linked differences are visible amongst the Passiflora species concerning callus production, total phenolics, and antioxidant activity. The protocols developed for screening phytochemicals and antioxidants in P. tenuifila and P. setacea callus extracts have shown their potential for phenolic production and antioxidant activity. The high level of phenolic compounds seems to account for the antioxidant activity of methanolic extracts of P. tenuifila derived from 45-day-old immature seed callus. The methanolic extracts of callus derived from P. setacea seedling leaf node and cotyledonary node explants have shown the highest antioxidant activity despite their lower content of phenolics, as compared to cotyledon callus extracts. The optimized micropropagation and callus culture protocols have great potential to use cell culture techniques for further vegetative propagation, in vitro germplasm conservation, and secondary metabolite production using biotic and abiotic elicitors.

  2. 不同磁场环境和培养基对拟南芥种子诱导愈伤组织的影响%EFFECTS OF VARIOUS MAGNETIC FIELDS AND CULTURE MEDIUMS ON CALLUS INDUCTION FROM Arabidopsis SEEDS

    Institute of Scientific and Technical Information of China (English)

    吕毅; 商澎

    2012-01-01

    为探明磁场强度和培养基成份在拟南芥愈伤组织诱导中的作用,以野生型拟南芥(Columbia型)种子为材料,研究3种磁场环境(亚磁场、地磁场和中强磁场)、4种诱导培养基(NB1、NB2、MS1、MS2)条件下愈伤组织的诱导情况。结果显示,与地磁场相比,亚磁场对接种后第3~6天愈伤组织的发生有明显的抑制作用,中强磁场有明显的促进作用;在接种后第3~7天,NB2、MS2比NB1、MS1培养基促进了愈伤组织形成,且差异显著。愈伤组织净重反映了细胞增殖的状态,统计结果显示,与地磁场相比,亚磁场对细胞增殖没有明显影响,而中强磁场有显著的抑制作用;NB2、MS2比NB1、MS1培养基促进了细胞增殖,且差异显著。以上试验结果说明,磁场强度和培养基成份的变化对拟南芥种子诱导愈伤组织过程中第3~7天的愈伤组织形成和细胞增殖有不同程度影响,但2因素不存在交互作用,且2种因素对15d后的愈伤组织总出愈率无显著影响。6-BA对早期的愈伤组织诱导和细胞增殖均有促进作用,而且在地磁和亚磁场中,添加有6-BA的NB2和MS2培养基所诱导的愈伤组织色泽鲜亮,致密度适中,利于后续继代培养。%Three magnetic fields(hypomagnetic fields,HMF;geomagnetic fields,GMF;moderate static magnetic fields,MSMF) and four culture mediums(NB1,NB2,MS1 and MS2) were applied to investigate the effects of magnetic field intensity and culture medium ingredient on callus induction in Arabidopsis(Columbia type).The results indicated that magnetic intensity and culture medium ingredient had different effects on callus induction in whole process.In the first 3~6 days after inoculation,the callus induction rate under HMF was inhibited,but the rate under MSMF was promoted significantly contrasting with the GMF condition.As far as the culture medium was concerned,in the first 3~7 days after inoculation,the callus induction rate on NB2 and

  3. Tea Callus Inducing Culture and Catechins Synthesis%茶树愈伤组织诱导培养及儿茶素合成的研究

    Institute of Scientific and Technical Information of China (English)

    杜鸿标; 王坤波; 陈勤操; 李银花; 刘仲华

    2014-01-01

    以茶鲜叶不同部位为外植体,在不同消毒方法下诱导产生愈伤组织。同时研究了不同茶树品种、不同培养基和不同激素组合对儿茶素合成的影响。通过试验优化,以茶树第二叶作为外植体,35%次氯酸钠,浸泡18min,可获得较好的诱导效率。在继代培养中,金牡丹愈伤组织中儿茶素含量最高。除黄玫瑰外,其它五种茶树品种使用B5培养基儿茶素含量均高于MS培养基。以金牡丹茶鲜叶为材料,使用B5培养基,2,4-D浓度为0.5mg/L,KT浓度为1.0mg/L时,茶树愈伤组织中儿茶素含量最高。%Different parts of fresh tea leaves were induced in the different disinfection methods to produce callus. Meanwhile, The effects of different tea varieties, different medium and different hormone combinations on catechins synthesis were investigated. Results showed that the second tea leaves as explants, 35%sodium hypochlorite, soak for 18 minutes, could be obtained better efficiency of the induction. In the subculture, the catechin content of Camellia sinensis var. Jinmudan callus was the highest among tea callus. Except Camellia sinensis var. Huangmeigui, the catechin content in the other five tea varieties using B5 medium was higher than that using the MS medium. We obtained the maximum catechins production,when the B5 medium used for Camellia sinensis var. Jinmudan callus cultivation was supplemented with 2,4-D 0.5 mg/L and KT 1.0mg/L.

  4. In vitro production of gymnemic acid from Gymnema sylvestre (Retz) R. Br. ex roemer and schultes through callus culture under abiotic stress conditions.

    Science.gov (United States)

    Ali Ahmed, Abdul Bakrudeen; Rao, Adhikarla Suryanarayana; Rao, Mandali Venkateswara

    2009-01-01

    Plant secondary metabolites have enormous potential for research and new drug development. Many secondary metabolites have a complex and unique structure and their production is often enhanced by biotic and abiotic stress conditions. Gymnemic acid (C(43)H(68)O(14)), a pentacyclic triterpenoid isolated from the leaves of Gymnema sylvestre, exhibits potent inhibitory effect on diabetes. The gymnemic acid content is determined by chromatographic methods: Camag HPTLC system equipped with a sample applicator Linomat IV and TLC scanner and integration software CAT 4.0. In HPLC C(18) (ODS) reverse phase column; water 486 UV detector; mobile phase, water/methanol (35:65, HPLC grade) + 0.1% acetic acid are used. Sample (20 microL) is applied with a flow rate of 1 mL/min and read at 230 nm with UV detector. The production of gymnemic acid is significantly higher in callus treated with 2,4-dichloro phenoxy acetic acid (2,4-D) and kinetin (KN). The blue light increases gymnemic acid accumulation upto 4.4-fold as compared with fluorescent light treatment and out of which 2.8 is found in leaves. Gymnemic acid is isolated from callus, grown under stress conditions followed by preparative TLC, simple and reproducible character based on HPTLC and high performance liquid chromatography.

  5. Callus Induction and Establishment of Cell Suspension Culture System for Thymus vulgaris L.%百里香愈伤组织的诱导及细胞悬浮培养体系的建立

    Institute of Scientific and Technical Information of China (English)

    徐世千; 李晓东; 张建国

    2011-01-01

    The stems and leaves of aseptic seedling of Thymus vulgaris L. Were taken as explants to study the optimum culture conditions of callus induction and proliferation. In addition, the growth characteristics of suspension cell were investigated and the system of thymus cell suspension culture was established preliminarily. The results showed that stem explants were the most suitable for callus induction, the optimal medium for callus induction was MS + NAA 1. 0 mg/L + 6-BA 0. 5 mg/L+ 2,4-D 0. 5 mg/L, the optimal formula for callus proliferation and suspension culture was MS + 6-BA 1. 0 mg/L+ 2,4-D 0. 1 mg/L and MS + 6-BA 0. 5 mg/L+ 2,4-D 0. 1 mg/L respectively. The growth of suspension cells exhibited an S-shaped curve and growth mass reached the maximum after 20 days' culture, the optimal subculture cycle was 18 - 20 d. In the process of suspension culture, the same change trends of pH value and conductivity were showed; firstly decreased then increased and then became stable, and the suspension cell number exhibited the change trend: firstly increased then decreased.%以普通百里香无菌苗的茎段和叶片为材料,对百里香愈伤组织诱导和继代增殖的最佳培养条件进行研究,并对悬浮细胞的生长特性进行监测,初步建立细胞悬浮培养体系.结果表明,茎段是诱导愈伤组织和进行悬浮培养的理想外植体,愈伤组织最佳诱导培养基为MS+NAA 1.0 mg/L+6-BA 0.5 mg/L+2,4-D0.5 mg/L,最佳增殖培养基为MS+6-BA 1.0 mg/L+2,4-D 0.1 mg/L,悬浮培养适宜培养基为MS+6-BA 0.5 mg/L+2,4-D 0.1 mg/L.所建立的悬浮细胞系生长呈“S”型曲线,培养20 d达到最大生长量,最适继代周期为18~20 d,在悬浮培养过程中,培养液的pH和电导率变化趋势均为先下降后上升最后趋于稳定,细胞数量则呈现先上升后下降的趋势.

  6. Ziziphus mauritiana (masau) fruits fermentation in Zimbabwe: from black-box to starter culture development

    NARCIS (Netherlands)

    Nyanga, L.K.

    2012-01-01

    This thesis reports on studies of microbiological and biochemical properties of masau (Ziziphus mauritiana) fruit fermentation and the development of starter cultures for the production of masau beverages. A survey to document the traditional processing techniques was conducted using a

  7. Optimization of callus and cell suspension cultures of Barringtonia racemosa (Lecythidaceae family for lycopene production Otimização de culturas de suspensões de calos e células de Barringtonia racemosa (família Lecythidaceae para produção de licopeno

    Directory of Open Access Journals (Sweden)

    Mandana Behbahani

    2011-02-01

    Full Text Available Lycopene is present in a range of fresh fruits and vegetables, especially in the leaves of Barringtonia racemosa. The traditional lycopene extraction from the plant is being employed instead of an easy propagation technique like cell culture process from the leaf explants. We intend to assess how lycopene could be extracted via tissue culture under light (illuminance: 8,200 lux under white fluorescent lamps, photoperiod 16 h per day at 25ºC and dark. Leaf explants of Barringtonia racemosa were cultured on modified Murashige and Skoog (MS, Woody Plant Medium (WPM and B5 media, supplemented with different concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D. Optimal conditions for callus induction and maintenance under both dark and light were investigated, and growth and lycopene accumulation were evaluated. Among media with different concentrations of 2,4-D, fast growing, friable callus initiated within three weeks after culturing on WPM basal medium supplemented with 2.0 mg L-1 (weight per volume of 2,4-D, whereas callus induction in explants cultured on all other media started only after five weeks. Calli were subcultured once every fortnight. Pale yellow and green calli developed under conditions of dark and light respectively were then selected for evaluation of their lycopene contents. An improved reversed phase of high performance liquid chromatography (HPLC method was used for a selective chemical determination of the lycopene content. Light induced lycopene production; and likewise maximum lycopene level incubated in light was higher than those incubated in darkness. The best growth rates of callus and cell suspension were achieved in WPM and B5 media respectively. The production of lycopene was growth-dependent through analysis of growth and lycopene content of both callus and cell suspension cultures.O licopeno está presente numa série de frutas frescas e hortaliças principalmente na folhas de Barringtonia racemosa. A extra

  8. The role of 2,4-D and auxin-binding proteins during the induction of embryogenic and non-embryogenic callus in Zea mays (L.).

    NARCIS (Netherlands)

    Bronsema, F.

    1998-01-01

    This thesis deals with the influence of the growth regulator 2,4-dichlorophenoxyacetic acid (2,4-D) on the induction of callus in cultured immature embryos of Zea mays (L). In maize, two types of embryogenic callus can be induced in immature zygotic embryos.Type I callus, which is compact in appeara

  9. Effects of auxins and cytokinins on tomato callus from anthers

    Directory of Open Access Journals (Sweden)

    Janina H. Rogozińska

    2015-01-01

    Full Text Available An investigation was carried out on growth substance requirements of tomato callus derived from anthers for culture in vitro. Linsmaier and Skoog (1965 medium was used with various levels of auxins (IAA and NAA and cytokinins (K and BAP. The results show that cytokinin is an absolute requirement for callus growth irrespective of the auxin level. The optimum concentration of auxin in combination with cytokinin was found to be 5 μM of NAA or 25 μM of IAA, with 5 μM of K or BAP. Callus growth on media with NAA and cytokinin was superior to that on IAA, amounting to 6.05 g per piece on medium with 5 μM of NAA and BAP. Tissues grown on this medium have the highest water content. At the onset of culture the tissue is characterized by weak growth and attains its maximal increase in fresh weight after 6 weeks.

  10. Tissue culture and top-fruit tree species.

    Science.gov (United States)

    Ochatt, S J; Davey, M R; Power, J B

    1990-01-01

    The commercial cultivation of rosaceous fruit trees (e.g., pear, apple, cherry, peach, plum) relies heavily upon the quality and performance of the rootstocks. This is even more the case now that self-rooted scions produce larger trees with a longer juvenile phase (1). It would, therefore, be of special interest for the fruit breeder to have general purpose rootstocks with a wide ecophysiological adaptation and high compatibility coupled with early cropping. In addition, many of the older and highly adapted scion varieties of fruit trees could benefit greatly from the introduction of stable, yet minor changes in their genome. Fruit trees are generally highly heterozygous, outbreeding, and thus are asexually propagated (see Chapter 10 , this vol.). Consequently, genetic improvement is likely to be based on protoplast technology, and achieved mainly through somatic methods, such as somaclonal variation or somatic hybridization.

  11. Effect Of Explant Source And Different Medium Culture On Friable Embryogenic Callus Induction Of Four Cultivars Of Cassava Manihot Esculenta Crantz

    Directory of Open Access Journals (Sweden)

    Simplice Prosper Yandia

    2015-08-01

    Full Text Available In order to obtain Friable Embryogenic Callus FEC for protoplast isolation we have evaluated in this research the competance for Friable Embryogenic Callus FEC of four cassava cultivars M61033 Rendre Yalipe and Six-mois in media containing MS supplemented with 8mgl 24-D MS supplemented with 10 mgl BAP and GD supplemented with 12mgl picloram using apical bud AB and immature leaves lobes ILL as explants. In general in the medium GD12mgl picloram the highest efficiencies of FEC ranged from 58 to 87 and the highest score of FEC ranged from 4.2 to 5.4 with explants AB however we have observed with explants ILL the efficiencies of somatic embryos ranged form 41 to 75 and the score ranged from 4.1 to 4.4. The mediums MS28 mgl 24-D have induced with explants AB the efficiencies of FEC ranged from 43 to 57 and the score ranged from 3.1 to 3.8 however with ILL explants the efficiencies of FEC ranged from 39 to 49 and the score ranged from 2.9 to 3.7. The least FEC were observed in the medium MS210 mgl BAP with BA explants however the efficiencies ranged from 6 to 11 and the score ranged from 1.1 to 1.8. Whereas the efficiencies of FEC with ILL explants ranged from 4 to 7 and the score ranged from 0.5 to 0.8. All of four cultivars showed capability of producing FEC although their efficiency varied according to gonotype donors explants and medium taking into acount. Abbreviations GD Gressoff and Doy MS Murashige and Skoog 24-dichloro phenoxyacetic acid BAP Benzylamino-purin-Acid AB Apical Bud ILL Immature Leaves lobes

  12. 氮素对印楝愈伤组织和悬浮细胞培养的影响%Effects of Nitrogen on Azadirachta indica A. Juss Callus and Suspension Culture

    Institute of Scientific and Technical Information of China (English)

    韩广建; 李兴林; 别振宇; 张国运

    2015-01-01

    通过改变印楝组织培养基中硝态氮与铵态氮物质的量比,并与培养物的生物量、可溶性蛋白含量、印楝素、柠檬苦素含量相关联,探讨了 MS 培养基中氮素对印楝愈伤组织和悬浮细胞培养的影响.实验结果表明:在印楝的愈伤组织培养过程中,当硝态氮与铵态氮物质的量比为 4:1 时,生物量、可溶性蛋白含量、印楝素以及柠檬苦素含量均达到最大积累量;在悬浮细胞培养的过程中,当硝态氮与铵态氮物质的量比为 3:1 时,可溶性蛋白含量、印楝素以及柠檬苦素含量达到最高,而在 2:1 时生物量达到最大.因此,印楝组织培养的不同阶段应该采用不同形式氮素比例的培养基,只有这样才能获得最佳的目标培养物.%To study the effects of nitrogen in MS medium on neem(Azadirachta indica A. Juss)tissue culture,we deter-mined the biomass,soluble protein content,azadirachtin and limonoid contents of neem callus and suspension cells in the end of their cultures by changing the ratio between nitrate and ammonium. The results indicate,that while culturing callus, the four measurement indicators reached the maximum accumulation when the ratio of nitrate to ammonium was 4:1;while culturing suspension cells,the soluble protein contain,azadirachtin and limonoids contains got their maximum whenthe ratio of nitrate to ammonium was 3:1,but the maximum biomass was obtained when the ratio was 2:1. In conclusion,the ratio between nitrate and ammonium in the MS medium should be corresponded with the objects being cultured at different peri-ods.Only in this way,can we get the best target cultures.

  13. 紫果西番莲种子萌发及根部诱导愈伤组织研究%Seed Germination of Passion fruit edulis and Callus Induction from Root

    Institute of Scientific and Technical Information of China (English)

    张婷婷; 吴田; 张瑞丽; 蓝增全

    2013-01-01

    The germination rate ofPassiflora edulis seed and the callus induction of root were studied in different seed soaking time with gibberellin and different seed treatment conditions.The results showed that the shucked Passiflora edulis seed on MS medium was the optimal way for seed germination.The optimal medium of root callus induction was MS + 0.5 mg/L 2,4-D,and the induced callus were mostly light green,granular and loose texture.%以西番莲种子为试验材料,在不同的赤霉素浸种时间及不同种子处理方式条件下,进行西番莲种子萌发试验,并利用发芽种子的根进行愈伤组织诱导.结果表明,将西番莲种子剥壳后置于MS培养基中萌发,是西番莲种子萌发的最适方式.根部愈伤组织诱导的最适培养基为MS +0.5 mg/L2,4-D,诱导出的愈伤组织多为浅绿色,颗粒状,质地疏松.

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

    African Journals Online (AJOL)

    RAMA

    2013-05-01

    May 1, 2013 ... Key words: Callus culture, medicinal plant, root induction, shoot organogenesis. INTRODUCTION ... media fortified with 2 mg/l 2,4-dichlorophenoxyacetic acid. The pH ... salts, vitamins, 0.5 to 2.0 mg/l BAP or Kn and in different.

  15. Anionic peroxidase production by Arnebia euchroma callus.

    Science.gov (United States)

    Farhadi, Sahar; Haghbeen, Kamahldin; Marefatjo, Mohammad-Javad; Hoor, Marjan Ghiyami; Zahiri, Hossein Shahbani; Rahimi, Karim

    2011-01-01

    Arnebia euchroma callus, obtained from the root cell culture of an Iranian native specimen, has gained a doubling time of 63 H after regular subculturing on Linsmaier-Skoog (LS) medium containing sugar (50 g/L), 2,4-dichlorophenoxyacetic acid (10(-6) M), and kinetin (10(-5) M) under darkness at 25°C. Despite the observed somaclonal variations, peroxidase production by the A. euchroma calli has been stable over 4 years under the aforementioned conditions. Isoelectric focusing experiments revealed that the partially purified A. euchroma peroxidases (AePoxs) are mainly anionic with pI values of about 5.5 and 6.6. AePox reaches its optimal activity at 55°C and pH 7.5. Results of the various kinetic studies suggest that AePox belongs to the type III plant peroxidases with no activity for the oxidation of 3-indoleacetic acid, but seems to play a role in the lignin biosynthesis and H(2) O(2) regulation during the proliferation of the A. euchroma cells on LS medium. Comparing the biochemical properties of AePox with horseradish peroxidase and in view of the ease of solid cell culture, the A. euchroma callus could be considered as a source of plant peroxidase for some biotechnological applications. Copyright © 2011 International Union of Biochemistry and Molecular Biology, Inc.

  16. Effects of growth regulators on callus induction and secondary metabolite production in Cuminum cyminum.

    Science.gov (United States)

    Farvardin, Atefeh; Ebrahimi, Asa; Hosseinpour, Batul; Khosrowshahli, Mahmood

    2017-01-03

    Cumin (Cuminum cyminum) is an annual plant from Apiaceae family that is cultivated in Iran as landraces. The most important chemical composition of the cumin essential oil was cuminaldehyde. In this research, the effect of different landraces and growth regulators was evaluated on callus induction, and best callus was used for amount of cuminaldehyde content. Node, root, leaf and hypocotyl explant from seedlings of Birjand and Qaen landraces were cultured on MS and MS5 medium supplemented with different concentrations of 2, 4-D and Kin. This experiment has been carried out in a completely randomised design with 3 replications. Percentage of callogenesis, callus volume, fresh and dry weight were measured. The best treatment for callus induction was 2.5 mg/L 2, 4-D and 0.5 mg/L Kin in MS5 medium. The best callus result was evaluated for cuminaldehyde content. An amount of 5.7% cuminaldehyde was measured using hydrodistillation method.

  17. Callogenesis in stem explants of Eucalyptus grandis (Hill ex Maiden showing resilience in oxide calluses

    Directory of Open Access Journals (Sweden)

    Regina Paula Willemen Pereira

    Full Text Available Callogenesis was achieved using growth regulators at different concentrations to obtain the best characteristics regarding the calluses texture aspect with the main objective to produce friable calluses from stem explants of Eucalyptus grandis. Stem segments of approximately 1 cm were excised from 30-day seedlings. The treatments were prepared based on the MS medium added with TDZ alone or combined with AIA; 2,4-D; ANA. Each treatment consisted of five repetitions with five explants per plate. After 210 days of in vitro culture, the calluses were evaluated visually and by manipulation for texture. The explants that did not produce calluses were those exposed to treatments with 2,4-D 50.0 µM; TDZ 3.0 µM and in absence of a growth regulator (control.The best protocols for maintenance of friable calluses of stem explants of Eucalyptus grandis under the conditions performed in this work were those using only TDZ.

  18. Cultural Characteristics and Fruiting Body Production in Cordyceps bassiana

    Science.gov (United States)

    Lee, Je-O; Shrestha, Bhushan; Sung, Gi-Ho; Han, Sang-Kuk; Kim, Tae-Wong

    2010-01-01

    Single ascospore isolates of Cordyceps bassiana were observed for their colony pigmentation on Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates and were inoculated in a brown rice medium for production of fruiting bodies. Colony pigmentation did not show any relationship with perithecial stromata formation. The isolates were also grown on opposite sides of SDAY agar plates and were observed for vegetative compatibility. Neither vegetative compatibility nor perithecial stromata could be found to be related to each other. It was concluded that fertile fruiting body production was independent of colony pigmentation and vegetative compatibility. Synnemata formation was found to be more common than perithecial stromata formation. This might be due to its highly conidiogenous anamorphic stage, i.e., Beauveria bassiana. PMID:23956638

  19. LATERAL ORGAN BOUNDARIES DOMAIN transcription factors direct callus formation in Arabidopsis regeneration

    Institute of Scientific and Technical Information of China (English)

    Mingzhu Fan; Chongyi Xu; Ke Xu; Yuxin Hu

    2012-01-01

    The remarkable regeneration capability of plant tissues or organs under culture conditions has underlain an extensive practice for decades.The initial step in plant in vitro regeneration often involves the induction of a pluripotent cell mass termed callus,which is driven by the phytohormone auxin and occurs via a root development pathway.However,the key molecules governing callus formation remain unknown.Here we demonstrate that Arabidopsis LATERAL ORGAN BOUNDARIES DOMAIN (LBD)/ASYMMETRIC LEAVES2-LIKE (ASL) transcription factors are involved in the control of callus formation program.The four LBD genes downstream of AUXIN RESPONSE FACTORs (ARFs),LBD16,LBD17,LBD18 and LBD29,are rapidly and dramatically induced by callus-inducing medium (CIM) in multiple organs.Ectopic expression of each of the four LBD genes in Arabidopsis is sufficient to trigger spontaneous callus formation without exogenous phytohormones,whereas suppression of LBD function inhibits the callus formation induced by CIM.Moreover,the callus triggered by LBD resembles that induced by CIM by characteristics of ectopically activated root meristem genes and efficient regeneration capacity.These findings define LBD transcription factors as key regulators in the callus induction process,thereby establishing a molecular link between auxin signaling and the plant regeneration program.

  20. Callus formation from leaf mesophyll protoplasts of Malus Xdomestica Borkh. cv. Greensleeves.

    Science.gov (United States)

    Doughty, S; Power, J B

    1988-05-01

    Large yields (1.85 × 10(7)/g.f.wt.) of viable protoplasts were obtained from leaves of axenic shoot cultures of Malus Xdomestica Borkh. cv. 'Greensleeves'. Protoplasts cultured in liquid or agarose semi-solidified KM8P medium underwent cell wall regeneration and colony formation.Protoplast-derived cell colonies developed to callus on semi-solid KM8 medium. This is the first report of callus formation from mesophyll protoplasts of apple.

  1. INDUCTION OF CALLUS FORMATION FROM DIFFERENT PARTS OF Citrus grandis (OSBECK FLOWERS

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    Zarina Binti Zakaria

    2010-01-01

    Full Text Available The objectives of the present work were to study the capability of pomelo's floral tissues to produce callus and to investigate the influence of plant growth regulators on callus induction and development. Various parts of flower namely petal, sepal, style, ovary, pistil and cup base were cultured onto Murashige and Skoog (MS basal medium supplemented with different levels of 2,4-dichlorophenoxyacetic acid (2,4-D, 6-benzylaminopurine (BAP and 6-Furfurylaminopurine (kinetin. It was found that different parts of flowers favored different levels of hormone for callus induction. The highest formation of callus were obtained from petal and sepal cultured on MS media supplemented with 1.5 mg/l kinetin. A 50 ± 2.3%explants from style and pistil had initiated callus when cultured on MS media supplemented with 0.05 mg/l and 0.10 mg/l BAP, respectively. On the other hand, sucrose agar (SA media alone had managed to induce callus formation from almost every part of the flowers with a success rate between 7.69 ± 0.32 to 50 ± 3.18%. The uncut part of flowers initiated high percentage of callus (14.6 ± 0.35% as shown in the study on petal.

  2. Effect of salicylic acid on callus formation in three Theobroma cacao L. clones

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    Lillien Fajardo Rosabal

    2015-10-01

    Full Text Available Cocoa is a crop of great economic importance and it has interest as a source of naturally occurring compounds. The tissue culture using metabolic elicitation is an alternative for large scale production of secondary metabolites. This study aimed to determine the effect of salicylic acid as elicitor for the production of cell biomass of Theobroma cacao L. in three clones. For callus formation staminodes of ‘UF-613’, ‘UF-650’ and ‘Pound-7’ clones were used. It were added 0.02, 0.05, 0.10 and 0.15 mg l-1 salicylic acid to the culture medium. At 15 and 28 days of culture the number of explants formed callus were quantified and the percentage of callus formation was calculated. In the best results clone the secondary metabolites in callus and culture medium were identified by phytochemical screening. The addition of salicylic acid accelerated callus formation in the cocoa clones ‘UF-613’, ‘UF-650’ and ‘Pound-7’. The results depended of the clone and the concentrations used. The identification of secondary metabolites in callus of ‘UF-650’ clone and in the culture medium is the starting point for future research.   Key words: tissue culture, phytochemical screening, secondary metabolites.

  3. Impact of gamma rays exposure and growth regulators on Oryza sativa L. c.v MR269 callus induction

    Science.gov (United States)

    Kadhimi, Ahsan A.; Zain, Che Radziah Che Mohd; Alhasnawi, Arshad Naji; Isahak, Anizan; Ashraf, Mehdi Farshad; Mohamad, Azhar; Doni, Febri; Yusoff, Wan Mohtar Wan

    2016-11-01

    The study is aimed to evaluate in vitro somatic embryogenesis to gamma ray exposure and the use of growth regulators to mature embryos explants for rice callus induction. Seeds of local rice genotype (MR269) were exposed to gamma rays at 350 Gy (Source: Caesium-137). Matured embryos were cultured to induce callus on Murashige and Skoog (MS) supplemented at different concentrations of 1.0, 2.0, 3.0 mg/L 2.4-D and 0.1 and 0.2 mg/L Kinetin for 4 weeks. Callus induction and callus fresh weight were decreased after exposure to gamma ray. The most efficient response to callus induction and callus fresh weight was found at 3 mg/L 2,4-D and 0.1 mg/L kinetin.

  4. Electrostimulation of rat callus cells and human lymphocytes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Aro, H.; Eerola, E.; Aho, A.J.; Penttinen, R.

    1984-01-01

    Asymmetrical pulsing low voltage current was supplied via electrodes to cultured rat fracture callus cells and human peripheral blood lymphocytes. The (/sup 3/H)thymidine incorporation of the callus cells and 5-(/sup 125/I)iodo-2'-deoxyuridine incorporation of the lymphocytes were determined. The growth pattern of callus cells (estimated by cellular density) did not respond to electrical stimulation. However, the uptake of (/sup 3/H)thymidine was increased at the early phase of cell proliferation and inhibited at later phases of proliferation. The (/sup 3/H)thymidine uptake of confluent callus cell cultures did not respond to electrical stimulation. Lymphocytes reacted in a similar way; stimulated cells took up more DNA precursor than control cells at the early phase of stimulation. During cell division, induced by the mitogens phytohemagglutinin and Concanavalin-A, the uptake of DNA precursor by stimulated cells was constantly inhibited. The results suggest that electrical stimuli affect the uptake mechanisms of cell membranes. The duality of the effect seems to be dependent on the cell cycle.

  5. Tissue culture of Cecropia glaziovii Sneth (urticaceae: vegetative micropropagation and plant regeneration from callus Cultura de tecidos de Cecropia glaziovii Sneth (Urticaceae: micropropagação vegetativa e regeneração de plantas via calos

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    Marcos Nopper Alves

    2010-10-01

    Full Text Available Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS medium supplemented with 6-benzylaminopurine alone (BAP (1.0, 5.0 or 10.0 mg L-1 or combined with -naphthalene acetic acid (NAA (1.0 or 2.0 mg L-1, after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D (5.0 mg L-1 combined with BAP (1.0 mg L-1. Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA (0.1 mg L-1 alone or combined with 2,4-D (1.0 or 5.0 mg L-1 was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA (1.0 mg L-1. Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.Cecropia glaziovii é uma planta lenhosa, popularmente usada no Brasil como medicinal. Métodos que visem a sua propagação clonal podem ser de grande utilidade na preservação de seus genótipos de elite. Foram examinados efeitos de diferentes reguladores de crescimento e explantes na forma

  6. Impact of exogenous ascorbic acid on biochemical activities of rice callus treated with salt stress

    Science.gov (United States)

    Alhasnawi, Arshad Naji; Zain, Che Radziah Che Mohd; Kadhimi, Ahsan A.; Isahak, Anizan; Mohamad, Azhar; Ashraf, Mehdi Farshad; Doni, Febri; Yusoff, Wan Mohtar Wan

    2016-11-01

    The application of in vitro systems can lead to new methods of crop amelioration. This method has been widely utilized for breeding tenacities, particularly for stress tolerance selection. Salinity causes oxidative stress in callus by enhancing the production of Reactive Oxygen Species (ROS), resulting in an efficient antioxidant system. The exogenous application of ascorbic acid (AsA) is an important requirement for tolerance. The present study aimed to examine in vitro selection strategy for callus induction in rice mature embryo culture on MS culture medium and to produce salt-tolerant callus under sodium chloride (NaCl) and AsA conditions in callus rice variety, MR269. This study also highlights changes in the activities of proline and antioxidants peroxidase (POD), catalase (CAT) and superoxide dismutase (SOD) of callus under NaCl stress to understand their possible role in salt tolerance. However, various levels of exogenously applied AsA under saline conditions improved callus, and the antioxidant enzyme activities of AsA are related to resistance to oxidative stress. Our results provide strong support for the hypothesis that AsA-dependent antioxidant enzymes play a significant role in the salinity tolerance of callus rice.

  7. Cell dedifferentiation, callus induction and somatic embryogenesis in Crataegus spp.

    Science.gov (United States)

    Taimori, N; Kahrizi, D; Abdossi, V; Papzan, A H

    2016-09-30

    The present study describes the effects of light conditions, different kinds and concentrations of auxins [Naphthylacetic acid (NAA) and dichlorophenoxyacetic acid (2,4-D)] with cytokinin (Kin) in MS medium on callus induction and embryogenesis in Crataegus pseudoheterophylla, C. aronia and C.meyeri. At first leave explants sections were cultured on different combinations of plant growth regulators in dark and light for callus initiation and light conditions to evaluation the percentage and duration of survival, callus diameter, callus fresh weight and dry. Results of effects of plant growth regulators and light conditions on callus initiation revealed that highest percentage of callus initiation leaves in treatment (0.5 mg/l 2.4-D+0.5 mg/l KIN) for species C.pseudoheterophylla in dark conditions (100%). Dark conditions (100%) were more effective on callogenesis than light conditions (Photoperiodicity of 16-h and at light intensity of 40 µmol m-2 s-1). The callus induction of in vitro (64-100%) leaves was better than the ex vitro ones (0-100%). The combination of 2,4-D and Kin of in vitro leaves callogenesis has been indicated faster (one weeks) than the other combinations. The results also showed that the highest percentage (100%) and survival duration (6 months) was found in species C. pseudoheterophylla and C. meyeri in 0.1 mg/l 2,4.D + 0.5 mg/l KIN and 0.5 mg/l 2,4.D + 0.5 mg/l Kin. The minimum survival (0%) was absorbed in species C. aronia in 1 mg/l NAA. Maximum callus (10.63 and 10.00 mm respectively) was shown in 0.1 mg/l 2,4.D + 0.5 mg/l Kin and 0.5 mg/l 2,4.D + 0.5 mg/l Kin and was not significant differences after five week among species. The results showed that the highest fresh (1081.49 mg) and dry weight (506.88 and 506.98 mg respectively) was absorbed in species C. pseudoheterophylla in 0.1 mg/l 2,4.D + 0.5 mg/l Kin and 0.5 mg/l 2,4.D + 0.5 mg/l Kin. The embryogenesis was not occurred in any plant growth regulator combinations and species. The

  8. Induction of callus from axillary buds of taro (Colocasia esculenta var. esculenta, Araceae) and subsequent plantlet regeneration.

    Science.gov (United States)

    Yam, T W; Young, J L; Fan, K P; Arditti, J

    1990-12-01

    Axillary buds of taro (Colocasia esculenta var. esculenta, Araceae) cultured on half strength Murashige-Skoog medium (HMS) containing taro extract (HMSTE) and 2, 4, 5-trichlorophenoxyacetic acid produce a compact, hard, slow growing callus which is not very active morphogenetically and produces only a few plantlets. When cultured on HMSTE plus 5 mg 1(-1) each of naphthaleneacetic acid and benzyl adenine (HMSNB) the buds produce a fast growing, friable and morphogenetically active callus. Meristematic regions form on the friable callus after 30 days on HMSNB. If transferred to HMSTE at this point the callus gives rise to plantlets. Addition of taro extract to the media is required for the culture of buds, induction of callus and plantlet regeneration.

  9. Ziziphus mauritiana (masau) fruits fermentation in Zimbabwe: from black-box to starter culture development

    NARCIS (Netherlands)

    Nyanga, L.K.

    2012-01-01

    This thesis reports on studies of microbiological and biochemical properties of masau (Ziziphus mauritiana) fruit fermentation and the development of starter cultures for the production of masau beverages. A survey to document the traditional processing techniques was conducted using a questionnair

  10. Label-Free Quantitative Proteomics of Embryogenic and Non-Embryogenic Callus during Sugarcane Somatic Embryogenesis.

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    Angelo Schuabb Heringer

    Full Text Available The development of somatic cells in to embryogenic cells occurs in several stages and ends in somatic embryo formation, though most of these biochemical and molecular changes have yet to be elucidated. Somatic embryogenesis coupled with genetic transformation could be a biotechnological tool to improve potential crop yields potential in sugarcane cultivars. The objective of this study was to observe somatic embryo development and to identify differentially expressed proteins in embryogenic (E and non-embryogenic (NE callus during maturation treatment. E and NE callus were cultured on maturation culture medium supplemented with different concentrations (0.0, 0.75, 1.5 and 2.0 g L(-1 of activated charcoal (AC. Somatic embryo formation and differential protein expression were evaluated at days 0 and 21 using shotgun proteomic analyses. Treatment with 1.5 g L(-1 AC resulted in higher somatic embryo maturation rates (158 somatic embryos in 14 days in E callus but has no effect in NE callus. A total of 752 co-expressed proteins were identified through the SUCEST (The Sugarcane EST Project, including many housekeeping proteins. E callus showed 65 exclusive proteins on day 0, including dehydrogenase, desiccation-related protein, callose synthase 1 and nitric oxide synthase. After 21 days on maturation treatment, 14 exclusive proteins were identified in E callus, including catalase and secreted protein. NE callus showed 23 exclusive proteins on day 0 and 10 exclusive proteins after 21 days on maturation treatment, including many proteins related to protein degradation. The induction of maturation leads to somatic embryo development, which likely depends on the expression of specific proteins throughout the process, as seen in E callus under maturation treatment. On the other hand, some exclusive proteins can also specifically prevent of somatic embryos development, as seen in the NE callus.

  11. Early fruit setting from tissue culture-derived mangosteen tree

    OpenAIRE

    Sompong Te-chato; Mongkol Lim

    2004-01-01

    Vitro-plantlets of mangosteen derived from culturing young leaves were acclimatized in 1993. Small and large polybag seedlings were carefully raised under controlled environmental conditions until 1994 when they were ready to be transferred to the field. During this stage, morphological abnormalities of the seedlings were recorded. After transferring to the field for 5-6 years (1994-1999) at Yi Ngo District, Narathiwat Province and Klong Hoi Khong District, Songkhla Province, morphological ch...

  12. Exploration of English Idioms about Fruit and the Frequent Usage in Different Language Culture

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    Guilan Jiao

    2015-08-01

    Full Text Available English idioms are commonly found in the oral and written English, which are quite vivid, concise and carried with a large amount of information. It makes up the major part of English language. In English idioms, fruit ones are a unique expression, which are widely accepted and applied in the daily life of English-speaking countries and carried with heavy cultural connotations. This study made an exploration of the historical origins of English idioms about fruit based on Greek mythology and Bible of Christianity. Besides, it made a detailed analysis of their frequent usage from the perspective of daily life. With the deeper understanding of culture of English-speaking countries, there will be gradually enhanced skills of interpersonal communication between different cultures.

  13. In vitro isoflavonoid production and analysis in natural tetraploid Trifolium pratense (red clover calluses

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    Tugba Ercetin

    2012-10-01

    Full Text Available Isoflavones are polyphenolic phytoestrogens, predominantly found in leguminous plants. Trifolium pratense L., Fabaceae (red clover, is rich in isoflavones that possess estrogenic activity due to their similar molecular structure and effectiveness in preventing health conditions such as menopause, osteoporosis, cardiovascular disease, hypertension and hormone-dependent cancers. In this study, presence and amount of various phytoestrogens in the tetraploid plant and in the calluses derived from the plants were investigated. Calluses were generated from explants obtained from natural tetraploid T. pratense seedlings. The best callus formation was obtained from hypocotyl explants cultured in Phillips Collins and Gamborg B5 media containing different plant growth regulators. Flowers of plants and calluses were analysed for formononetin, biochanin A, genistein and daidzein contents using HPLC. In HPLC analysis, high levels of formononetin (0.249 µg/mg were determined in natural tetraploid T. pratense flowers in addition to genistein and biochanin A. In calluses, highest isoflavone content (1.15 µg/mg formononetin was observed in modified Gamborg B5 medium. Biochanin A content of calluses and the plant were found to be nearly the same. But formononetin and genistein contents of the calluses in this medium were found to be respectively 4.62 and 21.39 folds higher than the tetraploid plant.

  14. An efficient protocol devised for rapid callus induction from leaf explants of Biophytum sensitivum (lDC.

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    Sirigiri Chandra Kala

    2014-03-01

    Full Text Available The Cell cultures are used extensively for in vitro secondary metabolite productions were obtained from callus tissue through cell suspension culture.  The establishment of callus cultures has considerable potential for the production of known and novel secondary metabolites. The objective of the study was to scientifically assess callus culture of Biophytum sensitivum (L DC. was established from leaf explants with different growth regulators greatly influenced the growth of callus cultures. The callus from leaf explants is induced by inoculating the young leaf bits on MS medium supplemented with various auxins (2, 4- Dichlorophenoxyacetic acid (2, 4-D, α-Naphthalene Acetic Acid (NAA and Indole Buteric Acid (IBA, cytokinins (6-Benzyladenine (BA and Kinetin (KN and cytokinin-auxin combination (BA+NAA in different concentrations were (0.5 to 5.0 mg/l used. BA 1mg/l, in combination with NAA (1.0 mg/l also produced maximum amount of callus.  So, this research is concluded that the plant leaf explants cultured on MS medium with 1 mg/ l  BA with 0.5 mg/l NAA was found most efficient for callus induction, provided calli with quite good in texture and friable in nature.

  15. Analysis of indole compounds in fruiting bodies and in mycelia from in vitro cultures of Calocera viscosa (Basidiomycota)

    OpenAIRE

    Bożena Muszyńska; Katarzyna Sułkowska-Ziajda

    2013-01-01

    Calocera viscosa (Pers.: Fr.) Fr. (Basidiomycota) from Dacrymycetaceae family is a widespread species of mushroom in Poland. The aim of this study was to investigate the content of indole compounds in fruiting bodies and in mycelium cultured in vitro on solid and liquid medium of this species. Fruiting bodies of Calocera viscosa were collected in coniferous forests in south Poland and were used to derive in vitro cultures. The optimal medium composition for cultures was determined. Fresh mate...

  16. Effects of Auxins and Media on Callus Induction of Chinese Spring Wheat( Triticum aestivum L.)

    Institute of Scientific and Technical Information of China (English)

    WU Li-min; WEI Yu-ming; ZHENG You-liang

    2005-01-01

    The effects of auxins and media on callus induction from the mature and immature embryos of Chinese spring wheat(Triticum aestivum L.) varieties were investigated. It was found that genotype, medium, auxin source and concentration had the significant effects on the induction of embryogenic callus, explants germination and the increment of callus fresh weight. For immature embryos cultured on MS medium, 2 mg L-1of 2, 4-D was optimal, and the highest frequency of embryogenic callus (33.50%) was observed. For the mature embryos on N6 medium, 4 mg L-1 of 2, 4-D was optimal. The frequency of embryogenic callus and increment of callus fresh weight on 2, 4, 5-T media were higher than those on 2, 4-D media, and in the presence of 2, 4, 5-T the precocious germination of explants for all genotypes were significantly suppressed. These results indicated that 2, 4, 5-T was superior to 2, 4-D and NAA in the culture of immature embryos. This is the first report about the effect of 2, 4, 5-T and NAA on wheat tissue culture, particularly in comparison with 2, 4-D in detail.

  17. In vitro Accumulation of Polyphenols in Tea Callus Derived from Anther

    Science.gov (United States)

    Chevala, Naga Pavan Kumar; Chevala, Naga Thirumalesh; Dhanakodi, Kirubakaran; Nadendla, Rama Rao; Nagarathna, Chandrashekara Krishnappa

    2016-01-01

    Background: Tea is an economic important crop with high medicinal value due to rich polyphenols content. In the present research we studied the accumulation of polyphenols of in vitro regenerated callus from anthers. Objective: Callus induction of tea anthers and in vitro accumulation of phenolic compounds from the anther-derived callus. Materials and Methods: Standardization of callus induction for tea anthers. In vitro generated callus was screened for in vivo accumulation of catechins and its isomers were screened by FC reagent staining technique. The methanol extract of dry and green callus obtained were estimated qualitatively by Fourier transform infrared spectroscopy (FTIR)–alternative total reflection (ATR) and quantitatively by HPLC method. Results: Anthers inoculated on half strength MS media fortified with 2,4-dichloro acetic acid (2 mg/L), Kn (1 mg/L), and BAP (1 mg/L) induced callus under photoperiod of 9:15 h light. The in vivo histochemical studies revealed the accumulation of polyphenols in the callus. The in vitro generated fresh and dry callus were used for extraction and screened for accumulated polyphenols [galic acid, (+)-catechin (C), (−)-epicatechin, (−)-epigallocatechin, (−)-epigallocatechin gallate, (−)-gallocatechins, (−)-epicatechin gallate] were estimated qualitatively by FTIR–ATR method and quantitatively by HPLC method. Conclusion: The FC staining technique used here helps in localization of polyphenol compounds accumulation in the tissues by instant microscopic studies. The study have scope in large-scale isolation of various medicinally important flavonol by using anther culture. Abbreviations used: HPLC: high pressure liquid chromatography; FTIR: Fourier transform infrared spectroscopy; 2,4-D: 2,4-dichloro acetic acid; BAP: N6-benzyl amino purine; kn: kinetin

  18. Optimising Sterilisation Techniques and Callus Induction of Nodes Durio Zibethinus Murr in Vitro Method with Various Media

    Science.gov (United States)

    Hermayani, N.; Retnoningsih, A.; Rahayu, E. S.

    2017-04-01

    The application of in vitro propagation method needs an aseptic or sterile condition. The objective of the study was to get an optimal sterilisation techniques and medium of callus induction of Durio zibethinus. Sterilisation treatments studied were NaClO and Ca(ClO)2. The three kind of callus induction medium studied were Gamborg (B5), Woody Plant Medium (WPM), and Murashige and Skoog (MS) with the addition of auxin and cytokinin. The experiment unit was three bottles with nodes as explant. Cultures were kept in the culture room for 16 hours daily by LED light intensity of 1000 lux. Parameters investigated of sterilisation technique development was the percentage of contamination, the percentage of explant browning, and the percentage of life explants; whereas the callus induction measured were the rate of callus formation, percentage of callus covered, callus texture, colour and diameter of callus. The results showed that the code of C3 is soaking in calcium hypochlorite (Ca(ClO)2) 50% for 3 minutes, calcium hypochlorite 40% for 2 minutes, and 70% alcohol for 30 seconds separately were suitable for sterilisation of nodes explant. The development of callus on B5 medium with the addition of auxin (2,4-D) 2 ppm and cytokinin (Thidiazuron) 1 ppm was best compared to the other.

  19. The effects of cultural practice methods on fruit orchard rehabilitation after flooding crisis in Songkhla province

    Directory of Open Access Journals (Sweden)

    Chanaweerawan, S.

    2004-01-01

    Full Text Available To rehabilitate the fruit orchards exposed to flooding crisis that occurred in year 2000 in Songkhla province, an investigation of the effects of cultural practice methods was done in the farmers’ orchards at 2 experimental sites (at Tumbol Kho Hong and Tumbol Kuan Lang, Amphur Hat Yai. The first site, at Tumbol Kho Hong, was mixed cropping (durian, longkong and mangosteen. The second site, at Tumbol Kuan Lang, was a monocrop of pummelo orchard. The experiment was comprised a stratified sampling method with 3 treatments: 1 control, 2 fertilization (15-15-15, 8-24-24 and 13-13-21 and soil improvement with humic acid (S and 3 foliar application (glucose was applied with 16-12-0+micronutrients+extracted seaweed and 7-13-34+12.5Zn+extracted seaweed+Ca-B spraying with fertilization and soil improvement (F+S. The results from the both experimental sites showed that the F+S treatment exhibited the best result. This promoted the plant growth and yield of fruit trees. In addition, the other orchards surrounding the experimental sites were surveyed. It was noticeable that fruit trees grown in raising-beds could recover and exhibit normal fruit bearing. This pointed out that the impact of flooding on fruit orchards would be possibly alleviated by a drainage system.

  20. Aplicação de técnicas de biotecnologia à cultura e melhoramento do maracujazeiro Application of biotechnological processes to passion fruit culture

    Directory of Open Access Journals (Sweden)

    Daniela Macedo de Lima

    2000-04-01

    Full Text Available A tecnologia da cultura de protoplastos, células e tecidos de plantas in vitro constitui-se em uma das áreas de maior êxito na biotecnologia. No gênero Passiflora, poucas espécies têm sido utilizadas com fins biotecnológicos nos estudos de cultura de tecido. Do pouco já realizado, obteve-se sucesso na regeneração de brotos, por via indireta, a partir de calos, ou por via direta, a partir dos explantes cotiledonares, hipocotiledonares ou foliares. A regeneração in vitro é facilmente induzida em entrenós e segmentos de gavinha, com a adição de citocinina ao meio de cultivo. Recentes avanços na regeneração de protoplastos de outras frutíferas permitiram a aplicação da hibridização somática no maracujá. O uso da engenharia genética, aproveitando-se a habilidade de regeneração das plantas de P. edulis f. flavicarpa, torna-se relevante na tentativa de reduzir a devastação imposta por certas doenças e pragas, e também na adição de outras características de importância agronômica.The technology of cells, protoplast and tissue culture of plants is one of the most successful biotechnological areas. In the genus Passiflora, a few species have been used with biotechnological ends on the studies concerning tissue cultures. Regeneration of shoots can be indirect, from callus, or direct, by cotiledonary, hipocotiledonary and leaf explants. It is easy to induce in vitro regeneration of internodes and axillary tendrils if cytokinin is added to the culture medium. Early advances in the regeneration of other fruit tree protoplasts have allowed the use of the somatic hybridization process in the passion fruit. Since P. edulis f. flavicarpa has a natural regeneration ability, the application of genetic engineering techniques becomes necessary in order to reduce the infestation by some diseases or insects, and to add relevant agricultural properties to this species.

  1. Control mechanisms operating for lipid biosynthesis differ in oil-palm (Elaeis guineensis Jacq.) and olive (Olea europaea L.) callus cultures.

    Science.gov (United States)

    Ramli, Umi S; Baker, Darren S; Quant, Patti A; Harwood, John L

    2002-01-01

    As a prelude to detailed flux control analysis of lipid synthesis in plants, we have examined the latter in tissue cultures from two important oil crops, olive (Olea europaea L.) and oil palm (Elaeis guineensis Jacq.). Temperature was used to manipulate the overall rate of lipid formation in order to characterize and validate the system to be used for analysis. With [1-14C]acetate as a precursor, an increase in temperature from 20 to 30 degrees C produced nearly a doubling of total lipid labelling. This increase in total lipids did not change the radioactivity in the intermediate acyl-(acyl carrier protein) or acyl-CoA pools, indicating that metabolism of these pools did not exert any significant constraint for overall synthesis. In contrast, there were some differences in the proportional labelling of fatty acids and of lipid classes at the two temperatures. The higher temperature caused a decrease in polyunsaturated fatty acid labelling and an increase in the proportion of triacylglycerol labelling in both calli. The intermediate diacylglycerol was increased in olive, but not in oil palm. Overall the data indicate the suitability of olive and oil-palm cultures for the study of lipid synthesis and indicate that de novo fatty acid synthesis may exert more flux control than complex lipid assembly. In olive, diacylglycerol acyltransferase may exert significant flux control when lipid synthesis is rapid. PMID:12023881

  2. In vitro Induction and Proliferation of Callus from Immature Cotyledons and Embryos of Juglans regia cv. 'Xiangling'

    Directory of Open Access Journals (Sweden)

    Xiao-Dong CAI

    2013-12-01

    Full Text Available Callus holds great potential for biotechnology applications in plants. In this study, several experiments were performed to establish a protocol for in vitro induction of good quality callus by using immature cotyledons and embryos of Juglans regia cv. ‘Xiangling’ as the explants. DKW basal media supplemented with different concentrations of plant growth regulators were applied to determine the induction rate, texture and color of the recovered callus. Results showed that the optimum culture medium for the callus induction from these explants was a DKW basal medium consisting of 1 mg/L BA, 2 mg/L KT 250 mg/L glutamine, 500 mg/L casein hydrolysate, 200 mg/L Vc, 50 g/L sucrose and 2.8 g/L Gelrite (i.e., the T8 medium. The average rate of callus induction on the medium reached 80.7% for the immature cotyledons and 66.7% for the immature embryos, and all the callus was characterized by a friable, creamy or yellow appearance. In vitro proliferation of callus was not very successful as a result of the fact that the friable callus with creamy or yellow color grew very slowly, whereas the fast-growing callus was easy to turn brown and died eventually. Even though further investigation will be required to overcome the problem of browning and assess the regeneration ability of the callus, this approach for the production of friable callus may have valuable applications for walnut and other woody trees.

  3. The Tolerance Ability to Low Temperature of Radopholus similis Reduced Cultured Longterm on Carrot Callus and Recovered Rejuvenated on Anthurium andraeanum%胡萝卜愈伤组织上长期培养和红掌上复壮繁殖对相似穿孔线虫耐低温能力的影响

    Institute of Scientific and Technical Information of China (English)

    武玉环; 谢辉; 徐春玲; 张超; 金惺惺

    2013-01-01

    The enduring fitness to low temperature of six isolates of Radopholus similis from ornamental plants was studied through longterm culture propagation on carrot callus and rejuvenation culturing on An-thurium andraeanum. The tolerance to low temperature of R. similis was determined by culturing on carrot callus. The results showed that the tolerance to low temperature of R. similis reduced after successive culturing on carrot callus for six generations, and the survival and fecundity decreased significantly. After rejuvenation on A. andraeanum, the survival and fecundity of R. similis could recover in different degrees, namely the resistant capacity to low temperature was enhanced. The tolerance to low temperature of R. similis was different in longterm cultured carrot callus according to the varying nematode groups. Similarly, the degree of recovery of the resistance to low temperature was also different after being cultured on A. andraeanum.%利用从观赏植物上截获的相似穿孔线虫Radopholus similis6个种群,研究在胡萝卜愈伤组织上长期培养繁殖和在红掌Anthurium andraeanum上接种繁殖复壮后,相似穿孔线虫耐低温能力的变化.将经2种方式培养繁殖的相似穿孔线虫接种在胡萝卜愈伤组织上,测定其耐低温能力.结果表明,在胡萝卜愈伤组织上长期(6代)连续培养后,相似穿孔线虫的耐低温能力降低,在低温下的生存能力和繁殖能力明显降低;通过在自然寄主植物红掌根系上接种繁殖后,相似穿孔线虫在低温下的生存和繁殖能力又可得到不同程度的恢复,即耐低温能力得到复壮.胡萝卜愈伤组织上长期培养繁殖对相似穿孔线虫不同群体耐低温能力的影响不同,相似穿孔线虫不同种群在红掌上繁殖后耐低温能力的恢复程度也有所不同.

  4. An improved culturing method for opiine fruit fly parasitoids and its application to parasitoid monitoring in the field.

    Science.gov (United States)

    Masry, Ayad; Furlong, Michael J; Clarke, Anthony R; Cunningham, John Paul

    2016-09-21

    Good culturing methods play an important role in the study of insect behavior and its application to pest management. Here, we describe and validate a new method for rearing the parasitoid wasp, Diachasmimorpha kraussii, which attacks some of the world's worst fruit fly pests and is an internationally used biological control agent. Our method differs from standard culturing approaches by presenting adult wasps with host-infested artificial media within a "culturing bag," which mimics a natural (fruit) oviposition substrate. In laboratory trials using wild collected D. kraussii, the culturing bag method was compared to the use of host-infested nectarines, and a commonly used laboratory method of presenting host-infested artificial media within Petri dishes. The culturing bag method proved to be a significant improvement on both methods, combining the advantages of high host survival in artificial media with parasitism levels that were the equivalent to those recorded using host-infested fruits. In our field study, culturing bags infested with the Queensland fruit fly, Bactrocera tryoni, and hung in a mixed peach and nectarine orchard proved to be effective "artificial fruits" attracting wild D. kraussii for oviposition. Significantly more adult wasps were reared from the culturing bags compared to field collected fruits. This was shown to be due to higher fruit fly larval density in the bags, as similar percentage parasitism rates were found between the culturing bags and ripe fruits. We discuss how this cheap, time-efficient method could be applied to collecting and monitoring wild D. kraussii populations in orchards, and assist in maintaining genetic variability in parasitoid laboratory cultures.

  5. Protocol for callus induction and somatic embryogenesis in Moso Bamboo.

    Science.gov (United States)

    Yuan, Jin-Ling; Yue, Jin-Jun; Wu, Xiao-Li; Gu, Xiao-Ping

    2013-01-01

    Moso bamboo [Phyllostachys heterocycla var. pubescens (Mazel ex J. Houz.) Ohwi] is one of the most important forest crops in China and the rest of Asia. Although many sympodial bamboo tissue culture protocols have been established, there is no protocol available for plantlet regeneration as indicated by callus induction for monopodial bamboos, such as Moso bamboo. In the present report, embryogenic callus induction, embryoid development, and germination were established for Moso bamboo from zygotic seed embryos. Callus was initiated from zygotic embryos after 10-20 d culture on MS media supplemented with 4.0 mg/L 2, 4-D and 0.1 mg/L zeatin (ZT). About 50% of the explants produced calli, and nearly 15% of the calli were found to be embryogenic in nature. These embryogenic calli can be subcultured for proliferation in the Murashige and Skoog media (MS) supplemented with 0.5-2.0 mg/L 2, 4-D. These calli were found to have maintained their capacity for regeneration even after one year of subculture. The viable somatic embryoids regenerated in medium containing 5.0-7.0 mg/L ZT. Nearly 5% of the calli were found capable of regenerating into plantlets directly in MS medium containing 5.0-7.0 mg/L ZT. Root growth was more pronounced when the plantlets were transferred to medium containing 2.0 mg/L NAA. After 30 days of subculture, the plantlets were transferred to a greenhouse.

  6. Protocol for callus induction and somatic embryogenesis in Moso Bamboo.

    Directory of Open Access Journals (Sweden)

    Jin-Ling Yuan

    Full Text Available Moso bamboo [Phyllostachys heterocycla var. pubescens (Mazel ex J. Houz. Ohwi] is one of the most important forest crops in China and the rest of Asia. Although many sympodial bamboo tissue culture protocols have been established, there is no protocol available for plantlet regeneration as indicated by callus induction for monopodial bamboos, such as Moso bamboo. In the present report, embryogenic callus induction, embryoid development, and germination were established for Moso bamboo from zygotic seed embryos. Callus was initiated from zygotic embryos after 10-20 d culture on MS media supplemented with 4.0 mg/L 2, 4-D and 0.1 mg/L zeatin (ZT. About 50% of the explants produced calli, and nearly 15% of the calli were found to be embryogenic in nature. These embryogenic calli can be subcultured for proliferation in the Murashige and Skoog media (MS supplemented with 0.5-2.0 mg/L 2, 4-D. These calli were found to have maintained their capacity for regeneration even after one year of subculture. The viable somatic embryoids regenerated in medium containing 5.0-7.0 mg/L ZT. Nearly 5% of the calli were found capable of regenerating into plantlets directly in MS medium containing 5.0-7.0 mg/L ZT. Root growth was more pronounced when the plantlets were transferred to medium containing 2.0 mg/L NAA. After 30 days of subculture, the plantlets were transferred to a greenhouse.

  7. [Effect of methyl jasmonate on salidroside and polysaccharide accumulation in Rhodiola sachalinensis callus].

    Science.gov (United States)

    Li, Yang; Lian, Mei-Lan; Shao, Chun-Hui; Jin, Chan; Piao, Xuan-Chun

    2014-11-01

    To provide a new material for producing the Rhodiolasachalinensis products, the effect of methyl jasmonate (MeJA) on callus biomass and effective compound accumulation of Rhodiolasachalinensis was studied. The calluses-cultured in 3 L-air lift balloon type bioreactor were treated with MeJA after 20 d of bioreactor culture and the effect of MeJA concentration and treatment days on callus biomass, salidroside or polysaccharide accumulation and superoxide dismutase (SOD) and peroxidase (POD) activities were investigated. The callus biomass was not significantly different after MeJA treatment (125) for 0-6 d but obviously decreased after 6 d treatment. The maximum salidroside or polysaccharide contents and SOD or POD activities were found after 4 d treatment of MeJA. MeJA concentration significantly affected callus biomass and effective compound accumulation, biomass decreased at MeJA concentrations higher than 125 μmol x L(-1). However, the effective compound contents were determined at higher MeJA concentration, and the highest salidroside and polysaccharide accumulation was found at 225 and 275 μmol x L(-1) MeJA, respectively and the maximum SOD and POD activities was found at 225 μmol x L(-1) MeJA. The effective compound contents in callus were compared with field-grown plants. Salidroside contents in calluses were 1.1-fold and 2. 4-fold more than in plant roots and stem or leave, respectively. Polysaccharide content in calluses were 3. 6-fold and 8.0-fold more than in plant roots and stem or leave, respectively. Salidorside and polysaccharide in Rhodiolasachalinensiscalluses improved by MeJA treatment, 225 μmol x L(-1) MeJA and 4 d treatment were optimal. The effective compound contents in callus were obviously higher than in field-grown plants. Therefore, bioreactor culture is efficient for obtaining mass effective compounds of Rhodiolasachalinensis by culturing calluses. This method could provide an alternative material source for production of

  8. Effect of Hypergravity and Phytohormones on Isoflavonoid Accumulation in Soybean ( Glycine max. L.) Callus

    Science.gov (United States)

    Downey, Peter J.; Levine, Lanfang H.; Musgrave, Mary E.; McKeon-Bennett, Michelle; Moane, Siobhán

    2013-02-01

    The objective of this study was to explore the potential interaction between gravity and growth hormones on isoflavonoid accumulation. Soybean callus ( Glycine max (L.) Merr. cv. `Acme') was grown in the dark for 16 days at 22 °C in a growth medium supplemented with four different combinations of phytohormones and subjected to 4- g and 8- g forces simulated in a centrifuge and 1- g in an adjacent stationary control. Isoflavonoid aglycones and their glycoside concentrations (daidzein, genistein, daidzin, 6″-O-malonyl-7-O-glucosyl daidzein, genistin, 6″-O-malonyl-7-O-glucosyl genistein) were determined in the resulting tissues. Although gravity had no significant impact on callus growth, increasing gravity reduced isoflavonoid accumulation in three out of the four phytohormone-supplemented culture media. The ratio of the auxin naphthalene acetic acid (NAA) to the cytokinin benzylaminopurine (BAP) was found to have profound effect on both callus growth and isoflavonoid accumulation. The cytokinin BAP promoted callus tissue growth, but reduced callus isoflavonoid suggesting the isoflavonoid accumulation was not keeping pace with the cell growth in the elevated concentration of BAP. On the other hand, NAA had little or no effect on callus growth, but greatly enhanced isoflavonoid accumulation. Interactive effects of gravity and hormone on isoflavonoid accumulation were evident and its implication to the mechanism by which gravity exerts the effect on plant secondary metabolites is discussed.

  9. Efficient callus formation and plant regeneration are heritable characters in sugar beet (Beta vulgaris L.).

    Science.gov (United States)

    Kagami, Hiroyo; Taguchi, Kazunori; Arakawa, Takumi; Kuroda, Yosuke; Tamagake, Hideto; Kubo, Tomohiko

    2016-01-01

    Obtaining dedifferentiated cells (callus) that can regenerate into whole plants is not always feasible for many plant species. Sugar beet is known to be recalcitrant for dedifferentiation and plant regeneration. These difficulties were major obstacles for obtaining transgenic sugar beets through an Agrobacterium-mediated transformation procedure. The sugar beet line 'NK-219mm-O' is an exceptional line that forms callus efficiently and is easy to regenerate, but the inheritance of these characters was unknown. Another concern was whether these characters could coexist with an annual habitat that makes it possible to breed short life-cycle sugar beet suitable for molecular genetic analysis. Five sugar beet lines including NK-219mm-O were crossed with each other and subjected to in vitro culture to form callus. F1s with a NK-219mm-O background generally formed callus efficiently compared to the others, indicating that efficient callus formation is heritable. The regeneration potential was examined based on the phenotypes of calli after placement on regeneration medium. Five phenotypes were observed, of which two phenotypes regenerated shoots or somatic embryo-like structures. Vascular differentiation was evident in regenerable calli, whereas non-regenerable calli lacked normally developed vascular tissues. In a half-diallel cross, the callus-formation efficiency and the regeneration potential of reciprocal F1s progeny having a NK-219mm-O background were high. Finally, we crossed NK-219mm-O with an annual line that had a poor in vitro performance. The callus-formation efficiency and the regeneration potential of reciprocal F1 were high. The regenerated plants showed an annual habitat. Efficient callus formation and the high plant regeneration potential of NK-219mm-O were inherited and expressed in the F1. The annual habitat does not impair these high in vitro performances.

  10. Isolation, quantification and anti-microbial screening of flavonoids from callus of Vitex negundo Linn

    Directory of Open Access Journals (Sweden)

    Keerti Gautam

    2013-01-01

    Full Text Available Context: Natural habitats for medicinal plant are disappearing fast and together with environmental instabilities, it is increasingly difficult to acquire plant-derived compounds. This has prompted scientists to consider the possibilities of using cell cultures as an alternative supply for the production of plant natural products. Aim: The present investigation deals with the establishment of callus cultures from leaf explants of Vitex negundo, to extract flavonoids from the raised callus and to assess their anti-microbial potential. Materials and Methods: Callus culture of Vitex negundo has been established on Murashige and Skoog′s medium supplemented with 2, 4-D using leaf explants. Growth index (GI was calculated for the tissues obtained after 2, 4, 6 and 8 weeks, respectively. Calli of 2, 4, 6 and 8 weeks old were used for extraction of total flavonoids (free and bound using standard method. Flavonoid extract of 6 weeks old callus with maximum GI and maximum flavonoid content was then tested against Staphylococcus aureus, Agrobacterium tumefaciens (bacteria, Candida albicans (yeast for their anti-microbial potential. Minimum inhibitory concentration, minimum bactericidal/fungicidal concentrations and total activity were also evaluated. Results: GI was found maximum in 6 weeks old tissue (0.76. Total flavonoids from 6 week old tissue showed maximum zone of inhibition against A. tumefaciens. Kaempferol was identified from total flavonoids of 6 week old callus through, thin layer chromatography (TLC, preparative TLC, Melting point and Infra red spectral studies. Content of kaempferol in 6-week-old calli was also estimated using standard Spectrophotometric method, which came out to be 54.25 μg/mg d.wt (dry weight of calli. Conclusion: The protocol of callus induction of V. negundo opens new vistas that could facilitate phytochemical production, extraction of pharmaceuticals and their screening against different human pathogens from the callus

  11. Analysis of indole compounds in fruiting bodies and in mycelia from in vitro cultures of Calocera viscosa (Basidiomycota

    Directory of Open Access Journals (Sweden)

    Bożena Muszyńska

    2013-12-01

    Full Text Available Calocera viscosa (Pers.: Fr. Fr. (Basidiomycota from Dacrymycetaceae family is a widespread species of mushroom in Poland. The aim of this study was to investigate the content of indole compounds in fruiting bodies and in mycelium cultured in vitro on solid and liquid medium of this species. Fruiting bodies of Calocera viscosa were collected in coniferous forests in south Poland and were used to derive in vitro cultures. The optimal medium composition for cultures was determined. Fresh material: fruiting bodies and mycelium from culture in vitro was frozen and then dried by lyophilization. The crushed dry biomass was extracted with petroleum ether to remove oil fraction, material was dried and extracted with methanol. Analysis of indole compounds was performed in methanol extracts using chromatographic methods: TLC, UV Vis, EIMS and HPLC. This analysis presented in all three extracts the following indole compounds: L-tryptophan, 5-hydroxytryptophan, 5-methyltryptophan, melatonin and indole (contents fluctuated in the range: 0.37 to 11.88 mg/100 g d.w.. 5-hydroxytryptophan contents in all extracts were significant and amounted to 11.88 mg/100 g d.w. in fruiting bodies, and 11.42 in mycelium from liquid cultures and and 10.59 in mycelium from solid cultures. In addition, the fruiting bodies and mycelium from cultures on liquid medium revealed the presence of serotonin (0.39 and 3.19 mg/100 g d.w. respectively.

  12. Laboratory culture of the myxomycetes: formation of fruiting bodies of Didymium bahiense and its plasmodial production of Makaluvamine A.

    Science.gov (United States)

    Ishibashi, M; Iwasaki, T; Imai, S; Sakamoto, S; Yamaguchi, K; Ito, A

    2001-01-01

    Laboratory cultures of more than 100 strains of myxomycetes were investigated, and the spore germinations of six strains were observed. The plasmodium of the myxomycete Didymium bahiense was cultured on oatmeal agar plates in a laboratory. The formation of fruiting bodies was observed in a plate culture. From the cultured organisms, a marine sponge metabolite, makaluvamine A (1), was isolated and identified on the basis of spectral data.

  13. Ethanol Metabolism in Calluses of Several Selected Plant Species on Two Typical Plant-Growth-Regulator Balanced Media

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    For investigation on the characteristics of ethanol metabolism in tissues of different plant species,calluses from eight selected plant species were cultured on medium supplemented with ethanol in tightly sealed culture flasks. Changes of the ethanol level were detected by gas chromatography. During the culture period, the calluses of tobacco, potato and petunia were able to catabolize exogenous ethanol, resulting in the prominent decline of the ethanol level in the medium. The calluses of melon and peanut were also able to ca-tabolize ethanol but with lower efficiency. The other three calluses of carrot, soybean and rice did not catabo-lize ethanol but instead produced small to large amount of ethanol, resulting in the increase of the ethanol level in the media. It was also found that changing the balance between auxin and cytokinin could influence only the ethanol metabolism efficiency but could not change the metabolism patterns on ethanol of the cul-tured calluses. It can be concluded that, ethanol metabolism pattern of calluses in cultures is an innate physi-ological characteristic of the respective plant species.

  14. The Effect of Chitosan on Organogenesis of Oil Palm Embryo-Derived Callus

    Directory of Open Access Journals (Sweden)

    Kantamaht KANCHANAPOOM

    2010-06-01

    Full Text Available Zygotic embryos of oil palm (Elaeis guineensis Jacq. var. tenera were excised and cultured on MS medium containing 3 mg/l 2, 4-D either with or without 0.05% activated charcoal (AC. Improved growth of embryos was obtained on MS medium supplemented with 0.05% AC. Callus cultures were initiated from embryos, young leaves and roots on MS medium containing 2, 4-D, NAA and 0.05% AC. On these media, two morphologically distinct types of white and yellow compact calluses were produced. Green shoots regenerated after several transfers of the yellow compact calluses from zygotic embryos to MS medium supplemented with 15 mg/l chitosan either with or without 5 mg/l 2, 4-D. Histological sectioning revealed that regenerated shoots originated from a clump of meristematic cells that had dense cytoplasm. Regenerated shoots rooted when transferred to MS medium in the presence of 0.05% AC. Transfer of plantlets to soil was achieved. Callus from young seedling leaves and roots did not regenerate shoots or roots in medium containing 2, 4-D or TDZ, with or without chitosan. This finding shows that chitosan can initiate organogenesis in oil palm callus.

  15. Comparação de diferentes metodologias para obtenção de cultura de calos de Stevia rebaudiana (Bertoni Bertoni = Comparison of different methodologies for obtaining callus cultures from Stevia rebaudiana (Bertoni Bertoni

    Directory of Open Access Journals (Sweden)

    Ana Paula Patrão

    2007-07-01

    Full Text Available As folhas de Stevia rebaudiana (Bertoni Bertoni (Asteraceae contêm glicosídeos diterpenóides (GDS, que são cerca de 300 vezes mais doce que a sacarose a 4%. O objetivo deste estudo foi avaliar a formação de calos, a partir de folhas obtidas in vivo e in vitro de S. rebaudiana em dois meios já descritos na literatura: Murashige e Skoog (MS, suplementado com 3 mg L-1 de ácido 2,4-diclorofenóxiacético (2,4-D, e o MS suplementado com 1 mg L-1 de ácido naftalenoacético (ANA e 0,5 mg L-1 de 6-benzilaminopurina 6-BAP e um desenvolvido em nosso laboratório o Woody Plant Medium (WPM, suplementado com 6 mg L-1 de ANA e 4 mg L-1 de cinetina (CIN. Os explantes obtidos in vitro iniciaram a formação de calos um pouco mais rapidamente que os das folhas de plantas advindas da natureza. A utilização dos nutrientes do meio WPM, associada a uma combinação de fitorreguladores adequada, proporcionou velocidade de indução e multiplicação de calos bem maiores que as apresentadas nos meios que empregaram os nutrientes do MS. Novos experimentos serão realizados, depois de alcançada a estabilidade genética dos calos, visando avaliar a capacidade destes em biossintetizar os GDS.The leaves from Stevia rebaudiana (Bertoni Bertoni (Asteraceae contain diterpenoid glycosides (GDS, which are almost 300 times sweeter than sucrose at 4%. The subject of this study was to evaluate the callus-formation from in vivo and in vitro leaves of Stevia rebaudiana in two already described in literature: Murashige and Skoog (MS supplemented with 3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D; MS supplemented with 1 mg L-1 of naphthaleneacetic acid (NAA and 0.5 mg L-1 of 6-benzylaminopurine (6-BAP and other developed in our laboratory the Woody Plant Medium (WPM with 6 mg L-1 of NAA and 4 mg L-1 of Kinetin (KIN. The explants obtained in vitro initiated callus formation faster than leaves from natural plants. The utilization of WPM nutrients, associated with an

  16. Fruit extract of the medicinal plant Crataegus oxyacantha exerts genotoxic and mutagenic effects in cultured cells.

    Science.gov (United States)

    de Quadros, Ana Paula Oliveira; Mazzeo, Dania Elisa Christofoletti; Marin-Morales, Maria Aparecida; Perazzo, Fábio Ferreira; Rosa, Paulo Cesar Pires; Maistro, Edson Luis

    2017-01-01

    Crataegus oxyacantha, a plant of the Rosaceae family also known "English hawthorn, haw, maybush, or whitethorn," has long been used for medicinal purposes such as digestive disorders, hyperlipidemia, dyspnea, inducing diuresis, and preventing kidney stones. However, the predominant use of this plant has been to treat cardiovascular disorders. Due to a lack of studies on the genotoxicity of C. oxyacantha, this investigation was undertaken to determine whether its fruit extract exerts cytotoxic, genotoxic, or clastogenic/aneugenic effects in leukocytes and HepG2 (liver hepatocellular carcinoma) cultured human cells, or mutagenic effects in TA100 and TA98 strains of Salmonella typhimurium bacterium. Genotoxicity analysis showed that the extract produced no marked genotoxic effects at concentrations of 2.5 or 5 µg/ml in either cell type; however, at concentrations of 10 µg/ml or higher significant DNA damage was detected. The micronucleus test also demonstrated that concentrations of 10 µg/ml or higher produced clastogenic/aneugenic responses. In the Ames test, the extract induced mutagenic effects in TA98 strain of S. typhimurium with metabolic activation at all tested concentrations (2.5 to 500 µg/ml). Data indicate that, under certain experimental conditions, the fruit extract of C. oxyacantha exerts genotoxic and clastogenic/aneugenic effects in cultured human cells, and with metabolism mutagenicity occurs in bacteria cells.

  17. Availability of commonly consumed and culturally specific fruits and vegetables in African-american and Latino neighborhoods.

    Science.gov (United States)

    Grigsby-Toussaint, Diana S; Zenk, Shannon N; Odoms-Young, Angela; Ruggiero, Laurie; Moise, Imelda

    2010-05-01

    Although the importance of culture in shaping individual dietary behaviors is well-documented, cultural food preferences have received limited attention in research on the neighborhood food environment. The purpose of this study was to assess the availability of commonly consumed and culturally specific fruits and vegetables in retail food stores located in majority African-American and Latino neighborhoods in southwest Chicago, IL. A cross-sectional survey of 115 stores (15% grocery stores, 85% convenience/corner stores) in African-American neighborhoods and 110 stores (45% grocery stores, 55% convenience/corner stores) in Latino neighborhoods was conducted between May and August of 2006. chi(2) tests were used to assess differences in the availability (presence/absence) of commonly consumed (n=25) and culturally specific fruits and vegetables for African Americans (n=16 varieties) and Latinos (n=18 varieties). Stores located in neighborhoods in which the majority of residents were African American or Latino were more likely to carry fresh fruits and vegetables that were culturally relevant to the dominant group. For example, grocery stores located in Latino neighborhoods were more likely to carry chayote (82.0% vs 17.6%, P<0.05), whereas grocery stores located in African-American neighborhoods were more likely to carry black-eyed peas (52.9% vs 20%, P<0.05). Most stores, however, carried fewer than 50% of commonly consumed or culturally specific fruits and vegetables. Findings from this study highlight that limited availability of culturally specific as well as commonly consumed fruits and vegetables in the neighborhood may be a barrier to fruit and vegetable consumption among African Americans and Latinos.

  18. Preliminary Study of Embryogenic Callus Induction using Immature Banana Male Flowers and Stems of Tissue Culture Seedlings%香蕉雄花和组培苗假茎胚性愈伤组织诱导的初步研究

    Institute of Scientific and Technical Information of China (English)

    林志聪; 赵辉; 曾会才

    2012-01-01

    从外植体来源、生长调节剂种类及浓度等方面研究影响巴西蕉胚性愈伤组织诱导的因素。结果表明,以未成熟雄花为外植体,MS培养基+1 mg/L 2,4-D+1 mg/L IAA+1 mg/L NAA较适宜胚性愈伤组织诱导,26℃暗培养条件下,未成熟雄花的胚性愈伤组织诱导率为3%~4%。在MS培养基+0.7 mg/L 2,4-D+4 mg/L 6-BA中,以不同品种假茎为外植体的脱分化愈伤组织诱导率最高可达75%,但只有巴西(青杆)和海南本地米蕉可以诱导出胚性愈伤组织,诱导率分别为0.1%和0.5%.%An attempt was made to study factors which affect induction of embryogenic callus derived from the banana cultivar Brazil(Musa AAA Cavendish Subgroup cv.Brazil),including source of explants,plant hormones and concentration of plant hormones.Explants from immature male flowers were cultured on the MS at the presence of 2,4-D 4 mg/L + IAA 1 mg/L + NAA 1 mg/L and incubated in the dark at 26℃.The callus inducing rates were 3%~4% for immature male flowers.In the MS+2,4-D 0.7 mg/L+6-BA 4 mg/L,the different varieties of stems as explants for callus induction rate of up to 75%.However,only Banana Brazil(blue bar) and Hainan powder coke can induce embryogenic callus,induction rate was 0.1% and 0.5%.

  19. Protoplasts Culture Isolated from Friable Embryogenic Callus of Cassava and Plant Regeneration%木薯脆性胚性愈伤组织原生质体培养与植株再生

    Institute of Scientific and Technical Information of China (English)

    文峰; 肖诗鑫; 聂扬眉; 马秋香; 张鹏; 郭文武

    2012-01-01

    [Objective] The objective of this study is to establish an efficient system of protoplast regeneration for further developing protoplast fusion and transformation in cassava. [ Method ] Protoplasts were isolated from suspension cultures derived from friable embryogenic callus (FEC) of cassava genotype TMS60444. The highest protoplast yield obtained was 3.5x 106 protoplasts/g fresh weight. Viabilities of the protoplasts assessed by the fluorescein diacetate (FDA) were approximately 90%. Protoplasts were cultured in TM2G medium with liquid thin layer culture at densities of 5x105p/mL or 2x105p/mL. During the first 30 d, the medium was refreshed by 0.3 mol-L"1 TM2G fresh medium every 10 d. After that, the medium was refreshed by 0.25 mol-L-1 TM2G fresh medium every 10 d. After cultured for 45 d, calli of 1-2 mm were picked out and separately developed into embryos on MSN medium, into mature embryos on CMM medium, into shoots on CEM medium and into roots on MS medium. [Result] K was showed that all protoplasts cultured at density of 5x105p/mL developed into compact calli (could develop into embryos), protoplasts cultured at density of 2xl05 p/mL developed into compact calli and vacuolar calli (could not develop into embryos). A total of 1 479 compact calli were picked out and developed into 757 cotyledon embryos and regenerated 186 plants in the experiment. [Conclusion] The yield and viability of isolated protoplasts had been greatly increased, the bottleneck of predecessors mentioned was improved, and the efficiency of plant regeneration from protoplasts was promoted.%[目的]建立有效的木薯原生质体再生体系,为原生质体融合以及原生质体转化等研究奠定技术基础.[方法]酶解木薯品种TMS60444的脆性胚性愈伤组织(FEC)的悬浮系,分离原生质体的产量最高达3.5×106个/g,FDA检测其活性约90%.用TM2C培养基以液体浅层法分别在5 ×105个/mL和2× 105个/mL密度下培养,培养过程中前30 d用0.3 mol.L-1

  20. (Catharanthus roseus) tissue culture

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-18

    Aug 18, 2008 ... indole alkaloids in plant tissue cultures of C. roseus have. *Corresponding ... alkaloids from C. roseus cell cultures have failed (review- ed by Van der ..... that vinblastine occur in callus culture with differentiated roots. Dimeric ...

  1. The Role of Nitrobenzoic Acid Derivatives on Callus Induction and Plant Regeneration in Paulownia shan tong

    Directory of Open Access Journals (Sweden)

    Roxana Luca

    2016-11-01

    Full Text Available Paulownia is a multipurpose tree species with a rapid growth which is cultivated in several temperate zones worldwide for its economic importance. The present study was undertaken to investigate the effects of different alkanolamine salts of 4-nitrobenzoic acid on in vitro callus induction and plant regenerating in Paulownia shan tong.The alkanolamine salts, namely methylethanolamine 4-nitrobenzoate (4-NO2BA MMEA, diethanolamine 4-nitrobenzoate (4-NO2BA DEA and triethanolamine 4-nitrobenzoate (4-NO2BA TEA are new synthesized compounds with biological activity and low toxicity. The influence of title compounds on sprout length, number of internodes, number of leaves and number of ramifications has been studied at two different concentrations: 1 mg/L respectively 0.5 mg/L. The experimental data show that new tested compounds have a better growth effect on plant vegetable parties in report to control, the most eloquent results being obtained for 4-NO2BA MMEA. For callus induction 2,4-D hormonal balances were attempted and the development of callus was examined at two different concentrations. An induced callus development was observed regarding callus dimensions for all explants inoculated on culture medium supplemented with 2,4-D.

  2. In Vitro Callus Induction and Embryogenesis of Oil Palm (Elaeis guineensis Jacq. from Leaf Explants

    Directory of Open Access Journals (Sweden)

    DWI HAPSORO

    2011-06-01

    Full Text Available This research was to study in vitro callus induction and somatic embryogenesis in oil palm from leaf explants. Young leaf segments from mature oil palm were cultured on MS medium supplemented with different concentrations of 2,4-D with or without addition of 2 g/l activated charcoal (AC or 2,4-D and picloram. Embryogenesis induction was done using MS medium containing 2,4-D 450 M and benziladenine 4.4 M with 3g/l activated charcoal. The treatment of 2,4-D 15 M resulted in the highest percentage of callus induction. The treatment of 2,4-D and AC showed that 2,4-D 450 M and AC led to higher percentage of callus induction than that of 2,4-D 400 M and 2 g/l AC. Embryogenesis occured in 27 out of 250 clumps of primary callus was occurred after 2-3 times subcultures. Somatic embryo development occurred when the embryogenic callus was transferred on the same basal medium supplemented with casein hydrolysate with 1 M BA or growth regulator free basal medium with 2 g/l activated charcoal.

  3. Isolation of lactic acid bacteria with potential protective culture characteristics from fruits

    Science.gov (United States)

    Hashim, Nurul Huda; Sani, Norrakiah Abdullah

    2015-09-01

    Lactic acid bacteria are also known as beneficial microorganisms abundantly found in fermented food products. In this study, lactic acid bacteria were isolated from fresh cut fruits obtained from local markets. Throughout the isolation process from 11 samples of fruits, 225 presumptive lactic acid bacteria were isolated on MRS agar medium. After catalase and oxidase tests, 149 resulted to fit the characteristics of lactic acid bacteria. Further identification using Gram staining was conducted to identify the Gram positive bacteria. After this confirmation, the fermentation characteristics of these isolates were identified. It was found that 87 (58.4%) isolates were heterofermentative, while the rest of 62 (41.6%) are homofermentative lactic acid bacteria. Later, all these isolates were investigated for the ability to inhibit growth of Staphylococcus aureus using agar spot assay method. Seven (4.7%) isolates showed strong antagonistic capacity, while 127 (85.2%) and 8 (5.4%) isolates have medium and weak antagonistic capacity, respectively. The other 7 (4.7%) isolates indicated to have no antagonistic effect on S. aureus. Results support the potential of LAB isolated in this study which showed strong antagonistic activity against S. aureus may be manipulated to become protective cultures in food products. While the homofermentative or heterofermentative LAB can be utilized in fermentation of food and non-food products depending on the by-products required during the fermentation.

  4. Effect of UV-C on Drought Tolerance of Alfalfa (Medicago sativa Callus

    Directory of Open Access Journals (Sweden)

    Ali A. Ehsanpour

    2005-01-01

    Full Text Available Water stress and unfavorable climate decrease the growth and development of plant globally. Medicago is a valuable plant as a source of food for animal, foliage and for medicine. Since, Iran is located in a dry area; consequently, the study of plant responses to water stress is important. In this research UV-C radiation was used to induce physiological and genetic changes in alfalfa callus. In vitro grown calluses of M. sativa were exposed to UV-C for 0, 15, 30 and 60 min and then, were transferred to MS medium containing 0, 2, 5, 10, 20, 30% PEG. All explants were kept in the dark in the culture room. Calluses tolerate to osmotic stress were selected according to fresh weight. Results showed that radiation of UV-C for 60 min increased the osmotic tolerance and decreased the effect of drought stress.

  5. [Fresh fruit and occultism as ways to salvation: conversions in Leipzig's alternative culture at around 1900].

    Science.gov (United States)

    Bigalke, Bernadett

    2008-01-01

    During the time of the Wilhelmine Empire, there were multiple interdependencies between adherents of the life reform movement (vegetarians, naturopathists, nudists, etc.) and new religious movements such as esoteric groups like the theosophists in the alternative cultural milieu around 1900. These networks became visible in the form of double memberships in associations. However, there were also ambiguous affiliations, migration between groups and syncretistic beliefs without institutional belonging. The similarity between patterns of argumentation for this specific lifestyle and the congruence of chosen goals, ways and goods of salvation become particularly clear in this context. These forms of "methodical lifestyle" may lead to the development of a specific ethos or habitus (Max Weber). To illustrate these processes, this article analyses the report of a Leipzig lady who ate raw fruits and vegetables only, and examines her broader social context. Thereby the analysis will employ sociological theories of conversion to explain the case of Hedwig Bresch.

  6. Mature seed-derived callus of the model indica rice variety Kasalath is highly competent in Agrobacterium-mediated transformation.

    Science.gov (United States)

    Saika, Hiroaki; Toki, Seiichi

    2010-12-01

    We previously established an efficient Agrobacterium-mediated transformation system using primary calli derived from mature seeds of the model japonica rice variety Nipponbare. We expected that the shortened tissue culture period would reduce callus browning--a common problem with the indica transformation system during prolonged tissue culture in the undifferentiated state. In this study, we successfully applied our efficient transformation system to Kasalath--a model variety of indica rice. The Luc reporter system is sensitive enough to allow quantitative analysis of the competency of rice callus for Agrobacterium-mediated transformation. We unexpectedly discovered that primary callus of Kasalath exhibits a remarkably high competency for Agrobacterium-mediated transformation compared to Nipponbare. Southern blot analysis and Luc luminescence showed that independent transformation events in primary callus of Kasalath occurred successfully at ca. tenfold higher frequency than in Nipponbare, and single copy T-DNA integration was observed in ~40% of these events. We also compared the competency of secondary callus of Nipponbare and Kasalath and again found superior competency in Kasalath, although the identification and subsequent observation of independent transformation events in secondary callus is difficult due to the vigorous growth of both transformed and non-transformed cells. An efficient transformation system in Kasalath could facilitate the identification of QTL genes, since many QTL genes are analyzed in a Nipponbare × Kasalath genetic background. The higher transformation competency of Kasalath could be a useful trait in the establishment of highly efficient systems involving new transformation technologies such as gene targeting.

  7. In vitro callus induction and in vivo antioxidant activity of Passiflora foetida L. leaves

    Directory of Open Access Journals (Sweden)

    Rasool SN *

    2011-03-01

    Full Text Available Summary: Callus was induced efficiently from the leaf explants of Passiflora foetida L. (Passion fruit on Murashige and Skoog (MS medium containing 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D and 0.5 mg/l kinetin (KN after 16 days. Maximum callus (65% response, 610 mg fresh weight and 55 mg dry weight was formed on MS medium supplemented with low sucrose and salt concentration than that of high sucrose and salt concentration. The crude ethanolic extract prepared from the callus of Passiflora foetida L. leaf showed significant (PIndustrial relevance: CCl4 is a well established hepatotoxin. It induces chronic and acute injuries not only in liver also in kidneys, heart and brain. In this study, the hepatoprotective effects of ethanolic extract from the callus of Passiflora foetida leaves were studied and results were found very promising. To day a vast number of drugs are developed from plants possessing excellent therapeutic potential and less side effects compared to synthetic drugs which cause toxicity in various organs. Isolation and characterization of active principle from this extract could lead to the development of effective therapeutic drug in industry for the treatment of hepatic injury.

  8. Transcriptome analysis of Carica papaya embryogenic callus upon De-etiolated 1 (DET1 gene suppression

    Directory of Open Access Journals (Sweden)

    Diyana Jamaluddin

    2017-06-01

    Full Text Available Papaya is considered to be one of the most nutritional fruits. It is rich in vitamins, carotenoids, flavonoids and other phytonutrient which function as antioxidant in our body [1]. Previous studies revealed that the suppression of a negative regulator gene in photomorphogenesis, De-etiolated 1 (DET1 can improve the phytonutrient in tomato and canola without affecting the fruit quality [2,3]. This report contains the experimental data on high-throughput 3′ mRNA sequencing of transformed papaya callus upon DET1 gene suppression.

  9. Comparison of fracture site callus with iliac crest bone marrow as the source of plastic-adherent cells

    Directory of Open Access Journals (Sweden)

    Achmad Zaki

    2013-05-01

    Full Text Available Background: Red marrow has been described as the main source of mesenchymal stem cells although its aspiration and isolation from bone marrow was reported to have significant donor site morbidity. Since secondary bone healing occurs through formation of callus as the result of proliferation and differentiation of mesenchymal stem cells, callus may become alternative source for mesenchymal stem cells. In this study, we compared the number of plastic-adherent cells from fracture site callus and bone marrow of iliac crest after two and four weeks of culture.Methods: Sixteen New Zealand rabbits were fracturized at the femoral shaft. Then, these rabbits were taken care. After two weeks of fracturization, 3 mL iliac crest bone marrow aspiration and callus extraction of eight rabbits were cultured (group I. The other eight rabbits were treated equally after four weeks of fracturization (group II. Simultaneously, the cultures were observed after one and two weeks. Four weeks later, they were harvested. Cells were counted using Neubauer hemocytometer. The average number of cells between the sources and groups were statistically analyzed using the unpaired t-test. Results: In group I, there were 2.6 ± 0.1 x 104 cells in the culture of iliac crest bone marrow aspirate and 2.5 ± 0.1 x 104 cells in culture of callus extract from fracture site (p = 0.34. In group II, there were 2.7 ± 0.1 x 104 cells and 2.1 ± 0.1 x 104 cells, respectively (p < 0.001.Conclusion: Fracture site callus at the second week post-fracturization may be potential as source of plastic-adherent cells compared with iliac crest bone marrow. (Med J Indones. 2013;22:70-5Keywords: Bone marrow, fracture site callus, iliac crest, long bone, mesenchymal stem cell, plastic-adherent cells

  10. Meios de cultura, reguladores de crescimento e fontes de nitrogênio na regulação da calogênese do pau-brasil (Caesalpinia echinata Lam. Culture media, growth regulators and nitrogen sources in callus formation regulation of Brazilwood (Caesalpinia echinata Lam.

    Directory of Open Access Journals (Sweden)

    Elias Terra Werner

    2010-12-01

    Full Text Available A regulação da calogênese é o primeiro passo para a embriogênese somática indireta in vitro. Nesse sentido, foram investigados os efeitos de diferentes meios de cultura, reguladores de crescimento e fontes nitrogenadas no desenvolvimento de calos em tecidos foliares de Caesalpinia echinata (pau-brasil. Explantes foliares foram cultivados em meios de cultura MS, B5, White e WPM. Testou-se, também, o efeito dos compostos nitrogenados (NH4NO3, KNO3 e glutamina e a interação entre auxinas (2,4-D, AIA e AIB e citocininas (BAP e KIN na desdiferenciação celular indicado pela formação de calos. Os meios MS, B5 e White não foram significantemente diferentes e proporcionaram melhores resultados. Em meio MS contendo as fontes nitrogenadas 2,4 g L-1 NH4NO3 e 1,35 g L-1 glutamina + 4,11 g L-1 de KNO3 na presença de 1,0 mg L-1 2,4-D e 5,0 mg L-1 BAP ocorreu maior proliferação celular indicada pela massa fresca dos calos. As combinações dos reguladores 0,5 mg L-1 2,4-D + 5 mg L-1 BAP e 0,5 mg L-1 AIB + 5,0 mg L-1 BAP suplementado com 2,4 g L-1 NH4NO3 em meio MS estimularam o crescimento dos calos. De maneira geral, os calos apresentaram aspectos friáveis, não embriogênicos, com acúmulo de fenóis e presença de áreas meristemáticas (meristemóides.The regulation of callus formation is the first step to indirect somatic embryogenesis in vitro. Therefore, we investigated hormonal and nutrient factors on development of callus in leaf tissues of Caesalpinia echinata (Brazilwood. Explants were cultivated in culture media MS, B5, WPM, and White. The effect of nitrogen sources (NH4NO3, KNO3 and glutamine and the interaction between auxins (2,4-D, IAA and IBA and cytokinins (BAP and KIN were also evaluated. Media MS, B5 and White were not significantly different and provided better results. MS medium containing 2.4 g L-1 NH4NO3, 1.35 g L-1 glutamine, and 4.11 g L-1 of KNO3 in the presence of 1 mg.L-1 2,4-D and 5 mg.L-1 BAP increased cellular

  11. Callus Growth Kinetics of Physic Nut (Jatropha curcas L.) and Content of Fatty Acids from Crude Oil Obtained In Vitro.

    Science.gov (United States)

    da Luz Costa, Jefferson; da Silva, André Luís Lopes; Bier, Mário César Jucoski; Brondani, Gilvano Ebling; Gollo, André Luiz; Letti, Luiz Alberto Junior; Erasmo, Eduardo Andrea Lemus; Soccol, Carlos Ricardo

    2015-06-01

    The callus growth kinetics allows identifying the appropriate moment for callus pealing and monitoring the accumulation of primary and secondary metabolites. The physic nut (Jatropha curcas L.) is a plant species used for biofuel production due to its high oil content; however, this plant presents a great amount of bioactive compounds which can be useful for industry. The aim of this research was to establish a calli growth curve and to evaluate the fatty acid profile of crude oil extracted from callus. The callus growth kinetics presented a sigmoid standard curve with six distinct phases: lag, exponential, linear, deceleration, stationary, and decline. Total soluble sugars were higher at the inoculation day. Reducing sugars were higher at the inoculation day and at the 80th day. The highest percentage of ethereal extract (oil content) was obtained at the 120th day of culture, reaching 18 % of crude oil from the callus. The calli produced medium-chain and long-chain fatty acids (from 10 to 18 carbon atoms). The palmitic acid was the fatty acid with the highest proportion in oil (55.4 %). The lipid profile obtained in callus oil was different from the seed oil profile.

  12. Efeito de diferentes componentes no meio de cultura na produção de alcalóides em tecidos de calos de Cereus peruvianus (Cactaceae - DOI: 10.4025/actascibiolsci.v27i1.1355 Effect of different culture medium components on production of alkaloid in callus tissues of Cereus peruvianus (Cactaceae - DOI: 10.4025/actascibiolsci.v27i1.1355

    Directory of Open Access Journals (Sweden)

    Claudete Aparecida Mangolin

    2005-03-01

    Full Text Available Tecidos de calos em cultura prolongada de Cereus peruvianus foram utilizados para investigar a produção de alcalóides pelos calos quando mantidos em meio de cultura suplementados com diferentes concentrações de tirosina, com diferentes níveis de 2,4-D/cinetina, e contendo NaCl. Foi analisado também, o padrão de isozimas α- e β-esterases, para investigar a expressão diferencial de genes nos calos subcultivados com as diferentes concentrações de tirosina. A maior quantidade de alcalóides foi obtida a partir de calos mantidos em meio contendo 200 mg/L de tirosina. A adição de tirosina no meio induziu uma expressão gênica diferencial para a síntese de isozimas α- e β-esterases. Duas α-carboxilesterases (EC 3.1.1.1 e uma α/β-arilesterase (EC 3.1.1.2 foram induzidas nos calos, uma α/β-acetilesterase (EC 3.1.1.6 e uma α-carboxilesterase foram detectadas como bandas mais intensamente coradas, enquanto duas α/β-carboxilesterases não foram detectadas ou foram fracamente coradas após a adição de tirosina como precursor no meio de culturaThe production of alkaloids from Cereus peruvianus callus cultured in medium supplemented with different tyrosine concentrations, different 2,4-D/kinetin levels, and with NaCl was investigated using long-term callus culture. α- and β-Esterase isozyme patterns were also analyzed to investigate the differential gene expression in callus subcultured with the different tyrosine concentrations. The greatest amounts of alkaloids were obtained from callus maintained in medium containing 200 mg/L of tyrosine. Tyrosine induced a differential gene expression for the synthesis of specific α- and β -esterase isozymes. Two α-carboxylesterases (EC 3.1.1.1 and one α/β-arylesterase (EC 3.1.1.2 were induced in callus, one α/β-acetylesterase (EC 3.1.1.6 and two α-carboxylesterase were detected as more intensely stained while two α/β-carboxylesterases were absent or detected as more weakly stained

  13. Plant Regeneration by Callus-Mediated Protocorm-Like Body Induction of Anthurium andraeanum Hort.

    Institute of Scientific and Technical Information of China (English)

    YU Yi-xun; LIU Ling; LIU Juan-xu; WANG Jing

    2009-01-01

    This research aims at developing a plant regeneration system from leaf and petiole explants of Anthurium andraeanum Hort.,thereby establish a foundation for mass production and transformation.Using tissue culture technique,the conditions for callus induction,protocorm-like body (PLB) formation and plant regeneration from leaf explants and petiole of A.andraeanum,such as basal medium and plant growth regulator,were investigated.Totipotent callus was induced on a 1/2-strength MS medium containing 0.90 μmol L-1 2,4-dichlorophenoxyacetic acid (2,4-D) and 8.88 μmol L-1 N6-benzyladenine (BA).The callus exhibited complete hormone autonomy for growth and differentiation of PLBs.This callus proliferated well and was maintained by subculturing on 1/2 MS medium containing 0.90 μmol L-1 2,4-D and 4.44 μmol L-1 BA.On average,8 protocorm-like bodies could be obtained from a piece of 4 mm callus after being transferred to the 1/2 MS medium with 4.44 μtmol L-1 BA after 8 wk of culture.The regenerated PLBs formed shoots and roots on 1/2 MS medium.After 24 wk of culture on these medium,well-developed plantlets for potting were produced.An efficient micropropagation method was established for indirect PLB formation and plant regeneration from leaf and petiole ofA.andraeanum.

  14. Plant tissue culture techniques

    Directory of Open Access Journals (Sweden)

    Rolf Dieter Illg

    1991-01-01

    Full Text Available Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus or organized tissues or organs put in culture, under controlled sterile conditions.

  15. Plant tissue culture techniques

    OpenAIRE

    Rolf Dieter Illg

    1991-01-01

    Plant cell and tissue culture in a simple fashion refers to techniques which utilize either single plant cells, groups of unorganized cells (callus) or organized tissues or organs put in culture, under controlled sterile conditions.

  16. Embryogenic callus proliferation and regeneration conditions for genetic transformation of diverse sugarcane cultivars.

    Science.gov (United States)

    Basnayake, Shiromani W V; Moyle, Richard; Birch, Robert G

    2011-03-01

    Amenability to tissue culture stages required for gene transfer, selection and plant regeneration are the main determinants of genetic transformation efficiency via particle bombardment into sugarcane. The technique is moving from the experimental phase, where it is sufficient to work in a few amenable genotypes, to practical application in a diverse and changing set of elite cultivars. Therefore, we investigated the response to callus initiation, proliferation, regeneration and selection steps required for microprojectile-mediated transformation, in a diverse set of Australian sugarcane cultivars. 12 of 16 tested cultivars were sufficiently amenable to existing routine tissue-culture conditions for practical genetic transformation. Three cultivars required adjustments to 2,4-D levels during callus proliferation, geneticin concentration during selection, and/or light intensity during regeneration. One cultivar gave an extreme necrotic response in leaf spindle explants and produced no callus tissue under the tested culture conditions. It was helpful to obtain spindle explants for tissue culture from plants with good water supply for growth, especially for genotypes that were harder to culture. It was generally possible to obtain several independent transgenic plants per bombardment, with time in callus culture limited to 11-15 weeks. A caution with this efficient transformation system is that separate shoots arose from different primary transformed cells in more than half of tested calli after selection for geneticin resistance. The results across this diverse cultivar set are likely to be a useful guide to key variables for rapid optimisation of tissue culture conditions for efficient genetic transformation of other sugarcane cultivars.

  17. Investigations on some metabolites of Tecoma stans Juss. callus tissue. Part II. Chromatographical analysis of alkaloid and quinone compounds

    Directory of Open Access Journals (Sweden)

    Barbara Dohnal

    2015-01-01

    Full Text Available Tecoma stans Juss. callus tissue grown on Murashige-Mei Lie Lin (M-L and Murashige-Skoog (RT-k medium supplemented with Tecoma alkaloid precursors like lysine, mevalonic acid lactone and quinolinic acid, were investigated for their alkaloid content by thin-layer (TLC and paper (PC chromatography methods. The results were compared with those obtained by parallel analysis of greenhouse plant leaves. Seven alkaloid spots were detected in the leaf extracts, namely: actinidine, 4-noractinidine, boschniakine, tecomanine, two spots of skytanthine derivatives and one unidentified spot. Only l spot corresponding to skytanthine derivatives appeard in the extracts of callus tissues cultured on basal M-L medium or supplemented with lysine or mevalonic acid lactone. However, 3 alkaloid spots were revealed in extracts of homogeneous parenchyma-like callus tissues obtained o-n, RT-k medium. Beside the skytanthine derivatives rnentioned above, actinidine and tecomanine were identified, and moreover, in callus tissues growing on the same medium (RT-k supplemented with quinolinic acid, boschniakine was found. It was found that T. stans leaves as well as callus tissues cultured on RT-k medium contained traces of lapachol and another quinone-type compound.

  18. Suicidal tomato cells : programmed cell death in suspension-cultured tomato cells and ripening fruit

    NARCIS (Netherlands)

    Hoeberichts, F.A.

    2002-01-01

     Tomato fruit ripening involves a series of highly organised biochemical, physiological and structural changes that are under strict genetic control. The plant hormone ethylene (C 2 H 4 ), in synergy with certain developmental cues, regulates fruit ripening b

  19. 海南龙血树基于愈伤组织诱导途径的组培快繁技术体系优化%Optimization of Tissue Culture and Rapid Propagation Technical System Based on Callus Induction of Dracaena cambodiana

    Institute of Scientific and Technical Information of China (English)

    罗冠勇; 钟云芳; 宋希强; 戴好富; 杨冬华

    2012-01-01

    Used the lateral and top buds of Dracaena cambodiana as the explants to induce callus tissue with that differentiated to seedling and induced dragon's blood at the same time. The results are as follows: The optimized culture medium for callus tissue induction and differentiation: 1. 1/2MS+6-BA 2.0 mg/L+NAA 0.1 mg/L+ CM 50 ml/L+sugar 30 g/L+arrageenan 8.6 g/L, buds of induction were more and neat, induction rate was more than 90%; 2. The Optimized culture medium for subculture proliferation of buds: modified MS+6-BA 2.0 mg/L+NAA 0.1 mg/L+CM 50 ml/L+sugar 30 g/L+arrageenan 8.6 g/L, proliferation coefficient was up to 5~6; 3. he Optimized culture medium for rooting: without strong seedling stage directly from seedling to rooting stages, 1/2MS+NAA 0.4 mg/L+GA 1.0 mg/L+sugar 30 g/L+arrageenan 8.6 g/L, rooting rate was up to 100%. (4) After dragon's blood induction, separation and extraction, using thin—layer chromatography and HPLC—control, the study found that 14 ingredients had higher similarity between induced and natural dragon's blood in HPLC peak shape. The callus tissue culture technique gave consideration to the production of seedling and dragon's blood, and provided new ideas for an efficient, fast, convenient scale production in the future.%以海南龙血树侧芽和顶芽为外植体,诱导愈伤组织进而分化成苗,同时利用愈伤组织诱导血竭.研究结果如下:①诱导愈伤组织与分化最适培养基为:1/2 MS+6-BA 2.0mg/L+NNA 0.1 mg/L+CM 50 mL/L+糖30 g/L+卡拉胶8.6 g/L,诱导的愈伤组织结构致密、结实,分化的芽多,诱导达90%以上;②芽继代增殖最适培养基为:改良MS+6-BA 2.0 mg/L+NNA 0.1 mg/L+CM 50 mL/L+糖30 g/L+卡拉胶8.6 g/L,芽的长势好且整齐,增殖系数达5~6;③生根最适培养基为:不经过壮苗阶段直接从小苗进入生根阶段,1/2 MS+NNA 0.4 mg/L+GA 1.0 mg/L+糖30 g/L+卡拉胶8.6 g/L,根多且均匀,生根率达100%;④诱导出血竭后分离、提取龙血竭,经

  20. Synthetic effects of heavy metal ions on the phytochelatin induction in rice callus; Ine no phytochelatin yudo ni oyobosu jukinzoku ion no kyozon koka

    Energy Technology Data Exchange (ETDEWEB)

    Takatera, K.; Ono, Y.; Watanabe, T. [The University of Tokyo, Tokyo (Japan). Institute of Industrial Science

    1998-03-01

    This paper describes the result of investigating synergistic effects of heavy metal ions on phytochelatin (PC) in rice callus. The following matters were made clear: with regard to rice callus cultured in culture media added with Cd, Cu, Zn and Hg independently, measurements were carried out on heavy metals contained in SH group of PC and in PC by using a method that combines high pressure liquid chromatography (HPLC) with induction bonded plasma mass analysis; PC induction can be recognized in those cultured under presence of Cd and Cu, but PC incorporating heavy metals is not induced in those cultured under presence of Hg and Zn; and when PC synthesis amount was measured by gel filtration - HPLC with regard to rice callus cultured in a culture medium in which Cd coexists with any of Zn, Cu, Pb, Hg and Ni, the effect of impeding PC synthesis increases in the order of Zn, Pb, Cu, Ni and Hg. 7 figs., 1 tab.

  1. The Influence of salinity and nitrogen on tomato fruit quality and micronutrients concentration in hydroponic culture

    Directory of Open Access Journals (Sweden)

    S. Safarzadeh Shirazi

    2010-12-01

    Full Text Available Salinity plays an important role in the reduction of tomato growth, especially in arid and semi-arid regions. Nitrogen (N may increase tomato tolerance to salt stress by increasing plant growth. In order to investigate the interaction effect of salinity and nitrogen on tomato growth, fruit quality, and micronutrient concentration in tomato plants, a hydroponic experiment was conducted in a completely randomized design with three replications. Treatments included 3 salinity levels (0, 30 and 60 mM, and 3 nitrogen rates (0, 1.5, and 3 mM. Results indicated that salinity decreased tomato height, shoot and fruit fresh weight, and increased citric acid in tomato fruit and consequently caused blossom-end rot in tomato fruit. However, salinity improved fruit quality (flavor. The use of N increased plant height, shoot, and fruit fresh weight compared to control. By application of N to saline nutrient solution, plant height increased. Salinity increased concentration of iron (Fe, manganese (Mn, zinc (Zn, and copper (Cu in roots, as`well as Fe and Cu in fruit. Increasing N rates in nutrient solution increased micronutrients concentration in tomato roots. Interaction of 60 mM salinity and N decreased micronutrients concentration in root, compared to control. Our tentative conclusion shows that the addition of N to the nutrient solution may decrease detrimental effect of salinity on the growth of tomato plants.

  2. In vitro callus induction and in vivo antioxidant activity of Passiflora foetida L. leaves

    Directory of Open Access Journals (Sweden)

    A N Rasool

    2011-03-01

    Full Text Available Summary: Callus was induced efficiently from the leaf explants of Passiflora foetida L. (Passion fruit on Murashige and Skoog (MS medium containing 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D and 0.5 mg/l kinetin (KN after 16 days. Maximum callus (65% response, 610 mg fresh weight and 55 mg dry weight was formed on MS medium supplemented with low sucrose and salt concentration than that of high sucrose and salt concentration. The crude ethanolic extract prepared from the callus of Passiflora foetida L. leaf showed significant (P< 0.05 in vivo antioxidant activity in CCl4 induced hepatotoxicity model in rats. After 14 days oral feeding of the ethanolic extract (200 and 500 mg/kg b.wt resulted in the significant (P< 0.05 reduction of serum aspartate aminotransferase(AST and alanine aminotransferase (ALT and hepatic thiobarbutiric acid reacting substances (TBARS levels which were elevated by CCl4 challenge in rats. Antioxidant enzymes superoxide dismutase (SOD, catalase (CAT and glutathione peroxidase (GPX levels and reduced glutathione (GSH levels in liver were increased significantly (P< 0.05 which were reduced by CCl4 challenge in rats. Industrial relevance: CCl4 is a well established hepatotoxin. It induces chronic and acute injuries not only in liver also in kidneys, heart and brain. In this study, the hepatoprotective effects of ethanolic extract from the callus of Passiflora foetida leaves were studied and results were found very promising. To day a vast number of drugs are developed from plants possessing excellent therapeutic potential and less side effects compared to synthetic drugs which cause toxicity in various organs. Isolation and characterization of active principle from this extract could lead to the development of effective therapeutic drug in industry for the treatment of hepatic injury.

  3. CALLUS INDUCTION AND PLANT REGENERATION IN PUNICA GRANATUM L. ?NANA' FROM LEAF EXPLANTS

    Directory of Open Access Journals (Sweden)

    Alireza Bonyanpour

    2013-09-01

    Full Text Available ABSTRACT In this investigation, leaf explants of a local cultivar of dwarf pomegranate were placed on Murashige and Skoog (1962 (MS medium supplemented with various concentrations of 6-benzyl adenin (BA and naphthalene acetic acid (NAA for callus induction. After 40 days, maximum callus induction was observed on a media containing 1 mg L-1 BA and 0.2 to 0.4 mg L-1 NAA. However, the highest callus growth was obtained on a medium containing 1 mg L-1 BA and 1 mg L-1 NAA. The highest number of shoots (7 shoots per explants was obtained by transferring the calli to the media containing 5 mg L-1 BA with 0.1 mg L-1 NAA. Maximum shoot proliferation was observed when shoots were cultured on woody plant medium (WPM supplemented with 5 mg L-1 kinetin (Kin. In this treatment, after 4 subcultures, 36 shoots were produced from one original explant. Among treatments used in rooting experiments, shoots cultured on WPM medium containing 0.2 mg L-1 indol butyric acid (IBA had the maximum root percentage (100% and good root growth (2.06 cm mean length and 2 roots in each explants. Rooted plantlets were cultured in a soil mixture containing vermiculite (60%, perlite (30% and coco peat (10% v/v. After 2 months, 80% of plants survived and transferred to the greenhouse.

  4. Green and non-green callus induction from excised rice (Oryza sativa) embryos: effects of exogenous plant growth regulators

    Science.gov (United States)

    Kim, D.; Brock, T. G.; Kaufman, P. B.

    1992-01-01

    Calli were induced either from excised rice embryos or from whole seeds in the presence of 1 to 5 mg l-1 NAA. After 12 days of culture, calli were induced only from excised rice embryos. We found that excised embryos accumulated NAA up to 6 times higher concentration than did whole seeds. In the presence of 1 to 5 mg l-1 NAA and 2 to 10 mg l-1 kinetin, chlorophyllous calli were induced from excised rice embryos. Chlorophyll contents in the callus tissue increased with increasing kinetin concentration while percent callus induction decreased. The total chlorophyll content was linearly correlated with the ratio of kinetin to NAA in the medium.

  5. Callus induction and plant regeneration from mature zygotic embryos of a tetraploid Alstroemeria (A. pelegrina × A. psittacina).

    Science.gov (United States)

    Hutchinson, M J; Tsujita, J M; Saxena, P K

    1994-12-01

    A simple procedure was developed to induce callus growth and whole plant regeneration for a tetraploid cultivar of Alstroemeria. The callus, induced from mature zygotic embryos cultured on a medium supplemented with 20 μM kinetin with 10 or 20 μM NAA, could be maintained for one year without any loss of regeneration potential. Maximum frequency of regeneration (40%) was obtained with calli maintained on the medium containing 20 μM kinetin and 20 μM NAA. Whole plant regeneration occurred via somatic embryogenesis in the absence of growth regulators and the plantlets grew to maturity and flowered in the greenhouse conditions.

  6. Amino Acid and Secondary Metabolite Production in Embryogenic and Non-Embryogenic Callus of Fingerroot Ginger (Boesenbergia rotunda.

    Directory of Open Access Journals (Sweden)

    Theresa Lee Mei Ng

    Full Text Available Interest in the medicinal properties of secondary metabolites of Boesenbergia rotunda (fingerroot ginger has led to investigations into tissue culture of this plant. In this study, we profiled its primary and secondary metabolites, as well as hormones of embryogenic and non-embryogenic (dry and watery callus and shoot base, Ultra Performance Liquid Chromatography-Mass Spectrometry together with histological characterization. Metabolite profiling showed relatively higher levels of glutamine, arginine and lysine in embryogenic callus than in dry and watery calli, while shoot base tissue showed an intermediate level of primary metabolites. For the five secondary metabolites analyzed (ie. panduratin, pinocembrin, pinostrobin, cardamonin and alpinetin, shoot base had the highest concentrations, followed by watery, dry and embryogenic calli. Furthermore, intracellular auxin levels were found to decrease from dry to watery calli, followed by shoot base and finally embryogenic calli. Our morphological observations showed the presence of fibrils on the cell surface of embryogenic callus while diphenylboric acid 2-aminoethylester staining indicated the presence of flavonoids in both dry and embryogenic calli. Periodic acid-Schiff staining showed that shoot base and dry and embryogenic calli contained starch reserves while none were found in watery callus. This study identified several primary metabolites that could be used as markers of embryogenic cells in B. rotunda, while secondary metabolite analysis indicated that biosynthesis pathways of these important metabolites may not be active in callus and embryogenic tissue.

  7. Comparative Effects of Auxins, Jasmonic Acid and Salicylic Acid on Callus Initiation and Organogenesis in Vigna mungo (L. Hepper Using Hypocotyl Explant

    Directory of Open Access Journals (Sweden)

    K. Lingakumar

    2014-03-01

    Full Text Available An attempt has been made to compare the effects of conventional phytohormones like IAA, NAA, BAP to the recent hormones viz., JA and SA in callus induction in blackgram using hypocotyl as explant source. Instead of testing the hormone individually, a combination of hormones was used to induce callus and organogenesis. A significant callusing response was noticed in MS medium supplemented with 0.5 ppm of 2,4-D and 1 ppm of IAA. Likewise, Salicylic acid at 1 ppm induced high percentage of callus induction proving its efficiency in inducing the root formation. Jasmonic acid induced a high percentage of callus induction at 1.5 ppm and greater rooting response than SA. Organogenic callus was observed at JA and SA supplementation. The combination of NAA, BAP and SA showed that 0.75 & 1.5 ppm of NAA, 1 ppm of BAP and 0.5 ppm of SA showed a better response in culture medium. Thus, the combination of these hormones seems serve as efficient growth supplements for in vitro culture of many agricultural crops.

  8. Factors influencing callus induction and plant regeneration of ...

    African Journals Online (AJOL)

    ajl yemi

    2012-01-12

    Jan 12, 2012 ... order to optimize the callus induction and plant rege- neration from ... To determine the role of basal medium for callus induction and plant .... Chu CC, Wang CS, Sun CC, Hsu C, Yin KC, Chu CY, Bi FY (1975). Establishment of ...

  9. Determination of Emodin and Chrysophanol Contents. in Callus of Cassia tora L. Leaf%决明子叶愈伤组织中大黄素与大黄酚含量的测定

    Institute of Scientific and Technical Information of China (English)

    周嘉裕; 廖海; 李利平; 刘乐; 贺葵邦; 马丽

    2008-01-01

    [Objective] Through inductive culture, emedin and ehrysophanol contents in cassia seed were increased. [Method]MS culture medium was used to bourgcon seedlings of cassia seed and to induce cotyledon callus of cassia seed while HPLC method was adopted to determine emodin and chry-sophanol contents. [Result] Emedin and chrysophanol contents in cotyledon callus were 0.099% and 0.312%, respectively, while they were 0.029% and 0.190% respectively in cassia seed. [Conclusion]The method of inducing cotyledon callus was helpful for increasing emedin content in cassia seed.

  10. Polyamines as salinity biochemical marker in callus of eucalyptus urograndis

    Directory of Open Access Journals (Sweden)

    Giuseppina Lima Pace Pereira

    2003-01-01

    Full Text Available Biochemical markers have been used for the analysis of plant cells submitted to several types of stress, among them salinity. This work aimed at analyzing the effect of saline stress in callus of Eucalyptus urograndis on polyamine contents. Explants (hypocotyls obtained from seeds were inoculated in callus inductive medium, submitted to different levels of NaCl and analyzed at 10, 20 and 30 days after the inoculation. The free polyamines were extracted, isolated and quantified using TLC (Thin-Layer Chromatography. Putrescine content was higher and a fall in the spermidine content was observed in callus submitted to salinity condition. The results showed that polyamine accumulation is related to NaCl exposure in callus of Eucalyptus urograndis. The decrease in spermine content could be used as a biochemical marker for Eucalyptus callus subjected to salinity.

  11. Vitamin C and total phenols quantification in calli of native passion fruit induced by combinations of Picloram and Kinetin

    Directory of Open Access Journals (Sweden)

    Fabiane Aparecida Artioli-Coelho

    2015-08-01

    Full Text Available Brazil is one of the center of origin of passion fruit and has an important natural variability of the genus Passiflora. Several wild species of this genus are resistant to some pests and diseases and many are considered as medicinal. The aim of this research was to induce callus from in vitro Passiflora gibertii leaf explants for quantification of vitamin C and total phenols. Once the appropriate auxin/cytokine balance promotes callus formation and may optimize the production of secondary compounds and vitamins, calli were induced using a half-strength MS medium with a combination of the auxin Picloran (PIC and the cytokine Kinetin (KIN. The vitamin C and total phenols were quantified by colorimetric methods from calli after different culture periods. The calli induction was strongly dependent of the combination PIC/KIN. It was observed high vitamin C content (94.8mg 100g-1 during the calli induction period in MS+4.14µM PIC+ 0.207µM KIN. Higher PIC/KIN concentrations promoted an increase in the vitamin C content after three subcultures. The higher PIC (8.28µM/KIN (0.828µM concentration was the higher was the total phenols production (66mg tannic acid 100g-1 of fresh callus during the calli induction period.

  12. Propagation of jarrah (Eucalyptus marginata) by organ and tissue culture

    Energy Technology Data Exchange (ETDEWEB)

    Bennett, M.J.; McComb, J.A.

    1982-01-01

    Micropropagation methods are described for the production of clonal lines from Eucalyptus marginata (jarrah) seedlings. Nodal explants from mature trees can also yield shoot cultures, but a high frequency of contamination occurs among such explants. Uncontaminated callus cultures can be produced from mature trees by culturing stamen filaments and shoots can subsequently be regenerated from this callus. The rooting percentage of shoot cultures from either nodes or stamen callus of mature trees is low compared with that from seedling explants. Considerable variation was observed between trees in the ability of stamen callus to regenerate shoots and in the frequency of rooting. (Refs. 27)

  13. Comparative study of antibacterial and antifugal activity of callus culture and adult plants extracts from Alternanthera maritima (Amaranthaceae Estudo comparativo da atividade antibacteriana e antifúngica de extratos obtidos da cultura de calos e da planta adulta de Alternanthera maritima (Amaranthaceae

    Directory of Open Access Journals (Sweden)

    Marcos J. Salvador

    2004-06-01

    Full Text Available The aim of this study was to evaluate the antibacterial and antifungal activity of callus culture (two different hormonal combination culture medium and adult plants (two collect extracts from Alternanthera maritima (Amaranthaceae investigating the maintenance of antimicrobial activity in vivo and in vitro. The antibacterial and antifungal activity was determined by the agar-well diffusion method against thirty strains of microorganisms including Gram-positive and Gram-negative bacteria, yeasts and dermatophytes. All the organic crude extracts studied were bioactive. Extracts of aerial parts and roots of adult plants collected during the same period of years of 1995 and 1998 (Restinga de Maricá (RJ, collect 1 and 2 inhibited the growth of several microorganisms (bacteria, yeasts and dermatophytes with inhibition halo between 6 and 20 mm. Plant cell callus culture extracts obtained from two culture conditions were also bioactive. Thus, the positive results suggest that the A. maritima extracts should be further studied to determine the bioactive chemical compounds as well as to understand the possible mechanisms of action and evaluate their toxicity looking toward a pharmaceutical employment.Neste estudo procedeu-se a avaliação da atividade antibacteriana e antifúngica dos extratos brutos de Alternanthera maritima (Amaranthaceae planta in natura de duas coletas distintas e obtidos por cultura de células buscando-se averiguar a manutenção da atividade antimicrobiana dos extratos obtidos da planta in vivo e in vitro. A ação antibacteriana e antifúngica foi determinada pelo método de difusão em ágar (técnica do poço utilizando-se trinta cepas de microrganismos indicadores (bactérias Gram-positivas e Gram-negativas, leveduras e dermatófitos. Todos os extratos obtidos com solventes orgânicos avaliados apresentaram-se bioativos com halos de inibição de 6 a 20 mm. Os extratos da planta in natura das duas coletas (Restinga de Marica

  14. The effect of organic manures in soilless culture on shelf-life extension of strawberry fruit (Fragaria × ananassa Duch. cv. Camarosa

    Directory of Open Access Journals (Sweden)

    S. Bidaki

    2014-07-01

    Full Text Available   Growing media could affect quality and shelf life of strawberry fruits. In the present study, effect of seven growing media including a control medium (50% cocopeat + 50% perlite v/v , and media supplemented with either 10, 20 and 40% of vermicompost or cattle manure in a completely randomized design with three replications was studied on postharvest quality and shelf life extension of Camarosa strawberry fruits . Freshly picked strawberry fruits were submerged in 1 mM putrescine solution for 5 minutes and after being air-dried at room temperature, were kept in plastic containers at 4 ºC refrigerator. After 15 days, total soluble solids ( TSS, total acidity (TA, TSS/TA ratio, fruit firmness and the percentage of fruit weight loss were determined . The results of analysis of variance showed that growing media had significant effect on fruit firmness, TA, TSS, TSS/TA ratio and percentage of fruit weight loss. The lowest fruit weight loss (16.2% belonged to 10% cattle manure medium and the highest weight loss (36% was observed in control medium. The highest fruit TSS content (19.13% was observed in the growing medium containing 40% vermicompost. Since soilless culture growing media of strawberry are pathogen-free, no fruit was rotted before or after the harvest. Therefore, by using organic fertilizers in growing media, not only there is no need to use chemical fungicides, but also it would be possible to increase postharvest shelf life of strawberry fruits.

  15. Regeneração de plantas híbridas entre Lycopersicon esculentum e L. peruvianum a partir de calos com dois anos de cultura in vitro Regeneration of Lycopersicon esculentum X L. peruvianum hybrid plants from two year old callus culture

    Directory of Open Access Journals (Sweden)

    Walter José Siqueira

    1988-01-01

    Full Text Available Calos obtidos da cultura in vitro de embrião imaturo do cruzamento interespecífico L. esculentum x L. peruvianum, praticamente perderam a capacidade morfogenética, após dois anos de subcultura. Na tentativa de recuperação do processo de organogênese desses calos, realizaram-se dois experimentos, utilizando-se os fitorreguladores ácido indolacético (IAA e 6-benziladenina (6-BA, cujas concentrações foram combinadas em dialélicos de 5 x 5 e 3 x 3. A composição de sais minerais e vitaminas baseou-se no meio de Murashige e Skoog, adicionando-se sacarose a 3% e ágar a 0,8%, e ajustando-se o pH final dos meios de cultura para 5,5. As condições para o dialélico 5 x 5 foram fotoperíodo de 16 horas de luz a 600 lux e temperatura de 25 ± 3°C. No dialélico 3 x 3, os tratamentos foram mantidos em câmara de crescimento a 2.000 lux, sob a mesma variação de temperatura e fotoperíodo. Em cada frasco, inoculou-se um calo com cerca de 1 cm³, totalizando quinze repetições. Avaliaram-se o desenvolvimento de calos, atribuindo-se uma escala de notas de 1 a 5, e a presença de plantas (organogênese após 30 dias de cultura. Observou-se o número total de plantas por tratamento, bem como o desenvolvimento das plantas em centímetro. No dialélico 5 x 5, a organogênese foi apenas incipiente em três tratamentos, porém as melhores combinações para o desenvolvimento dos calos foram de 0,5, 2,5 e 5,0µM de IAA com 2,5µM de 6-BA. No dialélico 3 x 3, houve a indução de plantas em sete tratamentos, sendo mais eficientes 25 e 50µM de 6-BA, sem auxina. O tratamento de 0,5 e 10,0µM de IAA e 6-BA, respectivamente, permitiu simultaneamente o crescimento de calos e a regeneração de plantas. Nota-se a influência das condições ambientais de manutenção das culturas, principalmente da intensidade de luz.Callus from Lycopersicon esculentum x L. peruvianum interespecific hybrids cultured in vitro lost their morphogenetic abilities after two

  16. Acetylcholine suppresses shoot formation and callusing in leaf explants of in vitro raised seedlings of tomato, Lycopersicon esculentum Miller var. Pusa Ruby.

    Science.gov (United States)

    Bamel, Kiran; Gupta, Rajendra; Gupta, Shirish C

    2016-06-02

    We present experimental evidence to show that acetylcholine (ACh) causes decrease in shoot formation in leaf explants of tomato (Lycopersicon esculentum Miller var Pusa Ruby) when cultured on shoot regeneration medium. The optimum response was obtained at 10(-4) M ACh-enriched medium. ACh also causes decrease in percentage of cultures forming callus and reduces the callus mass. Inhibitors of enzymatic hydrolysis of ACh, neostigmine and physostigmine, also suppresses callogenesis and caulogenesis. On the other hand, the breakdown products of Ach, choline and acetate, do not alter the morphogenic response induced on the shoot regeneration medium. Neostigmine showed optimal reduction in shoot formation at 10(-5) M. The explants cultured on neostigmine augmented medium showed decline in the activity of ACh hydrolyzing enzyme acetylcholinesterase. ACh and neostigmine added together showed marked reduction in callus mass. These results strongly support the role of ACh as a natural regulator of morphogenesis in tomato plants.

  17. Effect of Different Plant Growth Regulators on Callus Induction and in vitro Rapid Propagation of Wild Petunia Juss.

    Institute of Scientific and Technical Information of China (English)

    Yan ZHAO; Na XU; Zhongyou MA; Wei LIU

    2012-01-01

    In this study, the seeds of wild Petunia Juss. were used as explants to in- vestigate the optimal condition for tissue culture. Several different kinds and concen- trations of growth regulators were adopted to produce more multiple bud clumps, callus or roots in this study. The experiments may provide experimental foundation for the rapid propagation technology and establishment of tissue culture system for wild Petunia Juss.

  18. 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.

  19. In vitro callus induction and micropropagation of Thymus persicus (Lamiaceae, an endangered medicinal plant

    Directory of Open Access Journals (Sweden)

    Ziba Bakhtiar

    2016-03-01

    Full Text Available This is the first attempt towards an efficient regeneration protocol for an endangered and valuable medicinal plant, Thymus persicus using in vitro callus induction and indirect organogenesis. Callus induction was performed on MS medium supplemented with different concentrations of NAA and 2,4-D, alone or in combination with BAP and KN. Maximum callus induction (100% was achieved from internode explants cultured on MS medium fortified with 2.0 mg L–1 NAA and 0.5 mg L–1 KN. The highest frequency of shoot multiplication (96% was observed with 2.0 mg L–1 BAP+1.0 mg L–1 NAA. The maximum number of rootlets (16.6 ± 1.4 was induced on half-strength MS medium with 0.5 and 1.0 mg L–1 IBA. Rooted plantlets were then successfully grown and acclimatized in the greenhouse with a 70-85% survival rate. The benefits of the protocol described here include all-year-round application, germplasm conservation, suitability for commercial production and also for the biotechnological production of pentacyclic triterpenoids.

  20. Hyperplastic callus formation in osteogenesis imperfecta. A case report

    Energy Technology Data Exchange (ETDEWEB)

    Burchardt, A.J. (Depts. of Radiology and Pediatric Orthopedic Surgery, Rigshospitalet, Copenhagen Univ. (Denmark)); Wagner, A.A. (Depts. of Radiology and Pediatric Orthopedic Surgery, Rigshospitalet, Copenhagen Univ. (Denmark)); Basse, P. (Depts. of Radiology and Pediatric Orthopedic Surgery, Rigshospitalet, Copenhagen Univ. (Denmark))

    1994-09-01

    We report a case of bilateral hyperplastic callus formation as a complication of fracture in a patient with osteogenesis imperfecta. The clinical and radiographic findings and the differential diagnosis are discussed. (orig.).

  1. Effects of plant growth regulators on callus induction from Cananga ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-06-17

    Jun 17, 2009 ... the dried flowers are used against malaria, and the fresh flowers are pounded into a .... were terminated due to several problems such as browning, low callus ... Innovation of Malaysia (MOSTI) for Post-Graduate. Research ...

  2. CALLUS INDUCTION FROM 15 CARNATION (DIANTHUS CARYOPHYLLUS L.) CULTIVARS

    OpenAIRE

    Teixeira da Silva, Jaime A.

    2014-01-01

    Plant growth regulators (PGRs) were used to induce callus in 15 carnation (Dianthus caryophyllus L.; Caryophyllaceae) cultivars: Orange Sherbert, Avalanche, Magenta, La France, Stripe Red, Marie, Concerto PVP, Snap, Lucky Pierot, Cinnamon Tea, White Love, Siberia, Magesta, Spark Bruno, and Honono no Estejo. Seeds were initially sown on autoclaved moistened filter paper and internodes of surface-sterilized seedlings were used as explants. Most callus was induced in the presence of 0.5 mg/L α-n...

  3. Improvement of friable callus production of Boerhaavia paniculata Rich and the investigation of its lipid profile by GC/MS.

    Science.gov (United States)

    Souza, Joanne M M; Berkov, Strahill; Santos, Alberdan S

    2014-09-01

    In this study, a protocol to induce high amount of friable callus of Boerhaavia paniculata RICH and a lipidomics technique were applied to investigate the profile of lipids to relate to those present in the roots of this plant that presented anti-inflammatory activity in the crude hexane extract. The callus culture was induced from seeds in solidified Murashige and Skoog medium containing different amounts of glucose and different concentrations of 2,4-Dichlorophenoxyacetic acid. The explants were kept in a germination chamber at 30±2°C with a photoperiod of 16 h under light intensity of 27 µmol m-2 s-1 for 4 weeks. The best results for friable callus formation and development of the biomass were obtained in the treatment containing 2.26 µM 2.4-D and glucose (1.5 %; w/v). Lipidomics techniques were applied in hexane fraction showing higher concentrations of the steroids β-sitosterol (3.53 mg/100 g dc-dry cells), and fatty acids, especially 2-hydroxy-tetracosanoic acid (0.34 mg/100 g dc), eicosanoic acid (86.25 mg/100 g dc), stearic acid (420.83 mg/100 g dc), tetradecanoic acid (10.74 mg/100 g dc) and linoleic acid (100.61 mg/100 g dc). The lipid profile of callus versus that found in the roots of wild plant is described in this work.

  4. Improvement of friable callus production of Boerhaavia paniculata Rich and the investigation of its lipid profile by GC/MS

    Directory of Open Access Journals (Sweden)

    JOANNE M.M. SOUZA

    2014-09-01

    Full Text Available In this study, a protocol to induce high amount of friable callus of Boerhaavia paniculata RICH and a lipidomics technique were applied to investigate the profile of lipids to relate to those present in the roots of this plant that presented anti-inflammatory activity in the crude hexane extract. The callus culture was induced from seeds in solidified Murashige and Skoog medium containing different amounts of glucose and different concentrations of 2,4-Dichlorophenoxyacetic acid. The explants were kept in a germination chamber at 30±2°C with a photoperiod of 16 h under light intensity of 27 µmol m–2 s–1 for 4 weeks. The best results for friable callus formation and development of the biomass were obtained in the treatment containing 2.26 µM 2.4-D and glucose (1.5 %; w/v. Lipidomics techniques were applied in hexane fraction showing higher concentrations of the steroids β-sitosterol (3.53 mg/100 g dc–dry cells, and fatty acids, especially 2-hydroxy-tetracosanoic acid (0.34 mg/100 g dc, eicosanoic acid (86.25 mg/100 g dc, stearic acid (420.83 mg/100 g dc, tetradecanoic acid (10.74 mg/100 g dc and linoleic acid (100.61 mg/100 g dc. The lipid profile of callus versus that found in the roots of wild plant is described in this work.

  5. Selection of yeast starter culture strains for the production of marula fruit wines and distillates.

    Science.gov (United States)

    Fundira, M; Blom, M; Pretorius, I S; van Rensburg, P

    2002-03-13

    Juice of the Sclerocarya birrea subsp. caffra (marula) fruit was fermented by indigenous microflora and different commercial Saccharomyces cerevisiae yeast strains at different temperatures, namely, 15 and 30 degrees C. Volatile acids, esters, and higher alcohols were quantified in the wine and distillates, and the results were interpreted using a multivariate analysis of variance and an average linkage cluster analysis. Significant differences between 15 and 30 degrees C and also among yeasts with respect to volatile compounds were observed. Yeast strains VIN7 and FC consistently produced wines and final distillates significantly different from the other strains. A panel of tasters and marula and brandy producers was asked to select wines and distillates that had an acceptable and typical marula "nose". They were also asked to detect the differences among wines and distillates fermented with the same yeast strain at different temperatures.

  6. Determination of Suitable Microspore Stage and Callus Induction from Anthers of Kenaf (Hibiscus cannabinus L.

    Directory of Open Access Journals (Sweden)

    Ahmed Mahmood Ibrahim

    2014-01-01

    Full Text Available Kenaf (Hibiscus cannabinus L. is one of the important species of Hibiscus cultivated for fiber. Availability of homozygous parent lines is prerequisite to the use of the heterosis effect reproducible in hybrid breeding. The production of haploid plants by anther culture followed by chromosome doubling can be achieved in short period compared with inbred lines by conventional method that requires self pollination of parent material. In this research, the effects of the microspore developmental stage, time of flower collection, various pretreatments, different combinations of hormones, and culture condition on anther culture of KB6 variety of Kenaf were studied. Young flower buds with immature anthers at the appropriate stage of microspore development were sterilized and the anthers were carefully dissected from the flower buds and subjected to various pretreatments and different combinations of hormones like NAA, 2,4-D, Kinetin, BAP, and TDZ to induce callus. The best microspore development stage of the flower buds was about 6–8 mm long collected 1-2 weeks after flower initiation. At that stage, the microspores were at the uninucleate stage which was suitable for culture. The best callus induction frequency was 90% in the optimized semisolid MS medium fortified with 3.0 mg/L BAP + 3.0 mg/L NAA.

  7. In vitro callus induction and plantlet regeneration of Achyranthes aspera L., a high value medicinal plant

    Institute of Scientific and Technical Information of China (English)

    Monokesh Kumer Sen; Shamima Nasrin; Shahedur Rahman; Abu Hena Mostofa Jamal

    2014-01-01

    Objective:To study callus induction from different explants (internode, leaf, root) and in vitro plantlets propagation from medicinally important plant Achyranthes aspera L. Methods:Sterilized explants were prepared by using 0.1%HgCl2 and 0.5%Bavistin and callus was obtained when cultured onto Murashige Skoog’s (MS) medium by using different concentrations and combination of 2,4-D, NAA, BAP, IAA, IBA with 3%sucrose and 0.8%agar. Induced callus was immediately transferred to MS medium containing at different concentrations of phytohormones for shootlets and rootlets induction respectively. Results:Sterilization treatment of 0.1%HgCl2 for 2-3 min and Bavistin 0.5%for 10-12 min showed the highest percentage of asepsis and survival rate. Maximum induction of callus was obtained from a combination of 2.0 mg/L 2,4-D and 0.5 mg/L NAA from leaf. Highest shootlets number (4.83±0.17) and length (3.8±0.16) cm were observed on full strength MS medium when fortified with BAP 4.0 mg/L and KIN 0.5 mg/L. Concerted efforts of BAP 2.0 mg/L and NAA 0.5 mg/L on full strength MS medium showed highest leaf number (6.77±0.94). In vitro raised shoots were allowed to root on different strengths of MS medium fortified with IAA and IBA at different concentrations. Experimentally, 3.0 mg/L IBA was enabled to induce maximum rootlets number (10.0±9.82) on full strength MS medium. Afterwards, regenerated shoots with well developed roots were successfully subjected to hardening process and were acclimatized. The survived plantlets showed 66.67%survival frequency without any morphological abnormality. Conclusions: The results demonstrated that different explants were good source of callus induction, morphology analysis as well as indirect plantlets regeneration.

  8. Callus patterns in femur lengthening using a monolateral external fixator

    Energy Technology Data Exchange (ETDEWEB)

    Isaac, Dileep; Fernandez, Harry; Song, Hae-Ryong; Kim, Tae-Young; Kumar Shyam, Ashok [Korea University Medical Centre, Guro Hospital, Department of Orthopaedic Surgery, Rare Diseases Institute, Seoul (Korea); Lee, Seok-Hyun [Dongguk University International Hospital, Department of Orthopaedic Surgery, Ilsan (Korea); Lee, Jong-Chan [Korea University, Department of Statistics, Seoul (Korea)

    2008-04-15

    We studied the callus pattern seen in femoral lengthening using monolateral external fixator to determine whether callus types and shapes can predict the final outcome of the procedure. The radiographs of 41 cases of femoral lengthening (33 unilateral and 8 bilateral) in 33 patients with a mean age 11.9 years (range 4-22 years) were retrospectively analysed by four observers and classified into different shapes and types in concordance with the Ru Li classification. The classification was tested for concurrence and reproducibility by inter-observer studies. An average of 6.2 cm of lengthening (range 3-10.8) was achieved with an external fixator index of 36.5 (range 20.9-55.3). The fusiform type of callus (fixator index 32.04, maturation index 21.6) showed the best result followed by the cylinder type of callus (fixator index 35.7, maturation index 22.3) and the lateral type of callus (fixator index 33.2, maturation index 21.5). However, the concave type of callus showed poor results with a fixator index of 49.4 and a maturation index of 37.1. The homogeneous path showed the best results (fixator index 30.4, maturation index 18.6). The heterogeneous path also showed good results (fixator index 36.4, maturation index 23.9). The mixed path (fixator index 42.5, maturation index 30.8) and the lucent path (fixator index 45.1, maturation index 32.8) showed poor results. Analysis of the callus pattern helps the surgeon to predict the outcome of the procedure and guide him in planning any additional interventions if necessary. (orig.)

  9. Effect of colchicine applied to callus of mangosteen on morphological changes of regenerated plants

    Directory of Open Access Journals (Sweden)

    Te-chato, S.

    2000-07-01

    Full Text Available Calli of mangosteen raised on callus induction medium (CIM at 20 days of culture were submerged in various concentrations of colchicine. The cultures were incubated on a rotary shaker and shaken at 80 rpm for 2 hours. In the case of high concentrations of colchicine (500 and 750 mg/l, the calli were cultured directily on the colchicine-containing media for 90 days. The results showed that leaves of regenerated shoots contained more chlorophyll b than those of control. However, shoot size, leaf area and leaf number tended to decrease while number and lenght of roots increased. A high concentration of colchicine, 500 mg/l, produced shoots with brown-callused leaves. However, roots could be induced from these shoots to from complete plantlets. Some of the intact leaves produced a large number of shoots. Colchicine at 750 mg/l gave some morphological abnormalities with 5 roots per shoot. A small shoot could produce a large healthy root. Some shoots produced 3 leaves per whorl.

  10. RAP-011 augments callus formation in closed fractures in rats.

    Science.gov (United States)

    Morse, Alyson; Cheng, Tegan L; Peacock, Lauren; Mikulec, Kathy; Little, David G; Schindeler, Aaron

    2016-02-01

    ACE-011 is a bone anabolic agent generated by fusing the extracellular domain of the Activin Type 2A receptor (ActRIIA) to an IgG-Fc. The orthopedic utility of ACE-011 was investigated using a murine analogue, RAP-011. Initially, a rat closed fracture model was tested using bi-weekly (biw) 10 mg/kg RAP-011. RAP-011 significantly increased callus length and callus bone volume (BV, +43% at 6w, p RAP-011(p)] and "sustained" [RAP-011(s)] regimens of biw 0.6mg/kg × 2, 0.35mg/kg × 3 and 0.18mg/kg × 2, 0.1mg/kg × 7 respectively, compared with PTH(1-34) (25 μg/kg/d) and vehicle controls. RAP-011 treatments gave modest increases in callus length and callus BV at 6w (p RAP-011 is effective in promoting bone formation during repair, but optimizing callus bone quality will require further investigation. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  11. Primary structures of two ribonucleases from ginseng calluses - New members of the PR-10 family of intracellular pathogenesis-related plant proteins

    NARCIS (Netherlands)

    Moiseyev, GP; Fedoreyeva, LI; Zhuravlev, YN; Yasnetskaya, E; Jekel, PA; Beintema, JJ

    1997-01-01

    The amino acid sequences of two ribonucleases from a callus cell culture of Panax ginseng were determined, The two sequences differ at 26% of the amino acid positions, Homology was found with a large family of intracellular pathogenesis-related proteins, food allergens and tree pollen allergens from

  12. Primary structures of two ribonucleases from ginseng calluses - New members of the PR-10 family of intracellular pathogenesis-related plant proteins

    NARCIS (Netherlands)

    Moiseyev, GP; Fedoreyeva, LI; Zhuravlev, YN; Yasnetskaya, E; Jekel, PA; Beintema, JJ

    1997-01-01

    The amino acid sequences of two ribonucleases from a callus cell culture of Panax ginseng were determined, The two sequences differ at 26% of the amino acid positions, Homology was found with a large family of intracellular pathogenesis-related proteins, food allergens and tree pollen allergens from

  13. Forskolin synthesis in in vitro cultures of Coleus forskohlii Briq transformed with Agrobacterium tumefaciens.

    Science.gov (United States)

    Mukherjee, S; Ghosh, B; Jha, S

    1996-05-01

    Agrobacterium tumefaciens mediated tumor tissue and shooty teratomas of Coleus forskohlii were cultured in vitro. Forskolin was detected in tumorous callus (0.002%), rhizogenic callus (0.011%) and root cultures (0.014%), but not in shooty teratomas. Forskolin synthesis and accumulation in tumorous C. forskohlii cultures may permit the elucidation of diterpene metabolism in this species.

  14. Cytokinin Activity in Avocado Seeds during Fruit Development.

    Science.gov (United States)

    Blumenfeld, A; Gazit, S

    1970-08-01

    The soybean callus bioassay was used to determine levels of cytokinin activity in avocado (Persea americana) seeds.In the embryo, levels are high during the early stages of development, but diminish as the fruit grows. The level of cytokinin activity in the endosperm is very high throughout the period that this tissue exists. The seed coats have very high activity levels while the fruit is young, reaching values comparable with those found in the endosperm. The activity level falls as the rate of fruit growth slows down and disappears completely by the time the seed coats shrivel at approximately the same time the fruit reaches "horticultural maturity".

  15. CALLUS INDUCTION FROM 15 CARNATION (DIANTHUS CARYOPHYLLUS L. CULTIVARS

    Directory of Open Access Journals (Sweden)

    Jaime A. TEIXEIRA DA SILVA

    2014-12-01

    Full Text Available Plant growth regulators (PGRs were used to induce callus in 15 carnation (Dianthus caryophyllus L.; Caryophyllaceae cultivars: Orange Sherbert, Avalanche, Magenta, La France, Stripe Red, Marie, Concerto PVP, Snap, Lucky Pierot, Cinnamon Tea, White Love, Siberia, Magesta, Spark Bruno, and Honono no Estejo. Seeds were initially sown on autoclaved moistened filter paper and internodes of surface-sterilized seedlings were used as explants. Most callus was induced in the presence of 0.5 mg/L α-naphthaleneacetic acid used together with 1 mg/L 6-benzyladenine or 1 mg/L 2,4-dichlorophenoxyacetic acid on basal Murashige and Skoog medium. Callus is not a desirable method to clonally propagate important germplasm but can serve as one possible way of deriving periclinal mutants as a result of somaclonal variation.

  16. Callus formation and organogenesis of tomato (Lycopersicon ...

    African Journals Online (AJOL)

    USER

    2010-07-12

    Jul 12, 2010 ... In vitro culture response was assessed in tomato (Lycopersicon esculentum Mill. c. v. Omdurman) for ... successful application of plant tissue culture presupposes the establishment of ..... Kino (Pterocarpus marsupium Roxb.) ...

  17. CALOGÊNESE EM ÓVULOS DE ESPÉCIES E VARIEDADES DE CITRUS CALLUS INDUCTION FROM CITRUS SPECIES AND VARIETIES

    Directory of Open Access Journals (Sweden)

    ROBERTO PEDROSO DE OLIVEIRA

    2001-08-01

    used as protoplast sources: Cleopatra, Sunki and 'Swatow' mandarins, Rangpur lime selections Santa Cruz and Santa Bárbara, Volkamer lemon and Mazoe rough lemon, 'Hamlin' sweet orange selections CNPMF 04 and CNPMF 20, sour orange selections Common and Narrow Leaf, Troyer citrange, Citrus amblycarpa and Microcitrus papuana. The study focused on obtaining and cultivating embryogenic calli. Ovules taken from immature fruits were used in a modified MT culture medium, containing 50g.L-1 of sucrose and 500 mg.L-1 of malt extract, solidified with 7 g.L-1 of agar. In general, callus induction in the studied varieties was observed between the 6th and the 8th weeks of cultivation. In the Rangpur lime, 'Hamlin' sweet orange, Cleopatra and 'Swatow' mandarins and Troyer citrange, the percentage of callus formation was equal or superior to 50%. The Cleopatra mandarin stood out with a percentage near 70% of callus formation.

  18. EFFECT OF SYNTHETIC AUXIN LIKE GROWTH REGULATORS ON CALLUS REGENERATIVE ABILITY OF COMMON WHEAT VC. ZYMOYARKA

    Directory of Open Access Journals (Sweden)

    I. R. Gorbatyuk

    2015-02-01

    Full Text Available The aim of the study was to determine the dependence of morphogenetic reactions of wheat callus tissues to content of syntetic growth regulators of auxin nature (picloram, dicamba in the nutrient medium. Apical meristems of Triticum aestivum wheat were the primary explants for callusogenesis. Basic culture medium MS supplemented by vitamins of Gamborg, dicamba at different concentrations (0.2, 0.4, 0.6 mg/l, and picloram (0.16; 0.25; 0.5 mg/l was used for regeneration. It was established that dicamba at a concentration of 0.2 mg/l is the most effective for production of regenerants. It was also observed that at the concentration of 0.16 mg/l picloram there are the formation of the greatest number of morphogenic zones (60% and a significant amount of plant-regenerants. Increased concentrations of picloram to 0.25 mg/l and 0.5 mg/l caused a decrease in the number of morphogenic islands: in the first case, 10%, and the second – 36.4%. Among the described options the MS medium supplemented with 0.5 mg/l 6-benzylaminopurine and 0.16 mg/l picloram was the most effective. Shoots obtained from callus culture were capable to form roots in vitro and adapt to septic conditions. Regenerated plants when cultivated in greenhouse showed high viability (over 75% and reached the generative phase.

  19. Somatic embryogenesis from leaf derived callus of Tylophora indica (Burm. f.) Merrill.

    Science.gov (United States)

    Manjula, S; Job, A; Nair, G M

    2000-10-01

    Mature leaf explant derived callus of Tylophora indica (Burm. f.) Merrill yielded somatic embryos on MS medium supplied with BA(1-2 mg/L) or kinetin(1-5 mg/L) or kinetin/BA (1-2 mg/L) used along with IAA(0.1-1 mg/L). Maximum somatic embryos (30) could be recovered from 100 mg of embryogenic callus within 60 days at an optimum concentration of 2 mg/L of BA which was also best suited for providing the maximum conversion rate (90%) of embryoids to plantlets. Kinetin (1-5 mg/L), used as the sole growth hormone, induced the development of embryoids showing either shoot or root primordia in 30% of the cultures. However, embryoids with shoot primordia developed roots upon transfer to medium containing IAA(0.1 mg/L) and kinetin(2 mg/L). Embryoids from all cultures germinated in the initiation medium and were transplanted to sterile vermiculite for hardening. After two weeks of hardening, the plantlets were transferred to the green house where they grew and established well showing a high rate of survival (90%).

  20. Hyperplastic callus formation in osteogenesis imperfecta: CT and MRI findings

    Energy Technology Data Exchange (ETDEWEB)

    Rieker, O.; Kreitner, K.F. [Klinik fuer Radiologie, Johannes-Gutenberg-Univ. Mainz (Germany); Karbowski, A. [Orthopaedische Abtl., Krankenhaus der Augustinerinnen, Koeln (Germany)

    1998-09-01

    Hyperplastic callus formation is a noteworthy condition in patients with osteogenesis imperfecta because it often mimicks osteosarcoma on radiography. The findings of CT and MRI in hyperplastic callus formation have not been reported. In the presented case, MRI demonstrated contrast enhancement and edema of the surrounding soft tisssue, consistent with benign as well as malignant disease. Computed tomography showed a calcified rim of the lesion which may be a useful feature to rule out osteosarcoma in this condition. (orig.) With 2 figs., 18 refs.

  1. Effect of gamma rays on different explants of callus treatment of multiple shoots in Cucumis melo cv. Bathasa.

    Science.gov (United States)

    Venkateshwarlu, M

    2008-09-01

    A mutagenesis programme was carried out using physical mutagens (gamma rays) on Cucumis melo cv. Bathasa. In irradiated seeds the number of shoots formed in the lower doses was significantly higher than the controls. Decrease in the number of shoots and shoot bud formation was observed with higher doses in all the explants studies. There was complete lethality in the 10, 15 and 20 kR doses. In irradiated stem cultures the maximum number of shoots were observed in 2 kR. The number of shoots decreased with increasing doses of irradiation. At higher doses of 4 and 5kR, light green compact callus was formed in almost all the explants. The effect of lower doses of gamma irradiation on shoot bud formation and rooting efficiency from nodal explants cultured on MS +2.0 mgl(-1) L-glutamic acid + 0.5 mgl(-1) BAP, stimulation of shoot and root induction were studied. The number of shoots and root lengths decreased with the increasing dosage of irradiation. The irradiated callus was grown on solidified MS medium with containing 2.0 mgl(-1) BAP + 1.0 mgl(-1) IAA. There was a significant stimulation of growth in the callus at lower doses. At higher doses like 15 and 20 kRs growth was drastically reduced.

  2. Experiments on tissue culture in the genus Lycopersicon miller : Shoot formation from protoplasts of tomato long-term cell cultures.

    Science.gov (United States)

    Koblitz, H; Koblitz, D

    1982-06-01

    Callus cultures from cotyledon explants were established and maintained in culture for more than two years. After several months callus cultures were transferred into liquid medium and cultured as cell suspensions. Protoplasts were isolated from these cell suspension cultures and cultured in a liquid medium. After formation of new cell walls the cells were further cultured in liquid medium and afterwards transferred to an agar-solidified medium to give a vigorously growing callus culture. In the case of the cultivar 'Lukullus' shoots were recovered from callus. All efforts to root these shoots failed and this, in addition to variations in appearence, suggests that the shoots are changed genetically possibly due to the prolonged culture period.

  3. Plant Tissue Cultures of Juniperus virginiana.

    Science.gov (United States)

    Kašparová, Marie; Spilková, Jirina; Cvak, Ladislav; Siatka, Tomáš; Martin, Jan

    2016-05-01

    Callus cultures of Juniperus virginiana L. (varieties 'Hetzii', 'Glauca', 'Grey Owl') were derived from fresh leaves of garden-grown trees on Schenk and Hildebrandt medium supplemented with 3.0 mg/L of α-naphthaleneacetic acid, 0.2 mg/L of kinetin and 15 mg/L of ascorbic acid. The growth characteristics of one-year-old and two-years-old cultures were determined. The maximum biomass in all varieties was achieved on the 35th day of the cultivation period. The increase in fresh weights of two-years-old callus cultures, when compared with one-year-old callus cultures, was as follows: variety 'Hetzii' by 25%, variety 'Glauca' by 29% and variety 'Grey Owl' by 49%. J. virginiana suspension cultures (varieties 'Hetzii', 'Glauca', 'Grey Owl') were derived from two-years-old callus cultures on Schenk and Hildebrandt medium supplemented with 3.0 mg/L of α-naphthaleneacetic acid, 0.2 mg/L of kinetin and 15 mg/L of ascorbic acid. The maximum biomass of all varieties was found on the 21st day of the cultivation period. These results indicate that a sub-cultivation interval of 35 days for callus cultures and of 21st days for suspension cultures can be recommended. The callus and suspension cultures of J. virginiana of the variety 'Glauca' have the best survivability and thus provide the most biomass.

  4. Analysis of indole compounds in methanolic extracts from the fruiting bodies of Cantharellus cibarius (the Chanterelle) and from the mycelium of this species cultured in vitro.

    Science.gov (United States)

    Muszyńska, Bożena; Sułkowska-Ziaja, Katarzyna; Ekiert, Halina

    2013-12-01

    Methanolic extracts obtained from the fruiting bodies of Cantharellus cibarius (the Chanterelle) and from the mycelium of this species cultured in vitro were analyzed for the qualitative and quantitative composition of non-hallucinogenic indole compounds. The extracts were found to contain eight indole compounds: L-tryptophan, 5-hydroxytryptophan, serotonin, melatonin, indole, kynurenine sulfate, 5-methyltryptophan, and indoleacetonitrile. The extract from the fruiting bodies also contained tryptamine. The amounts of individual compounds varied widely, ranging from 0.01 to 17.61 mg/100 g DW in the fruiting bodies, and from 0.01 to 35.34 mg/100 g DW in the biomass from in vitro cultures. The quantitatively dominating compounds included: serotonin (17.61 and 20.49 mg/100 g DW, respectively) and kynurenine sulfate (3.62 and 35.34 mg/100 g DW). In addition, the material from in vitro cultures contained a considerable amount of 5-hydroxytryptophan (12.52 mg/100 g DW). The levels of the remaining indole compounds under analysis: L-tryptophan, melatonin, indole, 5-methyltryptophan, and indoleacetonitrile in the material under study were low, below 1 mg/100 g DW.

  5. Biological and Cultural Control of Olive Fruit Fly in California---Utilization of Parasitoids from USDA-APHIS-PPQ, Guatemala

    Science.gov (United States)

    The parasitoid Psytallia cf. concolor (Szépligeti) was reared on sterile Mediterranean fruit fly larvae at the USDA-APHIS-PPQ, Petapa Quarantine Laboratory in Guatemala and shipped to the USDA-ARS, Parlier, for wide-spread release and biological control of olive fruit fly in California. As many as 3...

  6. Antioxidant Capacity and Total Phenolics Content of the Fruiting Bodies and Submerged Cultured Mycelia of Sixteen Higher Basidiomycetes Mushrooms from India.

    Science.gov (United States)

    Prasad, Rajendra; Varshney, Vinay K; Harsh, N S K; Kumar, Manoj

    2015-01-01

    The fruiting bodies and the submerged cultured mycelia of 16 higher Basidiomycetes mushrooms- Agaricus bisporus, Armillaria mellea, Auricularia auricula-judae, Ganoderma applanatum, G. lucidum, Laetiporus sulphureus, Lentinus tigrinus, Lycoperdon pyriforme, Phellinus linteus, Pleurotus ostreatus, P. sajor-caju, Polyporus arcularius, Russula brevipes, Schizophyllum commune, Sparassis crispa, and Spongipellis unicolor-from different taxonomic groups were examined for their antioxidant capacity (AOXC) and total phenolics content (TPC). Extraction of the freeze-dried and pulverized fruiting bodies and mycelia with methanol and water (8:2, v/v), followed by evaporation of the solvent under a vacuum, created their extracts, which were analyzed for their AOXC and TPC using a DPPH· scavenging assay and the Folin-Ciocalteu method, respectively. The fruiting bodies and the culture mycelia of all the mushroom species exhibited varied antioxidant capacity; however, the fruiting bodies had more potent DPPH· scavenging than the corresponding mycelia irrespective of the mushroom species, as evident by the effective concentrations of extract that scavenges 50% of DPPH· (EC50) of the former (0.56-1.24 mg mL-1) being lower than those of the latter (2.51-8.39 mg mL-1). TPC in the fruiting bodies (6.08-24.85 mg gallic acid equivalent [GAE] g-1) were higher than those in the mycelia (4.17-13.34 mg GAE g-1). AOXC of the fruiting bodies (r = -0.755) and the culture mycelia (r = -0.903) also was correlated to their TPC. Among the cultured mycelia, A. bisporus, A. mellea, L. tigrinus, P. ostreatus, and S. crispa were highly promising in terms of their highest TPC (10.55, 13.34, 11.00, 10.37, and 10.19 mg GAE g-1, respectively) and the lowest EC50 values (3.33, 2.85, 2.51, 3.65, and 3.17 mg mL-1, respectively) as they relate to the development of antioxidants.

  7. Comparação de diferentes metodologias para obtenção de cultura de calos de Stevia rebaudiana (Bertoni Bertoni - DOI: 10.4025/actascihealthsci.1090 Comparison of different methodologies for obtaining callus cultures from Stevia rebaudiana (Bertoni Bertoni - DOI: 10.4025/actascihealthsci.1090

    Directory of Open Access Journals (Sweden)

    Regina Aparecida Correia Gonçalves

    2008-03-01

    Full Text Available As folhas de Stevia rebaudiana (Bertoni Bertoni (Asteraceae contêm glicosídeos diterpenóides (GDS, que são cerca de 300 vezes mais doce que a sacarose a 4%. O objetivo deste estudo foi avaliar a formação de calos, a partir de folhas obtidas in vivo e in vitro de S. rebaudiana em dois meios já descritos na literatura: Murashige e Skoog (MS, suplementado com 3 mg L-1 de ácido 2,4-diclorofenóxiacético (2,4-D, e o MS suplementado com 1 mg L-1 de ácido naftalenoacético (ANA e 0,5 mg L-1 de 6-benzilaminopurina 6-BAP e um desenvolvido em nosso laboratório o Woody Plant Medium (WPM, suplementado com 6 mg L-1 de ANA e 4 mg L-1 de cinetina (CIN. Os explantes obtidos in vitro iniciaram a formação de calos um pouco mais rapidamente que os das folhas de plantas advindas da natureza. A utilização dos nutrientes do meio WPM, associada a uma combinação de fitorreguladores adequada, proporcionou velocidade de indução e multiplicação de calos bem maiores que as apresentadas nos meios que empregaram os nutrientes do MS. Novos experimentos serão realizados, depois de alcançada a estabilidade genética dos calos, visando avaliar a capacidade destes em biossintetizar os GDSThe leaves from Stevia rebaudiana (Bertoni Bertoni (Asteraceae contain diterpenoid glycosides (GDS, which are almost 300 times sweeter than sucrose at 4%. The subject of this study was to evaluate the callus-formation from in vivo and in vitro leaves of Stevia rebaudiana in two already described in literature: Murashige and Skoog (MS supplemented with 3 mg L-1 of 2,4-dichlorophenoxyacetic acid (2,4-D; MS supplemented with 1 mg L-1 of naphthaleneacetic acid (NAA and 0.5 mg L-1 of 6-benzylaminopurine (6-BAP and other developed in our laboratory the Woody Plant Medium (WPM with 6 mg L-1 of NAA and 4 mg L-1 of Kinetin (KIN. The explants obtained in vitro initiated callus formation faster than leaves from natural plants. The utilization of WPM nutrients, associated with an

  8. Effect of plant growth regulators on callus induction and plant ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-06-03

    Jun 3, 2009 ... Effect of plant growth regulators on callus induction and plant ... MS media supplemented with different levels of BA and TDZ were employed for shoot ... acre many times that of any grain crop (Burton, 1969) and are used in a ... plant regeneration from explants require the presence of .... light green. 2.50 ± ...

  9. Induction and multiplication of callus from endosperm of Cycas ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-12-03

    Dec 3, 2008 ... In the induction studies, 20 µM picloram was the most efficient formulation for .... three treatments, maximum percentage of callus forma- .... Ser. Bot. 45(2): 79-86. Bordallo PN, Silva DH, Maria J, Cruz CD, Fontes EP (2004).

  10. Phenotypic differentiation between wild and domesticated varieties of Crescentia cujete L. and culturally relevant uses of their fruits as bowls in the Yucatan Peninsula, Mexico.

    Science.gov (United States)

    Aguirre-Dugua, Xitlali; Pérez-Negrón, Edgar; Casas, Alejandro

    2013-11-14

    Selection criteria are important for analyzing domestication of perennial plant species, which experience a selection pressure throughout several human generations. We analyze the preferred morphological characteristics of Crescentia cujete fruits, which are used as bowls by the Maya of Yucatan, according to the uses they are given and the phenotypic consequences of artificial selection between one wild and three domesticated varieties. We performed 40 semi-structured interviews in seven communities. We calculated Sutrop's salience index (S) of five classes of ceremonial and daily life uses, and of each item from the two most salient classes. We sampled 238 bowls at homes of people interviewed and compared their shape, volume and thickness with 139 fruits collected in homegardens and 179 from the wild. Morphology of varieties was assessed in fruit (n = 114 trees) and vegetative characters (n = 136 trees). Differences between varieties were evaluated through linear discriminant analysis (LDA). Use of bowls as containers for the Day of the Dead offerings was the most salient class (S = 0.489) with chocolate as its most salient beverage (S = 0.491), followed by consumption of daily beverages (S = 0.423), especially maize-based pozol (S = 0.412). The sacred saka' and balche' are offered in different sized bowls during agricultural and domestic rituals. Roundness was the most relevant character for these uses, as bowls from households showed a strong selection towards round shapes compared with wild and homegarden fruits. Larger fruits from domesticated varieties were also preferred over small wild fruits, although in the household different sizes of the domesticated varieties are useful. LDA separated wild from domesticated trees (p people model the domestication process of this important tree in their culture.

  11. 金线兰愈伤组织诱导条件的优化%Optimization of Callus Induction Condition in Anoectochilus roxburghii

    Institute of Scientific and Technical Information of China (English)

    杨玉红; 林海

    2011-01-01

    The rhizome, young stem, leaf and in vitro embryo of Anoectochilus roxburghii were cultured on media with different hormone combinations to optimize callus induction condition for probing the technique of callus induction and cell culture in Anoectochilus roxburghii. The results showed that 6-BA and 2,4-D had the significant effect on its callus induction, the callus induction effect of four tested hormones was 6-BA>2,4-D>NAA>ZT, the optimum medium for callus induction was MS+ 0.2 mg/L 2,4-D+0.9 mg/L NAA + 1.0 mg/L 6-BA+0.25 mg/LZT, the callus induction rate of rhizome and young stem was higher than that of leaf and in vitro embryo, and the callus browning rate cultured under dark condition was lower than that cultured under light/dark condition.%为探索金线兰愈伤组织的诱导和细胞培养技术,以金线兰根状茎、幼茎、叶片和离体胚为外植体,2,4-D、NAA、6-BA和ZT 4种激素采用L9(34)正交试验设计,对外植体愈伤组织诱导的条件进行了研究.结果表明:6-BA和2,4-D在诱导金线兰愈伤组织中作用极为显著,4种激素对愈伤组织的诱导作用大小依次为6-BA>2,4-D>NAA>ZT,初步断定金线兰愈伤组织诱导的最佳激素配比为0.2 mg/L 2,4-D+0.9 mg/L NAA+1.0 mg/L 6-BA+0.25 mg/L ZT;根茎和幼茎作为外植体更易诱导愈伤组织形成;暗培养较光暗交替培养褐化率低.

  12. Embryogenic callus formation in Dioscorea rotundata Poir cv. `Blanco de Guinea'

    Directory of Open Access Journals (Sweden)

    Dayana Rodríguez González

    2014-07-01

    Full Text Available Yam contributes to energy and nutritional requirements of most of the population of developing countries. However, their extensive culture is constrained by the limited availability of planting material with physiological and sanitary quality, and also part of the harvesting is used as seed in the next planting. For this reason, it is necessary to establish a methodology for plant regeneration and somatic embryogenesis could facilitate their micropropagation and genetic improvement. This study aimed to form embryogenic callus in Dioscorea rotundata Poir cv. `White Guinea'. The effect of the addition of 2,4-dichlorophenoxyacetic acid (2,4-D (0, 1.0, 2.0 and 4.0 mg l-1 was determined, in combination with three types of explants from in vitro plants (leaves petiole, petiole segments and root sections. The highest percentage of embryogenic callus was obtained with 1.0 mg l-1 2,4-D and leaves with petiole as explants. These were characterized by the presence of compact whitish nodules. Key words: 2,4-dichlorophenoxyacetic acid, somatic embryogenesis, micropropagation, yam

  13. SUSPENSION CULTURE AND PLANT REGENERATION OF TYPHA LATIFOLIA

    Science.gov (United States)

    This study is the first reported attempt to generate a growth curve from Typha latifolia L. (broadleaf cattail) callus cells in suspension culture. Several media and hormone combinations were tested for their capacity to induce callus cell formation from T. latifolia leaf section...

  14. SUSPENSION CULTURE AND PLANT REGENERATION OF TYPHA LATIFOLIA

    Science.gov (United States)

    This study is the first reported attempt to generate a growth curve from Typha latifolia L. (broadleaf cattail) callus cells in suspension culture. Several media and hormone combinations were tested for their capacity to induce callus cell formation from T. latifolia leaf section...

  15. Establishment and optimization of cell growth in suspension culture of Papaver bracteatum: a biotechnology approach for thebaine production

    OpenAIRE

    FARJAMINEZHAD, Reza; Nasser ZARE; ASGHARI-ZAKARIA, Rasool; Farjaminezhad, Manoochehr

    2013-01-01

    Iranian poppy (Papaver bracteatum) is an important medicinal plant that is the main source of the opium alkaloids codeine, morphine, and thebaine. To establish an efficient protocol for cell suspension culture and growth, the effects of different plant growth regulators (2,4-D, NAA, BAP, and kinetin) on callus induction and cell suspension culture of Iranian poppy were evaluated. The maximum percentage of callus induction (86.67%) and fresh weight of callus were obtained in MS medium suppleme...

  16. A Comparative Study of Cultural--loaded Meaning of English and Chinese Fruit Words in View of Cultural Psychology%从文化心理角度看英汉水果类词的文化义

    Institute of Scientific and Technical Information of China (English)

    陈艳芳

    2012-01-01

    Cultural- loaded meaning is the reflection of culture. Different cultural language environments have some common cultural--loaded meaning, but also have particular cultural-- loaded meaning. Combining the living examples of fruit words, and with an eye to the relationship between cultural- loaded meaning and cultural psychology' this ,oaper analyses the cultural differences in different cultural environment of cultural--loaded meaning., and therefore grasp the language phenomenon from the view of culture.%文化义是文化色彩的再现。不同文化语境既存在共有的文化义,也存在特有的文化义。本文结合水果类词的实例,着眼于文化义与文化心理的关系,分析其文化义在不同文化语境下体现的文化差异,从文化的角度来把握语言现象。

  17. Influence of nutrient media on callus induction, somatic embryogenesis and plant regeneration in selected Turkish crocus species.

    Science.gov (United States)

    Verma, Sandeep Kumar; Das, Ashok Kumar; Cingoz, Gunce Sahin; Uslu, Emel; Gurel, Ekrem

    2016-06-01

    Callus induction, somatic embryogenesis and plant regeneration were initiated in selected five species of Turkish crocus using three diffrent explants (leaf, stem and corm) cultured on four different media (MS, GB5, LS and CHE). The highest frequencies of callus induction (100%) and shoot regeneration (70%, with 7.2 shoots/callus) were found in the crocus species Crocus oliveri ssp. Oliveri, using the MS medium containing 5% (w/v) sucrose supplemented with (4 mg/L NAA + 4 mg/L TDZ) and (2 mg/L IAA + 2 mg/L TDZ + 2 mg/L BAP). When the embryogenic calli were transferred into the four nutrient media containing (2 mg/L IAA + 2 mg/L TDZ) and 100 mg/L ABA, these further developed into cotyledonary embryos. Maximum number of somatic embryos (2.9 embryos per leaf explant, with a frequency 46.6%) was obtained in C. oliveri ssp. Oliveri. During subculture using the half strength media, cotyledonary embryos gradually developed into plantlets.

  18. Comparative metabolite profiling of the insecticide thiamethoxam in plant and cell suspension culture of tomato.

    Science.gov (United States)

    Karmakar, Rajib; Bhattacharya, Ramcharan; Kulshrestha, Gita

    2009-07-22

    The metabolism of thiamethoxam [(EZ)-3-(2-chloro-1,3-thiazol-5-yl-methyl)-5-methyl-1,3,5-oxadiazinan-4-ylidene (nitro) amine] was investigated in whole plant, callus, and heterotrophic cell suspension culture of aseptically and field grown tomato (Lycopersicon esculentum Mill.) plants. The structure of the metabolites was elucidated by chromatographic (HPLC) and spectroscopic (IR, NMR, and MS) methods. Thiamethoxam metabolism proceeded by the formation of a urea derivative, a nitroso product, and nitro guanidine. Both urea and nitro guanidine metabolites further degraded in plants, and a mechanism has been proposed. In the plant, organ-specific differences in thiamethoxam metabolism were observed. Only one metabolite was formed in whole plant against four in callus and eight metabolites in cell suspension culture under aseptic conditions. Out of six metabolites of thiamethoxam in tomato fruits in field conditions, five were similar to those formed in the cell suspension culture. In the cell suspension culture, thiamethoxam degraded to maximum metabolites within 72 h, whereas in plants, such extensive conversion could only be observed after 10 days.

  19. Primary studies on tissue culture from mature embryos in diploid and tetraploid wheat

    Institute of Scientific and Technical Information of China (English)

    Ruiming BI; Honggang WANG

    2008-01-01

    Using mature embryos (MEs) as the explants,the callus induction,embryogenic callus differentiation,plantlet regeneration and culture efficiency in diploid and tetraploid wheat of four genotypes were studied.The tested four genotypes wheat included cultivable emmer wheat ( Triticum dicoccum Schuble),durum wheat ( Triticum durum Desf.) and the common wheat progenitors Triticum dicoccoides and Triticum aegilopides.Results indicated that there were significant differences in the efficiency of callus induction,callus differentiation and plant regeneration among the tested genotypes.The efficiency of differentiation and regeneration shows strong genotype dependence.The rates of callus induction,embryogenic callus differentiation,plantlet regeneration and culture efficiency respectively were 95.00%,90.00%,32.40%,and 27.70% in cultivable emmer wheat,which were significantly higher than other tested genotypes.Therefore,this study has provided a basis for genetic transformation,gene cloning and molecular plant breeding in wheat and other related species.

  20. Effects of plant growth regulators, carbon sources and pH values on callus induction in Aquilaria malaccensis leaf explants and characteris-tics of the resultant calli

    Institute of Scientific and Technical Information of China (English)

    Shashita JAYARAMAN; Nurul Hazwani DAUD; Rasmina HALIS; Rozi MOHAMED

    2014-01-01

    The endangered tropical tree, Aquilaria malaccensis, produces agarwood for use in fragrance and medicines. Efforts are currently un-derway to produce valuable agarwood compoundsn tissue culture. The purpose of this study was to develop an optimal growth medium, specif-ically, the best hormone combination for callus suspension culture. Using nursery-grown A. malaccensis, sterilized leaf explants were first incu-bated on basic Murashige and Skoog (MS) gel medium containing 15g/L sucrose and at pH 5.7. Different auxin types including 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and indole-3-butyric acid (IBA), were tested at various concentrations (0.55, 1.1 and 1.65 µM) using the basic medium. Leaf explants were incubated for 30 days in the dark. Callus induced by 1.1 µM NAA had the highest biomass dry weight (DW) of 17.3 mg;however the callus was of a compact type. This auxin concentration was then combined with either 6-benzylaminopurine (BAP) or kinetin at 0.55, 1.1, 2.2 or 3.3 µM to induce growth of friable callus. The 1.1µM NAA+2.2µM BAP com-bination produced friable callus with the highest biomass (93.3mg DW). When testing the different carbon sources and pHs, sucrose at 15g/L and pH at 5.7 yielded highest biomasses at 87.7mg and 83mg DW, respec-tively. Microscopic observations revealed the arrangement of the friable cells as loosely packed with relatively large cells, while for the compact callus, the cells were small and densely packed. We concluded that MS medium containing 15 g/L sucrose, 1.1 µM NAA + 2.2 µM BAP hor-mone combination, and a pH of 5.7 was highly effective for inducing friable callus from leaf explants of A. malaccensis for the purpose of establishing cell suspension culture.

  1. Growth characteristics and dynamics of protein synthesis in callus cultures from Glycine wightii (Wight & Arn. Verdc. Características do crescimento e dinâmica da síntese de proteínas em cultura de calos de Glycine wightii (Wight & Arn. Verdc.

    Directory of Open Access Journals (Sweden)

    André Luis Coelho da Silva

    2005-12-01

    Full Text Available Cotyledon explants were first cultured on MS medium supplemented with 4.52 M 2,4-D and 0.46 mM kinetin. The development of the calli was followed (0, 4, 8, 12, 16, 20, 24, 28 and 32 days after and the growth curve was determined, based in fresh and dry weight. The growth curve presented sigmoidal form with four distinct phases. The highest growth percentage was observed at the exponential phase and the lowest at the stationary phase. These results indicated that cotyledon callus subculture should be performed 20 days after inoculation. The calli obtained after a period of 28 days were freeze dried, macerated and submitted to extraction with buffers of different pH values (2.6; 4.0; 6.0; 8.0 and 10.0 and the proteins in the extracts were determined by Bradford method. The pH 8.0 buffer was the most efficient to extract the largest amount of protein. The amino acid analyses calli showed a high content of aspartic acid and low content of metionin. The dynamics of protein synthesis in calli was followed by SDS-PAGE electrophoresis.Explantes cotiledonários foram primeiramente cultivados em meio MS suplementado com 2,4-D 4,52 mM e cinetina 0,46 mM. O desenvolvimento dos calos foi acompanhado (0, 4, 8, 12, 16, 20, 24, 28 até 32 dias após o subcultivo e uma curva de crescimento foi determinada com base nos pesos fresco e seco dos calos. A curva de crescimento apresentou um padrão sigmoidal com quatro fases distintas. A maior porcentagem de crescimento foi observada na fase exponencial e a menor na fase estacionária. Estes resultados indicam que o subcultivo dos calos pode ser realizado 20 dias após a inoculação. Os calos obtidos após um período de 28 dias foram liofilizados, macerados e submetidos à extração de proteínas solúveis, utilizando tampões com diferentes valores de pH (2,6; 4,0; 6,0; 8,0 e 10,0 e o teor protéico dos extratos foi determinado pelo método de Bradford. O tampão pH 8,0 foi o mais eficiente na extração de prote

  2. Effects of controlled-release fertilizer on leaf area index and fruit yield in high-density soilless tomato culture using low node-order pinching.

    Directory of Open Access Journals (Sweden)

    Takafumi Kinoshita

    Full Text Available To further development of a simplified fertigation system using controlled-release fertilizers (CRF, we investigated the effects of differing levels of fertilizers and plant density on leaf area index (LAI, fruit yields, and nutrient use in soilless tomato cultures with low node-order pinching and high plant density during spring-summer (SS, summer-fall (SF, and fall-winter (FW seasons. Plants were treated with 1 of 3 levels of CRF in a closed system, or with liquid fertilizer (LF with constant electrical conductivity (EC in a drip-draining system. Two plant densities were examined for each fertilizer treatment. In CRF treatments, LAI at pinching increased linearly with increasing nutrient supply for all cropping seasons. In SS, both light interception by plant canopy at pinching and total marketable fruit yield increased linearly with increasing LAI up to 6 m(2 · m(-2; the maximization point was not reached for any of the treatments. In FW, both light interception and yield were maximized at an LAI of approximately 4. These results suggest that maximizing the LAI in SS and FW to the saturation point for light interception is important for increasing yield. In SF, however, the yield maximized at an LAI of approximately 3, although the light interception linearly increased with increasing LAI, up to 4.5. According to our results, the optimal LAI at pinching may be 6 in SS, 3 in SF, and 4 in FW. In comparing LAI values with similar fruit yield, we found that nutrient supply was 32-46% lower with the CRF method than with LF. In conclusion, CRF application in a closed system enables growers to achieve a desirable LAI to maximize fruit yield with a regulated amount of nutrient supply per unit area. Further, the CRF method greatly reduced nutrient use without decreasing fruit yield at similar LAIs, as compared to the LF method.

  3. Effects of controlled-release fertilizer on leaf area index and fruit yield in high-density soilless tomato culture using low node-order pinching.

    Science.gov (United States)

    Kinoshita, Takafumi; Yano, Takayoshi; Sugiura, Makoto; Nagasaki, Yuji

    2014-01-01

    To further development of a simplified fertigation system using controlled-release fertilizers (CRF), we investigated the effects of differing levels of fertilizers and plant density on leaf area index (LAI), fruit yields, and nutrient use in soilless tomato cultures with low node-order pinching and high plant density during spring-summer (SS), summer-fall (SF), and fall-winter (FW) seasons. Plants were treated with 1 of 3 levels of CRF in a closed system, or with liquid fertilizer (LF) with constant electrical conductivity (EC) in a drip-draining system. Two plant densities were examined for each fertilizer treatment. In CRF treatments, LAI at pinching increased linearly with increasing nutrient supply for all cropping seasons. In SS, both light interception by plant canopy at pinching and total marketable fruit yield increased linearly with increasing LAI up to 6 m(2) · m(-2); the maximization point was not reached for any of the treatments. In FW, both light interception and yield were maximized at an LAI of approximately 4. These results suggest that maximizing the LAI in SS and FW to the saturation point for light interception is important for increasing yield. In SF, however, the yield maximized at an LAI of approximately 3, although the light interception linearly increased with increasing LAI, up to 4.5. According to our results, the optimal LAI at pinching may be 6 in SS, 3 in SF, and 4 in FW. In comparing LAI values with similar fruit yield, we found that nutrient supply was 32-46% lower with the CRF method than with LF. In conclusion, CRF application in a closed system enables growers to achieve a desirable LAI to maximize fruit yield with a regulated amount of nutrient supply per unit area. Further, the CRF method greatly reduced nutrient use without decreasing fruit yield at similar LAIs, as compared to the LF method.

  4. Efficient callus formation and plant regeneration of goosegrass [Eleusine indica (L.) Gaertn.].

    Science.gov (United States)

    Yemets, A I; Klimkina, L A; Tarassenko, L V; Blume, Y B

    2003-02-01

    Efficient methods in totipotent callus formation, cell suspension culture establishment and whole-plant regeneration have been developed for the goosegrass [ Eleusine indica (L.) Gaertn.] and its dinitroaniline-resistant biotypes. The optimum medium for inducing morphogenic calli consisted of N6 basal salts and B5 vitamins supplemented with 1-2 mg l(-1) 2,4-dichlorophenoxyacetic acid (2,4-D), 2 mg l(-1) glycine, 100 mg l(-1) asparagine, 100 mg l(-1) casein hydrolysate, 30 g l(-1) sucrose and 0.6% agar, pH 5.7. The presence of organogenic and embryogenic structures in these calli was histologically documented. Cell suspension cultures derived from young calli were established in a liquid medium with the same composition. Morphogenic structures of direct shoots and somatic embryos were grown into rooted plantlets on medium containing MS basal salts, B5 vitamins, 1 mg l(-1) kinetin (Kn) and 0.1 mg l(-1) indole-3-acetic acid (IAA), 3% sucrose, 0.6% agar, pH 5.7. Calli derived from the R-biotype of E. indica possessed a high resistance to trifluralin (dinitroaniline herbicide) and cross-resistance to a structurally non-related herbicide, amiprophosmethyl (phosphorothioamidate herbicide), as did the original resistant plants. Embryogenic cell suspension culture was a better source of E. indica protoplasts than callus or mesophyll tissue. The enzyme solution containing 1.5% cellulase Onozuka R-10, 0.5% driselase, 1% pectolyase Y-23, 0.5% hemicellulase and N(6) mineral salts with an additional 0.2 M KCl and 0.1 M CaCl(2) (pH 5.4-5.5) was used for protoplast isolation. The purified protoplasts were cultivated in KM8p liquid medium supplemented with 2 mg l(-1) 2,4-D and 0.2 mg l(-1) Kn.

  5. Indoleacetic Acid synthesis in soybean cotyledon callus tissue.

    Science.gov (United States)

    Black, R C; Hamilton, R H

    1976-03-01

    Growth of an auxin-requiring soybean cotyledon callus tissue (Glycine max L., Merr. var. Acme) was promoted by tryptophan, tryptamine, indole, indoleacetamide and, to a very slight degree, anthranilic acid. When tryptophan-3-(14)C was supplied in the growth medium, labeled indoleacetic acid (IAA) was found in both the tissue and the medium. Medium, from which the cells had been removed, was also found to convert labeled tryptophan to IAA. Soybean callus contained 0.044 mumole/g free tryptophan, but this is apparently not available for conversion to IAA. These results suggest that while exogenously supplied trytophan could elevate a specific internal pool where IAA synthesis occurs some of the growth on a tryptophan medium can be accounted for by external conversion.

  6. Indoleacetic Acid Synthesis in Soybean Cotyledon Callus Tissue 1

    Science.gov (United States)

    Black, Robert C.; Hamilton, Robert H.

    1976-01-01

    Growth of an auxin-requiring soybean cotyledon callus tissue (Glycine max L., Merr. var. Acme) was promoted by tryptophan, tryptamine, indole, indoleacetamide and, to a very slight degree, anthranilic acid. When tryptophan-3-14C was supplied in the growth medium, labeled indoleacetic acid (IAA) was found in both the tissue and the medium. Medium, from which the cells had been removed, was also found to convert labeled tryptophan to IAA. Soybean callus contained 0.044 μmole/g free tryptophan, but this is apparently not available for conversion to IAA. These results suggest that while exogenously supplied trytophan could elevate a specific internal pool where IAA synthesis occurs some of the growth on a tryptophan medium can be accounted for by external conversion. PMID:16659498

  7. Avocado fruit (Persea americana Mill) exhibits chemo-protective potentiality against cyclophosphamide induced genotoxicity in human lymphocyte culture.

    Science.gov (United States)

    Paul, Rajkumar; Kulkarni, Paresh; Ganesh, Narayan

    2011-01-01

    Diets rich in fruits and vegetables have been associated with reduced risks for many types of cancers. Avocado (Persea americana Mill.) is a widely consumed fruit containing many cancer preventing nutrients, vitamins and phytochemicals. Studies have shown that phytochemicals extracted from the avocado fruit selectively induce cell cycle arrest, inhibit growth, and induce apoptosis in precancerous and cancer cell lines. Our recent studies indicate that phytochemicals extracted with 50% Methanol from avocado fruits help in proliferation of human lymphocyte cells and decrease chromosomal aberrations induced by cyclophosphamide. Among three concentrations (100 mg, 150 mg and 200 mg per Kg Body Weight), the most effective conc. of extract was 200 mg/Kg Body Wt. It decreased significant level of numerical and structural aberrations (breaks, premature centromeric division etc. up to 88%, p < 0.0001)), and accrocentric associtation within D & G group (up to 78%, p = 0.0008). These studies suggest that phytochemicals from the avocado fruit can be utilized for making active chemoprotective ingredient for lowering the side effect of chemotherapy like cyclophosphamide in cancer therapy.

  8. Effect of Zinc and Manganese Nutrition on Fruit Yield and Nutrient Concentrations in Greenhouse Tomato in Hydroponic Culture

    Directory of Open Access Journals (Sweden)

    A. Tavassoli

    2010-06-01

    Full Text Available Abstract This research was performed in a completely randomized block design with four replications to investigate zinc (Zn and manganese (Mn nutrition effects on greenhouse tomato (Lycopersicon esculentum Mill. cv. HAMRA in a perlite-containing media. Experimental treatments were: (1 control (Mn and Zn-free nutrient solution, (2 application of Mn in a concentration equal to the full Hoagland’s nutrient solution (4.06 mg/l, (3 application of Zn in a concentration equal to the full Hoagland’s nutrient solution (4.42 mg/l, (4 application of Mn and Zn in concentrations equal to the 50% Hoagland’s nutrient solution (2.03 mg/l Mn + 2.21 mg/l Zn, and (5 application of Mn and Zn in concentrations equal to the full Hoagland’s nutrient solution (4.06 mg/l Mn + 4.42 mg/l Zn. Results showed that the highest fresh-fruit yield, fruit and leaf dry matter and content of Mn and Zn in fruit were obtained from single or combined application of Mn and Zn in concentrations equal to the full Hoagland’s nutrient solution. In addition, Zn and Mn nutrition significantly affected the fruit concentrations of crude protein, nitrogen and phosphorus, while the effect of these treatments on fruit size of tomato was not significant.

  9. Plant Tissue Culture and Its Application on Fruit Tree%植物组织培养及其在果树上的应用

    Institute of Scientific and Technical Information of China (English)

    刘书荣; 刘静; 吴曼

    2011-01-01

    Plant tissue culture refers to the use of totipotency cells in order to regeneration plant.It has important value with its application on fruit tree which could breeding rapidly,produce virus-free,cultivate new varieties,overcome such hybrids obstacles and save germplasm resources.This paper reviewed the progress of plant tissue culture on fruit tree while suggests references for further study of plant tissue culture.%植物组织培养是指利用细胞全能性培养再生植株的技术,由于其具有快速繁殖、产生脱毒苗、培育新品种、克服杂交不亲和障碍和保存种质资源等应用优点,在果树应用上有重要的价值。通过介绍植物组织培养在果树上的应用,以为今后研究提供参考。

  10. Production of taxoids in callus and cell suspensions of Taxus chinensis by using abiotic elicitors

    Directory of Open Access Journals (Sweden)

    Naivy Pérez

    2004-07-01

    Full Text Available Until now, several strategies have been developed to improve the production of secondary metabolites using plant cell cultures, manipulating the parameters of the environment, selecting high yielding cell clones and the use of elicitors. In the present work, callus and cell suspensions of Taxus chinensis lines SN and E15 were used, important specie for the production of metabolites with anticancer activity. The effect of different concentrations of the silver nitrate (1.0; 4.0 and 8.0mg.l-1 and the conditions of total dark and 12 h artificial light about the production of biomass and the taxoids accumulation were studied. The concentrations of AgNO3 used didn’t cause same effect under the different conditions of illumination to those that the cultivations were exposed. The AgNO3 increased the fresh weight under conditions of darkness in which superior values were obtained at 6.0g in all the treatments being bigger when using the concentration of 8.0mg (7.39g. In relation to the taxoids accumulation a bigger influence was observed with 4.0mg under conditions of light (0.93 mg/ gDW, treatment where smaller increment of biomass took place. Cell suspensions in both lines showed better growth that the cultivation of callus, while the biggest accumulation in Taxuyunnanine C was in the cultivation of callus in the line SN. It was observed that the cellular lines had a different behavior in conditions of light and darkness. As for the increment of the biomass the line E15 behaved from a superior way to the line SN as much in the darkness as to the light although the biggest value of fresh weight was obtained in the darkness (7.89g and the biggest accumulation in Taxuyunnanine C was under conditions of darkness with the line SN (0.98 mg/gDW. Keywords: cell cultures, elicitation, secondary metabolites, taxuyunnanine C Abbreviations: gDW - Grams of dry weight, gFW - grams of fresh weight, AgNO3 - Silver nitrate, Tc - Taxuyunnanine C

  11. Modified concentration method for the detection of enteric viruses on fruits and vegetables by reverse transcriptase-polymerase chain reaction or cell culture.

    Science.gov (United States)

    Dubois, Eric; Agier, Cécilia; Traoré, Ousmane; Hennechart, Catherine; Merle, Ghislaine; Crucière, Catherine; Laveran, Henri

    2002-12-01

    Fruits and vegetables may act as a vehicle of human enteric virus if they are irrigated with sewage-contaminated water or prepared by infected food handlers. An elution-concentration method was modified to efficiently detect, by reverse transcriptase-polymerase chain reaction (RT-PCR) or by cell culture, contamination by poliovirus, hepatitis A virus (HAV), and Norwalk-like virus (NLV) of various fresh and frozen berries and fresh vegetables. The protocol included washing the fruit or vegetable surface with 100 mM Tris-HCl, 50 mM glycine, and 3% beef extract, pH 9.5 buffer, which favors viral elution from acid-releasing berries, supplemented with 50 mM MgCl2 to reduce the decrease in viral infectivity during the process. The viral concentration method was based on polyethylene glycol precipitation. Copurified RT-PCR inhibitors and cytotoxic compounds were removed from viral concentrates by chloroform-butanol extraction. Viruses from 100 g of vegetal products could be recovered in volumes of 3 to 5 ml. Viral RNAs were isolated by a spin column method before molecular detection or concentrates were filtered (0.22-microm porosity) and inoculated on cell culture for infectious virus detection. About 15% of infectious poliovirus and 20% of infectious HAV were recovered from frozen raspberry surfaces. The percentage of viral RNA recovery was estimated by RT-PCR to be about 13% for NLV, 17% for HAV, and 45 to 100% for poliovirus. By this method, poliovirus and HAV RNA were detected on products inoculated with a titer of about 5 x 10(1) 50% tissue culture infectious dose per 100 g. NLV RNA was detected at an initial inoculum of 1.2 x 10(3) RT-PCR amplifiable units. This method would be useful for the viral analysis of fruits or vegetables during an epidemiological investigation of foodborne diseases.

  12. Plant regeneration through somatic embryogenesis from callus induced on immature embryos of Alstroemeria spp. L.

    Science.gov (United States)

    Van Schaik, C E; Posthuma, A; De Jeu, M J; Jacobsen, E

    1996-01-01

    The plant regeneration ability of callus obtained from zygotic embryos of the monocot Alstroemeria spp. was studied. The best explants for somatic embryogenesis were immature zygotic embryos in half-ovules when the endosperm was still soft and white. For 2 genotypes embryogenic callus was induced on callus induction medium with a success rate of 54%. The best callus induction period was 10 weeks. The morphology of embryogenic callus was nodular. Somatic embryos were formed after transfer of the callus to regeneration medium. These somatic embryos revealed later on the typical features of zygotic Alstroemeria embryos. The total duration of the plant regeneration protocol, from inoculation till rooted plantlets ready for transfer to the greenhouse, was 28 weeks.

  13. Accumulation dynamics and some cytogenetical tests at Chelidonium majus and Papaver somniferum callus under the magnetic liquid effect

    Science.gov (United States)

    Pavel, A.; Trifan, M.; Bara, I. I.; Creanga, D. E.; Cotae, C.

    1999-07-01

    In this paper we studied the capacity of magnetic liquids which are biocompatible (as previous literature reports showed) to influence germination, de-differentiation and re-differentiation processes in vitro, as well as to influence both the mitotic index and the chromosomal aberration frequency. In vitro cultures of two species of pharmaceutical interest, Chelidonium majus L. and Papaver somniferum L., on media supplemented with various concentrations of magnetic liquid, for various explant types, revealed a small but significant effect of the petroleum magnetic liquid on the callus accumulation and induced a considerable rate of chromosomal aberrations.

  14. Biological and Cultural Control of Olive Fruit Fly in California---Utilization of Parasitoids from USDA-APHIS-PPQ, Guatemala and Cultural Control Methods

    Science.gov (United States)

    The parasitoid Psytallia humilis = P. cf. concolor (Szépligeti) was reared on sterile Mediterranean fruit fly, Ceratitis capitata (Wiedemann), larvae at the USDA, APHIS, PPQ, Moscamed biological control laboratory in San Miguel Petapa, Guatemala and shipped to the USDA, ARS, Parlier, for biological ...

  15. Citrus tissue culture employing vegetative explants.

    Science.gov (United States)

    Chaturvedi, H C; Singh, S K; Sharma, A K; Agnihotri, S

    2001-11-01

    Citrus being a number one fruit of the world due to its high nutritional value, huge production of fruits and fruit products, the citrus industry may be considered a major fruit industry. Though citrus orchard area in India is comparable to USA, the produce is far less, while its export is nil. Biotechnology has played an outstanding role in boosting the citrus industry, e.g., in Spain, which is now the biggest exporter of citrus fruit with the application of micrografting. Amongst the fruit trees, perhaps the maximum tissue culture research has been done in citrus during the past four decades, however, the results of practical value are meagre. The shortfalls in citrus tissue culture research and some advancements made in this direction along with bright prospects are highlighted, restricting the review to vegetative explants only. Whilst utilization of nucellar embryogenesis is limited to rootstocks, the other aspects, like, regeneration and proliferation of shoot meristems measuring 200 microm in length--a global breakthrough--of two commercially important scion species, Citrus aurantifolia and C. sinensis and an important rootstock, C. limonia, improvement of micrografting technique, cloning of the same two scion species as well as some Indian rootstock species, employing nodal stem segments of mature trees, of immense practical value have been elaborated. A rare phenomenon of shift in the morphogenetic pattern of differentiation from shoot bud differentiation to embryoid formation occurred during the long-term culture of stem callus of C. grandis. Stem callus-regenerated plants of C. aurantifolia, C. sinensis and C. grandis showed variation in their ploidy levels and a somaclonal variant of C. sinensis, which produced seedless fruits was isolated. Tailoring of rooting in microshoots to a tap root-like system by changing the inorganic salt composition of the rooting medium, resulting in 100% transplant success, and germplasm preservation through normal growth

  16. Callus induction of leaf explant Piper betle L. Var Nigra with combination of plant growth regulators indole-3-acetic acid (IAA), benzyl amino purin (BAP) and kinetin

    Science.gov (United States)

    Junairiah, Zuraidassanaaz, Nabilah Istighfari; Izdihar, Fairuz Nabil; Manuhara, Yosephine Sri Wulan

    2017-09-01

    The purpose of this research was to determine the combination of plant growth regulators IAA, BAP and kinetin towards callus induction and growth of leaf explants Piper betle L. VarNigra. Explants from leaf of Piper betle L. VarNigra was cultured on MS medium with 24 treatment combinations of plant growth regulators IAA and BAP and 24 treatment combinations of plant growth regulators IAA and kinetin with 0.0;0.5;1.0;1.5;2.0 mg/L concentration respectively, the observed variable were the length of time the formation of callus, callus morphology, fresh and dry weight of callus. The results of this research showed that the combination of growth regulators IAA with BAP and kinetin had effects on leaf growth of Piper betle L. VarNigra. During 8 weeks observation, it indicated that the combination of concentration IAA 0.5 mg/L and BAP 2.0 mg/L showed fastest callus formation at 8.5 days. Combination of concentration IAA 1.0 mg/L and BAP 1.5 mg/L showed the highest of fresh weight at 0.6596 grams, and the highest dry weight was obtained from the combination of concentration IAA 0.5 mg/L and BAP 0.5 mg/L at 0.0727 grams. Combination of concentration IAA 1.0 mg/L and kinetin 1.5 mg/L had the highest of fresh weight at 0.2972 grams and the highest dry weight at 0.1660 grams. Callus of Piper betle L. VarNigra had two textures, that were compact and friable, and also showed various kind of colors, like white, greenish white, yellowish white, tanned white, brown and black. Based on this research, that concentration IAA 1.0 mg/L and 1.5 mg/L kinetin was the best combination for induction of callus from leaf of Piper betle L. Var Nigra.

  17. Occurrence of fruiting structures allows determination of Purpureocillium lilacinum as an inciting agent of pleuritis and pneumonia in a loggerhead sea turtle (Caretta caretta by histopathologic correlation to culture

    Directory of Open Access Journals (Sweden)

    V.L. Schumacher

    2014-10-01

    Full Text Available Purpureocillium lilacinum and Beauveria bassiana were isolated from lung sampled at necropsy of a 12 year-old female loggerhead sea turtle (Caretta caretta that had displayed abnormal buoyancy. Histopathologic evaluation revealed pleuritis and pneumonia with non-melanized, septate hyphae and fruiting structures identical to those of P. lilacinum. This case emphasizes the importance of a histological correlate to fungal culture when environmental fungi are isolated and demonstrates the infrequent phenomenon of fruiting or conidial production in tissue.

  18. Light-emitting diodes and their potential in callus growth, plantlet development and saponin accumulation during somatic embryogenesis of Panax vietnamensis Ha et Grushv.

    OpenAIRE

    Nhut, Duong Tan; Huy, Nguyen Phuc; Tai, Ngo Thanh; Nam, Nguyen Ba; Luan, Vu Quoc; Hien, Vu Thi; Tung, Hoang Thanh; Vinh, Bui The; Luan, Tran Cong

    2015-01-01

    In recent years, LED (light-emitting diode) has been the subject of research within the field of plant growth and development. However, there has been little discussion about using LED in vitro cultures of Panax vietnamensis, one of the important medicinal plants belonging to the Panax genus. This study examines the influence of various LED lamps on callus growth and plant formation of P. vietnamensis. Results show significant differences in growth and development, as various light conditions...

  19. CALLUS INDUCTION FROM FLORAL EXPLANTS OF CUPUASSU

    Directory of Open Access Journals (Sweden)

    MARIA DAS GRAÇAS RODRIGUES FERREIRA

    2013-01-01

    Full Text Available There are few studies related to the in vitro cultivation of plants from theTheobroma genus and no effective micropropagation protocols for T.grandiflorum. The aim of this study was to evaluate the calli formation in cupuassu floral explants, targeting their organogenic or embryogenicdevelopment. Experiments were conducted in the Plant Tissue Culture Laboratory of EMBRAPA, Porto Velho, Rondônia, Brazil. Floral parts from unopened immature flower buds taken from seedless cupuassu trees were sterilized and employed as a source of explants. These explants were cultivated in Petri dishes in an induction medium consisting of MS salts and vitamins, supplemented with glycine(3 mg.L-1, lysine (0,4 mg.L-1, leucine (0,4 mg.L-1, arginine (0,4 mg.L-1, tryptophan (0,2 mg.L-1, 2,4-D (1 mg.L-1, kinetin (0,25 mg.L-1, coconut water (50 ml.L-1, sucrose (40 g.L-1, Gelrite (2,2 g.L-1 and pH adjusted to 5,8. Cultures were maintained in the dark for 3 weeks at 27°C and then subcultured for six weeks in medium without growth regulators supplemented with glycine (1 mg.L-1, lysine (0,2 mg.L-1, leucine (0,2 mg.L-1, arginine (0,2 mg.L-1, tryptophan (0,1 mg.L-1, coconut water (100 ml.L-1, sucrose (40 g.L-1, Gelrite (2,2 g.L-1 and pH 5,8. We used a completely randomized design with 10 replications of 5 explants per plate and four different explant sources: staminode, petal, ligule and ovary. As a result, we obtained a highercalli formation in theinduction medium when ovaries were used as source of explants. However, there was no development of somatic embryosor organogenic response in medium without growth regulators and further studies are being conducted.

  20. Influence of organic supplements on production of shoot and callus biomass and accumulation of bacoside in Bacopa monniera (L.) Pennell.

    Science.gov (United States)

    Parale, Anuradha; Barmukh, Rajkumar; Nikam, Tukaram

    2010-04-01

    Production of valuable secondary metabolites through plant cell or organ culture is the best suited alternative to extraction of whole plant material and to increase production of secondary metabolites in in-vitro systems, feeding precursor or intermediate metabolites is an obvious and popular approach. The present investigation was aimed to study the influence of feeding of organic supplements, glycine (0-125 μM), ferulic acid (0-200 μM), phenylalanine (0-200 μM), α-ketoglutaric acid (0-200 μM) and pyruvic acid (0-200 μM) on production of bacoside-A (a triterpenoid type secondary metabolite responsible for cognition effects) in shoot and callus biomass of Bacopa monniera (L.) Pennell. The shoots were raised in liquid Murashige and Skoog's (MS) medium fortified with 5 μM 6-benzyladenine (BA) and callus biomass on agar solidified MS medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4 -D) in conjunction with 5 μM 1-napthaleneacetic acid (NAA). Among the organic supplements used, 100 μM pyruvic acid effectively enhanced the production of bacoside-A in shoot as well as callus biomass. The bacoside-A content in in-vitro raised shoot biomass was 4.0 and 1.2 times higher as compared to control and shoot biomass of naturally grown plants respectively. Inclusion of pyruvic acid in MS medium for in-vitro shoot cultures of B. monniera, can be adapted for enhanced production of bacoside-A.

  1. Study on Cell Suspension Culture of Floribunda Rose

    Institute of Scientific and Technical Information of China (English)

    ZHANG Chun'ai; WANG Jingang; FAN Jinping; GONG Shufang; CHE Daidi

    2008-01-01

    Friable callus was induced when immature seeds of floribunda rose were inoculated on MS medium supplemented with 2,4-D 3.0 mg-L-1.When transfered onto subculture media,fi-iable callus developed into embryogenic callus,which was used to establish cell suspension lines.Cell suspensions had to be subcultured at a interval of 4-5 days at the first several culture cycles.The best subculturing cycle for the stable cell suspensions was 8-10 days.The best inoculum quantity was 1 mL PCV(Packed Cell Volume) per 40 mL culture fluid.

  2. Tissue Culture Responses from Different Explants of Rice

    Institute of Scientific and Technical Information of China (English)

    WANG Xiu-hong; SHI Xiang-yuan; WU Xian-jun

    2005-01-01

    Different culture explants, including anther, young panicle, young embryo, and mature embryo, from 19 rice varieties were used for callus induction and green plantlet differentiation. The culture efficiency differed significantly among the four types of explants, and varied from genotype to genotype. Callus induction frequency presented significantly positive correlation each between anther and young panicle, anther and mature embryo, and young panicle and young embryo. Green plantlet differentiation showed no relationship between different types of explants. In addition, no relationship was found between callus induction frequency and green plantlet differentiation frequency.

  3. Isolation of protoplasts and culture and regeneration into plants in Alstroemeria

    NARCIS (Netherlands)

    Kim, J.B.; Bergervoet-van Deelen, J.E.M.; Raemakers, C.J.J.M.; Jacobsen, E.; Visser, R.G.F.

    2005-01-01

    An efficient system for the regeneration of plants from protoplasts was developed in Alstroemeria. Friable embryogenic callus (FEC) proved to be the best source for protoplast isolation and culture when compared with leaf tissue and compact embryogenic callus. Protoplast isolation was most efficient

  4. Studies on Callus Induction and Regeneration of Medicinal Plant Chicory (Cichorium intybus L. from Leaf and Petiole Explants

    Directory of Open Access Journals (Sweden)

    H. Hadizadeh

    2016-07-01

    Full Text Available Introduction: Chicory (Cichorium intybus L. belongs to Asteraceae family is commonly known as witloof chicory. The leaves and the roots of this medicinal plant are edible and commonly used as salad. Some varieties are also cultivated as coffee substitute after roasting the roots. All parts of the plant contain these volatile oils, with the majority of the toxic components concentrated in the plant's root. In folk medicine, the plant is used for the treatment of diarrhea, spleen enlargement, fever, and vomiting. Antihepatotoxic activity on damaged rat’s liver sections and anti-bacterial activity of this crop has been recently reported. In vitro regeneration from leaf explants with various hormonal combinations has been reported previously. Moreover, in vitro regeneration of Chicory from cotyledon explants using different combinations of plant growth regulators has been studied. Also, a protocol for the regeneration of plantlets from leaf and petiole explants of witloof chicory has been developed. The aim of the present investigation was optimization of callus induction and shoot regeneration from leaf and petiole tissues of Chicory (Esfahan genotype. Materials and Methods: In this investigation, Esfahan genotype was used for callus induction and direct shoot regeneration. Seeds were first washed with running tap water for 30 min then seeds were surface sterilized by dipping in 70% ethanol for 90 s and rinsed with sterile distilled water, followed by immersing in 5% sodium hypochlorite solution for 25 min and thereafter rinsed for 30 min with sterile distilled water. The basal medium used in this investigation was MS. For shoot regeneration, leaf and petiole explants (5 mm segments were excised from 4-week-old sterile seedlings and cultured on MS medium containing different combinations of NAA / BA and KIN / BA in two separate experiments. Experiments were performed factorial based on completely randomized design. Cultures were incubated at 25

  5. High capacity of plant regeneration from callus of interspecific hybrids with cultivated barley (Hordeum vulgare L.)

    DEFF Research Database (Denmark)

    Bagger Jørgensen, Rikke; Jensen, C. J.; Andersen, B.

    1986-01-01

    Callus was induced from hybrids between cultivated barley (Hordeum vulgare L. ssp. vulgare) and ten species of wild barley (Hordeum L.) as well as from one backcross line ((H. lechleri .times. H. vulgare) .times. H. vulgare). Successful callus induction and regeneration of plants were achieved from...

  6. Shear Stress-Normal Stress (Pressure Ratio Decides Forming Callus in Patients with Diabetic Neuropathy

    Directory of Open Access Journals (Sweden)

    Ayumi Amemiya

    2016-01-01

    Full Text Available Aim. Callus is a risk factor, leading to severe diabetic foot ulcer; thus, prevention of callus formation is important. However, normal stress (pressure and shear stress associated with callus have not been clarified. Additionally, as new valuables, a shear stress-normal stress (pressure ratio (SPR was examined. The purpose was to clarify the external force associated with callus formation in patients with diabetic neuropathy. Methods. The external force of the 1st, 2nd, and 5th metatarsal head (MTH as callus predilection regions was measured. The SPR was calculated by dividing shear stress by normal stress (pressure, concretely, peak values (SPR-p and time integral values (SPR-i. The optimal cut-off point was determined. Results. Callus formation region of the 1st and 2nd MTH had high SPR-i rather than noncallus formation region. The cut-off value of the 1st MTH was 0.60 and the 2nd MTH was 0.50. For the 5th MTH, variables pertaining to the external forces could not be determined to be indicators of callus formation because of low accuracy. Conclusions. The callus formation cut-off values of the 1st and 2nd MTH were clarified. In the future, it will be necessary to confirm the effect of using appropriate footwear and gait training on lowering SPR-i.

  7. Shear Stress-Normal Stress (Pressure) Ratio Decides Forming Callus in Patients with Diabetic Neuropathy

    Science.gov (United States)

    Noguchi, Hiroshi; Takehara, Kimie; Ohashi, Yumiko; Suzuki, Ryo; Yamauchi, Toshimasa; Kadowaki, Takashi; Sanada, Hiromi

    2016-01-01

    Aim. Callus is a risk factor, leading to severe diabetic foot ulcer; thus, prevention of callus formation is important. However, normal stress (pressure) and shear stress associated with callus have not been clarified. Additionally, as new valuables, a shear stress-normal stress (pressure) ratio (SPR) was examined. The purpose was to clarify the external force associated with callus formation in patients with diabetic neuropathy. Methods. The external force of the 1st, 2nd, and 5th metatarsal head (MTH) as callus predilection regions was measured. The SPR was calculated by dividing shear stress by normal stress (pressure), concretely, peak values (SPR-p) and time integral values (SPR-i). The optimal cut-off point was determined. Results. Callus formation region of the 1st and 2nd MTH had high SPR-i rather than noncallus formation region. The cut-off value of the 1st MTH was 0.60 and the 2nd MTH was 0.50. For the 5th MTH, variables pertaining to the external forces could not be determined to be indicators of callus formation because of low accuracy. Conclusions. The callus formation cut-off values of the 1st and 2nd MTH were clarified. In the future, it will be necessary to confirm the effect of using appropriate footwear and gait training on lowering SPR-i. PMID:28050567

  8. Proliferation Potential of 18-Month-Old Callus of Ananas comosus L. cv. Moris

    Directory of Open Access Journals (Sweden)

    A.E. De Silva

    2006-01-01

    Full Text Available Differential effect of plant growth regulators and additives in proliferation of 18-month-old calli of Ananas comosus L. cv. Moris were assessed in vitro. The proliferation of callus relied on the growth regulators and additives. Of the different auxins supplemented in the Murashige and Skoog (MS media, 32.22 μM α-naphthaleneacetic acid (NAA gave the highest mean fresh weight of callus (46.817 g. Medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D was inferior to NAA, while b-naphthoxy acetic acid (BNOA and p-chlorophenoxy acetic acid (4-CPA were not effective in proliferating 18-months old callus. Addition of casein hydrolysate and coconut water to NAA supplemented medium showed better proliferation and production of callus. However, in terms of callus production, NAA at 32.22 μM was economically better.

  9. Proliferation potential of 18-month-old callus of Ananas comosus L. cv. Moris.

    Science.gov (United States)

    De Silva, A E; Kadir, M A; Aziz, M A; Kadzimin, S

    2006-02-17

    Differential effect of plant growth regulators and additives in proliferation of 18-month-old calli of Ananas comosus L. cv. Moris were assessed in vitro. The proliferation of callus relied on the growth regulators and additives. Of the different auxins supplemented in the Murashige and Skoog (MS) media, 32.22 microM alpha-naphthaleneacetic acid (NAA) gave the highest mean fresh weight of callus (46.817 g). Medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) was inferior to NAA, while b-naphthoxy acetic acid (BNOA) and p-chlorophenoxy acetic acid (4-CPA) were not effective in proliferating 18-months old callus. Addition of casein hydrolysate and coconut water to NAA supplemented medium showed better proliferation and production of callus. However, in terms of callus production, NAA at 32.22 microM was economically better.

  10. Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

    Institute of Scientific and Technical Information of China (English)

    CHEN Jun-ying; YUE Run-qing; XU Hai-xian; CHEN Xin-jian

    2006-01-01

    To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars (Triticum aestivum L. cv.) were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm (endosperm-supported culture, ES), the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm (non-endosperm-supported culture, NES). This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.

  11. Development of an efficient callus proliferation system for Rheum coreanum Nakai, a rare medicinal plant growing in Democratic People’s Republic of Korea

    Directory of Open Access Journals (Sweden)

    Song-Chol Mun

    2016-07-01

    Full Text Available A clonal mass propagation to obtain mountainous sources of Rheum coreanum Nakai, a rare medicinal plant in Democratic People’s Republic of Korea was established by rhizome tissue culture. Whole plants were selected and collected as a vigorous individual free from blights and harmful insects among wild plants of R. coreanum grown on the top of Mt. Langrim (1.540 m above the sea situated at the northern extremity of Democratic People’s Republic of Korea. Induction of the callus was determined using four organs separated from the whole plant and different plant growth regulators. The callus was successfully induced from rhizome explant on MS medium containing 2.4-D (0.2–0.3 mg/l. In the MS medium supplemented with a combination of BAP (2 mg/l and NAA (0.2 mg/l, single NAA (0.5 mg/l, or IBA (0.5 mg/l, a higher number of shoot, root and plantlets was achieved. The survival rate on the mountainous region of the plantlets successfully acclimatized (100% in greenhouse reached 95%, and yields of crude drug and contents of active principles were higher than those obtained by sexual and vegetative propagation. This first report of R. coreanum tissue culture provides an opportunity to control extinction threats and an efficient callus proliferation system for growing resources rapidly on a large scale.

  12. Development of an efficient callus proliferation system for Rheum coreanum Nakai, a rare medicinal plant growing in Democratic People's Republic of Korea.

    Science.gov (United States)

    Mun, Song-Chol; Mun, Gwan-Sim

    2016-07-01

    A clonal mass propagation to obtain mountainous sources of Rheum coreanum Nakai, a rare medicinal plant in Democratic People's Republic of Korea was established by rhizome tissue culture. Whole plants were selected and collected as a vigorous individual free from blights and harmful insects among wild plants of R. coreanum grown on the top of Mt. Langrim (1.540 m above the sea) situated at the northern extremity of Democratic People's Republic of Korea. Induction of the callus was determined using four organs separated from the whole plant and different plant growth regulators. The callus was successfully induced from rhizome explant on MS medium containing 2.4-D (0.2-0.3 mg/l). In the MS medium supplemented with a combination of BAP (2 mg/l) and NAA (0.2 mg/l), single NAA (0.5 mg/l), or IBA (0.5 mg/l), a higher number of shoot, root and plantlets was achieved. The survival rate on the mountainous region of the plantlets successfully acclimatized (100%) in greenhouse reached 95%, and yields of crude drug and contents of active principles were higher than those obtained by sexual and vegetative propagation. This first report of R. coreanum tissue culture provides an opportunity to control extinction threats and an efficient callus proliferation system for growing resources rapidly on a large scale.

  13. Plant regeneration through callus organogenesis and true-to-type conformity of plants by RAPD analysis in Desmodium gangeticum (Linn.) DC.

    Science.gov (United States)

    Cheruvathur, Meena K; Abraham, Jyothi; Thomas, T Dennis

    2013-03-01

    An efficient plant regeneration protocol was established for an endangered ethnomedicinal plant Desmodium gangeticum (Linn.) DC. Morphogenic calli were produced from 96 % of the cultures comprising the immature leaf explants on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (4.0 mg l(-1)) in combination with 6-benzylaminopurine (BA; 0.8 mg l(-1)). For callus regeneration, various concentrations of BA (1.0-5.0 mg l(-1)) or thidiazuron (TDZ; 1.0-5.0 mg l(-1)) alone or in combination with indole-3-acetic acid (IAA; 0.2-1.0 mg l(-1)) were used. Highest response of shoot regeneration was observed on MS medium fortified with TDZ (4.0 mg l(-1)) and IAA (0.5 mg l(-1)) combination. Here, 100 % cultures responded with an average number of 22.3 shoots per gram calli. Inclusion of indole-3-butyric acid in half MS medium favored rooting of recovered shoots. Out of 45 rooted plants transferred to soil, 40 survived. Total DNA was extracted from the leaves of the acclimatized plants of D. gangeticum. Analysis of random amplified polymorphic DNA using 13 arbitrary decanucleotide primers showed the genetic homogeneity in all the ten plants regenerated from callus with parental plant, suggesting that shoot regeneration from callus could be used for the true-to-type multiplication of this plant.

  14. Translational Enhancer of Tobacco mosaic virus Enhancing Expression of Hepatitis B Surface Antigen in Transgenic Panax ginseng C. A. Meyer Callus

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The 5'-nontranslated leader(omega sequence) of Tobacco mosaic virus(TMV) was used as a translational enhancer sequence in the expression of the hepatitis B surface antigen(HBsAg) gene in transgenic ginseng callus cultures.The adr subtype HBsAg gene was placed under the control of the Cauliflower mosaic virus(CaMV) 35S promoter linking to the TMV leader sequence. The antisense omega sequence was used in a control construct. The resulting constructs cloned in the binary vector pBI121 were used to transform the ginseng callus tissue via the Agrobacterium-mediated procedure. The integration and expression of the HBsAg gene were evaluated by PCR and western blot, respectively. Enzyme-linked immunoassays(ELISA) using a monoclonal antibody directed against human serum-derived HBsAg revealed a three to four-fold enhanced expression of HBsAg in ginseng cells conferred by the TMV omega element.

  15. NAA与BA对康乃馨愈伤组织诱导的影响%Effects of NAA and BA on Callus Induction of Dianthus Caryophyllus

    Institute of Scientific and Technical Information of China (English)

    王允威; 刁毅

    2012-01-01

    In the paper, effects of NAA and BA on callus induction of Dianthus Caryophyllus was studied to provide the theoretical basis for tissue culture of Dianthus Cao'ophyllus. As a result, the media with NAA or BA was in favor of the induction of callu, while media adding NAA 0.2 mg/L and BA 2 mg/L was the best hormone combination, induction rate was best, and callus grew well.%文章主要研究了NAA与BA对康乃馨愈伤组织诱导的影响,结果表明:单独使用NAA或BA时,能够诱导出愈伤组织;NAA浓度为0.2mg/L,BA浓度为2mg/L时,产生愈伤组织比例最高,生长状态最好。

  16. Investigations on some metabolites of Tecoma stans Juss. callus tissue. Part III. Chromatographical search for iridoids, phenolic acids, terpenoids and sugars

    Directory of Open Access Journals (Sweden)

    Barbara Dohnal

    2015-01-01

    Full Text Available Tissus cultures of Tecoma stans Juss. cultivated on modified Murashige-Skoog medium (RT-k were phytochemically analysed by means of chromatographical methods (PC, TLC. The following products were found as metabolites: phenolic acids - chlorogenics, caffeic, ferulic, vanillic, o-coumaric and sinapic; steroids - β-sitosterol; triterpenes - ursolic and oleanolic acids, α-amyrine; sugars - glucose, fructose, sucrose, xylose. Meso-inositol was isolated in 0.8% yield. In intact plant leaves, some differences concerning the content and/or number of individual compounds were observed, namely: lack of sinapic acid and occurrence of p-coumaric acid, lower content of β-sitosterol, lack of oleanolic acid, occurrence of β-amyrine and of one unidentified triterpenoid, lack of xylose, occurrence of maltose, raffinose, and stachiose. The level of mesoinositol inn leaves was distincly lower than in the callus tissues. Neither in callus tissues nor in leaves iridoid glycosides were found.

  17. Efficient production of fermentable sugars from oil palm empty fruit bunch by combined use of acid and whole cell culture-catalyzed hydrolyses.

    Science.gov (United States)

    Li, Qingxin; Ng, Wei Ting; Puah, Sze Min; Bhaskar, Ravindran Vijay; Soh, Loon Siong; MacBeath, Calum; Parakattil, Pius; Green, Phil; Wu, Jin Chuan

    2014-01-01

    Empty fruit bunch (EFB) of oil palm trees was converted to fermentable sugars by the combined use of dilute acids and whole fungal cell culture-catalyzed hydrolyses. EFB (5%, w/v) was hydrolyzed in the presence of 0.5% H2 SO4 and 0.2% H3 PO4 at 160 °C for 10 Min. The solid fraction was separated from the acid hydrolysate by filtration and subjected to enzymatic hydrolysis at 50 °C using the whole cell culture of Trichoderma reesei RUT-C30 (2%, w/v), which was prepared by cultivation at 30 °C for 7 days to reach its maximal cellulase activity. The combined hydrolyses of EFB gave a total sugar yield of 82.0%. When used as carbon sources for cultivating Escherichia coli in M9 medium at 37 °C, the combined EFB hydrolysates were shown to be more favorable or at least as good as pure glucose for cell growth in terms of the higher (1.1 times) optical density of E. coli cells. The by-products generated during the acid-catalyzed hydrolysis did not seem to obviously affect cell growth. The combined use of acid and whole cell culture hydrolyses might be a commercially promising method for pretreatment of lignocellulose to get fermentable sugars.

  18. Hypolipidemic Effects of Biopolymers Extracted from Culture Broth, Mycelia, and Fruiting Bodies of Auricularia auricula-judae in Dietary-induced Hyperlipidemic Rats

    Science.gov (United States)

    Jeong, Hun; Yang, Byung-Keun; Jeong, Yong-Tae; Kim, Guk-Nam; Jeong, Yu-Sun; Kim, Sang-Min; Mehta, Pradeep

    2007-01-01

    Hypolipidemic effect of biopolymers extracted from culture broth (CP), mycelia (MP), and fruiting bodies (FP) of Auricularia auricula-judae was investigated in dietary-induced hyperlipidemic rats. The experimental animals were administrated (100 mg/kg body weight) with different biopolymers, daily for 4 weeks. Hypolipidemic effects were achieved in all the experimental groups, however, FP was proved to be the most potent one. The administration of the FP reduced the plasma triglyceride, total cholesterol, low-density lipoprotein cholesterol, and atherogenic index by 24.3, 28.5, 36.4, and 40.9%, respectively, while increased the high-density lipoprotein cholesterol level (9.0%), when compared to the saline (control) administered group. PMID:24015062

  19. The effect of electric field on callus induction with rape hypocotyls

    Energy Technology Data Exchange (ETDEWEB)

    Filek, M.; Holda, M. [Inst. of Plant Physiology, Polish Academy of Sciences, Krakow (Poland); Machackova, I.; Krekule, J. [Inst. of Experimental Botany, Academy of Sciences of the Czech Republic, Praha (Czech Republic)

    2005-12-01

    The influence of electric field treatment on dedifferentiation and calli formation on rape hypocotyls was investigated. Segments, 10 mm long, of the upper part of rape (Brassica napus L., cv. Gorcza{delta}ski) hypocotyls were stimulated by different combinations of voltage/time (1.5 V/120 h, 3 V/3 h, 10 V/15 min and 30 V/30 s) under in vitro conditions. With all electric field treatments, segments oriented with their apical part towards the cathode produced more calli as compared to control (non-treated with electric field). Under opposite orientation slight inhibition of callus growth was observed. As the strongest effect on callus growth was observed after treatment with 30 V/30 s, this electric field treatment was selected for following analyses: the incorporation of [{sup 14}C]-2,4-D (2,4-dichlorophenoxyacetic acid) and [{sup 14}C]-BAP (benzylaminopurine) from the culture medium, changes in ACC (1-aminocyclopropane-1-carboxylic acid) level and the redox activity in apical and bottom parts of hypocotyls during 18 d of culture. In contrast to changes in fresh weight, electric field treatment (30 V/30 s) stimulated a higher accumulation of 2,4-D and BAP in basal parts of hypocotyls than in apical ones. Moreover, orienting the apical part towards the cathode resulted in lower uptake of hormones as compared with the opposite orientation. The ACC concentration increased, especially in the basal parts of hypocotyls, independently on electric field application. However, the highest level was observed after electric field treatment with orientation of the apical part towards the anode. The distribution of oxidative substances (measured as the amount of ferric ions) between the apical and bottom part of hypocotyls was not changed when the apical parts were oriented towards the cathode. Under these conditions a decrease in apical and an increase in basal parts was observed during culture. Opposite orientation influenced the redistribution of oxidative substances from the

  20. Aplicações da cultura de tecidos vegetais em fruteiras do Cerrado Applications of tissue culture techniques in Brazilian Cerrado fruit trees

    Directory of Open Access Journals (Sweden)

    Hernane Fernandes Pinhal

    2011-07-01

    Full Text Available Atualmente, percebe-se uma preocupação em relação às plantas do cerrado, com grande enfoque nas fruteiras em função de suas características e usos. Apesar de ser uma área ainda pouco explorada, é crescente o número de estudos dessas espécies nativas, dentre eles, os que abrangem as técnicas de cultura de tecidos. Isso se deve uma vez que essa ferramenta biotecnológica permite a propagação de espécies com dificuldade de germinação, minimiza o problema de sementes recalcitrantes, promove a produção de mudas em larga escala, complementa bancos de germoplasma e facilita as trocas de materiais genéticos. Dessa maneira, esta revisão visa a sumarizar o histórico e panorama atual das aplicações da cultura de tecidos em fruteiras do cerrado, proporcionando sustentação para novos estudos.Currently, it's been given a huge concern to the cerrado plants, focusing on fruit trees due to their characteristics and uses. Despite being a fairly unexplored area, the number of studies on these native species has increased, especially those involving tissue culture techniques. That's because this biotechnological tool provides the propagation of species with germination difficulty, reduces problems of recalcitrant seeds, promotes large scale seedling production, complements germplasm banks and facilitates the exchange of genetic materials. Therefore, this review summarizes the history and current situation of tissue culture techniques applied to Brazilian Cerrado fruit trees, providing support to further studies.

  1. Fruit Juice.

    African Journals Online (AJOL)

    The Effect of Homogenization, Stabilizer and Amylase Treatment on Viscosity of Passion. Fruit Juice. ... viscosity during storage of sweetened, pasteurized passion fruit juice were investigated. .... minutes after which the temperature was.

  2. Callus formation in vitro and internodal stem apices in savory = Calogênese in vitro de segmentos apicais caulinares e internodais em segurelha (Satureja hortensis L.

    Directory of Open Access Journals (Sweden)

    Marcio Carlos Navroski

    2012-12-01

    Full Text Available We sought to evaluate with this work different growth regulators on callus formation in shoot apical and internodal stem segments of Satureja hortensis. The explants were isolated from in vitro seedlings and cultured on MS nutrient medium supplemented with NAA (0 and 1 μM and BAP (0, 5, 10, 15 and 20 μM. The presence of auxin NAA gave higher marks to the calluses at 30 days of evaluation, these notes also increased with the addition of BAP. There were significant interactions between factors in evaluating NAA and BAP for 60 days, both in apical stem segments as in internodal stem segments. To stemapices percentage of friable callus tends to decrease with increasing concentration of BAP. As for compact calluses increased BAP leads to an increase in the percentage of this type of callus. The presence of NAA increased callus formation in both friable and compact calluses on. This trend was also observed in internodal stem segments. The callus formation was highly rhizogenic observed in the presence of NAA, hardly occurs in the absence of auxin. The use of BAP is recommended in case of regeneration of plants through micropropagation, if the goal is the production of metabolites, the use of BAP can be harmful by reducing the production of friable callus.= Buscou-se com este trabalho avaliar diferentes reguladores de crescimento sobre a calogênese em segmentos caulinares apicais e internodais de Satureja hortensis L.. Os explantes foram isolados de plântulas germinadas in vitro e cultivados em meio nutritivo MS acrescido de ANA (0 e 1 μM e de BAP (0; 5; 10; 15 e 20 μM. A presença da auxina ANA proporcionou notas mais altas para os calos aos 30 dias de avaliação, estas notas também aumentaram com o acréscimo da citocinina BAP. Houve interações significativas entre os fatores ANA e BAP na avaliação aos 60 dias, nos dois tipos de segmentos caulinares(apicais e internodais. Nos segmentos apicais caulinares a porcentagem de calos fri

  3. Effects of Hormones on Callus Induction and Propagation of Pteridium aquilinum (L.) kuhn%不同浓度激素对蕨菜愈伤组织诱导与增殖的影响

    Institute of Scientific and Technical Information of China (English)

    方利娟; 苏仕林; 蒲仕明

    2012-01-01

    Using tender rhizomes of fern as explants, different concentration of 6-BA and IBA were added in callus induction and propagation mediums in order to study their effects on callus induction and propagation. The results indicated that 1/2 MS+0.2mg/L 6-BA+0.4 mg/L IBA was the best callus induction medium with an induction rate of 82%; 1/2 MS+0.2 mg/L IBA+0.2 mg/L 6-BA was the best callus proliferation medium as the callus was proliferated by 8.07 times.The results of this study could provide references for reasonable use of 6-BA and IBA in callus culture of Pteridium aquilinum.%以蕨菜[ Pteridium aquilinum(L.)kuhn]幼嫩根状茎为外植体,在愈伤组织诱导和增殖培养基中设置不同的6-BA和IBA浓度,研究6-BA和IBA对蕨菜愈伤组织诱导及增殖的影响.结果表明,1/2MS+0.2mg/L 6-BA+0.4 mg/L IBA为愈伤组织的最佳诱导培养基,诱导率为82%;1/2MS+0.2 mg/L IBA+0.2mg/L 6-BA为愈伤组织的最佳增殖培养基,增殖倍数为8.07.该研究结果为6-BA和IBA在蕨菜愈伤组织培养中的合理使用提供了参考.

  4. Callus induction from protoplasts of V. unguiculata, V. sublobata and V. mungo.

    Science.gov (United States)

    Gill, R; Eapen, S; Rao, P S

    1987-05-01

    Protoplasts were isolated from hypocotyl of V. mungo (L.) Hepper or hypocotyl-derived callus of V. sublobata (Phaseolus sublobata Roxb.) and V. unguiculata (L.) Walp (syn. V. sinensis (L.) Saviex Hassk) using an enzyme solution comprising Cellulase 2.5%, Macerozyme, Hemicellulase and Driselase each at a 0.5% level in 0.5 M sorbitol. Isolated protoplasts were cultured in Murashige and Skoog's (1962) basal liquid medium supplemented with BA, NAA, 2,4-D (1 mg/l each) and sucrose (14%). After four weeks, protoplast colonies were transferred to the same medium with a reduced level of sucrose (7%). Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones of V. unguiculata differentiated roots on auxin/cytokinin supplemented media. Alternative methods for shoot differentiation from protoplastderived cultures were tried by the use of Agrobacterium tumefaciens "shooter" strains pGV 2215 or pGV 2298 or wild type strain B6S3.

  5. Callus induction and plant regeneration from mature bluegrass (Poa pratensis L.) seeds

    Institute of Scientific and Technical Information of China (English)

    Sun Bi-po; Zhang Wan-jun; Dong Jiang-li; Jin Yong-sheng; Wang Tao

    2006-01-01

    A protocol was discussed for high efficient plant regeneration from seven bluegrass (Poa pratensis L.) cultivars via an indirect callus induction and somatic embryogenesis method. Mature seeds were used as explants for callus initiation. Callus induction and proliferation efficiencies were investigated on NB, modified MS (MMS) and MS media, supplemented with 2.0 mg.L-12,4-dichlorophenoxyacetic acid (2,4-D). The MMS medium performed best. Based on the MMS medium, direct and indirect callus the direct callus induction method, the most suitable 2,4-D concentrations varied among cultivars. Under the indirect callus induction method, a significantly high callus induction frequency (93.33%-98.33%) was obtained and there were barely any statistically significant differences among the tested genetically diverse cultivars. Somatic embryos were promoted on the MMS medium supplemedium containing different concentrations of thidiazuron (TDZ), and the differentiation frequencies varied in the range from 20.15% to 77.65%. The 0.25 mg·L-1 TDZ was generally the most suitable concentration for the tested cultivars.

  6. Femoral shaft fracture callus formation after intramedullary nailing: a comparison of interlocking and Ender nailing.

    Science.gov (United States)

    Yamaji, Tetsuo; Ando, Kenichi; Nakamura, Toshiyasu; Washimi, Ohsuke; Terada, Nobuki; Yamada, Harumoto

    2002-01-01

    Intramedullary nailing is widely used for the operative treatment of femoral fractures. Recently, the biologic healing of fractures has become better understood from fundamental investigations. However, there has been no clinical comparison between the fracture healing process with these two fixation methods. The purpose of this study was to use radiographs to compare callus formation with two types of intramedullary nailing for femoral shaft fractures: reamed interlocking (IL) nails and Ender nails. Femoral shaft type A fractures (AO classification) were studied. Twenty-seven fractures were treated with reamed IL nailing, and 81 fractures were treated with Ender nailing. The callus area was calculated from the maximum cross-sectional area on the anteroposterior and lateral radiographs. The callus appeared at a mean of 3.9 weeks after surgery in the IL group, and at a mean of 2.8 weeks in the Ender group ( P Ender groups, fracture healing was noted at a mean of 3.4 and 2.0 months, respectively. The mean area of callus formation in the IL and Ender nailing groups was 439.5 mm(2) and 699.4 mm(2), respectively ( P Ender nailing results in abundant callus, which forms at an earlier stage after the procedure than in patients treated with IL nailing. Dynamization at the fracture site is reported to increase external callus formation. Our results indicate that the elasticity of the fixation obtained with Ender nailing promotes callus formation.

  7. In vitro androgenetic cultures of Hyoscyamus niger L., H. albus L. and alkaloid content assay

    Directory of Open Access Journals (Sweden)

    Maria Wesołwska

    2014-01-01

    Full Text Available In vitro cultures of Hyoscyamus niger L. and H. albus L. anthers were initiated which resulted in obtaining androgenectic plants and callus cultures. The leaves of these pants and the callus cultures were subjected to analysis (TLC, GC for the presence of alkaloids, derivatives of tropane. In the studied material, alkaloids of different qualitative and quantitative composition from that of ground-grown plants were found.

  8. Compositional variation of fibrous callus and joint cartilage in different internal environments

    Institute of Scientific and Technical Information of China (English)

    SUN Xiao-tang; HU Yun-yu; ZHAO Li; L(U) Rong; WANG Jun; BAI Jian-ping

    2006-01-01

    Objective: To evaluate the compositional variation of fibrous callus in the fracture site and the joint cavity and joint cartilage after being transplanted in the muscle pouch.Methods: Thirty 2-month-old New Zealand white rabbits (weighing 1-1.5 kg) were randomly divided into two groups: a callus transplantation group (Group A, n =15) and a cartilage transplantation group ( Group B, n =15). In Group A, closed radius fracture was made and the autologous fibrous callus was transplanted in the right knee joint cavity at 12 days postoperatively. In Group B, the right knee joint cartilage of the animals was transplanted in the autologous back muscle pouches under anesthesia. Then all the animals were killed by overdose anesthetic 3 weeks after transplantation. And the transplanted fibrous callus,the healed bones in the fracture sites and the transplanted joint cartilage were obtained for assessment of compositional variation.Results: Pure fibrous composition was found in the callus at the fracture sites in Group A at 12 days postoperatively. And for 11 out of the 15 animals, the fibrous callus was transformed into cartilaginous tissues after 3 weeks of transplantation, but the fibrous callus was absent in the other 4 animals. The fibrous calluses at the original site and the fracture locus were differentiated into bony tissues. Bony tissue transformation was found in the transplanted joint cartilages in the muscle pouch of all the animals in Group B.Conclusions: The fracture sites or joint cavity may facilitate callus differentiation in different ways: the former is helpful for osteogenesis while the latter for the development and maintenance of cartilages, and the muscle pouch is inclined to induce the osteogenic phenotype for cartilages.

  9. Callus cell, shoot and stem proliferation data from pineapple crown and banana inflorescence in vitro: Biochemical and antioxidant properties

    Directory of Open Access Journals (Sweden)

    ABM Sharif Hossain

    2017-02-01

    Full Text Available The data article contains the experimental data and images on the callus cell, shoot and stem proliferation from pineapple crown slice and banana inflorescence in vitro. Investigated data are related to the research article “Effects of benzylaminopurine and naphthalene acetic acid on proliferation and shoot growth of pineapple (Ananas comosus L. Merr in vitro” Alsaif et al. (2011 [1] and “Plantlet Production through Development of Competent Multiple Meristem Cultures from Male Inflorescence of Banana, Musa acuminta cv. ׳Pisang Mas׳(AA” Wirakarnain et al. (2008 [2]. In the experimental data 1, physiological, (shoot weight, number length and stem proliferation biochemical (total sugar and chlorophyll and nutritional ((K+ and NO3− data using BAP, MS medium and NAA growth regulators in pineapple have been explored. In the experimental data 2, physiological, (callus weight, shoot number and length biochemical (total sugar, chlorophyll, total phenol, DPPH and nutritional (K+ and NO3− data employing BAP +IAA, MS medium and NAA growth regulators in banana have been exhibited. Overall quantitative measurement was observed by Spectrophotometer. In the experimental data, BAP was shown the best effective hormone for the both pineapple and banana explants regeneration.

  10. Exuberant callus formation misdiagnosed as osteosarcoma: a case report

    Institute of Scientific and Technical Information of China (English)

    Fariba Binesh; Mohammad Sobhan; Reza Nafisi Moghadam; Ali Akhavan

    2013-01-01

    Reactive lesions of bone and soft tissue can appear alarming on histologic examination because they are often cellular and have atypical cytologic features, such as distinct nucleoli, mild hyperchromasia, and mitotic activity. Reactive lesions of bone and periosteum also produce bone and cartilage matrix, resulting in confusion with osteosarcoma or chon-drosarcoma. Careful attention to key cytomorphological features such as the pattern of bone formation, uniform appearance of cells, and absence of atypical mitoses should help identify the reactive nature of a lesion. Correlation with clinical and radiological findings is also imperative to avoid misclassification of the tumor because reactive lesions often arise at sites where osteosarcoma and chondrosarcoma are rare (eg, the hand) and lack aggressive radiological features. Here we pres-ent a case of exuberant callus formation after avulsion fracture of tibia in a three year-old Iranian girl which misdiagnosed as osteosarcoma.

  11. Some Factors Affecting Callus Induction from Oil Palm Leaves%影响油棕叶片愈伤组织诱导因素研究

    Institute of Scientific and Technical Information of China (English)

    邹积鑫; 尤丽莉; 林位夫

    2014-01-01

    研究不同取材部位、激素组合配比等因素对3个不同油棕品系叶片愈伤组织培养的影响。结果表明:不同部位的油棕叶片以及不同品系的油棕愈伤组织诱导存在极显著差异;试验以MS培养基+5 mg/L 2,4-D+10 mg/L TDZ+6 g/L琼脂+30 g/L糖配比获得较好的愈伤诱导率;添加2%活性炭可以有效抑制油棕叶片愈伤组织褐化。%Different parts of leaves and hormone combinations involved in oil palm leave callus culture for three strains of oil palm were studied. The result indicated that different parts of leaves and different strains of oil palm have significant difference in callus induction. The optimum medium components for oil palm callus induction were Murashige-Skoog (MS) medium supplemented with 5 mg·L-12, 4-D and 10 mg/L TDZ and 6 g/L agar and 30 g/L sucrose. The supplying of 2% charcoal can prevent explants from browning efficiently.

  12. Which perceived characteristics make product innovations appealing to the consumer? A study on the acceptance of fruit innovations using cross-cultural consumer segmentation

    NARCIS (Netherlands)

    Onwezen, M.C.; Bartels, J.

    2011-01-01

    In general, fruit consumption in the EU does not meet governments' recommended levels, and innovations in the fruit industry are thought to be useful for increasing fruit consumption. Despite the enormous number of product innovations, the majority of new products in the market fail within the first

  13. Which perceived characteristics make product innovations appealing to the consumer? A study on the acceptance of fruit innovations using cross-cultural consumer segmentation

    NARCIS (Netherlands)

    Onwezen, M.C.; Bartels, J.

    2011-01-01

    In general, fruit consumption in the EU does not meet governments' recommended levels, and innovations in the fruit industry are thought to be useful for increasing fruit consumption. Despite the enormous number of product innovations, the majority of new products in the market fail within the first

  14. 新疆紫草毛状根愈伤组织诱导及其生长条件研究(Ⅰ)%Xinjiang Anti- freckle Hairy Root Callus Induction and Its Growing Conditions Research (Ⅰ)

    Institute of Scientific and Technical Information of China (English)

    敬碧; 毛拉艾麦提·阿不都卡迪尔; 王芳; 唐雪梅

    2011-01-01

    [目的]建立以新疆紫草毛状根愈伤组织为基础的细胞悬浮体系.[方法]以新疆紫草毛状根为外植体,探讨激素、培养基、蔗糖、钙离子等对愈伤组织诱导、生长的影响.[结果]MS附加2,4-D 1.0 mg/L+ KT0.5 mg/L的愈伤组织诱导率为100%;钙离子浓度为250 mg/L时的B5 +6- BA0.1+ NAA 0.5愈伤鲜重最多,愈伤增殖倍数为9.63倍;愈伤组织在P-1中的生长量均较小,但愈伤的颜色普遍较红,当钙离子浓度为250mg/L、P-1附加1.0 mg/L KT的处理愈伤最红.[结论]毛状根是诱导新疆紫草的最佳外植体.研究为利用新疆紫草毛状根进行细胞悬浮培养提供了理论依据.%[Objective] The purpose of this program was to establish cell suspension system based on Xinjiang Anti- freckle Hairy Root Callus. [Method]Hie study, by taking Xinjiang anti - freckle hairy root as explant, probes into the effects of hormones,culture medium, sucrose, calcium ion on callus induction and growth. [Result] Results show that: MS, annexed with 2,4 - D 1.0 mg/L + KT. 0.5 mg/L, had 100% callus induction; and when calcium ion concentration was 250 mg/L for the B5 + 6 - BA 0.1 + NAA 0.5 callus fresh weight was the most, and callus proliferation multiples 9.63 times; callus in P- 1 were smaller, but the growth of callus color was generally red, when calcium ion concentration was 250 mg/L, P for- 1 additional 1.0 mg/L KT processing callus was at large red. [Conclusion]This study concluded that Xinjiang hairy roots were optimal explant, which has provided theoretical basis for cultivating cell suspension by applying Xinjiang anti - freckle hairy roots.

  15. 藏红花愈伤组织的诱导及植株再生%Callus Induction and Plant Regeneration of Saffron

    Institute of Scientific and Technical Information of China (English)

    王寿芹; 赵永钦; 刘莉莎; 郭仰东

    2011-01-01

    以藏红花(Crocus sativus L.)当年新生小球茎为材料,在MS+0.5 mg/L BA+1.0 mg/L NAA+7%CM(椰乳)培养基上进行初代培养,再转入MS+5 mg/L BA+5 mg/L NAA中进行愈伤组织的诱导,黑暗条件下继代1-2次,51 d就可以获得高诱导串的愈伤组织,愈伤诱导率可达65%.将愈伤组织转入分化培养基MS+5 mg/L BA+5 mg/L NAA中分化丛生芽,丛生芽诱导率高达95%,将丛生芽分成单个植株,最终形成完整再生植株.%The corms of saffron were used as explanta to establiah a regeneration protocol for Cro cus sativus L. The corms incubated in MS + 0. 5 mg/L BA +1.0 mg/L NAA +7 % CM at first, then tranaferred to MS +5 mg/ L BA +5 mg/ L NAA for callus induction. Callus formation was obaened after 51 days , culture and the callus induction rate was 65 % . After four weeks callus induction, the shoot regeneration rate (95 % ) was obtained when 2 mg/L BA was combined with 1 mg/L NAA. The shoot buds were fragmented and transfered to root induction medium containing 0.5 mg/L IBA. Plantlets were then successfully transplanted to greenhouse.

  16. Effect of Plant Growth Regulators on Callus, Cell Suspension and Cell Line Selection for Flavonoid Production from Pegaga (centella asiatica L. urban

    Directory of Open Access Journals (Sweden)

    Suat H. Tan

    2010-01-01

    Full Text Available Problem statement: Considering pegaga medicinal properties and over-exploitation, the requirement for a tissue culture technique as an alternative production system was crucial. Approach: Investigation of cell suspension culture response to different plant growth regulators (PRGs for flavonoid production from elite cell line was carried out. Callus cultures were initiated from the leaf explants of Centella asiatica on Murashige and Skoog (MS medium containing B5 vitamins and 30 g L−1 sucrose supplemented with different concentrations (0.5-2.5 mg L−1 of 2,4-D, NAA, Dicamba, Picloram and IBA supplied singly and in combination with different concentrations (0.5-1.5 mg L−1 of kinetin, BAP and TDZ. Results: Callus induction was observed for all the PGRs tested. The highest callus induction frequency (86.67% was observed in MS medium containing 2.0 mg L−1 2,4-D while the combination of 2.0 mg L−1 2,4-D and 1 mg L−1 kinetin in MS medium gave the highest biomass yield (0.27 g dry weight culture−1. This combination was also found to be best for callus proliferation for all the accessions investigated. Among the four accessions tested, UPM03 was found to have the highest biomass yield (0.041 g DW culture−1 and hydrolysed flavonoid content (10.75 mg g−1 DW after the 12th day of culture. The flavonoids present in the four accessions were quercetin, kaempherol, luteolin and rutin based on high performance liquid chromatography (HPLC analysis. These results indicated that C. asiatica accession UPM03 was the potential elite cell line in mass production of flavonoid, especially luteolin. Coclusions/Recommendations: In the establishment of cell suspension culture, 2 mg L−1 2,4-D and 1 mg L−1 kinetin were the best PGRs in supporting the cell growth and flavonoid production. This is the first report on the use of PRGs on the establishment of cell suspension cultures in flavonoid production of C. asiatica.

  17. Further improvement of N6 medium for callus induction and plant ...

    African Journals Online (AJOL)

    user

    2011-03-02

    Mar 2, 2011 ... ... Sichuan Agricultural University, Chengdu, Sichuan, 611130, People's Republic of China. ... necessary to improve the media used for callus induction ..... relation to changes in photosynthesis, hormones and antioxidant.

  18. Bringing back the fruit into fruit fly-bacteria interactions.

    Science.gov (United States)

    Behar, A; Jurkevitch, E; Yuval, B

    2008-03-01

    Female Mediterranean fruit flies (Ceratitis capitata) oviposit in fruits, within which the larvae develop. This development is associated with rapid deterioration of the fruit, and frequently with invasion by secondary pests. Most research on the associations between medflies and microorganisms has focused on the bacteria inhabiting the digestive system of the adult fly, while the role of the fruit in mediating, amplifying or regulating the fruit fly microflora has been largely neglected. In this study, we examine the hypothesis that the host fruit plays a role in perpetuating the fly-associated bacterial community. Using direct and cultured-based approaches, we show that this community is composed in its very large majority of diazotrophic and pectinolytic Enterobacteriaceae. Our data suggest that this fly-associated enterobacterial community is vertically transmitted from the female parent to its offspring. During oviposition, bacteria are transferred to the fruit, establish and proliferate within it, causing its decay. These results show that the host fruit is indeed a central partner in the fruit fly-bacterial interaction as these transmitted bacteria are amplified by the fruit, and subsequently maintained throughout the fly's life. This enterobacterial community may contribute to the fly's nitrogen and carbon metabolism, affecting its development and ultimately, fitness.

  19. Role of brassinosteroids in alleviating toxin-induced stress of Verticillium dahliae on cotton callus growth.

    Science.gov (United States)

    Bibi, Noreen; Ahmed, Imrul Mosaddek; Fan, Kai; Dawood, Muhammad; Li, Feng; Yuan, Shuna; Wang, Xuede

    2017-03-29

    Brassinosteroids are well known to mitigate biotic stresses; however, their role to induce tolerance against Verticillium dahliae is unknown. The current study employed V. dahliae (Vd) toxin as pathogen-free model system to induce stress on cotton callus growth, and its amelioration was investigated using 24-epibrassinolide (EBR). Results revealed that EBR has ameliorative effects against Vd toxin with greater seen effect when callus was treated with EBR prior to its exposure to Vd toxin (pre-EBR treatment) than EBR applied along with Vd toxin simultaneously (co-EBR treatment). Pre-EBR-treated calli remained green, while 65 and 90% callus browning was observed in co-EBR- and Vd toxin-alone-treated callus, respectively. Likewise, the fresh weight of the pre-EBR-treated callus was 52% higher than Vd toxin-alone treatment, whereas this increase was only 23% in co-EBR-treated callus. Meanwhile, EBR treatment of the cotton callus has also increased the contents of chlorophylls a and b, carotenoids, total phenols, flavonoids, soluble sugars, and proteins and increased the activity of enzymes involved in secondary metabolism like polyphenol oxidase (PPO), phenylalanine ammonialyase (PAL), cinnamyl alchol dehydrogenase (CAD), and shikimate dehydrogenase (SKDH) over Vd toxin-alone treatment with higher increments being observed in pre-EBR-treated callus. Furthermore, EBR treatment mimicked the DNA damage and improved the structure of mitochondria, granum, stroma thylakoids, and the attachment of ribosomes with the endoplasmic reticulum. This EBR-mediated mitigation was primarily associated with substantially increased contents of photosynthetic pigments and regulation of secondary metabolism.

  20. Ethylene and nitric oxide are involved in maintaining ion homeostasis in Arabidopsis callus under salt stress.

    Science.gov (United States)

    Wang, Huahua; Liang, Xiaolei; Wan, Qi; Wang, Xiaomin; Bi, Yurong

    2009-07-01

    In the present study, the role of ethylene in nitric oxide (NO)-mediated protection by modulating ion homeostasis in Arabidopsis callus under salt stress was investigated. Results showed that the ethylene-insensitive mutant etr1-3 was more sensitive to salt stress than the wild type (WT). Under 100 mM NaCl, etr1-3 callus displayed a greater electrolyte leakage and Na(+)/K(+) ratio but a lower plasma membrane (PM) H(+)-ATPase activity compared to WT callus. Application of exogenous 1-aminocyclopropane-1-carboxylic acid (ACC, an ethylene precursor) or sodium nitroprusside (SNP, a NO donor) alleviated NaCl-induced injury by maintaining a lower Na(+)/K(+) ratio and an increased PM H(+)-ATPase activity in WT callus but not in etr1-3 callus. The SNP actions in NaCl stress were attenuated by a specific NO scavenger or an ethylene biosynthesis inhibitor in WT callus. Under 100 mM NaCl, the NO accumulation and ethylene emission appeared at early time, and NO production greatly stimulated ethylene emission in WT callus. In addition, ethylene induced the expression of PM H(+)-ATPase genes under salt stress. The recovery experiment showed that NaCl-induced injury was reversible, as signaled by the similar recovery of Na(+)/K(+) ratio and PM H(+)-ATPase activity in WT callus. Taken together, the results indicate that ethylene and NO cooperate in stimulating PM H(+)-ATPase activity to modulate ion homeostasis for salt tolerance, and ethylene may be a part of the downstream signal molecular in NO action.

  1. Evaluation of cytotoxicity of Moringa oleifera Lam. callus and leaf extracts on Hela cells

    OpenAIRE

    Abbas Jafarain; Gholamreza Asghari; Erfaneh Ghassami

    2014-01-01

    Background: There are considerable attempts worldwide on herbal and traditional compounds to validate their use as anti-cancer drugs. Plants from Moringaceae family including Moringa oleifera possess several activities such as antitumor effect on tumor cell lines. In this study we sought to determine if callus and leaf extracts of M. oleifera possess any cytotoxicity. Materials and Methods: Ethanol-water (70-30) extracts of callus and leaf of M. oleifera were prepared by maceration method...

  2. Effect of Gamma Irradiation on Regeneration Capacity of Mangosteen Nodular Callus

    Directory of Open Access Journals (Sweden)

    WARID ALI QOSIM

    2007-12-01

    Full Text Available The research was conducted to determine the effect of gamma irradiation on regeneration capacity of mangosteed nodular callus. Nodular calli derived from a leaf as explants and cultured on MS medium containing combination of 2.2 µM benzilaminopurin (BAP and 2.27 µM tidiazuron (TDZ. Nodular calli were irradiated with 0 (control 5, 10, 15, 20, 25, 30, 35, and 40 Gy doses of gamma irradiation. After the irradiation, the calli were generated on woody plant medium (WPM, supplemented with 1.39 µM polyvinilpirolidon (PVP, 8 g.l-1 agar, 30 g.l-1 sucrose and 2.2 µM BAP concentration. Results showed that the irradiation influence the plant regeneration. Response dose of 50% (RD that could promote the nodular calli of shoot formation was the 25 Gy while that of the shoot number per nodular calli was the 21 Gy. The shoot number irradiated with total dose 5 Gy (9.1 shoot was higher than that of 0 Gy (8.6 shoot.

  3. Global analysis of differentially expressed genes and proteins in the wheat callus infected by Agrobacterium tumefaciens.

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    Xiaohong Zhou

    Full Text Available Agrobacterium-mediated plant transformation is an extremely complex and evolved process involving genetic determinants of both the bacteria and the host plant cells. However, the mechanism of the determinants remains obscure, especially in some cereal crops such as wheat, which is recalcitrant for Agrobacterium-mediated transformation. In this study, differentially expressed genes (DEGs and differentially expressed proteins (DEPs were analyzed in wheat callus cells co-cultured with Agrobacterium by using RNA sequencing (RNA-seq and two-dimensional electrophoresis (2-DE in conjunction with mass spectrometry (MS. A set of 4,889 DEGs and 90 DEPs were identified, respectively. Most of them are related to metabolism, chromatin assembly or disassembly and immune defense. After comparative analysis, 24 of the 90 DEPs were detected in RNA-seq and proteomics datasets simultaneously. In addition, real-time RT-PCR experiments were performed to check the differential expression of the 24 genes, and the results were consistent with the RNA-seq data. According to gene ontology (GO analysis, we found that a big part of these differentially expressed genes were related to the process of stress or immunity response. Several putative determinants and candidate effectors responsive to Agrobacterium mediated transformation of wheat cells were discussed. We speculate that some of these genes are possibly related to Agrobacterium infection. Our results will help to understand the interaction between Agrobacterium and host cells, and may facilitate developing efficient transformation strategies in cereal crops.

  4. Micropropagation and callus induction of Lantana camara L. – A medicinal plant

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    Varaporn Veraplakorn

    2016-09-01

    Full Text Available Lantana camara L., a medicinal plant, exhibits antimicrobial, fungicidal, insecticidal and nematicidal properties. Effective in vitro micropropagation and callus induction aid plant material production for bioactive compound studies and plant resource conservation. Shoot multiplication, root induction and callus formation were investigated. The results indicated a high shoot multiplication rate on Murashige and Skoog (MS medium supplemented with 16.0 μmol/L N6-benzyladenine (BA. Shoots showed good root regeneration with healthy plantlets on MS medium supplemented with 7.5 μmol/L indole-3-butyric acid. A large callus size occurred with roots at the basal end of shoots on the media added with 20.0 μmol/L or 40.0 μmol/L of 1-naphthalene acetic acid (NAA combined with 40.0 μmol/L BA. Leaf explants were more suitable for callus induction. Media containing 0.0–40.0 μmol/L BA in combination with 20.0 μmol/L or 40.0 μmol/L NAA could efficiently induce callus formation with the same level of relative growth rate. These plants and callus will be suitable sources of plant material for further study in producing bioactive compounds.

  5. Comparison of Major Bioactive Compounds of the Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes), Fruiting Bodies Cultured on Wheat Substrate and Pupae.

    Science.gov (United States)

    Guo, Mingmin; Guo, Suping; Huaijun, Yang; Bu, Ning; Dong, Cai-Hong

    2016-01-01

    In this study, the main bioactive compounds of the fruit bodies of Cordyceps militaris-such as adenosine, cordycepin, polysaccharides, mannitol, superoxide dismutase (SOD), and carotenoids-were cultivated on wheat and pupae, as well as sclerotium (the pupae portion) and sclerotium with fruiting bodies. The amounts of adenosine and polysaccharide in all the tested samples (except for the polysaccharides of sclerotium) are higher than the quality standards (adenosine ≥0.055% and polysaccharide ≥2.5%) determined by the Ministry of Health of the People's Republic of China. As the most important bioactive compound in C. militaris, cordycepin is the highest in the fruiting bodies on pupae than in other samples, whereas it is the lowest in the sclerotium. The amounts of cordycepin, carotenoids, and SOD were higher in the fruiting bodies on pupae than that in the fruiting bodies on wheat, whereas the amounts of adenosine, polysaccharides, and mannitol were higher in the fruiting bodies on wheat than in the fruiting bodies on pupae. There was no significant difference in the amounts of cordycepin, carotenoids, and SOD in the sclerotium with fruiting bodies and the fruiting bodies on wheat. The adenosine, polysaccharide, and mannitol contents in the sclerotium with fruiting bodies were significantly lower than those of the fruiting bodies on wheat. Overall, the results of this evaluation could not distinguish which is better: the fruiting bodies on pupae or those on wheat; each has its own merits. The fruiting bodies of C. militaris cultivated on both wheat and pupae are important candidates for medicinal and tonic use for the welfare of humankind.

  6. Fruits and Photosynthesis

    Directory of Open Access Journals (Sweden)

    Yuli Alexandra Deaquiz-Oyola

    2014-08-01

    Full Text Available Growth and fruit development are conditioned by environmental factors such as solar radiation, temperature, relative humidity and rainfall, affecting phenology and metabolic processes, which are reflected in its quality and  size. Besides the variety, the age of the plant species, cultural practices, the amount of CO2, plant growth regulators and nutrition also influence this process of maturation. Moreover, the photosynthetic process occurs in immature fruits same manner as in the leaves, however, when the ripening process starts it changes because chlorophyll is degraded and other pigments intervene such as carotenoids, α-carotene and β-carotene, which contain antioxidants good for human health.

  7. Study on the introduction factors of callus embryoid of Medicago sativa anther%苜蓿花药愈伤组织胚状体诱导因素的研究

    Institute of Scientific and Technical Information of China (English)

    耿小丽; 魏臻武; 姚喜红

    2011-01-01

    以7个基因型的苜蓿花药愈伤组织为材料,进行了基因型、愈伤组织生长率、培养温度、培养基碳源等因素对苜蓿花药培养诱导胚状体影响的研究.结果表明:不同基因型苜蓿花药愈伤组织生长率、胚状体诱导率都存在显著性差异,两者的相关系数r=-0.326 1(P<0.01),两者没有明显的相关性;低温处理可促进花药愈伤组织的分化,4℃低温预处理48 h的效果最好;碳源对胚状体的诱导有较大影响,其中,蔗糖效果好于其他碳源.%Seven genotypes of callus of alfalfa anther were used to study the effects of genotype,growth rate of callus,temperature and carbon source of culture on introduction of embryoid of alfalfa anther. The results showed that different genotypes were significant factors to growth rate of anther callus and induction rate of embryoid. Low temperature of culture can improve the differentiation rate of anther callus. The treatment of 4 'C with the precondition of 48 hours was best approach. The carbon source of culture also influenced on the embryoid induction,and the effect of culture with sucrose was better than other cultures with lactose and glucose.

  8. Which perceived characteristics make product innovations appealing to the consumer? A study on the acceptance of fruit innovations using cross-cultural consumer segmentation.

    Science.gov (United States)

    Onwezen, Marleen C; Bartels, Jos

    2011-08-01

    In general, fruit consumption in the EU does not meet governments' recommended levels, and innovations in the fruit industry are thought to be useful for increasing fruit consumption. Despite the enormous number of product innovations, the majority of new products in the market fail within the first two years, due to a lack of consumer acceptance. Consumer segmentation may be a useful research tool to increase the success rates of new fruit products. The current study aims to identify consumer segments based on individual importance rankings of fruit choice motives. We conducted a cross-national, online panel survey on fresh fruit innovations in four European countries: the Netherlands (n=251), Greece (n=246), Poland (n=250), and Spain (n=250). Our cluster analysis revealed three homogeneous consumer segments: Average Joe, the Naturally conscious consumer, and the Health-oriented consumer. These consumer segments differed with respect to their importance ratings for fruit choice motives. Furthermore, the willingness to buy specific fruit innovations (i.e., genetically modified, functional food and convenience innovation) and the perceived product characteristics that influence this willingness differed across the segments. Our study could lead to more tailored marketing strategies aimed at increasing consumer acceptance of fruit product innovations based on consumer segmentation. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. Review of progress in tissue culture in stone fruit trees of Rosaceae%蔷薇科核果类果树组织培养研究进展

    Institute of Scientific and Technical Information of China (English)

    张悦; 傅建敏; 李福海; 杜红岩

    2011-01-01

    In order to provide reference for the tissue culture in the stone fruit trees of Rosaceae, the progress of tissue culture research was discussed, including culture of shoot tip, stem segment with bud, embryo culture protoplast and leaf. The effects of several major factors in procedure of tissue culture were reviewed, including basic media, carbon sources, hormone types and ratio and the other environmental conditions. Moreover, the existing questions in tissue culture of stone fruit trees were pointed out and corresponding solutions were also put forward.%为给核果类果树组织培养的深入研究提供参考,综述了有关胚培养、茎尖培养、茎段培养、叶片培养及原生质体培养的研究进展,重点分析了基本培养基、碳源、激素种类与配比和环境条件等影响因子对培养效果的影响情况,指出了核果类果树组织培养中存在的问题,并且提出了解决这些问题的途径.

  10. Production of Forskolin in in vitro Cultures of Coleus forskohlii.

    Science.gov (United States)

    Sen, J; Sharma, A K; Sahu, N P; Mahato, S B

    1992-08-01

    Shoot tip culture, root organ suspension, and callus culture producing roots were established from shoot tip, root tip, and hypocotyl segments, respectively, on Murashige and Skoog's and White's basal media with or without growth regulators. Forskolin was identified by TLC and HPLC in 60 days old shoot differentiating culture, 30 days old micropropagated plants, and root organ suspension but not in rhizogenic callus. Shoot differentiating culture established in the presence or absence of BAP revealed more or less similar amounts of forskolin as that of the micropropagated plants. Root organ suspension showed traces of forskolin production.

  11. Antibacterial activity of leaves and inter-nodal callus extracts of Mentha arvensis L

    Institute of Scientific and Technical Information of China (English)

    JohnsonM; WeselyEG; KavithaMS; UmaV

    2011-01-01

    Objective:To determine the anti-bacterial efficacy of chloroform, ethanol, ethyl acetate and water extracts of inter-nodal and leaves derived calli extracts from Mentha arvensis (M. arvensis) against Salmonella typhi(S. typhi), Streptococcus pyogenes(S. pyogenes), Proteus vulgaris(P. vulgaris) and Bacillus subtilis(B. subtilis). Methods: The inter-nodal and leaves segments of M. arvensis were cut into 0.5-0.7 cm in length and cultured on Murashige and Skoog solid medium supplemented with 3% sucrose, gelled with 0.7% agar and different concentration of 2, 4-Dichlorophenoxyacetie acid (2,4-D) either alone or in combinations. The preliminary phytochemical screening was performed by Brindha et al method. Antibacterial efficacy was performed by disc diffusion method and incubated for 24 h at 37 ℃. Results: Maximum percentage of callus formation (inter-nodal segments 84.3±0.78;leaves segments 93.8±1.27) was obtained on Murashige and Skoog’s basal medium supplemented with 3%sucrose and 1.5 mg/L of 2, 4-D. The ethanol extracts of leaves derived calli showed the maximum bio-efficacy than other solvents. The leaves and stem derived calli extracts on Proteus sp. showed that the plants can be used in the treatment of urinary tract infection associated with Proteus sp. Through the bacterial efficacy studies, it is confirmed that the in vitro raised calli tissue was more effective compared to in vivo tissue. Conclusions:The bio-efficacy study confirmed that the calli mediated tissues showed the maximum zone of inhibition. The present study paved a protocol to establish high potential cell lines by in vitro culture.

  12. Phenolic constituents and antioxidant capacities of Crataegus monogyna (Hawthorn) callus extracts.

    Science.gov (United States)

    Bahorun, Theeshan; Aumjaud, Esha; Ramphul, Hemlata; Rycha, Maheshwaree; Luximon-Ramma, Amitabye; Trotin, Francis; Aruoma, Okezie I

    2003-06-01

    Crataegus (Hawthorn) has long been used as a folk medicine and is widely utilized in pharmaceutical preparations mainly because of its neuro- and cardiosedative actions and its low toxicity. The pharmacological effects of Crataegus have mainly been attributed to the polyphenolic contents. In this study, the production of polyphenols by ten-year-old Crataegus monogyna calli was studied in relation to growth variation and antioxidant capacity within a subculture period. Assays based on the Trolox equivalent antioxidant capacity (TEAC), ferric reducing antioxidant power (FRAP) and stability in oil-in-water emulsion were used to characterize the antioxidant actions of the callus cultures. High TEAC (3.66 micromol/g dry weight) and FRAP (208.19 micromol Fe2+/g dry weight) values were observed when maximal growth was reached(days 30-35), and this seemed to be influenced by optimum total phenol (47.40 mg/g dry weight), proanthocyanidin (20.81 mg/g dry weight), flavonoid (7.01 mg/g dry weight), anthocyanin (6.18 mg/g dry weight), (-)-epicatechin (1.77 mgl/g dry weight), procyanidin B2 (3.97 mg/g dry weight), and chlorogenic acid (1.11 mg/g dry weight) production during that period. The TEAC values were strongly associated with total flavonoids and to a lesser extent with total phenols, anthocyanins and total proanthocyanidins. The FRAP antioxidant values correlated to total phenols, proanthocyanidins and flavonoids, respectively. The polyphenolic rich calli were as effective as butylated hydroxytoluene (BHT) in preventing hydroperoxide and conjugated diene formation in a 30% oil-in-water emulsion prepared with stripped sunflower oil, during 7days storage at 30 degrees C. Crataegus monogyna cell culture represents an important alternative source for natural antioxidants.

  13. Induction and Proliferation Conditions of Onion(Allium cepa L.) Embryogenic Callus%洋葱胚性愈伤组织的诱导及增殖条件的研究

    Institute of Scientific and Technical Information of China (English)

    代月; 吴雄; 朴炫春; 霍雨猛; 缪军; 廉美兰

    2012-01-01

    [目的]探讨洋葱离体培养的最佳条件,为建立其高频再生体系及转基因研究奠定技术基础.[方法]以洋葱(Allium cepa L.)种子为外植体,对洋葱愈伤组织的诱导、胚性愈伤组织获得及增殖的培养条件进行了初步研究.[结果]不同培养基种类对愈伤组织的诱导有影响,BDS优于MS和B5培养基;当培养基中2,4-D浓度为0.5 mg,/L时种子出愈率最高;适当降低生长素浓度,添加分裂素BA能够促进胚性愈伤组织的发生,且无分化现象产生,胚性愈伤组织诱导最佳培养基为BDS+2,4-D 0.25 mg/L+BA 2.0 mg/L,增殖最佳培养基为BDS+2,4-D O.25 mg/L+BA 1.0 mg/L.[结论]该试验建立的基因转化受体系统为洋葱基因转化奠定了良好基础.%[Objective] To explore the best conditions for the in vitro culture of onion, so as to lay technical foundation for its trangenic research and the establishment of high frequency regeneration system. [ Method] With the onion seeds as explants, the induction and proliferation conditions of onion (Alliiim r.epa L. ) callus and embryogenic callus were preliminarily studied. [ Result] The types of callus produced certain impact on the induction of callus, BDS was better than MS and B5; the callus rate was the highest at the concentrations of 0. 5 mg/L 2,4-D; the appropriate reduction of auxin concentration and addition of cytokinin BA could promote the induction of enbryogenic callus without differentiation, the best embryogenic callus induction medium was BDS +2,4-D 0. 25 mg/L + BA 2.0 mg/L and the best proliferation medium was BDS + 2,4-D 0.25 mg/L + BA 1.0 mg/L. [Conclusion] Hie transgenic receptor system established in this experiment lays good foundation for the genetic transformation of onion.

  14. Effect of applied currents to growth in oil palm (Elaeis guineensis Jacq. tissue cultures

    Directory of Open Access Journals (Sweden)

    Panote Thavarungkul

    2002-04-01

    Full Text Available External currents of ± 2 μA were applied to the calluses of oil palm in three different culture medium recipes containing different growth substances and the effects of the current were investigated. The three media were medium for callus growth, embryogenetic medium, and organogenetic medium. The stimulation of callus growth was found for both directions of current in one unit of experiments where the embryogenetic culture medium contained NAA. In other cases the use of current seemed to have no significant stimulation effect. The mechanism by which the current may cause the alignment of the cells to promote polar transport of NAA which then increase growth and the relationship of these findings to earlier reports of the stimulation of growth in tobacco callus cultures is discussed.

  15. Histology of embryoid development in oil palm (Elaeis guineensis Jacq. cell suspension culture

    Directory of Open Access Journals (Sweden)

    Songrat Tinnongjig

    2001-11-01

    Full Text Available Embryos of oil palm (Elaeis guineensis Jacq. variety tenera were cultured on Eeuwens or Y3 (1976; 1978 medium supplemented with 2 mg/l 2,4-D. Calluses were initiated from these embryos. The eight-weekold calluses derived from embryos were transferred to modified Y3 liquid medium devoid of 2,4-D and supplemented with NAA, BA and coconut water to establish cell suspension culture. After a period of culture,these cells were then subcultured to the same medium without plant growth regulators to induce embryoid formation. The calluses and embryoids were harvested at various times, fixed, sectioned, stained and examined microscopically. Histological study revealed that embryoid occurred from meristematic cells with dense cytoplasm along the callus clumps.

  16. High frequency embryoid and plantlet formation from tissue cultures of the Finger millet-Eleusine coracana (L.) Gaertn.

    Science.gov (United States)

    Sivadas, P; Kothari, S L; Chandra, N

    1990-07-01

    Compact nodulated embryogenic callus differentiated from cultured seeds of Eleusine coracana (Finger Millet) on Murashige and Skoog (1962) basal medium with 2,4-dichlorophenoxyacetic acid (1.0, 3.0 mg (l)). This embryogenic callus was maintained on a medium with a lower level of 2,4 - dichlorophenoxyacetic acid. At every subculture the embryogenic callus had some preexisting embryoids in it. With this method of subculture the callus has retained its morphogenic potential for four years. Following transfer to media with different levels of auxins and cytokinins, the callus showed varied patterns of growth and morphogenesis. Embryoids could be germinated in profusion to form plantlets which could be transferred to the field. Shoot buds also differentiated from the whole surface of the embryoid or from the flattened meristemoids.

  17. Evaluation of cytotoxicity of Moringa oleifera Lam. callus and leaf extracts on Hela cells

    Directory of Open Access Journals (Sweden)

    Abbas Jafarain

    2014-01-01

    Full Text Available Background: There are considerable attempts worldwide on herbal and traditional compounds to validate their use as anti-cancer drugs. Plants from Moringaceae family including Moringa oleifera possess several activities such as antitumor effect on tumor cell lines. In this study we sought to determine if callus and leaf extracts of M. oleifera possess any cytotoxicity. Materials and Methods: Ethanol-water (70-30 extracts of callus and leaf of M. oleifera were prepared by maceration method. The amount of phenolic compounds of the extracts was determined by Folin Ciocalteu method. The cytotoxicity of the extracts against Hela tumor cells was carried out using MTT assay. Briefly, cells were seeded in microplates and different concentrations of the extract were added. Cells were incubated for 48 h and their viability was evaluated by addition of tetrazolium salt solution. After 3 h medium was aspirated, dimethyl sulfoxide was added and absorbance was determined at 540 nm with an ELISA plate reader. Cytotoxicity was considered when more than 50% reduction on cell survival was observed. Results: Callus and leaf extracts of M. oleifera significantly decreased the viability of Hela cells in a concentration-dependent manner. However, leaf extract of M. oleifera were more potent than that of callus extract. Conclusion: As the content of phenolic compounds of leaf extract was higher than that of callus extract, it can be concluded that phenolic compounds are involved in the cytotoxicity of M. oleifera.

  18. Glass distilling collector applied for HCN recovery from submerged culture broth and fruiting body of Pleurotus eryngii for identification and quantification.

    Science.gov (United States)

    Chou, Pei-Yu; Hong, Chian-Huei; Chen, Wenlung; Li, Yu-Jang; Chen, Yen-Shang; Chiou, Robin Y-Y

    2006-03-08

    Detection and surveillance of food commodities containing cyanide is a crucial issue of food safety. In this study, five strains of Pleurotus eryngii (P. eryngii) were grown in submerged culture of yeast malt broth (YMB) with the suspected production of HCN. A safety-warranted U-bent glass distilling collector with three enlarged bulbs on each arm was designed to recover the broth vapor. When AgNO(3) solution was used as an absorbent to interact with the vapor, a white precipitate was formed. The precipitate was isolated and identified as AgCN by FT-Raman spectroscopic analysis. When the absorbent was substituted by KOH, after evaporation to dryness, dissolved in D(2)O, and followed by (13)C-NMR analysis, a KCN spectrum was achieved. Formation of AgCN and KCN confirmed HCN production in the broth by P. eryngii. When a sodium picrate solution (1.4%) was used as an absorbent and various authentic KCN solutions were applied for distillation and followed by absorbance determination at 510 nm, a linear dose-dependent relationship was obtained and the procedure was applied for HCN quantification of the marketed P. eryngii mushrooms (fruiting body). As estimated, 67.3% of the products contained HCN less than 1.0 mg/kg, 17.3% between 1.0 and 2.0 mg/kg, and 15.4% higher than 2.0 mg/kg. When the mushrooms were sliced and cooked in water at 95 degrees C for 6 min, 89.1% of the original HCN was lost. When the P. eryngii strains were respectively grown by submerged cultivation in YMB or YMB supplemented with 2.5% glycine for 16 days, HCN content was slightly higher in the latter than in the former for each strain.

  19. Construcción de la cultura de apreciación de la fruta: aporte de Las mil y una noches Building the culture of appreciation of the fruit: contribution of Arabian Nights

    Directory of Open Access Journals (Sweden)

    Pablo Lacoste

    2012-12-01

    Full Text Available Este artículo examina al aporte de los árabes al proceso de construcción de la cultura de apreciación de la fruta y los frutales. Se avanzó en esa dirección no sólo mediante la incorporación y propagación de las plantas, particularmente desde Al Andalus, sino también con su puesta en valor en la cultura universal, a través de Las mil y una noches. Si los botánicos y agrónomos andalusíes aportaron avances científicos sobre las plantas frutales y la forma de cultivarlas y aprovecharlas, los relatos literarios de la cultura islámica complementaron la acción de aquellos, al situar las frutas y los frutales en el centro del imaginario social, en los lugares de mayor prestigio (jardines, palacios, salones y asociarlos a la sensualidad, la belleza y el placer.This article examines what the Muslim world contributed to the materialization of a cultural appraisal of fruits and the trees that bear them. This contribution was not only carried out through the propagation and incorporation of these plants, particularly from Al Andalus, but also by providing them with considerable cultural significance with their inclusion in Arabian Nights. While Andalusian botanists and agronomists achieved significant scientific breakthroughs in the methods involved in the cultivation and proper utilization of fruit trees, Muslim literary tales complimented these developments by placing both fruits and fruit trees in the cultural spotlight, in highly prominent locations (gardens, palaces, halls and by bonding them with sensuality, beauty and pleasure. Muslims acted as biocultural intermediaries.

  20. Diabetic foot ulcer calluses show histological similarities to HPV infection without evidence of HPV involvement by nested PCR.

    Science.gov (United States)

    Li, Lucy; Iwamoto, Satori; Jisun Cha; Falanga, Vincent

    2010-06-01

    A number of studies have reported an association between callus formation and the development of foot ulcers in diabetic patients. However, it has been noted that calluses may continue to form in spite of bed rest and, presumably, excellent patient compliance with offloading. Additionally, the authors have noted that, histologically, calluses in the diabetic foot often resemble lesions induced by human papillomavirus (HPV). As diabetes is associated with immune depression, the authors hypothesized that HPV may play at least a partial role in the pathogenesis of calluses in diabetic patients. The objective of the reported study was to determine whether calluses around diabetic neuropathic foot ulcers are associated with HPV infection. The authors carried out biopsies on 11 independent calluses from 6 patients with diabetic foot ulcers and analyzed each sample by histology and by nested polymerase chain reaction (PCR), screening for the presence of DNA from HPV-1, -2, -3, -4, -6, -10, -11, -16, -18, -27, -28, -29, -31, -41, -50, -57, -60, -63, -65, and -77. The callus biopsy specimens showed histological evidence of koilocytes, papillary hyperplasia, hypergranulosis, and hyperkeratosis, a picture very similar to HPV cutaneous infection. However, nested PCR using positive and negative controls did not show detectable levels of HPV DNA. The authors therefore conclude that HPV infection is unlikely to play a significant role in diabetic foot callus pathogenesis, in spite of histological findings similar to those seen with verruca vulgaris.

  1. First Report of Plant Regeneration via Somatic Embryogenesis from Shoot Apex-derived Callus of Hedychium muluense

    Science.gov (United States)

    Plants were successfully regenerated via somatic embryogenesis from shoot apex-derived callus of Hedychium muluense R.M. Smith, an important monocotyledonous ornamental ginger plant. Callus was induced on a modified Murashige and Skoog (MS) medium supplemented with 9.05 µM 2-4, D and 4.6µM kinetin. ...

  2. Hyperplastic callus formation in osteogenesis imperfecta type V mimicking osteosarcoma: 4-year follow-up with resolution

    Energy Technology Data Exchange (ETDEWEB)

    Vieira, R.L.V.; Amaral, D.T. [Federal University of Sao Paulo, Department of Radiology, Sao Paulo (Brazil); Jesus-Garcia, Filho R. [Federal University of Sao Paulo, Department of Orthopedic Surgery, Sao Paulo (Brazil); Saraiva, G. [Federal University of Sao Paulo, Department of Endocrinology, Sao Paulo (Brazil); Fernandes, A.R.C. [University of California San Diego, Department of MSK Radiology, San Diego, CA (United States); Resnick, D.

    2006-06-15

    We report a case of hyperplastic callus formation that occurred in both femurs in a patient with type V osteogenesis imperfecta (OI), with 4-year follow-up and resolution. The clinical, histological and imaging aspects of this condition are discussed. Recognition of the hyperplastic callus formation in this particular type of OI is important in order to avoid misdiagnosis. (orig.)

  3. A quantitative method for determining relative colonization rates of maize callus by Fusarium graminearum for resistance gene evaluations

    Science.gov (United States)

    A quantitative PCR method was developed for detecting Fusarium graminearum growing in maize callus. Fungal DNA was found 12 hours after inoculation and was correlated with visual ratings. We demonstrated the efficacy of the method to quantify fungal growth in callus overexpressing a peroxidase gene ...

  4. Proliferation Potential of 18-Month-Old Callus of Ananas comosus L. cv. Moris

    OpenAIRE

    Silva,A.E.; Kadir, M.A.; Aziz, M A; S. Kadzimin

    2006-01-01

    Differential effect of plant growth regulators and additives in proliferation of 18-month-old calli of Ananas comosus L. cv. Moris were assessed in vitro. The proliferation of callus relied on the growth regulators and additives. Of the different auxins supplemented in the Murashige and Skoog (MS) media, 32.22 μM α-naphthaleneacetic acid (NAA) gave the highest mean fresh weight of callus (46.817 g). Medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) was inferior to NAA, while b-n...

  5. Structure of the osseous callus formed under electric stimulation. An experimental histological study.

    Science.gov (United States)

    Mehedinţi, T; Roşca, D; Cernăianu, E

    1985-01-01

    The study tried to evidence histologically, histometrically and mechanically the comparative qualities of the osseous callus experimentally formed in dogs and rabbits under the influence of an electric stimulation. The materials were decalcified or polished, and the collagen network and the Haversian systems analysed in the callus. The tearing strength in kg-power on the surface unity of the osseous tissue was measured using the EDZ 20 apparatus. The results indicated that the osseous tissue formed under electric stimulation had a structure and a tearing strength similar to that of controls, and that the consolidation time of fractures is reducing.

  6. [Effects of light intensity on associated enzyme activity and gene expression during callus formation of Vitis vinifera].

    Science.gov (United States)

    Liu, Rong; Yang, Guowei; Wu, Yueyan; Rao, Huiyun; Li, Xuefu; Li, Meiqin; Qian, Pingxian

    2015-08-01

    We analyzed the best light intensity for callus induction and maintenance in Vitis vinifera and explored the mechanism of grape callus browning. Tender stem segments of grape cultivar "gold finger" were used to study the effects of different light intensities (0, 500, 1 000, 1 500, 2 000, 2 500, 3 000 and 4 000 Lx) on the induction rate, browning rate and associated enzyme activity and gene expression during Vitis vinifera callus formation. The callus induction rate under 0, 500, 1 000 and 1 500 Lx was more than 92%, significantly higher than in other treatments (P Vitis vinifera was 1 000-1 500 Lx, higher or lower light intensities significantly impaired normal callus growth.

  7. Impact of application of zinc oxide nanoparticles on callus induction, plant regeneration, element content and antioxidant enzyme activity in tomato (Solanum lycopersicum Mill. under salt stress

    Directory of Open Access Journals (Sweden)

    Alharby Hesham F.

    2016-01-01

    Full Text Available The properties of nanomaterials and their potential applications have been given considerable attention by researchers in various fields, especially agricultural biotechnology. However, not much has been done to evaluate the role or effect of zinc oxide nanoparticles (ZnO-NP in regulating physiological and biochemical processes in response to salt-induced stress. For this purpose, some callus growth traits, plant regeneration rate, mineral element (sodium, potassium, phosphorous and nitrogen contents and changes in the activity of superoxide dismutase (SOD and glutathione peroxidase (GPX in tissues of five tomato cultivars were investigated in a callus culture exposed to elevated concentrations of salt (3.0 and 6.0 g L-1NaCl, and in the presence of zinc oxide nanoparticles (15 and 30 mg L-1. The relative callus growth rate was inhibited by 3.0 g L-1 NaCl; this was increased dramatically at 6.0 g L-1. Increasing exposure to NaCl was associated with a significantly higher sodium content and SOD and GPX activities. Zinc oxide nanoparticles mitigated the effects of NaCl, and in this application of lower concentrations (15 mg L-1 was more effective than a higher concentration (30 mg L-1. This finding indicates that zinc oxide nanoparticles should be investigated further as a potential anti-stress agent in crop production. Different tomato cultivars showed different degrees of tolerance to salinity in the presence of ZnO-NP. The cultivars Edkawy, followed by Sandpoint, were less affected by salt stress than the cultivar Anna Aasa.

  8. Genotoxicity assessment of high concentrations of 2,4-D, NAA and Dicamba on date palm callus (Phoenix dactylifera L. using protein profile and RAPD markers

    Directory of Open Access Journals (Sweden)

    Mohammed H. Abass

    2017-06-01

    Full Text Available Genetic stability and uniformity of in vitro-derived date palm plants has a major importance to ascertain true-to-typeness of produced plants. The goal of present study was to evaluate the genetic toxicity of different plant growth regulators on date palm callus at initiation stages using protein patterns and RAPD analysis. Date palm offshoots of Hillawii cultivar were dissected, apical meristems were divided into four segments and cultured on callus induction medium containing the plant growth regulators as 2,4-D at 50 and 100 mg/L; NAA at 30 mg/L and Dicamba at 10 mg/L. The changes occurred in protein profile of callus when treated with high concentration of 2,4-D (100 mg/L, including loss of normal fragments (19 and 66 KDa polypeptides in control, as well as, appearance of new fragments, while at low concentration of 2,4-D (50 mg/L and Dicamba treatment, the protein patterns showed no changes compared to control profile. Similar trends of polymorphisms were obtained with RAPD marker. The high concentration of 2,4-D produced more polymorphic fragments in comparison to control treatment. The DNA profile was identical between 2,4-D at low concentration and control. Dendrograms were generated using similarity indices of protein and RAPD results, and revealed that genetic similarity index was high between 2,4-D treatment at low concentration and control, as separated in one subcluster, followed by Dicamba and NAA, while, the highest genetic distance was obtained between 2,4-D at high concentration and control treatment and separated alone in one cluster.

  9. 辣椒花药培养影响因素的研究%Research on Factors Influencing Anther Culture of Hot Pepper (Capsicum annuum L.)

    Institute of Scientific and Technical Information of China (English)

    龚明霞; 何铁光; 董文斌; 赵坤; 王益奎; 王日升; 康德贤

    2013-01-01

    We analyzed the disinfection effect on six flower buds of hot pepper (Capsicum annuum L.), and studied the effects of different genotypes and inoculation amounts on anther culture. The results showed that, the disinfection method used in this study got good effect, and three genotypes were free of contamination, in addition, among the other three, the highest contamination rate was 23.08%. Embryoids could be induced from three genotypes, and the embryoid forming rates were 1.18%-1.38%, however embryoids could not be induced from hot pepper type with small fruits. Callus could be induced from all the six genotypes with callus forming rates of 17.65%-69.44%. Inoculation amount affected the callus induction but had no significant influence on embryoid forming, and as the inoculation amount increased, the callus forming rate was increased by 0.77%-13.67%. In callus subculture, inoculation amount had different effects on callus growth because the difference of genotypes, and neither embryiod nor adventitious bud were differentiated from the calluses of all six genotypes during subculturing for the first time.%以6种基因型辣椒的花蕾为材料,比较了不同基因型辣椒花蕾的消毒效果,辣椒基因型及花药接种量对花药培养的影响。研究结果表明,本试验的消毒方法获得了较好的消毒效果,3个基因型材料均未受污染,其他最高的污染率为23.08%;50%的基因型能诱导出胚状体,出胚率1.18%~1.38%,小果型的辣椒不能诱导出胚状体;所有基因型均能诱导出愈伤组织,出愈率17.65%~69.44%;花药接种量对花药愈伤组织诱导有影响,增大接种量,出愈率上升0.77%~13.67%,但对胚状体诱导影响不明显;在愈伤组织的继代培养中,接种量对其生长的影响因基因型不同而不同,所有基因型的愈伤组织在第一次继代培养中均不能分化出胚状体或不定芽。

  10. STUDIES REGARDING CULTURE MEDIUM INFLUENCE ON IN VITRO REGENERATION FROM WHEAT IMATURE EMBRYOS

    Directory of Open Access Journals (Sweden)

    M. DANCI

    2013-12-01

    Full Text Available Surnamed „embryos’ saving method”, embryos culture is an in vitro technique used for over half of the century for saving the distant hybridization products, that would have degenerate in other conditions. Immature embryos culture is used for initiation of in vitro cultures imposed by the impossibility of using other explants for some of the plant species. Wheat is one of the crops that immature embryos culture technique is suitable for. This methods principle is based on aseptic embryos excision and their inoculation to an adequate culture medium. In vitro culture results depend in a greater manner of the basic culture medium and the hormonal balance used. Immature embryos isolated from two Romanian wheat cultivars – Dropia and Lovrin 41 – were inoculated for callus production on two types of basic media added with 2,4 D. The selected calluses were transferred on MS basic medium and several parameters were registered. Both cultivars emphasized a good callusing capacity, in a different percentage depending on the culture media used, such as 71,09 – 94,45%.. big differences between the cultivars regarding embriogenic callus frequency, shooting callus frequency and regenerated plants percentage were registered.

  11. Substituted Indoleacetic Acids Tested in Tissue Cultures

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen

    1978-01-01

    Monochloro substituted IAA inhibited shoot induction in tobacco tissue cultures about as much as IAA. Dichloro substituted IAA inhibited shoot formation less. Other substituted IAA except 5-fluoro- and 5-bromoindole-3-acetic acid were less active than IAA. Callus growth was quite variable...

  12. Studies on suspension culture of virginia mallow

    Directory of Open Access Journals (Sweden)

    Anna Kasprzyk

    2014-04-01

    Full Text Available Virginia mallow (Sida hermaphrodita (L. Rusby belongs to the Malvaceae family. It is a very important industrial and energetic crop (Kasprzyk et al. 2013. In our studies, we used plant cell suspension cultures due to the fact that it is a useful tool to investigate biochemical, molecular and physiological aspects of many cellular functions (Dong et al. 2010. Virginia mallow seeds, obtained from Prof. Borkowska (University of Life Sciences in Lublin, Poland, were used in this investigation to obtain plants which were grown in sterile conditions in the Department of Plant Anatomy and Cytology, Maria Curie-Skłodowska University in Lublin, Poland. The seeds were surface sterilized and washed three times in sterile, distilled water. After 3 weeks of in vitro culture, young seedlings were used as a source of explants (to callus induction. Two types of explants were used to form callus culture: leaf and petiole. Callus tissues were then aseptically transferred to an Erlenmeyer flask with liquid medium and placed on an orbital shaker moving at 120 rpm. The observations of this suspense culture were conducted under light and confocal LSM microscopes. The authors observed that depending on the type of explants and composition of medium, callus tissue has varied in color and character of growth.

  13. Soybean protoplast culture and direct gene uptake and expression by cultured soybean protoplasts.

    Science.gov (United States)

    Lin, W; Odell, J T; Schreiner, R M

    1987-07-01

    A method was developed for culturing protoplasts freshly isolated from developing soybean (Glycine max L.) cotyledons. First cell divisions were observed within 5 days after protoplast isolation and microcalli, consisting of about 20 cells, were formed within 10 days. Thirty days after protoplast isolation, callus tissues were observed without the aid of a microscope. A 30 to 50% plating efficiency was consistently obtained. Using a polyethylene glycol-electroporation technique, DNA was introduced into these protoplasts. The protoplasts were then cultured to form callus. Chloramphenicol acetyltransferase (CAT) activity was detected in protoplast cultures 6 hours after introduction of a 35S-CAT-nopaline synthase 3' chimeric gene. The highest CAT activity was detected in 3-day-old electroporated protoplast cultures, indicating transient expression of the introduced gene. Some CAT activity was detected in 40-day-old callus cultures and in geneticin (G418) selected callus tissues which also received a chimeric neomycin phosphotransferase II gene, indicating the presence of stable transformants. A control chimeric gene with an inverted 35S promoter failed to produce any CAT activity in this system.

  14. Regulation of cell wall remodeling in grapevine (Vitis vinifera L.) callus under individual mineral stress deficiency.

    Science.gov (United States)

    Fernandes, João C; Goulao, Luis F; Amâncio, Sara

    2016-01-15

    Cell wall (CW) is a dynamic structure that determines the plant form, growth and response to environmental conditions. Vitis vinifera callus grown under nitrogen (-N), phosphorous (-P) and sulfur (-S) deficiency were used as a model system to address the influence of mineral stress in CW remodeling. Callus cells morphology was altered, mostly under -N, resulting in changes in cell length and width compared with the control. CW composition ascertained with specific staining and immuno-detection showed a decrease in cellulose and altered pattern of pectin methylesterification. Under mineral stress genes expression from candidate families disclosed mainly a downregulation of a glycosyl hydrolase family 9C (GH9C), xyloglucan transglycosylase/hydrolases (XTHs) with predicted hydrolytic activity and pectin methylesterases (PMEs). Conversely, upregulation of PMEs inhibitors (PMEIs) was observed. While methylesterification patterns can be associated to PME/PMEI gene expression, the lower cellulose content cannot be attributed to altered cellulose synthase (CesA) gene expression suggesting the involvement of other gene families. Salt extracts from -N and -P callus tissues increased plastic deformation in cucumber hypocotyls while no effect was observed with -S extracts. The lower endo-acting glycosyl hydrolase activity of -N callus extracts pinpoints a more expressive impact of -N on CW-remodeling.

  15. Entrapment of the Flexor Digitorum Profundus in the Callus after a Closed Distal Radial Fracture.

    Science.gov (United States)

    Thione, Alessandro; Cavadas, Pedro C; Rubi, Carlo G

    2016-04-01

    A 17-year-old boy sustained a closed distal radius fracture; a closed reduction and external fixation was performed. After failed rehabilitation for digital flexion restriction, a surgical exploration was decided, revealing entrapment of flexor digitorum profundus in the bony callus; tendons were freed, obtaining a full range of motion.

  16. Factors influencing induction, propagation and regeneration of mature zygotic embryo-derived callus from Allium cepa.

    NARCIS (Netherlands)

    Zheng, S.; Henken, B.; Sofiari, E.; Jacobsen, E.; Krens, F.A.; Kik, C.

    1998-01-01

    A systematic study on the effects of subspecies, cultivar, basal medium, sucrose concentration and 2,4-dichlorophenoxyacetic acid concentration on callus induction, propagation and subsequent plant regeneration in Allium cepa has been carried out. Mature zygotic embryos from two onion (cvs. Sturon a

  17. MRI and CT features of hyperplastic callus in osteogenesis imperfecta tarda

    Energy Technology Data Exchange (ETDEWEB)

    Dobrocky, I. [Diagnostic Center Meidling, Vienna (Austria); Seidl, G. [Diagnostic Center Meidling, Vienna (Austria)]|[Universitaetsklinik fuer Radiodiagnostik, Vienna (Austria); Grill, F. [Orthopaedisches Spital Wien Speising, Vienna (Austria)

    1999-05-01

    We describe the MRI and CT findings of hyperplastic callus formation simulating a tumour of pelvis in patient with osteogenesis imperfecta tarda. Possible differential diagnoses and the impact of different imaging techniques on the correct diagnosis are discussed. (orig.) With 3 figs., 5 refs.

  18. Callus induction and plant regeneration in the moss Aloina Aloides (Schultz Kindb. (Pottiaceae, Bryopsida

    Directory of Open Access Journals (Sweden)

    Bijelović Aneta

    2003-01-01

    Full Text Available Callus induction of moss species Aloina aloides (Schultz Kindb. was obtained on Murashige and Skoog (MS medium supplemented with 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D or with 1.0 mg/L 2,4-D and 1.0 mg/L kinetin (KIN or with 0.2 mg/L indole-3-butyric acid (IBA and 2.0 mg/L 6-benzylaminopurine (BAP or with 7.5 g/L of sucrose or with 15 g/L of sucrose or hormone - free and sugar free MS basal medium. The callus can be maintained for a long period of time without bud formation subcultured on the above media, at 16 h day/8 h night, 25 ± 2ºC, 60-70% air humidity and irradiance of 50 μmol m-2s-1. To obtain plant regeneration pieces, calli were transferred onto MS media supplemented with different concentrations of auxins and cytokinins (1.0 mg/L 2,4-D and 2 mg/L KIN; 0.2 mg/L IBA and 2 mg/L KIN; or 0.2 mg/L IAA and 2 mg/L BAP. In these media after subculturing, callus enlarges and turns to gametophytes with buds. Except for a smaller size, the plants obtained on the callus did not differ morphoanatomically from the shoots in the nature.

  19. Callus induction from leaves of different paulownia species and its plantlet regeneration

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The experiment was carried out on five different species of Paulownia for callus induction from leaves. MS medium was adopted as basic medium, and from different combinations of NAA and BA the suitable media were determined for callus induction, bud differentiation, and root differentiation of five different species. MS+0.5NAA+4BA, MS+0.3NAA+2BA, MS+0.5NAA+4BA, MS+0.3NAA+6BA, and MS+0.3NAA+8BA were suitable media of callus inductions of leaves, respectively, for Paulownia tomentosa, Paulownia australis, Paulownia fortunei, Paulownia elongata and P. tmentosa x P. fortunei, and MS+0.3NAA+12BA, MS+0.3NAA+12BA, MS+0.5NAA+12BA, MS+0.5NAA+12BA, and MS+0.7NAA+12BA were suitable media for bud differentiation from leaf callus respectively for above five species. The rooting media was determined as 2MS+0.1NAA, 1/2MS+0.1NAA, 1/2MS, 1/2MS+0.3NAA, and 1/2MS+0.5NAA. These results provide reference data for breeding new fine varieties with different kinds of Paulownia protoplasts fusions.

  20. 火龙果愈伤组织诱导与植株再生%Callus Induction and Plantlet Regeneration from Pitaya

    Institute of Scientific and Technical Information of China (English)

    黄红梅; 任希望; 刘清波

    2012-01-01

    为了建立火龙果愈伤组织诱导与植株再生体系,以火龙果茎段、幼苗和子叶为外植体进行离体培养试验.结果表明:茎段诱导愈伤组织的最优培养基为1/2MS+2,4-D 2.0 mg·L-16-BA 0.5 mg·L-1,诱导子叶愈伤组织的最适培养基是1/2MS+2,4-D 2.0 mg·L-1+6-BA 1.0 mg·L-1,诱导愈伤组织分化的最优培养基为1/2MS+6-BA 4.0 mg·L-1+NAA 0.5 mg·L-1,最佳生根培养基为1/2MS+6-BA 1 mg·L-1+NAA 0.3 mg·L-1.%To establish the system of callus induction and plantlet regeneration from pitaya,pitaya was in vitro cultured using stem segments,seedlings and cotyledons as explants. The results showed the optimum medium for the callus induction of stem segments was l/2MS+2,4-D 2.0 mg·L-1+6-BA 0.5 mg·L1. The most suitable medium to induce the callus from cotyledons was 1/2MS+2.4-D 2.0 mg·L-1+6-BA 1.0 mg·L-1. The optimal medium for inducing the callus differentiation was 1/2MS+6-BA 4.0 mg·L-1+NAA 0.5 mg·L-1. The greatest medium of root regeneration was 1/2MS+6-BA 1.0 mg·L-1+NAA0.3 mg·L-1.

  1. Biological Effects of Low Energy N+ Beams Implantation on Calluses of Autotetraploid Rice

    Institute of Scientific and Technical Information of China (English)

    ZHAO Shuai-peng; HUANG Qun-ce; LIANG Qiu-xia; ZHANG Shu-gen; JIAO Zhen; HUANG Wei-zhou

    2013-01-01

    Calluses of two autotetraploid rice (DPR (4) and Zijing (4)) were implanted with 30 KeV N+beams at doses ranging from 1.0×1015 to 9.0×1015 ions cm-2. The differentiation rates of calluses, the concentration of malondialdehyde (MDA), the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), and the change of esterase (EST) spectrum were measured and analyzed on the calluses stage. The results showed that differentiation rates of calluses under 3.0×1015 ions cm-2 ((46.12±2.01)%) were higher than the other treatment groups in DPR (4), and those of Zijing (4) were similar under the doses of 1.0×1015, 3.0×1015 and 5.0×1015 ions cm-2 ((40.55±2.74), (37.77±3.04) and (34.56±2.65)%) and were higher than that of 7.0×1015 and 9.0×1015 ions cm-2. When the doses were 3.0×1015 and 5.0×1015 ions cm-2, the activities of SOD, POD and CAT were notably enhanced, and the accumulation of MDA content was markedly alleviated in the implanted calluses of two materials, as well as the activity of EST was increased by the additional isforms DPR (4). Whereas the anti-oxidative systems and enzymes spectrum of EST were destroyed seriously, the accumulation of MDA was signiifcantly aggravated, also the differentiation rate of calluses reduced sharply and almost closed to zero when the implantation doses were 7.0×1015 and 9.0×1015 ions cm-2. It suggested that the proper dose of ion implantation effectively maintained the normal membrane structure and metabolism, and it would be also a feasible approach for autotetraploid rice to improve its genetic characteristics at calluses stage. Additionally, the differences of the radiation resistance between DPR (4) and Zijing (4) could be related to the different genotypes.

  2. Preliminary Screening of Antioxidant and Antibacterial Activities and Establishment of an Efficient Callus Induction in Curculigo latifolia Dryand (Lemba

    Directory of Open Access Journals (Sweden)

    Reza Farzinebrahimi

    2016-01-01

    Full Text Available Leaf, seed, and tuber explants of C. latifolia were inoculated on MS medium supplemented with various concentrations of BAP and IBA, alone or in combinations, to achieve in vitro plant regeneration. Subsequently, antioxidant and antibacterial activities were determined from in vitro and in vivo plant developed. No response was observed from seed culture on MS media with various concentrations of PGRs. The highest percentage of callus was observed on tuber explants (94% and leaf explants (89% when cultured on MS media supplemented with IBA in combination with BAP. A maximum of 88% shoots per tuber explant, with a mean number of shoots (8.8±1.0, were obtained on MS medium supplemented with combinations of BAP and IBA (2.5 mg L−1. The best root induction (92% and mean number (7.6±0.5 from tuber explants were recorded on 2.5 mg L−1 IBA alone supplemented to MS medium. The higher antioxidant content (80% was observed from in vivo tuber. However, tuber part from the intact plant showed higher inhibition zone in antibacterial activity compared to other in vitro and in vivo tested parts.

  3. Production of camptothecin in cultures of Chonemorpha grandiflora

    Directory of Open Access Journals (Sweden)

    A V Kulkarni

    2010-01-01

    Full Text Available Background : Chonemorpha grandiflora (Syn. Chonemorpha fragrans (Apocynaceae is an endangered medicinal plant. It is used in different preparations, such as sudarsanasavam and kumaryasavam used in Kerala Ayurvedic system. C. grandiflora is used for the treatment of fever and stomach disorders. Phytochemical investigations have revealed the presence of steroidal alkaloids, such as chonemorphine and funtumafrine in C. grandiflora. Camptothecin, a well-known anticancer alkaloid has been detected in ethanolic extracts of stem with bark and callus cultures derived from C. grandiflora. Methods: Callus cultures of C. grandiflora were raised on Murashige and Skoog′s medium supplemented with 2, 4-D. Stem with bark and callus were used for phytochemical analysis mainly the alkaloids. Detection and identification of camptothecin was carried out using thin-layer chromatography (TLC, high-performance thin-layer chromatography, (HPTLC and high-performance liquid chromatography (HPLC. Results: An important anticancer alkaloid, camptothecin was detected in ethanolic extracts of stem with bark and callus cultures of C. grandiflora. camptothecin content was 0.013 mg/g in stem with bark and 0.003 mg/g in callus. Conclusion : This is the first report on in vivo and in vitro production of camptothecin in C. grandiflora. Camptothecin is known to occur only in six plant sources so, alternative sources for camptothecin are needed. Thus of C. grandiflora could be a new promising alternative source of camptothecin.

  4. Influence of nutrient composition and plant growth regulators on callus induction and plant regeneration in glutinous rice (Oryza sativa L.).

    Science.gov (United States)

    Duangsee, K; Bunnag, S

    2014-01-01

    The potential for callus induction and regeneration depends on nutrient composition and plant growth regulators. The aim of the present study was to investigate the effect of nutrient composition and plant growth regulators on callus induction and plant regeneration in the glutinous rice cultivar Khunvang. The effect of 2,4-D concentrations (1, 2, 3, 4 and 5 mg L(-1)) on callus induction and growth were investigated. The results revealed that the highest percentage of callus induction (97%) was observed in MS medium supplemented with 5 mg L(-1) 2,4-D under 16 h Photoperiod. The effects of casein hydrolysate concentrations of casein hydrolysate (0, 300, 500, 700 and 900 mg L(-1)) and proline (0, 300, 500, 700 and 900 mg L(-1)) on callus induction and growth of Khunvang were also observed. The results indicated that the increasing casein hydrolysate and proline concentrations did not show a significant effect on callus growth. However, proline concentration of 900 mg L(-1) yielded 85.67% of callus growth.

  5. Mineral crystal alignment in mineralized fracture callus determined by 3D small-angle X-ray scattering

    Energy Technology Data Exchange (ETDEWEB)

    Liu Yifei; Manjubala, Inderchand; Fratzl, Peter [Department of Biomaterials, Max Planck Institute of Colloids and Interfaces, 14424 Potsdam (Germany); Roschger, Paul [4th Medical Department, Ludwig Boltzmann Institute of Osteology at Hanusch Hospital of WGKK and AUVA Trauma Centre Meidling, 1140 Vienna (Austria); Schell, Hanna; Duda, Georg N, E-mail: fratzl@mpikg.mpg.d [Julius Wolff Institut and Center for Musculoskeletal Surgery, Charite- University Medicine Berlin, Augustenburger Platz 1, 13353 Berlin (Germany)

    2010-10-01

    Callus tissue formed during bone fracture healing is a mixture of different tissue types as revealed by histological analysis. But the structural characteristics of mineral crystals within the healing callus are not well known. Since two-dimensional (2D) scanning small-angle X-ray scattering (sSAXS) patterns showed that the size and orientation of callus crystals vary both spatially and temporally [1] and 2D electron microscopic analysis implies an anisotropic property of the callus morphology, the mineral crystals within the callus are also expected to vary in size and orientation in 3D. Three-dimensional small-angle X-ray scattering (3D SAXS), which combines 2D SAXS patterns collected at different angles of sample tilting, has been previously applied to investigate bone minerals in horse radius [2] and oim/oim mouse femur/tibia [3]. We implement a similar 3D SAXS method but with a different way of data analysis to gather information on the mineral alignment in fracture callus. With the proposed accurate yet fast assessment of 3D SAXS information, it was shown that the plate shaped mineral particles in the healing callus were aligned in groups with their predominant orientations occurring as a fiber texture.

  6. Growth effect of culture conditions on the lactic acid bacteria in fermented fruit and vegetable products%培养条件对发酵果蔬制品乳酸菌生长的影响研究

    Institute of Scientific and Technical Information of China (English)

    杨永成

    2014-01-01

    乳酸菌是一类益生性生产菌株,广泛应用于各类果蔬、肉、乳制品中。尤其是在发酵果蔬制品中其生物特性,保健功效更深为人们所熟知。因此开发此类乳酸发酵产品具有很广泛的市场前景。因此,对这类源自发酵果蔬制品中的益酵益生乳酸菌最佳培养条件的研究具有深远的意义。故本课题通过研究培养条件对发酵果蔬制品乳酸菌生长的影响,旨在探索发酵果蔬制品中乳酸菌的最佳培养条件,为后继该类产品发酵菌剂制备培养乳酸菌提供理论依据及技术支撑。经试验得出:几种天然食材制成的培养基中以大米熬煮液最为适生长,食源乳酸菌在其上生长活菌数可达>106cfu/ml,其为源自果蔬发酵制品中的乳酸菌最佳天然培养基;并经L9(34)正交试验确立了该类乳酸菌株的最适培养条件为于大米熬煮液上,调pH值至6.5、添加2%葡萄糖,至温度38℃下培养可获得最高菌数级。%Lactobacillus is a type of probiotics, which are widely used in various kinds of fruits and vegetables, meat, dairy products. In particular in fermented fruit and vegetable products , their biological characteristics and health effects are well known by people. The development of such lactic acid fermentation products have a broad market prospect. It has a far-reaching significance to study the best culture cinditions for lactic acid bacteria which separated from the fermented fruit and vegetable products.This subject study the growth effect on culture conditions of lactic acid bacteria in fermented fruit and vegetable products, in order to explore the best culture condition on fermentated fruits and vegetables products and provide a theoretical basis and technological support for the fermentation agent preparation culture of lactic acid bacteria The results of experiments showed that : in Several mediums which are made of natural ingredients ,the water filter

  7. Insecticidal Activities of Extracts from Brucea javanica (L.) Merr. Callus%鸦胆子愈伤组织提取物的杀虫活性研究

    Institute of Scientific and Technical Information of China (English)

    曾宪儒; 曾涛; 黎柳锋; 韩美丽

    2008-01-01

    [Objective] The research aimed to lay a foundation for the screening of cell lines producing secondary metabolites of Brucea javanica(L.)Merr. [Method] The insecticidal activities of the extracts from branch and 3 different types of calluses of Brucea javanica(L.)Merr. was detected through methods of leaf disc and potted seedlings against the diamond back moth. [Result] Extracts from four kinds of Brucea javanica(L.)Merr. tissues assumed both the activities of antifeedant and oviposition deterrency against the diamond back moth. Antifeedant effect of extracts was in turn the callus C branch > callus B>callus C. The insecticidal activities of callus A and B were higher than that of the callus C. [Conclusion] The results show that insecticidal activity of callus and its growth rate is inversely proportional.

  8. Microstructure and homogeneity of distribution of mineralised struts determine callus strength

    Directory of Open Access Journals (Sweden)

    M Mehta

    2013-07-01

    Full Text Available Non-invasive assessment of fracture healing, both in clinical and animal studies, has gained favour as surrogate measure to estimate regain of mechanical function. Micro-computed tomography (µCT parameters such as fracture callus volume and mineralisation have been used to estimate callus mechanical competence. However, no in-depth information has been reported on microstructural parameters in estimating callus mechanical competence. The goal of this study is to use differently conditioned mice exhibiting good and impaired fracture healing outcomes and investigate the relationship between µCT imaging parameters (volume, mineralisation, and microstructure that best estimate the callus strength and stiffness as it develops over time. A total of 99 mice with femoral fracture and intramedullary stabilisation were divided into four groups according to conditioning: wild type, NF1 knock-out, RAG1 knock-out and macrophage depleted. Animals were sacrificed at 14, 21, 28 or 35 days and µCT parameters and torsional stiffness and strength were assessed post-sacrifice. Using linear regression for all groups and time points together, torsional stiffness could be estimated with strut thickness, strut number and strut homogeneity (R2 = 0.546, p < 0.0001; torsional strength could be estimated using bone mineral density, strut thickness and strut homogeneity (R2 = 0.568, p < 0.0001. Differently conditioned mice that result in different fracture healing outcomes have been shown to result in varying structural, material and volumetric µCT parameters which can be used to estimate regain of bone strength. This study is the first to demonstrate that microstructure and strut homogeneity influence callus stiffness and strength.

  9. Callus induction in papaya (Carica papaya L. and synseed production for low temperature storage and cryopreservation

    Directory of Open Access Journals (Sweden)

    Teixeira da Silva Jaime A.

    2014-12-01

    Full Text Available The mid- to long-term preservation of papaya (Carica papaya L. would allow for the safeguarding of important germplasm. In this study, soft friable callus (SFC and hard callus (HC were induced from the first two true leaves of 10-day-old seedlings containing a midrib derived from the germinated seed of two cultivars (‘Rainbow’ and ‘Sunrise Solo’. Following germination on a Murashige and Skoog (MS medium that contained 3% sucrose and was free of plant growth regulators (PGRs, sections of the first true leaves from 10-day-old seedlings were exposed to seven published callus or somatic embryogenesis protocols for zygotic embryos, leaves or hypocotyls. Optimal SFC and HC induction was carried out on a half-strength MS medium following the Fitch (1993 or the Ascêncio-Cabral et al. (2008 protocol, respectively. SFC formed shoots that could then convert to plants when transferred to a full-strength MS medium devoid of PGRs. Plantlets 10-cm tall were acclimatised in two steps: first by in vitro acclimatisation in aerated vessels, the Vitron, under CO2-enriched (3000 ppm CO2, then by the transfer of individually rooted plantlets in Rockwool® blocks to a substrate of soil: pine bark : perlite (1:1:1, v/v/v. SFC and HC were then encapsulated in alginate beads, which were exposed to low temperature storage (LTS at 10°C and 15°C, and also cryopreserved for 30 days. All encapsulated alginate beads that contained SFC, HC or leaf tissue that had been stored under LTS or cryopreserved were able to regenerate callus when placed on an optimal callus induction medium. Plants derived from the control, LTS and cryopreservation protocols, either from SFC or HC, were successfully acclimatised.

  10. Effects of. gamma. -irradiation on growth and embryogenesis of citrus callus

    Energy Technology Data Exchange (ETDEWEB)

    Nito, Nobumasa; Ling, Jing-Tian; Iwamasa, Masao; Katayama, Yukiyoshi (Saga Univ. (Japan). Faculty of Agriculture)

    1989-09-01

    The embryogenic callus of 'Valencia', orange, 'Yoshida' navel orange (Citrus sinensis (L.) Osbeck), Calamondin (C. madurensis Lour.), Yuzu (C. junos Sieb. ex Tan.) and 'Ishizuka Wase' satsuma (C. unshiu Marc.) were exposed to 1, 5, 10, 20 and 50 kiloroentgen (kR) of gamma irradiation at the dosage of 1 kR/h in Institute of Radiation Breeding, Ministry of Agriculture, Forestry and Fishery, Japan. The irradiation caused a retardation of callus growth. Callus growth was reduced to 50% or less of that in non-irradiated callus, at a dosage of 5 kR in Yuzu, 20 kR in 'Valencia' orange and 'Ishizuka Wase' satsuma, and 50 kR in 'Yoshida' navel orange and Calamondin. The irradiation stimulated embryoid formation from the callus. Optimum irradiation dosages to produce the maximum number of embryoids were 50 kR in 'Valencia' orange, 20 kR in Yuzu, 10 kR in Calamondin and 'Ishizuka Wase' satsuma. 'Yoshida' navel orange produced the maximum number of embryoids at the lowest irradiation of 5 kR. The ratios of embryoid number produced at the optimum irradiation to that at non-irradiation were, 16.4, 8.4, 7.8, 6.4 and 2.7 in Yuzu, 'Ishizuka Wase' satsuma, 'Valencia' orange, 'Yoshida' navel orange and Calamondin, respectively. (author).

  11. Relationship between endogenous hormonal content and somatic organogenesis in callus of peach (Prunus persica L. Batsch) cultivars and Prunus persica×Prunus dulcis rootstocks.

    Science.gov (United States)

    Pérez-Jiménez, Margarita; Cantero-Navarro, Elena; Pérez-Alfocea, Francisco; Le-Disquet, Isabel; Guivarc'h, Anne; Cos-Terrer, José

    2014-05-01

    The relationship between endogenous hormones content and the induction of somatic peach plant was studied. To induce multiple shoots from callus derived from the base of stem explants of the scion cultivars 'UFO-3', 'Flariba' and 'Alice Bigi', and the peach×almond rootstocks 'Garnem' and 'GF677', propagated plants were cultured on Murashige and Skoog salts augmented with 0.1mgL(-1) of indolebutyric acid, 1mgL(-1) of 6-benzylaminopurine and 3% sucrose. The highest regeneration rate was obtained with the peach×almond rootstocks. Endogenous levels of abscisic acid (ABA), indole-3-acetic acid (IAA), zeatin (Z), zeatin riboside (ZR), ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC), salicylic acid (SA), and jasmonic acid (JA) were analyzed in the organogenic callus. Lower levels of several hormones, namely Z, ZR, ABA, and ACC were found in the peach×almond rootstock compared to peach cultivars, while IAA and SA presented inconclusive returns. These results suggest that the difference in somatic organogenesis capacity observed in peach and peach×almond hybrids is markedly affected by the endogenous hormonal content of the studied genotypes.

  12. Callus Induction and Determination of Steroidal Saponin Content of Paris polyphylla Smith var. Chinensis (Franch.) Hara%华重楼愈伤组织培养及薯蓣皂苷含量测定

    Institute of Scientific and Technical Information of China (English)

    王跃华; 陈燕; 张珏; 张红玉; 游亚莉; 田孟良

    2014-01-01

    The multiple organs of Paris polyphylla Smith var. chinensis(Franch.)Hara were used as explants and were disinfect-ed in different way and induced in serious culture medium. The dioscin of subcultural callus was detected by TTC. The re-sults showed that the explants of cryptophytes, inner coleoptile, rhizome, pretreated rhizome and ovary could produce differ-ent levels of callus after inducing and subculturing. Among them ,the growth speed of callus by pretreated rhizome was fast, and the induction rate was highest to 33.5%. The several subcultured callus showed that the same dioscin or its analogue with rhizome and dioscin through detecting. The suitable explant and culture medium was screened out. Callus could be ac-quired after inducing culture. The several subcultured showed the same dioscin or its analogue with rhizome after detecting. It will provide an important way for building effective components in medicinal material by the use of biotechnology.%以华重楼[Paris polyphylla Smith var. chinensis (Franch.)Hara]植物多种器官为外植体,经不同消毒处理和培养基诱导培养后,观察其愈伤组织的诱导生长情况;并以TTC法检测诱导产生的愈伤组织中的薯蓣皂苷含量。结果表明,华重楼的隐芽、内层芽鞘、根茎、预处理后根茎和子房外植体经诱导培养后,均可不同程度地产生愈伤组织;其中以预处理后根茎外植体产生的愈伤组织生长速度最快,诱导率也最高,为33.5%。经检测结果显示,多次继代培养的华重楼愈伤组织中含有与华重楼对照药材和薯蓣皂苷对照品相同的薯蓣皂苷或其类似物。

  13. Establishment and content of sugars and phenols in Physalis callus obtained from different explants and concentrations of bap and naa - doi: 10.4025/actascibiolsci.v36i1.18074

    Directory of Open Access Journals (Sweden)

    Douglas Junior Bertoncelli

    2013-09-01

    Full Text Available Obtaining cells of Physalis pubescens is of interest for studies of primary and secondary metabolic pathways, in the search for new active molecules. Our objectives were to evaluate the regeneration potential of explants from different parts of the plant, growth regulators to be used, and the determination of the growth curve of the callus. We used explants of leaf, root, stem and petiole, cultured on Murashige and Skoog medium with different concentrations of 6-benzylaminopurine and a-naphthaleneacetic acid. The explants from stem and petiole had a higher regeneration potential of the shoot to the treatment with 0.5 mg L-1 6-benzylaminopurine, and the explants of leave and root emitted more roots, with lower production of callus. The tests showed that the regeneration of the whole plant should be done in two steps: cultivation for shoot regeneration and transplantation to a new rooting medium. The growth of callus showed five distinct phases, with accumulation of phenols in the final stages of growth. The levels of soluble sugars increased with age, while reducing sugars showed variations, with higher concentrations in the initial stages of cultivation, with fall and rise again at the final evaluation (28th day.

  14. 长心卡帕藻愈伤组织的诱导与其形态建成的初步研究%Callus induction and morphogenesis of callus in Kappaphycus alvarezii (Rhodophyta,Solieriaceae)

    Institute of Scientific and Technical Information of China (English)

    李虎; 刘建国; 庞通

    2015-01-01

    以 PES 液体培养基、固体培养基,采用正交设计,开展了长心卡帕藻(Kappaphycus alvarezii )棕色藻株组织培养和愈伤组织诱导实验,探索了蔗糖、光强、植物生长调节剂对新芽形成和愈伤组织形成的影响,结果表明,固体培养基、IBA、6-BA 处理可诱导该藻产生愈伤组织,同时固体培养基更利于诱导形成愈伤组织;显微跟踪观察显示,该藻愈伤组织与高等植物的疏松愈伤组织不同,由细丝状细胞组成,系藻枝段中心髓部细胞脱分化形成的致密型愈伤组织;另外,愈伤枝段比新芽枝段有更高的光合和呼吸速率。%The brown morphotype of red seaweed Kappaphycus alvarezii collected from Li’an Bay in Lingshui County,Hainan Province of China,was used as explants in order to generate new buds and induce callus.Effect of the PES (Provasoli enriched seawater)media (with or without gelling agent),light intensity,and phytoregulators (Indole-3-acetic acid (IAA),Indole-3-butyric acid (IBA),6-benzylaminopurine (6-BA))were tested on new bud generation and callus formation by orthogonality experiment.The callus forming was observed in microscope,and the changes of the respiration rate and photosynthesis rate in explants among callus forming were detected.The callus of Kappaphycus alvarezii was achieved successfully by the combination of PES solid medium (0.8% ager), IBA,6-BA,while callus was not observed in the combination of PES liqiud medium,IBA and 6-BA.That indicates the solid medium plays an important role in callus induction.Kappaphycus alvarezii callus which consists of com-pact filamentous cells was different from the loose callus of superior plants.And callus derived from the medullary cells.It was found that both photosynthetic rate and respiratory rate in the branch inducing callus were higher than ones of the branch germinating new bud by means of oxygen electrode.

  15. Effects of pigment glands andgossypol on somatic cell cul-ture of upland cotton (Gos-sypium hirsutum L.)

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The effects of pigment glands and gossypol on the somatic cell culture of upland cotton were studied, using the materials as follows: three pairs of glanded and glandless upland cotton near isogenic lines, TM-1, and Coker 312. The results showed that the pigment glands and gossypol contents in the explants had great inhibiting effect on the induction and growth of callus in somatic cell culture of upland cotton, and the induction rate of callus and the single callus weight of glandless cotton were much higher than those of their glanded near isogenic lines. It was easier to obtain regeneration plants from glandless cotton than from their glanded near isogenic lines. There was a significant inverse correlation between the gossypol contents in the explants and callus induction rate, with the correlation coefficient of ?0.84. The vitro gossypol in the medium had some inhibiting effect on the induction and growth of callus, especially for the glandless cotton. However, a certain concentration of vitro gossypol in the medium (0.1 mg/L) was an aid to the steadiness growth of callus in glandless cotton somatic cell culture, with a high rate of embryogenic cells which was in favor of plant regeneration, and it was also relatively easy to obtain regeneration plants when they were transferred into differentiation medium with 0.1 mg/L of vitro gossypol, even for some cultivars which are difficult in somatic cell culture. In addition, the gossypol content and its variation in the seedlings and callus during culture of Coker 312 were discussed, as well as the relationship between gossypol variation in the explants and its somatic cell culture. The probability of vitro gossypol used in cotton somatic cell culture for the improvement of somatic cell culture was suggested.

  16. Barley callus: a model system for bioengineering of starch in cereals

    Directory of Open Access Journals (Sweden)

    Carciofi Massimiliano

    2012-09-01

    Full Text Available Abstract Background Starch is the most important source of calories for human nutrition and the majority of it is produced by cereal farming. Starch is also used as a renewable raw material in a range of industrial sectors. It can be chemically modified to introduce new physicochemical properties. In this way starch is adapted to a variety of specific end-uses. Recombinant DNA technologies offers an alternative to starch industrial processing. The plant biosynthetic pathway can be manipulated to design starches with novel structure and improved technological properties. In the future this may reduce or eliminate the economical and environmental costs of industrial modification. Recently, many advances have been achieved to clarify the genetic mechanism that controls starch biosynthesis. Several genes involved in the synthesis and modification of complex carbohydrates in many organisms have been identified and cloned. This knowledge suggests a number of strategies and a series of candidate genes for genetic transformation of crops to generate new types of starch-based polymers. However transformation of cereals is a slow process and there is no easy model system available to test the efficiency of candidate genes in planta. Results We explored the possibility to use transgenic barley callus generated from immature embryo for a fast test of transgenic modification strategies of starch biosynthesis. We found that this callus contains 4% (w/w dw starch granules, which we could modify by generating fully transgenic calli by Agrobacterium-transformation. A Green Fluorescent Protein reporter protein tag was used to identify and propagate only fully transgenic callus explants. Around 1 – 1.5 g dry weight of fully transgenic callus could be produced in 9 weeks. Callus starch granules were smaller than endosperm starch granules and contained less amylose. Similarly the expression profile of starch biosynthesis genes were slightly different in callus

  17. Establishment of hairy root cultures of Ammi majus.

    Science.gov (United States)

    Królicka, A; Staniszewska, I; Bielawski, K; Malinski, E; Szafranek, J; L&z shtsls;ojkowska, E

    2001-01-01

    Axenically grown Ammi majus plantlets were inoculated with seven different Agrobacterium rhizogenes strains. Hairy root lines were established only after inoculation with the two agropine strains: A4 and LBA9402. The growth rate of hairy root cultures was about thirty times faster than that of callus and cell suspension cultures. Polymerase chain reaction with primers for the genes rolB and rolC confirmed the integration of the T-DNA fragment of Ri plasmid of A. rhizogenes to the genome of hairy roots obtained after transformation by both Agrobacterium strains. The furanocoumarins (psoralen, xanthotoxine, bergapten and imperatorin) usually found in seeds of A. majus were not detected in callus, cell suspension and hairy root cultures using Gas chromatography-mass spectrometry (GC-MS). However, umbelliferone, a precursor of furanocoumarins, was detected in callus, cell suspension and hairy root cultures. The umbelliferone content in extracts of hairy root cultures, obtained after transformation by A4, was similar to that determined in A. majus seeds (19 µg/g DW) and higher than those obtained for cell suspension and callus cultures (2 and 9 µg/g DW, respectively).

  18. Different Plant Growth Regulators on Callus Induction of Pinellia Ternata%不同植物生长调节剂对半夏愈伤组织诱导效应的研究

    Institute of Scientific and Technical Information of China (English)

    刘鑫欣; 崔晓星; 刘金欣; 孟繁蕴; 魏英勤

    2011-01-01

    目的 以半夏叶柄为外植体,研究不同植物生长调节剂对半夏愈伤组织形态的影响.方法 分别采用单一激素和多个激素正交试验,考察不同植物生长调节剂对半夏愈伤的诱导效果.结果 单一激素不同水平的愈伤组织诱导效果因激素种类和浓度的不同而有所差异;多种激素不同水平对半夏愈伤诱导的效果与诱导率和启动时间有关,同时还应考虑愈伤的质地、形态等;植物生长调节剂对半夏愈伤组织的诱导能力从大到小依次为6-苄基腺嘌呤、α-萘乙酸、2,4-二氯苯氧乙酸、蔗糖.结论 通过考察不同植物生长调节剂对半夏愈伤组织的诱导效应,确定了适宜做半夏悬浮培养材料的培养基和一次性成苗培养基.%Objective To study the influence of plant growth regulators on callus induction of Pinellia ternata, with the petioles as explants for tissue culture. Methods Single factor experiments and orthogonal experiments were used to study callus induction effects. Results. The induction effect of single plant growth regulators on Pinellia ternata callus were different depends on hind and concentration. The effect of multiple plant growth regulators was related with induction rate and starting time, as well as the texture and morphology of callus. The effective plant growth regulators were as follows: 6-BA, NAA, 2,4-D sucrose.Conclusion By examining the induction effect of different plant growth regulators on callus of Pinelliaternata, suitable mediums for suspension culture and seedling were determined.

  19. LC/MS/MS identification of glycosides produced by biotransformation of cinnamyl alcohol in Rhodiola rosea compact callus aggregates.

    Science.gov (United States)

    Tolonen, Ari; György, Zsuzsanna; Jalonen, Jorma; Neubauer, Peter; Hohtola, Anja

    2004-10-01

    Cinnamyl alcohol was added to the media of compact callus aggregates (CCA) of Rhodiola rosea for stimulating the production of cinnamyl glycosides. The biotransformation reaction produced high amounts of rosin, while only a very low amount of rosavin was produced. As the consumption rate of cinnamyl alcohol was much higher than production of rosin, the aqueous methanol extracts of compact callus aggregates were studied by liquid chromatography-mass spectrometric methods and four new unexpected biotransformation products of cinnamyl alcohol were identified.

  20. Pulp fruit added to culture medium for in vitro orchid developmentPolpa de frutos adicionada ao meio de cultivo no crescimento in vitro de orquídea

    Directory of Open Access Journals (Sweden)

    Gilberto Rostirolla Batista de Souza

    2013-06-01

    Full Text Available As an additive in in vitro culture media, fruits have a great potential for facilitating economical orchid production because of lower technology requirements and the ease of obtaining raw materials to formulate culture media. We studied the in vitro growth of Cattleya bicolor Lindl. grown in a simplified culture medium supplemented with different kinds of fruit pulp. The experimental design was completely randomised, with eight seedlings per replication and ten replications per treatment, for a total of 80 seedlings per treatment. The culture medium was made using 150 g L -1 of pulp (without peel or seed from the following fruits: ripe Santa Cruz tomatoes (Solanum lycopersicum L., dwarf bananas (Musa cavendishii L. of intermediate ripeness, light green chayote (Sechium edule (Jacq. Sw, ripe papaya (Carica papaya L. or green coconut (Cocos nucifera L..The treatment control was MS 50 %. The treatments and the control were kept in a growth chamber for seven months before evaluating seedling survival percentage, shoot height, number of leaves, rooting percentage, root number, root length and dry masses of shoot and roots. The highest percentages of seedling survival were obtained using MS 50 %, banana and coconut medium. The seedling survival and rooting percentages illustrate that it is possible to emphasise the culture medium MS 50% and the culture medium supplemented with coconut on the most traditional culture medium with banana or tomato pulp. For the in vitro development of Cattleya bicolor Lindl., a simplified culture medium supplemented with coconut pulp is the most suitable for use as an alternative to MS 50%. A simplified culture medium supplemented with papaya pulp is not recommended for the in vitro development of Cattleya bicolor Lindl. Os frutos apresentam potencial para serem utilizados na elaboração de meios de cultivo para facilitar a produção de orquídeas em pequenas propriedades, contribuindo para a rentabilidade do cultivo

  1. BIOTECHNOLOGY IN FRUIT GROWING

    Directory of Open Access Journals (Sweden)

    Z. Jurković

    2008-09-01

    Full Text Available Research studies in the area of biotechnologies in fruit growing started at the Agricultural Institute Osijek in 2006 with the establishment of the first experimental in vitro laboratory for micropropagation. The laboratory started an active research related to the Project "Biotechnological methods in fruit tree identification, selection and propagation" Project is part of program "Preservation and revitalization of grape and fruit autochthonous cultivars". The goal of this research is to determine genetic differences between autochthonous and introduced cultivars of cherry as well as cultivars and types of sour cherry, to find and optimize a method for fast recovery of clonal material. A great number of cherry cultivars and types within the population of cv. Oblacinska sour cherry exists in Croatia. A survey with the purpose of selecting autochthonous cultivars for further selection has been done in previous research. Differences have been found in a number of important agronomic traits within the populations of cv. Oblačinska sour cherry. Autochthonous cherry cultivars are suspected to be synonyms of known old cultivars which were introduced randomly and have been naturalized under a local name. Identification and description of cultivars and types of fruits is based on special visible properties which were measurable or notable. In this approach difficulties arise from the effect of non-genetic factors on expression of certain traits. Genetic-physiological problem of S allele autoincompatibility exists within cherry cultivars. Therefore it is necessary to put different cultivars in the plantation to pollinate each other. Apart form the fast and certain sort identification independent of environmental factors, biotechnological methods based on PCR enable faster virus detection compared with classical serologic methods and indexing and cover a wider range of plant pathogens including those undetectable by other methods. Thermotherapy and

  2. 木薯叶片愈伤组织对枯萎病病原菌的抗性筛选%Screening for Cassava Leaf Callus Resistance to Bacterial Wilt

    Institute of Scientific and Technical Information of China (English)

    陆荣生; 韩美丽; 霍秀娟; 马跃峰

    2012-01-01

    以木薯品种“GR911”、“面包木薯”试管苗叶片为材料,研究木薯枯萎病病原菌粗提物对愈伤组织增殖与分化的影响.以期建立枯萎病病原菌粗提物为选择压力的木薯离体抗病筛选体系。结果表明:所用品种幼叶愈伤诱导与增殖的适宜配方为改良的MS培养基+0.5-0.75mg/L2,4一D:愈伤分化适宜配方为的MS培养基+O.75—1.0mg/LZT。“GR911”与“面包木薯”愈伤增殖过程中。愈伤褐化率达到50%所需的细菌液粗提物浓度分别为5.5×、2.5×^6cfu/mE:愈伤组织分化过程中,愈伤褐化率达到50%所需的菌液粗提物浓度分别为7.0×10^6、4.0×10^6cfu/mL。通过建立木薯愈伤组织离体抗病筛选体系。为木薯无性系变异育种奠定基础。%This study was made to clarify the effects of bacterial crude toxin culture on proliferation and differentiation of callus in cassava, to screen system of resistance to bacterial wilt caused by Xanthomonas manihotis (Artheodet Bey.) Stars in vitro. The young leaves from plantlet were used as explants. On basal modified MS medium, the optimum hormone concentration is 0.5-0.75 mg/L 2 ,4-D for induction and proliferation of callus, and 0.75-1.0 mg/ L ZT for differentiation of callus. To achieve 50 % callus browning rate during callus proliferation, the concentration of bacteria crude toxin culture in medium should be 5.5×106 cfu/ml for species GR991 and 2.5× 106 cfu/ml for species MianBao. To get 50 % callus browning rate during callus differentiation, the concentration of bacteria crude toxin culture in medi-um should be 7.0×106 cfu/ml for species GR991 and 4.0× 106 cfu/ml for species MianBao. By this ex- periment, the screening system of resistance to bacterial wilt was established in vitro in cassava, and this system can be used for clone breeding studies.

  3. Influence of plant growth regulators on callus mediated regeneration and secondary metabolites synthesis in Withania somnifera (L.) Dunal.

    Science.gov (United States)

    Chakraborty, Nilanjan; Banerjee, Debarupa; Ghosh, Moumita; Pradhan, Prakash; Gupta, Namrata Shanu; Acharya, Krishnendu; Banerjee, Maitreyi

    2013-01-01

    Withania somnifera (L.) Dunal, is an important medicinal plant being the source of extremely important compounds like withanolides and withaferin. Influence of different plant growth regulators (PGRs) were evaluated for induction of callus, callus mediated regeneration and production of secondary metabolites in them. Explants for callusing were collected from plants grown in vitro and maximum callusing (98 %) was obtained on MS medium supplemented with a combination of 2,4-dichlorophenoxy acetic acid (2,4-D) (0.5 mg l(-1)) and kinetin (KN) (0.2 mg l(-1)). Among different types of calli, best shoot regeneration was observed on green, compact calli produced on MS medium with a combination of 6-benzylamino purine (BAP) and indole butyric acid (IBA). MS medium supplemented with BAP (2 mg l(-1)) showed highest frequency (98 %) of shoot bud regeneration. The micro-shoots were efficiently rooted on MS media supplemented with 0.5 mg l(-1) IBA. Rooted plants were transferred to soil-vermi-compost (1:3; w/w) medium in greenhouse for acclimatization. Presence of withanolide A and withaferin A in calli was validated through high performance thin layer chromatography (HPTLC). It was interesting to observe that the PGRs showed significant influence on the secondary metabolites production in callus and 2,4-D having the least effect. Histological studies revealed the origin of shoot tip in the callus during regeneration.

  4. Condensed tannins in the tissue culture of sainfoin (Onobrychis viciifolia Scop.) and birdsfoot trefoil (Lotus corniculatus L.).

    Science.gov (United States)

    Lees, G L

    1986-08-01

    Two forage legumes, birdsfoot trefoil (Lotus corniculatus L.) and sainfoin (Onobrychis viciifolia Scop.), containing condensed tannins in their leaves and stems were used as source material to study condensed tannins in tissue culture. More protoplasts were isolated from mesophyll tissue of a low tannin-containing strain of birdsfoot trefoil than from a high tannin-containing strain, but more tannin-filled protoplasts were observed in the latter. Growth rates of leaf explant-derived callus tissue were greater for the high-tannin than for the low-tannin strain. In sainfoin, callus cultures from leaf explants produced numerous tannin-filled cells by 21 days. Explants from sainfoin cotyledons and roots, tissues which normally do not contain tannins, also formed callus with tannin-filled cells in 21 days but in almost every case, a cytokinin was required for tannin formation to occur. The occurrence of tannin-filled cells in callus from root and cotyledon explants was variable and genotype specific. These results show that endogenous tannins can affect protoplast isolation and possibly callus growth in birds-foot trefoil, and that the formation of condensed tannins in sainfoin callus culture can be influenced by a growth regulator.

  5. In vitro cultures of Salvia officinalis L. as a source of antioxidant compounds

    Directory of Open Access Journals (Sweden)

    Izabela Grzegorczyk

    2011-01-01

    Full Text Available The concentrations of carnosic acid, carnosol and rosmarinic acid in different materials from differentiated (multiple shoot cultures and regenerated plants and undifferentiated (callus and cell suspension in vitro cultures of Salvia officinalis were determined by HPLC. The results suggested that diterpenoid (carnosic acid and carnosol production is closely related to shoot differentiation. The highest diterpenoid yield (11.4 mg g-1 for carnosic acid and 1.1 mg g-1 for carnosol was achieved in shoots of 10-week-old micropropagated plants. The levels were comparable to those found in shoots of naturally growing plants. Undifferentiated callus and cell suspension cultures produced only very low amounts of carnosol (ca. 0.05 mg g-1 of dry weight. In contrast, content of rosmarinic acid in callus and suspension cultures as well as shoots growing in vitro and in vivo was similar and ranged between 11.2 and 18.6 mg g-1 of dry weight.

  6. Purification and Analysis of Abscisic Acid-Specifically-Inducible Proteins from Rice Callus

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Two ABA-specifically-inducible proteins from rice callus were isolated and purified by precipitation with 65-100 % saturated(NH4)2SO4, followed by the DEAE-sepharose, TSK-gel, and two-dimension electrophoresis. Iso-electric points (pl) of the proteins with the same molecular mass (24.5 kD) were 6.1 and 6.9, respectively. The Western blot analysis indicated that the proteins expressed in different tissues were obviously different. The A1 (pl 6.1) protein was only detected in calli treated with ABA and seed embryos (SE).However, the A2 (pl 6.9) protein was found not only in the calli treated with ABA and SE, but also in the white dry callus. Thus it suggested that the two proteins might play some important roles in the processes of seed embryo (or somatic embryo) formation.

  7. Plant regeneration of transgenic China Rose (Rosa chinensis Jacq.) from organogenic callus

    Institute of Scientific and Technical Information of China (English)

    Chen Ji-ren; Liu Rong; Chen Shou-yi; Wang Hua-fang

    2006-01-01

    Different types of explants of China Rose (Rosa chinensis Jacq.) were placed on a Schenk and Hildebrandi (SH) medium containing L-proline and 2,4-dichlorophenoxyacefic acid (2,4-D). Organogenesis was observed on callus induced from both whole leaf and petiole and the high frequency of organogenesis was observed on the whole leaf. Shoot regeneration was obtained via organogenesis. The effects of pH and concentrations of antibiotics on maintenance of organogenesis capacity were investigated in subsequent subcultures. The pH value was found to play a critical role in retaining organogenesis capacity. The binary vector pBI121,carrying the gus gene coding for β-glucuronidase (GUS) and the npt Ⅱ gene mediated by Agrobacterium tumefaciens, was used for transformation of organogenic callus using 50 mg·L-1 geneficin for selection. Six regenerated lines showed GUS activity, of which five were verified for the presence of npt Ⅱ gene by PCR.

  8. Cytological characterization of Jatropha curcas callus in different periods of cultivation

    Directory of Open Access Journals (Sweden)

    Dalilhia Nazaré dos Santos

    2013-12-01

    Full Text Available The aimed was characterization and determining the time for Jatropha curcas callus subculture. The pattern of J. curcascallus development is sigmoidal with 5 distinct phases, and transplantation should be performed at the end of the deceleration phase(approximately day 28. During callus development, it was observed that at the onset of growth in the exponential phase the cells wereinternally disorganized; in the linear phase, respiratory metabolism was resumed through reorganization of the first mitochondria;and by the mid-deceleration phase, the cells were entirely active, and several organelles were detected. This organizational status wasmaintained throughout the stationary phase during which somatic pre-embryos were identified. At the end of the stationary phase, intracellulardisruption began, and the cells entered senescence, which characterized the decline phase for the J. curcas calli growth curve.

  9. [Molecular-genetic polymorphism of cellular lines of wheat resistant to cultural filtrate Gaeumannomyces graminis var. tritici and plant-regenerants from them].

    Science.gov (United States)

    Bavol, A V; Dubrovna, O V

    2009-01-01

    DNA polymorphism of wheat cellular lines resistant to culture filtrate of G. graminis var. tritici and regenerated plants has been investigated using ISSR-analysis. Specific changes in DNA sequence were revealed in resistant calluses. It was established, that all resistant cellular lines differed by the presence of specific amplicons of 2347 bp (primer 5'-TCTCTCTCTCTCTCTCG-3') and 1745 bp (primer 5'-AGAGAGAGAGAGAGAGTC-3'), as well as the absence of the amplicon of 1108 bp (primer 5'-ACACACACACACACACC-3') from the initial callus and the callus not exposed to the action of selective factor. These changes have been also revealed in R0 and R1 plants.

  10. Proline and Glutamine Improvein vitro Callus Induction and Subsequent Shooting in Rice

    Institute of Scientific and Technical Information of China (English)

    Bhausaheb PAWAR; Prashant KALE; Jyoti BAHURUPE; Ashok JADHAV; Anil KALE; Sharad PAWAR

    2015-01-01

    This study was conducted to evaluate the effects of proline and glutamine onin vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four ricegenotypes, viz. Pawana, Jaya, Indrayani and Ambemohar. Considerable variation in response to plant growth regulators and amino acid supplements used was observed in all the four genotypes. Medium supplemented with proline and glutamine was shown to be superior to medium without proline and glutamine. The best callusing from mature embryo was observed on Murashige and Skoog (MS) medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), 500 mg/L proline and 500 mg/L glutamine. Shoot induction was higher in the callus obtained from medium supplemented with 500 mg/L proline and 500 mg/L glutamine. The highest shoot regeneration frequency (83.2%) was observed on MS medium with 2.0 mg/L benzylaminopurine, 0.5 mg/L 1-naphthaleneacetic acid, 500 mg/L proline, and 500 mg/L glutamine in the callus obtained from MS medium supplemented with 2.0 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine. Among the four genotypes, Pawana has the highest regeneration efficiency (83.2%), whereas the regeneration efficiency of the rest three rice genotypes was in the range of 32.0% to 72.3%. This optimized regeneration protocol can be efficiently used forAgrobacterium mediated genetic transformation in rice.

  11. A pilot study investigating the utilization of crest pads for treatment of toe callus and ulceration.

    Science.gov (United States)

    Melo, Monica; Bernecker, Tricia; McCullough, James; Hong, John; Trumbauer, Jane Scott; Miller, Mary Ellen

    2015-12-01

    Patients with lesser-toe deformities are at increased risk of developing calluses and ulcers on the distal ends of the affected digits because of the increased pressures applied to these areas. The number of diabetic patients in the United States continues to increase, along with associated comorbidities such as peripheral vascular disease and peripheral neuropathy. These conditions predispose patients to developing foot ulcerations, especially if foot deformities are present. Crest pads are a simple-to-make, inexpensive option to treat calluses and ulcerations on the distal ends of digits; however, there is no research available that support their use. Crest pads consist of rolled gauze covered in moleskin, with a large opening that fits over several toes and lies on the dorsal aspect of the foot, with the padded portion resting under the toes. Over several days of use, the pad molds to the plantar aspect of the toes, offloading pressure from the distal end of the affected digit(s). The sample was obtained through a retrospective chart review of patients identified as having had at least one nail care visit and at least one follow-up visit at a vascular surgery practice between August 2011 and December 2014. Potential subjects with toe deformities who presented with callus or ulcer on the distal end of a digit were considered for inclusion, if they received a crest pad as part of their treatment plan. The scholarly project was a preintervention or postintervention design with subjects acting as their own controls. McNemar's test was used to analyze the results which were statistically significant (P ulcer improvement following the crest pad intervention. The results of this scholarly project support the use of crest pads in patients with lesser-toe deformities to treat distal toe calluses and/or ulcerations.

  12. Production and immunogenicity of Actinobacillus pleuropneumoniae ApxIIA protein in transgenic rice callus.

    Science.gov (United States)

    Kim, Mi-Young; Kim, Tae-Geum; Yang, Moon-Sik

    2017-04-01

    Actinobacillus pleuropneumoniae is a major etiological agent that is responsible for swine pleuropneumonia, a highly contagious respiratory infection that causes severe economic losses in the swine production industry. ApxIIA is one of the virulence factors in A. pleuropneumoniae and has been considered as a candidate for developing a vaccine against the bacterial infection. A gene encoding an ApxIIA fragment (amino acids 439-801) was modified based on a plant-optimized codon and constructed into a plant expression vector under the control of a promoter and the 3' UTR of the rice amylase 3D gene. The plant expression vector was introduced into rice embryogenic callus (Oryza sativa L. cv. Dongjin) via particle bombardment-mediated transformation. The integration and transcription of the ApxIIA439-801 gene were confirmed by using genomic DNA PCR amplification and Northern blot analysis, respectively. The synthesis of ApxIIA439-801 antigen protein in transgenic rice callus was confirmed by western blot analysis. The concentration of antigen protein in lyophilized samples of transgenic rice callus was 250 μg/g. Immunizing mice with protein extracts from transgenic plants intranasally elicited secretory IgA. These results demonstrate the feasibility of using a transgenic plant to elicit immune responses against A. pleuropneumoniae. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Effects of Salinity on growth and osmotic regulation substances of callus induced from Reaumuria soongorica

    Science.gov (United States)

    Tan, Huijuan; Li, Xinrong; Liu, Yubing; Zhao, Xin

    2014-05-01

    Reaumuria soongorica (Pall.) Maxim is the strong xerophils plant in the northwest arid and semiarid regions in China. It plays very important roles in stabilizing sand dunes and in construction of agricultural shelter belts in north-west China.The present study aimed to evaluate the response to salinity of R. soongorica, which is more salt-resistant than other valuable shrub species used for afforestation on saline and alkaline desert, at the cellular level. To this purpose, callus was induced from shoot segments of R. soongorica on Murashige and Skoog (MS) medium supplemented with 0.2 mgL-16-benzyladenine (BA) and 2.0 mg mgL-1 2,4-Dichlorophenoxyacetic acid (2 ,4-D). The relative growth rate of callus reached a maximum in the presence of 100 mmol L-1NaCl and growth was inhibited with increasing NaCl concentrations. Examination of the changes of osmotic substances under salt stress showed that accumulation of proline, trehalose, Glycine betain and flavonoids increased with increasing salt concentrations. The results indicate that the response of the callus of R. soongorica to salt stress is similar to that of the whole plant. .

  14. Rhizobium radiobacter conjugation and callus-independent shoot regeneration used to introduce the cercosporin export gene cfp from Cercospora into sugar beet (Beta vulgaris L.).

    Science.gov (United States)

    Kuykendall, L David; Stockett, Tammy M; Saunders, Joseph W

    2003-05-01

    Leaf spot disease caused by Cercospora is responsible for crop and profitability losses in sugar beet crops in the US and worldwide. The cfp gene that encodes a protein that exports phytotoxic cercosporins from Cercospora was conjugally transferred to sugar beet using Rhizobium radiobacter (Agrobacterium tumefaciens), to improve Cercospora-induced leafspot resistance. Conditions for shoot regeneration were optimized to increase regeneration/transformation efficiencies. Low-light and room-temperature conditions were favorable to sugar beet regeneration without callus when cytokinin had been added to the tissue culture medium. Using this procedure adventitious shoots from leaf pieces were obtained in a simple, one-step regeneration procedure. T7, a cfp-transgenic clone verified by PCR with gene-specific primers, is being propagated for leaf spot disease resistance evaluation.

  15. 人工培养蛹虫草C19提取液的体外抗氧化活性研究%In vitro Anti-oxidation of the Extraction from C19 Fruiting Bodies of Cultured Cordyceps militaris

    Institute of Scientific and Technical Information of China (English)

    毛穗芳; 巫光宏; 古清霞; 陈江磊; 张莉

    2011-01-01

    探讨人工培养的蛹虫草C19的生物学活性,采用化学比色法分别测定蛹虫草提取物的抗氧化活性.实验结果表明,蛹虫草C19提取液具有很强还原能力.提取液为0.12%时,对DPPH的清除率达到(49.7±1.83)%,对MDA生成的抑制率为(88.67±2.59)%;提取液为0.6%时,对O2-·的清除率为(83.22±3.07)%;提取液为1%时,对·OH的清除率为(69.59±2.24)%.表明人工培养的蛹虫草子C19可作为一种潜在的天然抗氧化剂.%The biological activity of fruiting bodies of cultured Cordyceps militaris C19 was examined and the antioxidant activity of the extract from Cordyceps militaris was investigated by using the chemical colorimetry. The experimental results showed that the extract from C19 fruiting bodies of cultured Cordyceps ,miilitaris had potent reducing power. When the concentration of the extract was 0.12%, its scavenging effect on DPPH reached (49.74±1.83)%, and its inhibitory effect on the generation of MDA was (88.67±2.59)%. When the concentration of the extract was 0.6%, its scavenging effect on O2-· reached (83.22±3.07)%. And its scavenging effect on·OH reached (69.59±2.24)% with its concentration being of 1%. It was indicated that the C19 fruiting bodies of cultured Cordyceps militaris was a potential natural antioxidant.

  16. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

    Full Text Available Es un estudio cualitativo que adoptó como referencial teorico-motodológico la antropología y la etnografía. Presenta las experiencias vivenciadas por mujeres de una comunidad en el proceso salud-enfermedad, con el objetivo de comprender los determinantes sócio-culturales e históricos de las prácticas de prevención y tratamiento adoptados por el grupo cultural por medio de la entrevista semi-estructurada. Los temas que emergieron fueron: la relación entre la alimentación y lo proceso salud-enfermedad, las relaciones con el sistema de salud oficial y el proceso salud-enfermedad y lo sobrenatural. Los dados revelaron que los moradores de la comunidad investigada tienen un modo particular de explicar sus procedimientos terapéuticos. Consideramos que es papel de los profesionales de la salud en sus prácticas, la adopción de abordajes o enfoques que consideren al individuo en su dimensión sócio-cultural e histórica, considerando la enorme diversidad cultural en nuestro país.

  17. In vitro callus and in vivo leaf extract of Gymnema sylvestre stimulate β-cells regeneration and anti-diabetic activity in Wistar rats.

    Science.gov (United States)

    Ahmed, A Bakrudeen Ali; Rao, A S; Rao, M V

    2010-11-01

    A methanol extract of Gymnema sylvestre leaf and callus showed anti-diabetic activities through regenerating β-cells. Optimum callus was developed under stress conditions of blue light with 2,4-D (1.5 mg/l) and KN (0.5 mg/l), which induced maximum biomass of green compact callus at 45 days, as determined by growth curve analysis. Leaf and optimum callus extracts contains gymnemic acid, which was analyzed using TLC, HPTLC and HPLC methods. The research reported here deals with leaf and callus extracts of G. sylvestre, which significantly increase the weight of the whole body, liver, pancreas and liver glycogen content in alloxan-induced diabetic rats (Wistar rats). The gymnemic acid of leaf and callus extracts significantly increases the regeneration of β-cells in treated rats, when compared with the standard diabetic rats. It could have potential as a pharmaceutical drug for insulin-dependent diabetes mellitus (IDDM).

  18. 虎榛子愈伤组织对土生空团菌菌丝生长的影响%Influence of Callus of Ostryopsis davidiana on the Mycelium Growth of Cenococcum geophilum Fr.

    Institute of Scientific and Technical Information of China (English)

    赵慧英; 薛丽宁; 姚庆智; 闫伟

    2012-01-01

    The latest research showed the obstacle of mycorrhizae formation was lack of some signal material that could induce mycorrhizal fungi mycelial to grow and branch. The co-cultures of callus of Ostryopsis davidiana and its mycorrhizal fungi were established for improving mycorrhizal fungi cultivation methods. And the secondary metabolites of the co-culture were studied. Using the dry weight method, the influence of the 0. davidiana callus and homogenate and secretion of callus on the mycelium growth of Cenococcum geophilum Fr. (Cg) was investigated in vitro. The results showed that, the two strains which co-cultured with the host callus could obviously promote the fungal hyphae germination and growth. At the same time, homogenate and secretion of callus could also facilitate early mycelia growth. While the most effective method was the co-cultures, and the single colony dry maximum biomass of each Cg strain reached to 63.5 mg and 45.2 mg. The research of preliminary got a good cultural method of ectomycorrhizal fungi mycelium which could evidently upgrade the growth of the hyphae. The O. davidiana callus cells surely secreted some biochemical substances that promoted the growth and branch of hyphae in the progress of the growth.%新研究表明植物不能形成菌根的障碍是缺失一种植物形成菌根的信号物质,这种信号物质可以诱导菌根真菌菌丝的生长和分枝,为下一步研究植物与外生菌根真菌互作的生化机制和互作过程中的信号传导途径提供理论基础,建立虎榛子愈伤组织与其外生菌根真菌的共培养体系,在纯培养条件下采用于重法,测定虎榛子愈伤组织、愈伤组织匀浆液和愈伤组织分泌物对2株土生空团菌[CenococcumgeophilumFr.(Cg)]菌株生长的影响.结果表明,供试Cg菌株与宿主的愈伤组织共同培养可以明显地促进菌丝的萌发和生长,而愈伤组织匀浆液和愈伤组织分泌物也可促进菌丝体早期

  19. IN-VITRO REGENERATION AND CALLUS FORMATION FROM DIFFERENT PARTS OF SEEDLING OF MUCUNA PRURIENS BAK ñ A VALUABLE MEDICINAL PLANT

    Directory of Open Access Journals (Sweden)

    SHALINI PATEL

    2007-01-01

    Full Text Available Callus proliferation was studied on cotyledon, leaf and stem explant of Mucuna pruriens Bakcultured on Murashige and Skoogís medium (MS supplemented with 2,4-D, IBA, NAA and BAPalone or in combination. Light brown callus formation was followed by formation of milky whitecallus on the surface of young excised shoots and leaf tissues of Mucuna pruriens. Sometimesgreen callus was also observed. Development of root and stem with leaves were investigated fromexcised stem, leaf and cotyledon tissues.

  20. Isolation of three dibenzocyclooctadiene lignans from in vitro cultures of Schisandra chinensis (Turcz.) Baill.--the first report.

    Science.gov (United States)

    Szopa, A; Kisiel, W; Ekiert, H; Szewczyk, A

    2015-05-01

    Three dibenzocyclooctadiene lignans: deoxyschizandrin (1), gomisin A (2) and schizandrin (3) were isolated from biomass extracts of Schisandra chinensis (Turcz.) Baill. shoot-differentiating callus cultures. The mentioned lignans were not isolated earlier from in vitro cultures of this plant species. This is the first report concerning on isolation of dibenzocyclooctadiene lignans from in vitro cultures of Schisandra chinensis.

  1. Anther culture of papaya (Carica papaya L.).

    Science.gov (United States)

    Tsay, H S; Su, C Y

    1985-02-01

    Papaya (Carica papaya L.) anther containing microspores in tetrad to early-binucleate stages were successfully cultured on 1/2 strength MS salts and vitamins with full strength Na-Fe-EDTA supplemented with 2 mg/l NAA, 1 mg/l BA and 6% sucrose for callus initiation and formation. Highest frequencies of callus induction were obtained when anthers at the uninucleate stage were cultured in the dark. Haploid plantlets and pollen-derived embryoids were obtained from anthers cultured at the uninucleate stage on solidified MS medium containing 3% sucrose without any growth regulators under a low light intensity (1,500 lux). Large quantities of embryoids were obtained when the original embryoids were transferred to MS medium with 3% sucrose and no growth regulators. Cytology of root tips of embryoid-derived plants confirmed the haploid chromosome number of 9 indicating that the embryoids originated from pollen.

  2. Isolation, culture and division of olive (Olea europaea L.) protoplasts.

    Science.gov (United States)

    Cañas, L A; Wyssmann, A M; Benbadis, M C

    1987-10-01

    Protoplasts from Olea europaea L. have been compared in terms of their yield, viability, cell division and callus differentiation. Viable protoplasts were isolated from in vitro cultured leaves and cotyledons by an overnight incubation in an enzyme solution containing 1-1.5% driselase and 0.5M sucrose. This method allowed high yield of purified protoplasts, which floated and formed a dark green band at the meniscus, after centrifugation. Purified protoplasts were diluted to 3×10(4) protoplasts·ml(-1) in culture medium. After cell wall regeneration, protoplasts gradually increased their volumes under appropriate conditions. The first divisions occurred during the second week in culture. Division efficiency ranged from 5.2 to 9.8% after 20 days in culture. Two weeks later visible microcolonies developed only from cotyledon protoplasts. After 6 weeks in culture, the microcalli were transferred to a solidified culture medium with 0.6% agarose, which induced active callus growth.

  3. Effects of Cerium on Accumulation of Anthocyanins and Expression of Anthocyanin Biosynthetic Genes in Potato Cell Tissue Cultures

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The effects of Ce (Ⅳ) on callus growth, anthocyanin content, and expression of anthocyanin biosynthetic genes in callus suspension cultures of Solanum tuberosum cv. Chieftain were studied by the measurement of fresh weight, spectrophotometric assays, and semiquantitative RT-PCR. The results indicate that 0.1 mmol·L-1 Ce (Ⅳ) can promote callus growth, increase the accumulation of anthocyanins, and enhance the expression of five anthocyanin biosynthetic genes (CHS, F3H, F3′5′H, DFR, and 3GT) most efficiently. At high concentrations of 1 mmol·L-1, Ce (Ⅳ) partially inhibits callus growth and at 2 mmol·L-1 eventually lends to cell death. The results show that Ce(Ⅳ) can induce the expression of anthocyanin biosynthetic genes to produce and accumulate anthocyanins and increase the yield of anthocyanins.

  4. Biological activities of indoleacetylamino acids and their use as auxins in tissue culture

    Energy Technology Data Exchange (ETDEWEB)

    Hangarter, R.P.; Peterson, M.D.; Good, N.E.

    1980-05-01

    The auxin activities of a number of indoleacetylamino acid conjugates have been determined in three test systems: growth of tomato hypocotyl explants (Lycopersicon esculentum Mill. cv. Marglobe); growth of tobacco callus cultures (Nicotiana tabacum L. cv. Wisconsin 38); and ethylene production from pea stems (Pisum sativum L. cv. Alaska). The activities of the conjugates differ greatly depending on the amino acid moiety. Indoleacetyl-L-alanine supports rapid callus growth from the tomato hypocotyls while inhibiting growth of shoots and roots. Indoleacetlyglycine behaves in a similar manner but is somewhat less effective in supporting callus growth and in inhibiting growth of shoots and roots. Indoleacetylglycine behaves in a similar manner but is somewhat less effective in supporting callus growth and in inhibiting shoot formation. The other amino acid conjugates tested (valine, leucine, aspartic acid, threonine, methionine, phenylalanine, and proline) support shoot formation without supporting root formation or much callus growth. The tobacco callus system, which forms abundant shoots in the presence or absence of free indoleacetic acid, produces only rapid undifferentiated growth in the presence of indoleacetyl-L-alanine and indoleacetylglycine. The other conjugates inhibit shoot formatin weakly if at all. Most of the conjugates induce sustained ethylene production from the pea stems but at rates well below the initial rates observed with free indoleacetic acid. Many, but not all of the effects of conjugates such as indoleacetyl-L-alanine can be mimicked by frequent renewals of the supply of free indoleacetic acid.

  5. Micropropagation of Dalbergia sissoo Roxb. through tissue culture technique.

    Science.gov (United States)

    Sahu, Jyoti; Khan, Shagufta; Sahu, Ram Kumar; Roy, Amit

    2014-04-01

    Multiple shoots of Dalbergia sissoo Roxb. (Sissoo) were incited from seeds through indirect somatic embryogenesis method. Seeds were inoculated in Murashige and Skoog's medium without any growth hormone. Than cotyledonary leaves were struck and used for callus induction on MS medium amplified with 2, 4-dichlorophenoxyacetic acid (0.5 to 4 mg mL(-1)). After 3 to 4 weeks the embryogenic callus clumps was transferred to medium supplemented with cytokinin (BAP 1 to 5 mg L(-1), kinetin 1-5.0 mg L(-1)) for embryo maturation and germination. The high-frequency shoot proliferation (82%) and maximum number of shoots per explants were recorded in MS medium containing NAA (0.5)+BAP (0.5). The findings of recent investigations have shown that, it is possible to induce indirect somatic embryogenesis in Dalbergia sissoo and plant regeneration from callus cultures derived from cotyledonary leaves as explants.

  6. The Pure Culture of Fruiting Bodies of Scorias spongiosa,an Edible Mushroom of Bamboo%竹类食用蕈菌——海绵胶煤炱菌子实体的纯培养

    Institute of Scientific and Technical Information of China (English)

    贺新生; 刘超洋; 郑俊娟; 商圆圆; 霍存录

    2012-01-01

    The study explored the pure culture condition and development law of the fruiting bodies of Scorias spongiosa, in order to provide technical solutions for large - scale production of this new type of mushroom. More than 100 specimen of wild fruitbodies were collected from Zhejiang, Sichuan and Jiangsu province. Pure strains were obtained through tissus separation and pure culturing on agar, liquid and solid medium. Mycelium directly secreted gelatinous material to form colloidal fruitbodies in the agar and solid culture medium. In liquid medium, only mycelium and the mycelium sphere were formed, but not fruitbodies. The development process of this fungus was: conidia spore germination→ hyphal→colloid tissue→ branches of fruitbodies → small fruitbodies → formation pycnidium and spores liquid group → releasing spores. Under pure culture conditions it was difficult to form large fruiting bodies.%探索海绵胶煤炱菌子实体形成的培养条件,研究该菌的子实体发育规律,为规模化生产这种新型食用蕈菌提供技术方案.在浙江、四川、江苏等地共采集到100多份野生子实体标本,经组织分离得到纯培养菌株,在多种琼脂、液体、固体培养基上进行纯培养.在琼脂和固体培养基上,菌丝体直接分泌胶质物质形成胶质的小型子实体;在液体培养基上只形成菌丝和胶质菌丝球,不形成子实体.该菌的形态发育过程:分生孢子萌发→菌丝→胶质组织→子实体分枝→小型子实体→形成分生孢子器和分生孢子→释放孢子.海绵胶煤炱菌在纯培养条件下难以形成大型子实体.

  7. Study Progress on Tissue Culture of Maize Mature Embryo

    Science.gov (United States)

    Wang, Hongzhen; Cheng, Jun; Cheng, Yanping; Zhou, Xioafu

    It has been paid more and more attention on maize tissue culture as it is a basic work in maize genetic transformation, especially huge breakthrough has been made in maize tissue culture utilizing mature embryos as explants in the recent years. This paper reviewed the study progress on maize tissue culture and plant regeneration utilizing mature embryos as explants from callus induction, subculture, plant regeneration and browning reduction and so on.

  8. 半夏疏松愈伤组织诱导条件的优化研究%Optimization of Inducing Conditions for Loose Callus of Pinellia ternata

    Institute of Scientific and Technical Information of China (English)

    朱艳芳; 毛春娜; 张爱民; 薛建平; 李佳娣; 宋运贤

    2011-01-01

    [Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of four plant growth substances(2,4-D,NAA,picloram and KT)and their mixture ratios on the formation of loose callus.[Result] The induction effect of 2,4-D and picloram on the petiole callus of Pinellia ternate was the most significant.Then,the second ones were KT and NAA.The optimal medium which induced Pinellia ternate petiole to form the loose callus was MS+0.5 mg/L of 2,4-D+1.0 mg/L of NAA+1.0 mg/L of picloram+1.5 mg/L of KT.[Conclusion] The research laid the foundation for extracting the active ingredient from the cell suspension of Pinellia ternate and producing the artificial seed.%[目的]建立半夏细胞悬浮培养体系。[方法]以半夏叶柄为外植体,采用正交试验设计研究了2,4-D、NAA、毒莠定和KT4种植物生长物质及其配比对疏松愈伤组织形成的影响。[结果]2,4-D和毒莠定对半夏叶柄愈伤组织诱导效果最显著,KT和NAA次之。诱导半夏叶柄形成疏松愈伤组织的适宜培养基为MS+0.5mg/L2,4-D+1.0mg/LNAA+1.0mg/L毒莠定+1.5mg/LKT。[结论]为通过半夏细胞悬浮提取有效成分和生产人工种子奠定了基础。

  9. Determination of fruit characteristics, fatty acid profile and total antioxidant capacity of Mespilus germanica L. fruit

    Directory of Open Access Journals (Sweden)

    Hale Seçilmiş Canbay

    2015-11-01

    Full Text Available Objective: To determine fruit characteristics, fatty acid profile and total antioxidant capacitiy of first cultured Mespilus germanica L. Methods: A total of 15 fruits were taken randomly from four directions of adult trees. Then the physical and chemical properties of first cultured medlar fruit (Istanbul/Turkey were measured by using refractometer, colorimeter, spectrophotometer and gas chromatograph mass spectrometer, respectivly. Results: In the fruit studied, the results showed that palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid and behenic acid were the most abundant fatty acids (FAs, and the main FA was palmitic acid [(35.35 ± 1.20%]. The percentage of linoleic acid and stearic acid in this fruit oil were (29.10 ± 1.70% and (8.53 ± 0.25%, respectively. As a result of the analysis, the total antioxidant capacity of medlar fruit was (1.1 ± 0.2 mmol trolox equivalents/L. Conclusions: The present study has demonstrated the concentrations of FAs and the antioxidantive capacity of first cultured Istanbul medlar fruits by using many tested methods. It is proved that in our daily life, medlar fruit plays a significant role with its nutrition and health effect.

  10. Evaluation of gelling agents on anther culture : response of two soybean cultivars

    OpenAIRE

    Milena Barcelos Cardoso; Maria Helena Bodanese-Zanettini; Elsa Cristina de Mundstock; Eliane Kaltchuk-Santos

    2007-01-01

    Anthers of two soybean cultivars were cultured in B5 long basal culture media gelled with agarose or PhytagelTM. Cytological examinations of the anthers were carried out during the first 45 days of culture to assay the viability and developmental stage of microspores. Frequency of callus formation was recorded at 45 days of culture. The analysis of variance of the microspore viability assay showed significant Cultivar X Gelling Agent X Day of Culture interactions. The frequencies of viable mi...

  11. Callus, shoot and hairy root formation in vitro as affected by the sensitivity to auxin and ethylene in tomato mutants.

    Science.gov (United States)

    Lima, Joni Esrom; Benedito, Vagner Augusto; Figueira, Antonio; Peres, Lázaro Eustáquio Pereira

    2009-08-01

    We analyzed the impact of ethylene and auxin disturbances on callus, shoots and Agrobacterium rhizogenes-induced hairy root formation in tomato (Solanum lycopersicum L.). The auxin low-sensitivity dgt mutation showed little hairy root initiation, whereas the ethylene low-sensitivity Nr mutation did not differ from the control Micro-Tom cultivar. Micro-Tom and dgt hairy roots containing auxin sensitivity/biosynthesis rol and aux genes formed prominent callus onto media supplemented with cytokinin. Under the same conditions, Nr hairy roots did not form callus. Double mutants combining Rg1, a mutation conferring elevated shoot formation capacity, with either dgt or Nr produced explants that formed shoots with little callus proliferation. The presence of rol + aux genes in Rg1 hairy roots prevented shoot formation. Taken together, the results suggest that although ethylene does not affect hairy root induction, as auxin does, it may be necessary for auxin-induced callus formation in tomato. Moreover, excess auxin prevents shoot formation in Rg1.

  12. Análise bioquímica de calos de pinhão-manso Biochemical analysis of callus from physic nut

    Directory of Open Access Journals (Sweden)

    Dalilhia Nazaré dos Santos

    2010-11-01

    Full Text Available O pinhão-manso (Jatropha curcas L. vem ganhando importância por seu potencial como insumo ao biodiesel. Estudos visam à adequação agronômica e a cultura de tecidos contribui indiretamente com este aspecto, auxiliando em pesquisas aplicadas como em biotecnologia, em situações que visam à regeneração de plantas. Nesse intuito, é fundamental avaliar, durante os períodos de cultivo dos calos, o comportamento dos metabólitos primários. A massa calogênica, por ocasião da inoculação, não apresentou reserva na forma de carboidratos. Aos 42 dias iniciou a absorção da sacarose do meio, enquanto o acúmulo no explante ocorreu a partir dos 98 dias de cultivo. O teor de aminoácidos foi alto no dia da inoculação e, por volta dos 98 dias, tanto os níveis de aminoácidos quanto os de proteínas decresceram.The physic nut (Jatropha curcas L. is gaining importance because of its potential as raw material for biodiesel. Studies aiming the adequacy of agronomic techniques and tissue culture contribute, indirectly, to this aspect, helping applied research as biotechnology, in situations that aim plant regeneration. Therefore, it is essential to evaluate, the behavior of primary metabolites during the callus growing season. The callus mass at the inoculation time showed no carbohydrates reserve. At the 42nd day the sucrose absorption started in the medium, while the accumulation in the explant occurred from the 98th day of cultivation. Amino acid content was high at the inoculation's day and around the 98th day, both amino acids and protein levels decreased.

  13. Chemical composition and nutritional and medicinal value of fruit bodies and submerged cultured mycelia of culinary-medicinal higher Basidiomycetes mushrooms.

    Science.gov (United States)

    Cohen, Nachshol; Cohen, Jacob; Asatiani, Mikheil D; Varshney, Vinay K; Yu, Hui-Tzu; Yang, Yi-Chi; Li, Yu-Hsuan; Mau, Jeng-Leun; Wasser, Solomon P

    2014-01-01

    This research gives the results of a proximate analysis (moisture, ash, crude protein, fat, total carbohydrates, and total energy); a bioactive compounds analysis (γ-aminobutyric acid [GABA], ergothioneine, lovastatin, and cordycepin); fatty acid and amino acid analysis; and an analysis of macro- and microelement content of fruit bodies and mycelia of 15 higher Basidiomycetes medicinal mushroom strains belonging to 12 species. The results obtained demonstrate that almost all investigated mushrooms were found to be good sources of proteins and carbohydrates, with content varying in the ranges of 8.6-42.5% and 42.9-83.6%, respectively. Different species exhibited distinct free amino acid profiles. The total amino acid content was highest in Ophiocordyceps sinensis (MB) (23.84 mg/g) and Cordyceps militaris (FB) (23.69 mg/g). The quantification of the identified fatty acids indicated that, in general, palmitic acid, oleic acid, stearic acid, and linoleic acid were the major fatty acids. The micro- and macroelement compositions were studied, and the highest results were (as milligrams per kilogram) 224-7307 for calcium, 1668-38564 for potassium, 1091-11676 for phosphorus, and 5-97 for zinc. Bioactive components were lovastatin, GABA, and ergothioneine, which are commonly found in most mushrooms. C. militaris (FB), Pleurotus ostreatus (FB), and Coprinus comatus (FB) were most abundant and contained a high amount of GABA (756.30 μg/g, 1304.99 μg/g, 1092.45 μg/g, respectively) and ergothioneine (409.88 μg/g, 2443.53 μg/g, 764.35 μg/g, respectively). The highest lovastatin content was observed in Hericium erinaceus (FB) (14.38 μg/g) and Ganoderma lucidum (FB) (11.54 μg/g). In contrast to C. militaris (FB), cordycepin was not detected in O. sinensis (MB). The fruit body biomass of C. militaris cordycepin content reached 1.743 mg/g dry weight. The nutritional values of the mushroom species studied here could potentially be used in well-balanced diets and as sources

  14. In vivo tracking of segmental bone defect healing reveals that callus patterning is related to early mechanical stimuli

    Directory of Open Access Journals (Sweden)

    M Mehta

    2012-11-01

    Full Text Available This study addresses the hypothesis that callus formation, patterning, and mineralisation are impaired during the early phase of critical sized bone defect healing, and may relate to inter-fragmentary tissue strains within the bone defect area. Twenty four 12 week old Sprague Dawley rats were used for this study. They were divided into two groups defined by the femur bone defect size: (i 1 mm resulting in normal healing (NH, and (ii a large sized 5 mm defect resulting in critical healing (CH. Callus formation, patterning, and mineralisation kinetics in both groups were examined in the periosteal and osteotomy gap regions using a novel longitudinal study setup. Finite element analyses on µCT generated tomograms were used to determine inter-fragmentary tissue strain patterns and compared to callus formation and patterning over the course of time. Using a novel longitudinal study technique with µCT, in vivo tracking and computer simulation approaches, this study demonstrates that: (i periosteal bone formation and patterning are significantly influenced by bone defect size as early as 2 weeks; (ii osteotomy gap callus formation and patterning are influenced by bone defect size, and adapt towards a non-union in critical cases by deviating into a medullary formation route as early as 2 weeks after osteotomy; (iii the new bone formation in the osteotomy gap enclosing the medullary cavity in the CH group is highly mineralised; (iv inter-fragmentary strain patterns predicted during the very early soft callus tissue phase (less than 2 weeks are concurrent with callus formation and patterning at later stages. In conclusion, bone defect size influences early onset of critical healing patterns.

  15. 光照和氮源对朱砂根愈伤组织生长及岩白菜素含量的影响%Effect of light and nitrogen source on the growth of callus and the content of bergenin in Ardisia crenata

    Institute of Scientific and Technical Information of China (English)

    何雪娇; 赖钟雄

    2012-01-01

    通过调节光照条件及培养基中铵态氮与硝态氮的比例来研究光照和氮源对朱砂根愈伤组织生长及岩白菜素含量的影响.结果表明,遮光和强白光都有利于朱砂根愈伤组织的生长,不同光照度和光质对愈伤组织的生长形态和褐化程度有一定的影响.愈伤组织中的岩白菜素含量随着光照度的不断增强而减少,当光照度为100 lx(遮光状态)时,岩白菜素含量达到最高,为0.0169%.光质对岩白菜素含量影响程度的大小为:绿光>蓝光>白光>红光,绿光下的岩白菜素含量最高,为0.0235%.培养基中的KNO3有利于愈伤组织生长,NH4NO3有利于岩白菜素的合成.当KNO3含量加倍但无NH4NO3时,愈伤组织的增殖系数会增大;当KNO3含量为3800 mg·L-1时,愈伤组织的增殖系数达到最大值,为19.9614;当培养基中仅有NH4NO3时,随着含量的增加愈伤组织的生长量和岩白菜素的含量都随之减少;当NH4NO3含量为1650 mg · L-1时,岩白菜素含量达到最高,为0.0125%.%This experiment researches into the effect of light and nitrogen source on the growth of callus and the content of bergenin in Ardisia crenata Sims by adjusting the proportion of ammonium and nitrate nitrogen in culture medium. The results showed that shading and strong white light were beneficial to the growth of the callus, light intensity and light type had certain effect on the growthform and degree of browning of the callus. The content of bergenin reduced with the increasing of light intensity . When the light intensity was 100 lx (shading) , the content of bergenin in the callus was highest(0.0169% ). The influence degree of light type on the content of bergenin was in the following order; green, blue, white, red. Under green light, the content of bergenin in the callus was highest(0.0235% ). The KNO3 was beneficial to the growth of the callus, and NH4NO3 was beneficial to the synthesis of bergenin. Under the condition of

  16. Effect of UV-C on Drought Tolerance of Alfalfa (Medicago sativa) Callus

    OpenAIRE

    Ali A. Ehsanpour; Roya Razavizadeh

    2005-01-01

    Water stress and unfavorable climate decrease the growth and development of plant globally. Medicago is a valuable plant as a source of food for animal, foliage and for medicine. Since, Iran is located in a dry area; consequently, the study of plant responses to water stress is important. In this research UV-C radiation was used to induce physiological and genetic changes in alfalfa callus. In vitro grown calluses of M. sativa were exposed to UV-C for 0, 15, 30 and 60 min and then, were trans...

  17. Isolation of protoplast from callus of Populus euphratica and H+ fluxes across plasma membrane under NaCl stress

    Institute of Scientific and Technical Information of China (English)

    Gao Zhun; Dai Song-xiang; Chen Shao-liang; Shen Xin; Wang Rui-gang

    2007-01-01

    We used callus of Populus euphratica Olive to isolate protoplasts, and H+ fluxes across plasma membrane were investigated. The concentration of enzymes for protoplast isolation, e.g. cellulase, pectolyase, macerozyme, hemicellulase, and sorbitol content, incubation time were systemically studied. High yield and viability of protoplast was achieved after 6-8 hours incubation of P. euphratica callus in enzyme solution containing 1.5% (w:v) cellulase R-10, 0.1% (w:v) pectolyase Y-23, 0.2% (w:v) macerozyme membrane of P. euphratica cells. The shift of H+ flux response to NaCl shock and the relevance to salt tolerance were discussed.

  18. Somatic embryogenesis in Lolium multiflorum suspension culture

    Directory of Open Access Journals (Sweden)

    Margarita Pavlova

    2014-01-01

    Full Text Available The embryogenic cell suspension was obtained from immature embryos of Lolium multiflorum through a callus culture. Somatic embryogenesis was induced by addition of 2,4-D, dicamba and picloram in 0,5 mg/l concentrations in MS liquid nutrient medium. It was shown that somatic embryos arised from single cells. In globular embryoids, the meristematic cells are characterized by the presence of phytoferritin inclusions in the leucoplasts.

  19. Methyl jasmonate differentially affects tocopherol content and tyrosine amino transferase activity in cultured cells of Amaranthus caudatus and Chenopodium quinoa.

    Science.gov (United States)

    Antognoni, F; Faudale, M; Poli, F; Biondi, S

    2009-03-01

    Tocopherols are lipid-soluble compounds synthesised exclusively by photosynthetic organisms. In this study, in vitro callus cultures were established from two plants that are naturally rich in tocopherols, Amaranthus caudatus and Chenopodium quinoa, in order to examine whether callus cultures were able to produce these compounds at levels comparable to those observed in planta. In both species, cotyledon explants produced the best callus induction and, once established, callus cultures were grown under two different hormonal treatments to check for effects of growth and to induce chloroplast differentiation in the cells. A rapid differentiation of chloroplasts occurred only in C. quinoa cell aggregates grown in the presence of benzyladenine, leading to the production of a homogeneous green callus. In both species, only alpha-tocopherol was produced by callus cultures, although levels were much lower than in planta, and the production was not influenced by the hormonal conditions. Interestingly, cell cultures of the two species responded in different ways to methyl jasmonate (MJ). In A. caudatus cultures, treatment with 100 mum MJ increased the production of alpha-tocopherol up to fivefold, and the inductive effect was influenced by the hormonal composition of the medium. This increase in alpha-tocopherol was associated with a proportional increase in tyrosine aminotransferase (TAT) activity, one of the key enzymes involved in tocopherol biosynthesis. By contrast, in C. quinoa cultures, elicitation with MJ did not have any effect, neither on tocopherol production, nor on TAT activity. These results are discussed in relation to chloroplast differentiation and the interplay between jasmonates and phytohormones.

  20. 黄瓜初花期与结瓜期叶片可培养内生细菌多样性研究%Diversity of culturable entophytic bacteria in cucumber leaves at blossoming and fruiting stages

    Institute of Scientific and Technical Information of China (English)

    孙占斌; 袁行方; 王音娴; 张辉; 张文会; 冯永君

    2012-01-01

    [目的]研究黄瓜初花和结瓜两个生长期叶片可培养内生细菌的多样性.[方法]采用叶片表面消毒、菌种分离、16S rDNA序列扩增和系统发育分析进行了系统研究.[结果]两时期黄瓜内生细菌的种类、数量及优势菌的种类都有明显差异.初花期叶片含菌量为(2.6±0.18)×106 CFU/g鲜重,分离出的38株内生细菌分别属于短小杆菌属(Curtobacterium sp.)、节杆菌属(Arthrobacter sp.)、微杆菌属(Microbacterium sp.)等14个已知属,其中优势种类为短小杆菌属菌株;结瓜期叶片含菌量为(5.2±0.42)×105 CFU/g鲜重,分离出的43株内生菌分别属于泛菌属(Pantoea sp.)、假单胞菌属(Pseudomonas sp.)、芽孢杆菌属(Bacillus sp.)等11个已知属,其中优势种类为泛菌属菌株.[结论]初花期内生菌含量是结瓜期的5倍,两时期内生菌的种类表现出很强的差异,体现出黄瓜不同生育期可培养内生菌数量和种类的多样性,相关研究为黄瓜促生内生细菌的理论探索和生产应用提供一定的研究基础.%[Objective) The aim of this study is to explore the quantity and diversity of cultur-able endophytic bacteria in cucumber leaves sampled at blossoming and fruiting stages. [Methods] Leaf surface disinfection, strain isolation, 16S rDNA amplification and phyloge-netic analysis were used in this study. [Results] It is found that there is a significant difference in the bacterial quantity and species at two stages. At blossoming stage, the isolated bacteria belong to 14 genera, including Curtobacterium sp., Arthrobacter sp., Microbacterium sp., etc. The isolating quantity is (2.6±0.18)×l06 CFU/g fw, with Curtobacterium sp. being the dominant strain. However, at fruiting stage, the isolated bacteria belong to 11 genera, including Pantoea sp., Pseudomonas sp., Bacillus sp., etc. The isolating quantity is (5.2±0.42)xl05 CFU/g fw, with Pantoea sp. being the dominant strain. [Conclusion] The species of endophytic

  1. Pepper Weevil (Coleoptera: Curculionidae Preferences for Specific Pepper Cultivars, Plant Parts, Fruit Colors, Fruit Sizes, and Timing

    Directory of Open Access Journals (Sweden)

    Dakshina R. Seal

    2016-03-01

    Full Text Available Peppers (Capsicum spp. are an important crop in the USA, with about 32,000 ha cultivated in 2007, which resulted in $588 million in farm revenue. The pepper weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae, is the most troublesome insect pest of peppers in the southern United States. It is therefore urgent to find different vulnerabilities of pepper cultivars, fruit and plants parts, fruit colors and sizes, and timing to infestation by A. eugenii. Also relevant is testing whether fruit length and infestation state affect fruit numbers, weights, and proportions of fruit that are infested. Counts of A. eugenii adults and marks from oviposition and feeding suggested that C. chinense Jacquin “Habanero” was least susceptible, and C. annuum L. cultivars “SY” and “SR” were most susceptible. Comparison of plant parts and fruit sizes revealed that A. eugenii preferred the peduncle, calyx, and top of pepper fruits over the middle, bottom, leaves, or remainder of flowers. Anthonomus eugenii does not discriminate between green or yellow fruit color nor vary diurnally in numbers. Based on adult counts, medium to extra-large fruits (≥1.5 cm long attracted more weevils than small fruits (<1.5 cm. However based on proportions of fruit numbers or fruit weights that were infested, there were no differences between large and small fruits. Choice of pepper cultivar can thus be an important part of an IPM cultural control program designed to combat A. eugenii by reduced susceptibility or by synchronous fruit drop of infested fruits. Our results are potentially helpful in developing scouting programs including paying particular attention to the preferred locations of adults and their sites of feeding and oviposition on the fruit. The results also suggested the potential value of spraying when the fruits are still immature to prevent and control infestation.

  2. Pulsed electromagnetic field treatment enhances healing callus biomechanical properties in an animal model of osteoporotic fracture.

    Science.gov (United States)

    Androjna, Caroline; Fort, Brian; Zborowski, Maciej; Midura, Ronald J

    2014-09-01

    Delayed bone healing has been noted in osteoporosis patients and in the ovariectomized (OVX) rat model of estrogen-depletion osteopenia. Pulsed electromagnetic field (PEMF) devices are clinically approved as an adjunct to cervical fusion surgery in patients at high risk for non-fusion and for the treatment of fracture non-unions. These bone growth stimulating devices also accelerate the healing of fresh fracture repair in skeletally mature normal rats but have not been tested for efficacy to accelerate and/or enhance the delayed bone repair process in OVX rats. The current study tested the hypothesis that daily PEMF treatments would improve the fracture healing response in skeletally mature OVX rats. By 6 weeks of healing, PEMF treatments resulted in improved hard callus elastic modulus across fibula fractures normalizing the healing process in OVX rats with respect to this mechanical property. Radiographic evidence showed an improved hard callus bridging across fibula fractures in OVX rats treated with PEMF as compared to sham treatments. These findings provide a scientific rationale for investigating whether PEMF might improve bone-healing responses in at-risk osteoporotic patients.

  3. Somatic embryogenesis and plant regeneration from cell suspension and tissue cultures of mature himalayan poplar (Populus ciliata).

    Science.gov (United States)

    Cheema, G S

    1989-02-01

    Somatic embryogenesis and plantlet formation were obtained from callus and cell suspension cultures of 40-year- old Himalayan Poplar (Populus ciliata Wall ex Royle). Callus and cell suspensions were obtained by transfer of inoculum of semiorganized leaf cultures, which were maintained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP), to MS with 2,4-dichlorophenoxyacetic acid (2,4-D). Reduction of 2,4-D concentration during subsequent subculture of cell suspensions resulted in the formation of embryoids. These embryoids developed further only after being transferred to agar-based MS medium supplemented with BAP and naphthalene acetic acid. Loss of embryogenic potential was observed in cell suspensions after 6 subcultures. However, callus cultures retained the embryogenic potential even after repeated subcultures for more than a year. Plantlets could be successfully hardened and grown in natural outdoor conditions.

  4. Chemical Composition and Medicinal Value of Fruiting Bodies and Submerged Cultured Mycelia of Caterpillar Medicinal Fungus Cordyceps militaris CBS-132098 (Ascomycetes).

    Science.gov (United States)

    Chan, Jannie Siew Lee; Barseghyan, Gayane S; Asatiani, Mikheil D; Wasser, Solomon P

    2015-01-01

    In this paper, we report the results of a proximate analysis (i.e., moisture, ash, protein, fat, carbohydrates, and energy); a bioactive compounds analysis (i.e., cordycepin and ergothioneine); fatty and amino acid analysis; and analyses of vitamin content, macro- and microelement composition of fruiting body (FB), and mycelial biomass (MB) of medicinal caterpillar fungus Cordyceps militaris strain CBS-132098. These results demonstrate that the FB and MB of C. militaris are good sources of proteins: 59.8% protein content in the FB and 39.5% in the MB. The MB was distinguished by its carbohydrate content (39.6%), which was higher than that of the FB (29.1% carbohydrate). In the FB of C. militaris, the total amino acid content was 57.39 mg/g and in the MB it was 24.98 mg/g. The quantification of the identified fatty acids indicated that palmitic acid, oleic acid, linoleic acid, and linolenic acid were the major fatty acids. The micro- and macroelement compositions were studied. The highest results were calcium (797 mg/kg FB; 11 mg/kg MB); potassium (15,938 mg/kg FB 12,183 mg/kg MB); magnesium (4,227 mg/kg FB; 3,414 mg/kg MB); sodium (171 mg/kg FB; 1,567 mg/kg MB); phosphorus (7,196 mg/kg FB; 14,293 mg/kg MB); and sulfur (5,088 mg/kg FB; 2,558 mg/kg MB). The vitamin composition was studied, and the most abundant vitamins were vitamin A, vitamin B3, and vitamin E. The bioactive components were cordycepin, cordycepic acid (D-mannitol), and ergothioneine. There were differences in cordycepin and ergothioneine contents between the FB and the MB. The cordycepin concentration was 0.11% in the FB and 0.182% in the MB, the cordycepic acid was 4.7 mg/100g in the FB and 5.2 mg/100 g in the MB, and the ergothioneine content was 782.37 mg/kg in the FB and 130.65 mg/kg in the MB. The nutritional values of the FB and the MB of C. militaris detected indicate its potential use in well-balanced diets and sources of bioactive compounds.

  5. A protocol for axenic liquid cell cultures of a woody leguminous mangrove, Caesalpinia crista, and their amino acids profiling.

    Science.gov (United States)

    Inoue, Aya; Ogita, Shinjiro; Tsuchiya, Shinpei; Minagawa, Reiko; Sasamoto, Hamako

    2015-05-01

    Callus induction, maintenance and protoplast cultures were achieved from immature seeds of a woody leguminous mangrove, Caesalpinia crista. Axenic cultures were possible during 1.5 months of pod storage in 0.1% benzalkonium chloride solution. Callus induction was achieved using 1 mL liquid medium in a 10 mL flat-bottomed culture tube. Protoplasts were isolated using Cellulase R10, Hemicellulase, and Driselase 20 in 0.6 M mannitol solution and sub-culturable calluses were obtained in 50 μL liquid medium using a 96-microplate method. The optimal hormonal concentration was 10 μM each of 2,4-dichlorophenoxyacetic acid and benzyladenine in liquid Murashige and Skoog's basal medium for both callus induction and maintenance, and protoplast cultures. Similarities and differences in amino acid profiles and culture conditions are discussed among woody mangrove species and non-mangrove leguminous species. Caesalpinia crista cultures were unique as they secreted a large amount of amino acids, including proline, into the liquid culture medium.

  6. PROPAGATION OF CLEOME SPINOSA JACQ. THROUGH TISSUE CULTURE

    Directory of Open Access Journals (Sweden)

    Cuiqin Yang

    2012-04-01

    Full Text Available The tissue culture and rapid propagation of Cleome spinosa Jacq. was explored by investigating the effects of different plant growth regulators on callus induction, bud differentiation, and root formation of three types of explants. The results showed that hypocotyls and stem segments regenerated buds directly on growth regulator-free Murashige and Skoog (MSmedium. The highest callus induction rates of hypocotyls, stem segments, and leaves reached 100% and were obtained on the culture medium of MS + (1.0 to 2.0 mg/L kinetin (KT + 0.02 mg/L α-naphthalene acetic acid (NAA, on which the leaves produced the best quality of calluses. The leaf-derived calluses were subcultured on MS + 0.5 mg/L KT +0.5 mg/L 6-benzylaminopurine (BAP and achieved the highest differentiation rate of 100%, producing an average of 7.5 buds per explant. Inoculation with MS + 0.5 mg/L indole-3-butyric acid (IBA resulted in the production of a number of thick roots by 66.7% of the regenerated buds. After transplanting, plantlets with more roots survived easily and grew well.

  7. 枸杞花药培养的初步研究%Preliminary Study on the Anther Culture in Lycium barbarum L.

    Institute of Scientific and Technical Information of China (English)

    段丽君; 周军; 曹有龙

    2008-01-01

    To investigate the culture technique in anther of Chinese wolfberry, we optimized the culture medium(including hormone combination) and cul-ture conditions. The results showed that calluses were induced from all the six tested Chinese wolfberry materials, hut the induction rate of callus varied to-ward the materials with different genotypes. When the experimental materials were cuhured on medium appended with 2,4-D 1.0 mg/L and KT 1.0 mg/L un-der dark, the callus induction rate reached 20.0 % in this study, and this hormone combination should be the optimum for anther culture of Chinese wolf-berry. With MS appended with 6-BA 0.5 mg/L and NAA 0.1 mg/L as differentiation medium and that appended with NAA 0.1 mg/L, the plants could be yielded in 20 days.

  8. Barley callus

    DEFF Research Database (Denmark)

    Carciofi, Massimiliano; Blennow, Per Gunnar Andreas; Nielsen, Morten M.;

    2012-01-01

    Background Starch is the most important source of calories for human nutrition and the majority of it is produced by cereal farming. Starch is also used as a renewable raw material in a range of industrial sectors. It can be chemically modified to introduce new physicochemical properties...... and cloned. This knowledge suggests a number of strategies and a series of candidate genes for genetic transformation of crops to generate new types of starch-based polymers. However transformation of cereals is a slow process and there is no easy model system available to test the efficiency of candidate...... suggest that this method can be used as a time-efficient model system for fast screening of candidate genes for the generation of modified starch or new types of carbohydrate polymers....

  9. Comparison of Harpagoside Yield of Callus, Adventitious Root and Endophytes in Scrophulariae Radix%玄参愈伤、不定根和内生菌产哈巴俄苷的比较

    Institute of Scientific and Technical Information of China (English)

    张林甦; 赵德刚

    2012-01-01

    Objective: Inducing callus and adventitious root from Scrophulariae Radix; isolating endophytes; and comparing Hapagoside yield of the different parts of Scrophulariae Radix. Method; Tender leafs of Scrophulariae Radix were cut into small pieces and sterilized to inoculate on MS, N6 mediums with different plant hormones to induce callus; Liquid medium were used in adventitious root inducing and medium transformed according to growing phase during the culture process; general method was used in endophytes isolated. Hapagoside detected with UV spectrophotometry at the wave length 255 nm. Result; Moderate density callus were induced with MS + NAA 0. 05 or 0. 2 or 2 mg -L-1 + 6-BA 2 mg -L-1' medium; mass adventitious roots obtained after 30 d ' s vibrate cultivation; 4 strains of endophytes produce hapagoside were isolated from Radix Scrophulariae root; hapagoside content of callus, adventitious root and fermentation liquor of the four endophyte straints in orderly are 0. 411 , 0. 099 5 , 0. 451, 0. 444 , 0. 489 , 0. 440 g -L-1. Conclusion: Content of hapagoside in callus is 4 times than that of adventitious root; endophytes can yield almost the same amount hapagoside as callus, and have great potential in producing secondary metabolites.%目的:诱导玄参愈伤、不定根产生,分离内生菌,并比较其中有效成分哈巴俄苷的含量.方法:玄参嫩叶片消毒切成小块接种于含不同激素水平的MS,N6培养基诱导愈伤;不定根诱导采用液体培养:将愈伤小块先转入不含激素MS液体培养基,100 r·min-1室温震荡培养,待开始出现不定根后转入含0.05 mg·L-1 NAA+2 mg·L-1 6-BA的MS液体培养基继续震荡培养,内生菌分离采用常规方法.哈巴俄苷含量测定采用紫外分光光度法,测定波长255 nm.结果:MS培养基+NAA0.05,0.2,2 mg·L-1 +6-BA 2 mg·L-1均能诱导出质地较疏松、生长较快的愈伤组织;接种1.5g愈伤30 d左右可得到100 mL满瓶不定根;从玄参鲜块根分离出4

  10. Influence of salt stress on growth and antioxidant responses of two malus species at callus and plantlet stages

    Science.gov (United States)

    Soil salinization has become a significant factor limiting apple (Malus domestica L.) production in some areas of China. This study was carried out to investigate the effects of salt stress i.e. 150 mmol/L NaCl in Murashige and Skoog medium on growth and antioxidant responses of 15-d old callus and ...

  11. Oxygen regulation of uricase and sucrose synthase synthesis in soybean callus tissue is exerted at the mRNA level

    DEFF Research Database (Denmark)

    Xue, Z T; Larsen, K; Jochimsen, B U

    1991-01-01

    The effect of lowering oxygen concentration on the expression of nodulin genes in soybean callus tissue devoid of the microsymbiont has been examined. Poly(A)+ RNA was isolated from tissue cultivated in 4% oxygen and in normal atmosphere. Quantitative mRNA hybridization experiments using nodule...

  12. Collagen structure regulates fibril mineralization in osteogenesis as revealed by cross-link patterns in calcifying callus

    NARCIS (Netherlands)

    Wassen, M.H.M.; Lammens, J.; Tekoppele, J.M.; Sakkers, R.J.B.; Liu, Z.; Verbout, A.J.; Bank, R.A.

    2000-01-01

    Although >80% of the mineral in mammalian bone is present in the collagen fibrils, limited information is available about factors that determine a proper deposition of mineral. This study investigates whether a specific collagen matrix is required for fibril mineralization. Calcifying callus from do

  13. The behaviour of tomato golden mosaic virus DNA in cultured cells isolated from systemically infected tobacco leaves.

    Science.gov (United States)

    Slomka, M J; Buck, K W; Coutts, R H

    1989-03-01

    When callus tissue was cultured from leaf pieces taken from a Nicotiana tabacum cv. Xanthi nc. plant systemically infected with tomato golden mosaic virus (TGMV), TGMV-specific DNA persisted for up to 6 months in culture. Analysis of TGMV-specific intracellular DNA forms indicated a decrease in double-stranded relative to single-stranded forms and an increase in sub-genomic relative to genomic single-stranded DNA species in the callus tissue compared to those in the original leaf explant. The implications of the results with regard to TGMV replication are discussed.

  14. Efficient production of a bioactive Bevacizumab monoclonal antibody using the 2A self-cleavage peptide in transgenic rice callus

    Directory of Open Access Journals (Sweden)

    Lei Chen

    2016-08-01

    Full Text Available Bevacizumab, a humanized monoclonal antibody (mAb targeting to the vascular endothelial growth factor (VEGF, has been widely used in clinical practice for the treatment of multiple cancers. Bevacizumab was mostly produced by the mammalian cell expression system. We here reported the first plant-derived Bevacizumab by using transgenic rice callus as an alternative gene expression system. Codon-optimized Bevacizumab light chain (BLC and heavy chain (BHC genes were designed, synthesized as a polyprotein with a 2A self-cleavage linker peptide from the Foot-and-mouth disease virus (FMDV, cloned into a plant binary vector under a constitutive maize ubiquitin promoter, and transformed into rice nuclear genome through Agrobacterium-mediated transformation. Southern blot and western blot analyses confirmed the integration and expression of BLC and BHC genes in transgenic rice callus. Enzyme linked immunosorbent assay (ELISA analysis indicated that the rice-derived Bevacizumab mAb was biologically active and the recombinant mAb was expressed at high levels (160.7-242.8 mg kg-1FW in transgenic rice callus. The mAb was purified by using protein A affinity chromatography and the purified antibody was tested for its binding affinity with its target hVEGF antigen by ELISA. Rice callus produced Bevacizumab and a commercial Bevacizumab (Avastin were shown to have similar binding affinity to hVEGF. These results indicated that rice callus produced Bevacizumab could have similar biological activity and might potentially be used as a cost-effective biosimilar molecule in future cancer treatment.

  15. Efficient Production of a Bioactive Bevacizumab Monoclonal Antibody Using the 2A Self-cleavage Peptide in Transgenic Rice Callus

    Science.gov (United States)

    Chen, Lei; Yang, Xiaoyu; Luo, Da; Yu, Weichang

    2016-01-01

    Bevacizumab, a humanized monoclonal antibody (mAb) targeting to the vascular endothelial growth factor (VEGF), has been widely used in clinical practice for the treatment of multiple cancers. Bevacizumab was mostly produced by the mammalian cell expression system. We here reported the first plant-derived Bevacizumab by using transgenic rice callus as an alternative gene expression system. Codon-optimized Bevacizumab light chain (BLC) and Bevacizumab heavy chain (BHC) genes were designed, synthesized as a polyprotein with a 2A self-cleavage linker peptide from the Foot-and-mouth disease virus, cloned into a plant binary vector under a constitutive maize ubiquitin promoter, and transformed into rice nuclear genome through Agrobacterium-mediated transformation. Southern blot and western blot analyses confirmed the integration and expression of BLC and BHC genes in transgenic rice callus. Enzyme-linked immunosorbent assay (ELISA) analysis indicated that the rice-derived Bevacizumab mAb was biologically active and the recombinant mAb was expressed at high levels (160.7–242.8 mg/Kg) in transgenic rice callus. The mAb was purified by using protein A affinity chromatography and the purified antibody was tested for its binding affinity with its target human VEGF (hVEGF) antigen by ELISA. Rice callus produced Bevacizumab and a commercial Bevacizumab (Avastin) were shown to have similar binding affinity to hVEGF. These results indicated that rice callus produced Bevacizumab could have similar biological activity and might potentially be used as a cost-effective biosimilar molecule in future cancer treatment. PMID:27555853

  16. Anther development stage and gamma radiation effects on tomato anther-derived callus formation; Efeitos do estadio de desenvolvimento da antera e da radiacao gama na formacao de calos derivados de anteras de tomate

    Energy Technology Data Exchange (ETDEWEB)

    Brasileiro, Ana Christina R.; Willadino, Lilia [Universidade Federal Rural de Pernambuco, Recife, PE (Brazil). Lab. de Cultura de Tecidos Vegetais. E-mail: lilia@truenet.com.br; Guerra, Marcelo [Pernambuco Univ., Recife, PE (Brazil). Dept. de Botanica. Lab. de Citogenetica Vegetal; Colaco, Waldeciro [Pernambuco Univ., Recife, PE (Brazil). Dept. de Energia Nuclear. Lab. de Radioagronomia; Meunier, Isabelle [Universidade Federal Rural de Pernambuco, Recife, PE (Brazil). Dept. de Engenharia Florestal; Camara, Terezinha R. [Universidade Federal Rural de Pernambuco, Recife, PE (Brazil). Dept. de Quimica. Lab. de Cultura de Tecidos Vegetais

    1999-12-01

    Two experiments were carried (I) to determine tomato anther development stage influence on callus production; and (II) to investigate gamma radiation effects on anther culture. In the first experiment, anthers of a tomato hybrid (IPA 5 x Rotam 4-F{sub 1}) were grown on three media. Although calli were induced at all stages of anther development, varying from prophase I to mono nucleate microspore, callus frequency decreased as anther development progressed and calli induction were not significantly affected by all media tested. Anthers containing prophase I meiocytes produced the highest calli frequency. Anther and flower bud length both were significantly correlated with anther development stage. In the second experiment, seed and floral buds of tomato hybrids IPA 5 x Rotam 4 (F{sub 2}), IPA 6 x Rotam 4 (F{sub 2}) and IPA 8 x 217.1 (F{sub 2}) were submitted to gamma-ray and anthers were plated on two media described by Gresshoff and Doy (1972) supplemented with 2.0 mg L{sup -1} NAA + 5.0 mg L{sup -1} KIN and 2.0 mg L{sup -1} NAA + 1.0 mg L{sup -1} KIN. No significant differences for genotype and dosage testes were found for calli formation. (author)

  17. Effects of the suicide inhibitors of arginine and ornithine decarboxylase activities on organogenesis, growth, free polyamine and hydroxycinnamoyl putrescine levels in leaf explants of Nicotiana xanthi N.C. Cultivated in vitro in a medium producing callus formation.

    Science.gov (United States)

    Burtin, D; Martin-Tanguy, J; Paynot, M; Rossin, N

    1989-01-01

    We studied the effects of dl-alpha-difluoromethylarginine (DFMA) and dl-alpha-difluoromethylornithine (DFMO), specific, irreversible inhibitors of arginine decarboxylase (ADC) and ornithine decarboxylase (ODC), respectively, on organogenesis growth and titers of free polyamines and conjugated putrescines (hydroxycinnamoyl putrescines) in tobacco (Nicotiana tabacum cv Xanthi n.c.) calli. These results suggest that ADC and ODC regulate putrescine biosynthesis during early and later stages of tobacco callus development, respectively. ADC appears active in biosynthesis of large levels of free amines (agmatine and putrescine) while ODC appears active only in biosynthesis of large levels of putrescine conjugates (hydroxycinnamoyl putrescines). DFMA inhibits the fresh and dry weight increases of tobacco calli, whereas DFMO even promoted the fresh and dry weight increases, thus supporting the view that ADC is important for cell division and callus induction. Inhibition of ODC activity by DFMO resulting in an amide deficiency after 4 weeks of culture facilates the expression of differentiated cell functions. Formation of buds is associated with a significant decrease of hydroxycinnamoyl putrescines.

  18. Effect of Agrobacterium rhizogenes and elicitation on the asiaticoside production in cell cultures of Centella asiatica

    Directory of Open Access Journals (Sweden)

    Komar Ruslan

    2012-01-01

    Full Text Available Background: Centella asiatica (L. Urb. (Apiaceae is an important medicinal plant, and it has been using to prepare herbal medicines. The compounds responsible for the biological activity of C. asiatica are triterpenoids such as asiaticoside. Asiaticoside is also important as a marker for standardization of C. asiatica. Due to the low content, there is a need to enhance the production of asiaticoside of C. asiatica. The biotechnological approach is one of the methods that can be used to enhance its production. Objectives: This study was designed to enhance the production of asiaticoside from C. asiatica using A. rhizogenes and elicitation experiments. Materials and Methods : Callus cultures were initiated using Murashige and Skoog (MS medium supplemented with 1.0 mg/L indole-3-acetic acid (IAA and 1.0 mg/L 6-benzylaminopurin (BAP. All media were supplemented with 4% (w/w sucrose and solidified with 0.9% agar. Elicitations were done using pectin, methyl jasmonate, and Cu 2+ ions. Transformed hairy root cultures were performed using A. rhizogenes. Results: Callus culture of C. asiatica was successfully initiated. Enhancement of the production of asiaticoside in the callus culture by elicitors pectin was up to 31%; methyl jasmonate (50 ΅M in cell suspension cultures at day 14 was up to 171% compared to explant and 494% compared to control callus; copper ion (25 ΅M at day 21 was up to 144% compared to explant, and 676% compared to control cell suspension cultures. While enhancement by genetic transformation using A. rhizogenes was 166-172% compare to untransformed roots Conclusion: Elicitation and genetically transformed hairy root cultures of C. asiatica produced asiaticoside up to 172% higher than untreated callus.

  19. Efecto del tipo de explante y la concentración de ácido 2,4-diclorofenoxiacético en la formación de callos en Morus alba L. Effect of explant type and concentration of 2,4-dichlorophenoxyacetic acid on callus formation in Morus alba L.

    Directory of Open Access Journals (Sweden)

    A Espinosa

    2012-12-01

    Full Text Available El objetivo del trabajo fue evaluar el efecto del tipo de explante y la concentración de 2,4-D en la formación de callos, en Morus alba L. Como fuentes de explantes se utilizaron limbos foliares, tallos y pecíolos, obtenidos a partir de estacas que brotaron en condiciones de laboratorio. El medio de cultivo basal estuvo constituido por las sales y vitaminas MS (1962, con diferentes concentraciones de 2,4-D (0; 0,5; 1,0 y 2,0 mg.L-1. Se utilizó un diseño experimental completamente aleatorizado. La formación de callos en todos los tipos de explantes se inició en los primeros 15 días posteriores al establecimiento in vitro. La callogénesis solo ocurrió en los medios de cultivo con 2,4-D. La zona de formación del callo, su color y textura dependieron del tipo de explante utilizado. El aumento de la concentración de 2,4-D en el medio de cultivo incrementó el tamaño de los callos, con los mejores resultados en las concentraciones de 1,0 y 2,0 mg.L-1The objective of the work was to evaluate the effect of explant type and concentration of 2,4-dichlorophenoxyacetic acid on callus formation, in Morus alba L. As explant sources leaf blades, stems and petioles were used , obtained from cuttings which produced growths under laboratory conditions. The basal culture medium was constituted by MS salts and vitamins (1962, with different concentrations of 2,4-D (0; 0,5; 1,0 and 2,0 mg.L-1. A completely randomized experimental design was used. Callus formation in all explant types started in the first 15 days after the in vitro establishment. Callogenesis only occurred in the culture media with 2,4-D. The callus formation zone, callus color and texture depended on the explant type used. The increase of 2,4-D concentration in the culture medium increased callus size, with the best results in the concentrations 1,0 and 2,0 mg.L-1.

  20. Cutin plays a role in differentiation of endosperm-derived callus of kiwifruit.

    Science.gov (United States)

    Popielarska-Konieczna, Marzena; Kozieradzka-Kiszkurno, Małgorzata; Bohdanowicz, Jerzy

    2011-11-01

    Cutin fluorescence, after auramine O treatment, was detected on the surface of organogenic areas (protuberances) of endosperm derived callus induced on Murashige and Skoog medium with thidiazuron (0.5 mg l(-1)) in darkness. Electron micrographs of the protuberances revealed cuticle, visible as a dark-staining layer, and amorphous waxes on the cell wall. In some cases the cells of the epidermis-like layer and shoot buds at early stages of development showed thick and characteristically wavy cutin. This waviness corresponds with the wrinkled appearance of the cell wall as observed by scanning electron microscopy. The role of multivesicular bodies in cutin production and transfer to the plasma membrane is discussed.

  1. Effects of Exogenous Indole Butyric Acid and Callus Formation on the Anti-oxidant Activity, Total Phenolic, and Anthocyanin Constituents of Mulberry Cuttings

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    In order to evaluate the effects of exogenous indole butyric acid (IBA) and callus formation on the antioxidant activity, total phenolics, and anthocyanin constituents of Morus nigra L. and M. alba L. cuttings, we investigated the variations before and after the treatment. The results indicate that anti-oxidant ability, total phenolic, and anthocyanin constituents of the callus stems of both Morus species were higher than those of non-callus forming species. There were also increases observed in anti-oxidant ability, total phenolic,and anthocyanin constituents of calli treated with IBA (1 000-3 000 mg/L).

  2. Somatic Embryogenesis in Lily Bulb Scale Cultures

    Institute of Scientific and Technical Information of China (English)

    WANG Shasha; WANG Jingang; FAN Jinping; CHE Daidi

    2008-01-01

    Somatic embryogenesis from lily bulb scales has not been studied in details, although tissue culture methods have been applied to the propagation for decades. The effects of different kinds and concentration of auxins for oriental lily somatic embryogenesis were investigated (Lilium hybrida car. Sorbonne).2,4-dichlorophenoxyacetic acid (2,4-D), thidiazuron (TDZ) and α-naphthaleneacetic acid (NAA) media with benzyladenine(6-BA) and lactalbumin hydrolysate (LH) were used for embryogenic callus in the darkness. The best response onembryogenic callus formation was obtained on MS media supplemented 2, 4-D 2.0 mg·L-1,6-BA 0.5 mg·L-1 and LH 300 mg·L-1. Transfer embryogenic callus to the media with TDZ, 6-BA, kinetin (KT) supplemented 2, 4-D. The highest number of somatic embryos has been produced on medium with 0.5 mg.L-1 2, 4-D and 0.3 mg·L-1 KT. Germinated embryos with shoot axes were changed to MS media with 6-BA 0.5 mg· L-1. The results suggest that in vitro culture of somatic embryogenesis from lily bulb scales can be used for plant regeneration.

  3. Selection of bacterial wilt-resistant tomato through tissue culture.

    Science.gov (United States)

    Toyoda, H; Shimizu, K; Chatani, K; Kita, N; Matsuda, Y; Ouchi, S

    1989-06-01

    Bacterial wilt-resistant plants were obtained using a tomato tissue culture system. A virulent strain ofPseudomonas solanacearum secreted some toxic substances into the culture medium. Leaf explant-derived callus tissues which were resistant to these toxic substances in the culture filtrate were selectedin vitro and regenerated into plants. These plants expressed bacterial wilt resistance at the early infection stage to suppress or delay the growth of the inoculated bacteria. On the other hand, complete resistance was obtained in self-pollinated progeny of regenerants derived from non-selected callus tissues. These plants showed a high resistance when inoculated with this strain, and were also resistant when planted in a field infested with a different strain of the pathogen.

  4. EPO promotes bone repair through enhanced cartilaginous callus formation and angiogenesis.

    Directory of Open Access Journals (Sweden)

    Lin Wan

    Full Text Available Erythropoietin (EPO/erythropoietin receptor (EPOR signaling is involved in the development and regeneration of several non-hematopoietic tissues including the skeleton. EPO is identified as a downstream target of the hypoxia inducible factor-α (HIF-α pathway. It is shown that EPO exerts a positive role in bone repair, however, the underlying cellular and molecular mechanisms remain unclear. In the present study we show that EPO and EPOR are expressed in the proliferating, pre-hypertrophic and hypertrophic zone of the developing mouse growth plates as well as in the cartilaginous callus of the healing bone. The proliferation rate of chondrocytes is increased under EPO treatment, while this effect is decreased following siRNA mediated knockdown of EPOR in chondrocytes. EPO treatment increases biosynthesis of proteoglycan, accompanied by up-regulation of chondrogenic marker genes including SOX9, SOX5, SOX6, collagen type 2, and aggrecan. The effects are inhibited by knockdown of EPOR. Blockage of the endogenous EPO in chondrocytes also impaired the chondrogenic differentiation. In addition, EPO promotes metatarsal endothelial sprouting in vitro. This coincides with the in vivo data that local delivery of EPO increases vascularity at the mid-stage of bone healing (day 14. In a mouse femoral fracture model, EPO promotes cartilaginous callus formation at days 7 and 14, and enhances bone healing at day 28 indexed by improved X-ray score and micro-CT analysis of microstructure of new bone regenerates, which results in improved biomechanical properties. Our results indicate that EPO enhances chondrogenic and angiogenic responses during bone repair. EPO's function on chondrocyte proliferation and differentiation is at least partially mediated by its receptor EPOR. EPO may serve as a therapeutic agent to facilitate skeletal regeneration.

  5. EPO promotes bone repair through enhanced cartilaginous callus formation and angiogenesis.

    Science.gov (United States)

    Wan, Lin; Zhang, Fengjie; He, Qiling; Tsang, Wing Pui; Lu, Li; Li, Qingnan; Wu, Zhihong; Qiu, Guixing; Zhou, Guangqian; Wan, Chao

    2014-01-01

    Erythropoietin (EPO)/erythropoietin receptor (EPOR) signaling is involved in the development and regeneration of several non-hematopoietic tissues including the skeleton. EPO is identified as a downstream target of the hypoxia inducible factor-α (HIF-α) pathway. It is shown that EPO exerts a positive role in bone repair, however, the underlying cellular and molecular mechanisms remain unclear. In the present study we show that EPO and EPOR are expressed in the proliferating, pre-hypertrophic and hypertrophic zone of the developing mouse growth plates as well as in the cartilaginous callus of the healing bone. The proliferation rate of chondrocytes is increased under EPO treatment, while this effect is decreased following siRNA mediated knockdown of EPOR in chondrocytes. EPO treatment increases biosynthesis of proteoglycan, accompanied by up-regulation of chondrogenic marker genes including SOX9, SOX5, SOX6, collagen type 2, and aggrecan. The effects are inhibited by knockdown of EPOR. Blockage of the endogenous EPO in chondrocytes also impaired the chondrogenic differentiation. In addition, EPO promotes metatarsal endothelial sprouting in vitro. This coincides with the in vivo data that local delivery of EPO increases vascularity at the mid-stage of bone healing (day 14). In a mouse femoral fracture model, EPO promotes cartilaginous callus formation at days 7 and 14, and enhances bone healing at day 28 indexed by improved X-ray score and micro-CT analysis of microstructure of new bone regenerates, which results in improved biomechanical properties. Our results indicate that EPO enhances chondrogenic and angiogenic responses during bone repair. EPO's function on chondrocyte proliferation and differentiation is at least partially mediated by its receptor EPOR. EPO may serve as a therapeutic agent to facilitate skeletal regeneration.

  6. INDUKSI KALUS JERUK KASTURI (CITRUS MICROCARPA MENGGUNAKAN HORMON 2,4-D DAN BAP DENGAN METODE IN VITRO

    Directory of Open Access Journals (Sweden)

    Imam Mahadi

    2016-08-01

    Full Text Available Calamansi fruits (Citrus microcarpa or musk lime is one horticulture crops has been known as beverage and food flavorings. This research aimed to determine the effect of 2,4-D and BAP on callus induction of explants calamansi fruits (Citrus microcarpa. Parameters observed is percentage of explants, time appeared callus, callus width, and texture of the callus. Research results and a growing percentage of the width callus explants were analyzed by ANOVA and a further test DMRT at 5% level. The next to appear callus timing parameters and texture of callus analyzed in description. The result shows that a combination of hormones D4B1 and D4B2 produces a callus emerged fastest of 3.3 days after culture. The highest percentage of callus formation that is 100% contained in the treatment of D2B0, D2B0.5-D4B0. The mean width of callus was highest in treatment D4B2with a width of 0.97 cm. Embryogenic callus resulting by treatment of D2B1, D2B2, and D3B0. However, the best a combination of hormones is treatment D2B2 (2 mg/l, 2,4-D, dan 2 mg/l BAP producing embryogenic callus edvanced to suspension cultures.

  7. Protein and Glycoprotein Patterns Related to Morphogenesis in Mammillaria gracillis Pfeiff. Tissue Culture

    Directory of Open Access Journals (Sweden)

    Biljana Balen

    2002-01-01

    Full Text Available As plants with Crassulacean Acid Metabolism (CAM, cacti are highly affected by artificial environmental conditions in tissue culture. Plants of Mammillaria gracillis Pfeiff. (Cactaceae propagated in vitro produced callus spontaneously. This habituated callus regenerated normal and hyperhydric shoots without the addition of growth regulators. In order to compare habituated callus with the tumorous one, cactus cells were transformed with two strains of Agrobacterium tumefaciens: the wild strain B6S3 (tumour line TW and the rooty mutant GV3101 (tumour line TR. Gene expression in cactus plants, habituated callus, regenerated shoots and two tumour lines was analysed at the level of cellular and extracellular protein and glycoprotein profiles. Proteins were separated by SDS-polyacrylamide gel electrophoresis and 2-D PAGE electrophoresis and silver stained. Concavalin A-peroxidase staining detected glycoproteins with D-manose in their glycan component on protein blots. Developmentally specific protein patterns of Mammillaria gracillis tissue lines were detected. The 2-D PAGE electrophoresis revealed some tissue specific protein groups. The cellular glycoprotein of 42 kDa detected by ConA was highly expressed in undifferentiated tissues (habituated callus, TW and TR tumours and in hyperhydric regenerants. Tumours produced extracellular proteins of 33, 23 and 22 kDa. The N glycosylation of cellular and extracellular proteins was related to specific developmental stage of cactus tissue.

  8. Biological activities of indoleacetylamino acids and their use as auxins in tissue culture.

    Science.gov (United States)

    Hangarter, R P; Peterson, M D; Good, N E

    1980-05-01

    THE AUXIN ACTIVITIES OF A NUMBER OF INDOLEACETYLAMINO ACID CONJUGATES HAVE BEEN DETERMINED IN THREE TEST SYSTEMS: growth of tomato hypocotyl explants (Lycopersicon esculentum Mill. cv. Marglobe); growth of tobacco callus cultures (Nicotiana tabacum L. cv. Wisconsin 38); and ethylene production from pea stems (Pisum sativum L. cv. Alaska). The activities of the conjugates differ greatly depending on the amino acid moiety. Indoleacetyl-l-alanine supports rapid callus growth from the tomato hypocotyls while inhibiting growth of shoots and roots. Indoleacetylglycine behaves in a similar manner but is somewhat less effective in supporting callus growth and in inhibiting shoot formation. The other amino acid conjugates tested (valine, leucine, aspartic acid, threonine, methionine, phenylalanine, and proline) support shoot formation without supporting root formation or much callus growth. The tobacco callus system, which forms abundant shoots in the presence or absence of free indoleacetic acid, produces only rapid undifferentiated growth in the presence of indoleacetyl-l-alanine and indoleacetylglycine. The other conjugates inhibit shoot formation weakly if at all. Most of the conjugates induce sustained ethylene production from the pea stems but at rates well below the initial rates observed with free indoleacetic acid. Many, but not all of the effects of conjugates such as indoleacetyl-l-alanine can be mimicked by frequent renewals of the supply of free indoleacetic acid.

  9. 7 CFR 917.4 - Fruit.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Fruit. 917.4 Section 917.4 Agriculture Regulations of the Department of Agriculture (Continued) AGRICULTURAL MARKETING SERVICE (Marketing Agreements and... production area exhibiting the characteristics of a peach and subject to cultural practices common to...

  10. Pneumatic hydrodynamics influence transplastomic protein yields and biological responses during in vitro shoot regeneration of Nicotiana tabacum callus: Implications for bioprocess routes to plant-made biopharmaceuticals.

    Science.gov (United States)

    Barretto, Sherwin S; Michoux, Franck; Hellgardt, Klaus; Nixon, Peter J

    2017-01-15

    Transplastomic plants are capable of high-yield production of recombinant biopharmaceutical proteins. Plant tissue culture combines advantages of agricultural cultivation with the bioprocess consistency associated with suspension culture. Overexpression of recombinant proteins through regeneration of transplastomic Nicotiana tabacum shoots from callus tissue in RITA(®) temporary immersion bioreactors has been previously demonstrated. In this study we investigated the hydrodynamics of periodic pneumatic suspension of liquid medium during temporary immersion culture (4 min aeration every 8 h), and the impact on biological responses and transplastomic expression of fragment C of tetanus toxin (TetC). Biomass was grown under a range of aeration rates for 3, 20 and 40-day durations. Growth, mitochondrial activity (a viability indicator) and TetC protein yields were correlated against the hydrodynamic parameters, shear rate and energy dissipation rate (per kg of medium). A critical aeration rate of 440 ml min(-1) was identified, corresponding to a shear rate of 96.7 s(-1), pneumatic power input of 8.8 mW kg(-1) and initial 20-day pneumatic energy dissipation of 127 J kg(-1), at which significant reductions in biomass accumulation and mitochondrial activity were observed. There was an exponential decline in TetC yields with increasing aeration rates at 40 days, across the entire range of conditions tested. These observations have important implications for the optimisation and scale-up of transplastomic plant tissue culture bioprocesses for biopharmaceutical production.

  11. Production of justicidin B, a cytotoxic arylnaphthalene lignan from genetically transformed root cultures of Linum leonii

    NARCIS (Netherlands)

    Vasilev, Nikolay; Elfahmi, [No Value; Bos, Rein; Kayser, Oliver; Momekov, Georgi; Konstantinov, Spiro; Ionkova, Iliana

    2006-01-01

    Callus and hairy root cultures of Linum leonii were established. The genetic transformation in hairy roots was proven by PCR analysis, which showed integration of rol A and rol C genes into the plant genome. Calli and hairy roots accumulate the arylnaphthalene lignan justicidin B as a major constitu

  12. Analysis of soybean tissue culture protein dynamics using difference gel electrophoresis

    Science.gov (United States)

    Excised hypocotyls from developing soybean (Glycine max (L.) merr. cv. Jack) were cultivated on agar-solidified medium until callus formed. The calli were then propagated in liquid medium until stable, relatively uniform, finely-divided suspension cultures were obtained. Cells were typically transfe...

  13. Tissue culture-induced DNA methylation polymorphisms in repetitive DNA of tomato calli and regenerated plants

    NARCIS (Netherlands)

    Smulders, M.J.M.; Rus-Kortekaas, W.; Vosman, B.

    1995-01-01

    The propagation of plants through tissue culture can induce a variety of genetic and epigenetic changes. Variation in DNA methylation has been proposed as a mechanism that may explain at least a part of these changes. In the present study, the methylation of tomato callus DNA was compared with that

  14. Calogênese em Cissus sicyoides L. a partir de segmentos foliares visando à produção de metabólitos in vitro Calluses from Cissus sicyoides L. leaves

    Directory of Open Access Journals (Sweden)

    F.R. Rodrigues

    2010-09-01

    faz-se necessário a adição de 6,0 mg L-1 de BAP ao meio de cultivo. Identificou-se a presença de heterosídeos cardiotônicos em calos de Cissus sicyoides L.Secondary metabolites are essentially produced and extracted from plants grown in the field under influence of seasonal variations. The use of biotechnological techniques is an alternative resource for drug production. Among these techniques, tissue culture through callus genesis is highlighted, since callus growth is desirable to induce somaclonal variation and physiological studies, especially when the presence of secondary metabolites can be related to cell growth. The aim of this work was to establish a protocol for Cissus sicyoides L. callus genesis from leaf segments in order to produce metabolites in vitro. Thus, leaf segments removed from adult plants grown in the field were used as explants. After disinfestation, the material was inoculated into MT medium + 1.0 mg L-1 NAA and kept in a BOD chamber, with controlled temperature and luminosity. After 30 days, the percentage of surviving explants and the percentage of contamination were evaluated. For culture, MT medium + 1.0 mg L-1 NAA was used, varying BAP concentrations: 2.0, 4.0, 6.0 and 12.0 mg L-1. In the cultivation, the number of compact and friable calluses was counted. For the first and second subculture, the material was introduced into MT medium + 1.0 mg L-1 NAA, varying the same BAP concentrations; the number of friable calluses formed and the size of callus mass were described. The number of replicates formed during subcultures, and fresh and dry matter (g were also obtained. Then, phytochemical tests were done in order to identify some compounds. The adopted time and concentration of sodium hypochlorite proved to be inefficient for disinfestation. For Cissus sicyoides L. callus genesis from leaf segments, the addition of 6.0 mg L-1 BAP to the culture medium is needed. Cardiotonic heterosides were detected in Cissus sicyoides L. calluses.

  15. Induction of bulb organogenesis in in vitro cultures of tarda tulip (Tulipa tarda Stapf.) from seed-derived explants.

    Science.gov (United States)

    Maślanka, Małgorzata; Bach, Anna

    2014-01-01

    A protocol for obtaining bulbs via in vitro organogenesis was developed for tarda tulip (Tulipa tarda Stapf). Scale explants were obtained from bulbs formed at the base of seedlings or from adventitious bulbs that developed from callus tissue forming on stolons or on germinating seeds. Some explants were subjected to chilling at 5°C for 12 wk. The culture media contained 3 or 6% sucrose and was supplemented with either no growth regulators, either 0.5 μM 6-benzyl-aminopurine (BAP) or 18.9 or 94.6 μM abscisic acid (ABA). Cultures were maintained in the dark at 20°C. Callus tissue developed mainly on media without growth regulators or with BAP. Callus was formed from up to 96% of explants derived from non-chilled adventitious bulbs that were treated with 3% sucrose and 0.5 μM BAP. Less callus was formed from chilled explants compared with non-chilled explants. Newly formed adventitious bulbs appeared on the explants via direct and indirect organogenesis. The media with BAP promoted the formation of adventitious bulbs at a rate of 56-92% from non-chilled explants, whereas a maximum rate of 36% was observed from chilled explants. ABA inhibited the induction of adventitious bulbs and callus. The adventitious bulbs obtained in these experiments contained a meristem, which was evidence that they had developed properly.

  16. Production of haploids from anther culture of banana [Musa balbisiana (BB)].

    Science.gov (United States)

    Assani, A; Bakry, F; Kerbellec, F; Haïcour, R; Wenzel, G; Foroughi-Wehr, B

    2003-02-01

    We report here, for the first time, the production of haploid plants of banana Musa balbisiana (BB). Callus was induced from anthers in which the majority of the microspores were at the uninucleate stage. The frequency of callus induction was 77%. Callus proliferation usually preceded embryo formation. About 8% of the anthers developed androgenic embryos. Of the 147 plantlets obtained, 41 were haploids (n=x=11). The frequency of haploid production depended on genotypes used: 18 haploid plants were produced from genotype Pisang klutuk, 12 from Pisang batu, seven from Pisang klutuk wulung and four from Tani. The frequency of regeneration was 1.1%, which was based on the total number of anthers cultured. Diploid plants (2n=2x=22) were also observed in the regenerated plants. The haploid banana plants that were developed will be important material for the improvement of banana through breeding programmes.

  17. Micro-propagation of Lepedium meyenii Walp. (Maca) by shoot culture

    Institute of Scientific and Technical Information of China (English)

    ChengHua; YuLongjiang; HuQiongyue; SunYouping; JinWenwen; ChenShancai; LiMaoteng

    2004-01-01

    The callus induction and plant regeneration system for an important plant, Lepedium meyenii Walp., has been established. Calli were induced from cotyledons petioles of Lepedium meyenii Walp within 4 weeks in a modified MS medium supplemented with BA plus NAA. The highest percentage of callus formation (57. 1% ) was found on MS medium supplemented with 0. 5 mg l-1 BA and 0. 5 mg l-1 NAA. During subculture on the shoot formation medium, most of calli proliferated and 50% -60% formed shoots. About 66.7% of shoots formed into roots on 1/2 strength MS containing 0.5mg l-1 IBA after 4 weeks in culture. Chromosome count confirmed the number of the regeneration Maca plantlet was the same as that of the native plant(8x=64) .For regeneration of plantlets, from seedling via primary callus production, a four-step process of organogenes is required about 16 weeks.

  18. 细辛组织培养研究%Tissue Culture of Asarum siboldii

    Institute of Scientific and Technical Information of China (English)

    南桂仙; 金光德

    2009-01-01

    以细辛叶片为外植体,研究了其组织培养技术.结果表明:愈伤组织继代1次,可增殖10~15倍,每块愈伤组织经过再分化能长出5~8株试管苗,反复继代3~4次,每个叶片可产出约500株试管苗.%Used the leaf of Asarum siboldii as explants,the tissue culture technique was studied.The results showed that the the callus could proliferate 10-15 times after subculturing for one time.5-8 plantlets grew out when the callus dedifferentiated.When the callus was subcultured for 3-4 times,about 500 plantlets could be obtained.

  19. In vitro culture of Cucumis sativus L. VI. Histological analysis of leaf explants cultured on media with 2, 4-D or 2, 4, 5-T

    Directory of Open Access Journals (Sweden)

    Anna Nadolska-Orczyk

    2014-01-01

    Full Text Available The developmental sequence of callus initiation and somatic embryogenesis in leaf explants of Cucumis sativus cv. Borszczagowski was analysed and compared on media containing two different auxin phenoxy-derivatives (2,4-D and 2,4,5-T and cytokinin (BAP or 2iP. During the first 20 days of culture on media with 2,4,5-T proliferation of parenchymatic tissue occurred mainly and only small meristematic centers were observed. There was an intensive detachment of parenchymatic cells and dissociation of their cell walls near vessels and in the lower part of the explant adjacent to the medium. These cells were strongly plasmolysed. On the 2,4-D containing medium mostly meristematic tissue developed, proliferating around vascular bundles and forming meristematic centers or promeristem-like structures. After 35-50 days of culture, secondary callus was formed by separation of meristematic cells from the meristem surface in explants cultured on the 2,4-D containing medium. On medium supplemented with 2, 4, 5-T the detachment of parenchymatic and meristematic cells occurred, along with formation of a gel-like substance. The gel-like callus contained multi-cellular aggregates, proembryoids and embryoids. This type of callus tissue was initiated more intensively on medium with 2, 4, 5-T, but the frequency of somatic embryogenesis was much lower. The periferial cells of aggregates, proembryoids and embryoids showed the tendency to separate from the surface of the tissue. Many embryoids formed adventitious embryos.

  20. Reprogramming of H3K27me3 is critical for acquisition of pluripotency from cultured Arabidopsis tissues.

    Directory of Open Access Journals (Sweden)

    Chongsheng He

    2012-08-01

    Full Text Available In plants, multiple detached tissues are capable of forming a pluripotent cell mass, termed callus, when cultured on media containing appropriate plant hormones. Recent studies demonstrated that callus resembles the root-tip meristem, even if it is derived from aerial organs. This finding improves our understanding of the regeneration process of plant cells; however, the molecular mechanism that guides cells of different tissue types to form a callus still remains elusive. Here, we show that genome-wide reprogramming of histone H3 lysine 27 trimethylation (H3K27me3 is a critical step in the leaf-to-callus transition. The Polycomb Repressive Complex 2 (PRC2 is known to function in establishing H3K27me3. By analyzing callus formation of mutants corresponding to different histone modification pathways, we found that leaf blades and/or cotyledons of the PRC2 mutants curly leaf swinger (clf swn and embryonic flower2 (emf2 were defective in callus formation. We identified the H3K27me3-covered loci in leaves and calli by a ChIP-chip assay, and we found that in the callus H3K27me3 levels decreased first at certain auxin-pathway genes. The levels were then increased at specific leaf genes but decreased at a number of root-regulatory genes. Changes in H3K27me3 levels were negatively correlated with expression levels of the corresponding genes. One possible role of PRC2-mediated H3K27me3 in the leaf-to-callus transition might relate to elimination of leaf features by silencing leaf-regulatory genes, as most leaf-preferentially expressed regulatory genes could not be silenced in the leaf explants of clf swn. In contrast to the leaf explants, the root explants of both clf swn and emf2 formed calli normally, possibly because the root-to-callus transition bypasses the leaf gene silencing process. Furthermore, our data show that PRC2-mediated H3K27me3 and H3K27 demethylation act in parallel in the reprogramming of H3K27me3 during the leaf-to-callus transition

  1. Reprogramming of H3K27me3 is critical for acquisition of pluripotency from cultured Arabidopsis tissues.

    Science.gov (United States)

    He, Chongsheng; Chen, Xiaofan; Huang, Hai; Xu, Lin

    2012-08-01

    In plants, multiple detached tissues are capable of forming a pluripotent cell mass, termed callus, when cultured on media containing appropriate plant hormones. Recent studies demonstrated that callus resembles the root-tip meristem, even if it is derived from aerial organs. This finding improves our understanding of the regeneration process of plant cells; however, the molecular mechanism that guides cells of different tissue types to form a callus still remains elusive. Here, we show that genome-wide reprogramming of histone H3 lysine 27 trimethylation (H3K27me3) is a critical step in the leaf-to-callus transition. The Polycomb Repressive Complex 2 (PRC2) is known to function in establishing H3K27me3. By analyzing callus formation of mutants corresponding to different histone modification pathways, we found that leaf blades and/or cotyledons of the PRC2 mutants curly leaf swinger (clf swn) and embryonic flower2 (emf2) were defective in callus formation. We identified the H3K27me3-covered loci in leaves and calli by a ChIP-chip assay, and we found that in the callus H3K27me3 levels decreased first at certain auxin-pathway genes. The levels were then increased at specific leaf genes but decreased at a number of root-regulatory genes. Changes in H3K27me3 levels were negatively correlated with expression levels of the corresponding genes. One possible role of PRC2-mediated H3K27me3 in the leaf-to-callus transition might relate to elimination of leaf features by silencing leaf-regulatory genes, as most leaf-preferentially expressed regulatory genes could not be silenced in the leaf explants of clf swn. In contrast to the leaf explants, the root explants of both clf swn and emf2 formed calli normally, possibly because the root-to-callus transition bypasses the leaf gene silencing process. Furthermore, our data show that PRC2-mediated H3K27me3 and H3K27 demethylation act in parallel in the reprogramming of H3K27me3 during the leaf-to-callus transition, suggesting a

  2. Tissue culture-induced DNA methylation polymorphisms in repetitive DNA of tomato calli and regenerated plants.

    Science.gov (United States)

    Smulders, M J; Rus-Kortekaas, W; Vosman, B

    1995-12-01

    The propagation of plants through tissue culture can induce a variety of genetic and epigenetic changes. Variation in DNA methylation has been proposed as a mechanism that may explain at least a part of these changes. In the present study, the methylation of tomato callus DNA was compared with that of leaf DNA, from control or regenerated plants, at MspI/HpaII sites around five middle-repetitive sequences. Although the methylation of the internal cytosine in the recognition sequence CCGG varied from zero to nearly full methylation, depending on the probe used, no differences were found between callus and leaf DNA. For the external cytosine, small differences were revealed between leaf and callus DNA with two probes, but no polymorphisms were detected among DNA samples of calli or DNA samples of leaves of regenerated plants. When callus DNA cut with HindIII was studied with one of the probes, H9D9, most of the signal was found in high-molecular-weight DNA, as opposed to control leaf DNA where almost all the signal was in a fragment of 530 bp. Also, an extra fragment of 630 bp was found in the callus DNA that was not present in control leaf DNA. Among leaves of plants regenerated from tissue culture, the 630-bp fragment was found in 10 of 68 regenerated plants. This 630-bp fragment was present among progeny of only 4 of these 10 plants after selfing, i.e. it was partly inherited. In these cases, the fragment was not found in all progeny plants, indicating heterozygosity of the regenerated plants. The data are interpreted as indicating that a HindIII site becomes methylated in callus tissue, and that some of this methylation persists in regenerated plants and is partly transmitted to their progeny.

  3. Zinc tolerance and accumulation in stable cell suspension cultures and in vitro regenerated plants of the emerging model plant Arabidopsis halleri (Brassicaceae).

    Science.gov (United States)

    Vera-Estrella, Rosario; Miranda-Vergara, Maria Cristina; Barkla, Bronwyn J

    2009-03-01

    Arabidopsis halleri is increasingly employed as a model plant for studying heavy metal hyperaccumulation. With the aim of providing valuable tools for studies on cellular physiology and molecular biology of metal tolerance and transport, this study reports the development of successful and highly efficient methods for the in vitro regeneration of A. halleri plants and production of stable cell suspension lines. Plants were regenerated from leaf explants of A. halleri via a three-step procedure: callus induction, somatic embryogenesis and shoot development. Efficiency of callus proliferation and regeneration depended on the initial callus induction media and was optimal in the presence of 1 mg L(-1) 2,4-dichlorophenoxyacetic acid, and 0.05 mg L(-1) benzylaminopurine. Subsequent shoot and root regeneration from callus initiated under these conditions reached levels of 100% efficiency. High friability of the callus supported the development of cell suspension cultures with minimal cellular aggregates. Characterization of regenerated plants and cell cultures determined that they maintained not only the zinc tolerance and requirement of the whole plant but also the ability to accumulate zinc; with plants accumulating up to 50.0 micromoles zinc g(-1) FW, and cell suspension cultures 30.9 micromoles zinc g(-1) DW. Together this work will provide the experimental basis for furthering our knowledge of A. halleri as a model heavy metal hyperaccumulating plant.

  4. 玉米自交系愈伤组织诱导及植株再生体系的建立%Establishment of Callus Induction and Plant Regeneration System of Maize Elite Inbred Lines

    Institute of Scientific and Technical Information of China (English)

    张颖君; 刘茜; 胡梦芸; 高慧敏; 李辉

    2011-01-01

    玉米的组织培养比较困难,其再生体系还不完善,建立骨干自交系稳定、高效的遗传转化体系是玉米转基因技术的必要前提。以玉米常用自交系齐319、478、502、黄C和7922不同大小(1.0、1.5和2.0 mm)的幼胚为外植体,探讨不同基因型、胚大小和植物生长调节剂浓度对幼胚再生的影响,从而建立玉米高频再生体系。结果表明:以1.5 mm大小的齐319幼胚为外植体,愈伤诱导率最高,达到了86.2%;1.5 mg/L的24-D有利于胚性愈伤的形成和胚性的保持;两步法分化培养可以提高愈伤组织分化、再生成苗。确定了玉米自交系齐319较适合的培养程序为:将1.5 mm大小的幼胚盾片向上接于诱导培养基(N6+24-D 1.5 mg/L+L-脯氨酸0.7 g/L+蔗糖30 g/L+琼脂粉8 g/L+硝酸银5μmol/L),在25℃下暗培养20 d;挑选胚性愈伤组织在温度25℃、暗培养条件下进行继代培养,继代培养基与诱导培养基相同,每14 d继代1次;将愈伤组织继代42 d后转移至分化培养基1(MS+肌醇100 mg/L+蔗糖60 g/L+gelrite 3 g/L),继续暗培养10 d左右,待愈伤组织形成象牙白色的块状物且有绿点出现时,将其转移到分化培养基2(MS+肌醇100 mg/L+蔗糖30 g/L+gelrite3 g/L)进行光培养,2~3 d即可分化出苗;待幼苗长到4~5 cm高时,移至生根培养基(1/2 MS+IBA0.8 mg/L+蔗糖30 g/L),14~21 d幼苗长出大量的根系,形成完整的植株。%Tissue culture of maize is relatively difficult,and establishing a stable and efficient transformation system is the premise of maize transgene.Using elite inbred lines(Qi 319,478,502,Huang C and 7922)as explants,the effects of different genotypes,embryo sizes and plant growth regulator concentrations on callus induction and regeneration was studied.The results showed that the frequency of callus induction of 1.5 mm Qi 319 embryos was highest,which reached 86.2%;2,4-D 1.5 mg/L solution was

  5. Determination of fruit characteristics, fatty acid profile and total antioxidant capacity ofMespilus germanica L. fruit

    Institute of Scientific and Technical Information of China (English)

    Hale Seilmi Canbay; Ersin Atay; Serdal Ot

    2015-01-01

    Objective:To determine fruit characteristics, fatty acid profile and total antioxidant capacitiy of first culturedMespilus germanica L. Methods: A total of15 fruits were taken randomly from four directions of adult trees.Then the physical and chemical properties of first cultured medlar fruit (Istanbul/Turkey) were measured by using refractometer, colorimeter, spectrophotometer and gas chromatograph mass spectrometer, respectivly. Results: In the fruit studied, the results showed that palmitic acid, stearic acid, oleic acid, linoleic acid, linolenic acid, arachidic acid and behenic acid were the most abundant fatty acids (FAs), and the main FA was palmitic acid [(35.35 ± 1.20)%]. The percentage of linoleic acid and stearic acid in this fruit oil were (29.10 ± 1.70)% and(8.53 ± 0.25)%, respectively. As a result of the analysis, the total antioxidant capacity of medlar fruit was (1.1 ± 0.2) mmol trolox equivalents/L. Conclusions: The present study has demonstrated the concentrations of FAs and the antioxidantive capacity of first cultured Istanbul medlar fruits by using many tested methods. It is proved that in our daily life, medlar fruit plays a significant role with its nutrition and health effect.

  6. Season and genotype influence in morphogenic and biochemical response of foliar explants of Coffea canephora P. var. Robusta used for callus formation

    Directory of Open Access Journals (Sweden)

    María Esther González

    2005-04-01

    Full Text Available The seasonal change is one from the factors to consider in the embryogenic answer of diverse crops, the variations of temperature, humidity and precipitations cause fluctuations in the plants that influence in the behavior in vitro of the explants. This study was carried out with the objective of evaluating the influence of the time of the year in which the explants were taken on the answer in vitro of Robusta genotypes. Samples were carried out during every month of the year, the explants were cultivated in a culture medium with the mineral salts MS, and 0.5 and 2.0 mg.l-1 of 2,4-D and kinetina, respectively. It was observed that the time of the year in that the samples were selected exercised a marked effect in the answer of the explants. For the studied conditions was favorable to take the leaves samples in the periods May - June, January - February and November - December, due to the few index of peroxidases activity, phenolic oxidation and fungous contamination, as well as a high percentage of callus formation and bigger content of proteins in the same ones. It was determined that the enzymatic peroxidases activity could constitute an important marker for the selection of samples in coffee. Key words: fungous contamination, peroxidases activity,phenolic oxidation

  7. Early Identification of Stable Transformation Events by Combined Use of Antibiotic Selection and Vital Detection of Green Fluorescent Protein (GFP) in Carrot (Daucus carota L.) Callus

    Institute of Scientific and Technical Information of China (English)

    Yuan-Yeu Yau; Seth J Davis; Ahmet Ipek; Philipp W Simon

    2008-01-01

    Genetic transformation is a useful technique to complement conventional breeding in crop improvement. Although carrot has been a model organism for in vitro embryogenesis study, genetic transformation of carrot is still lengthy and labor intensive. An efficient transformation and detection system is desirable. Direct infection of Agrobacterium to carrot calli has provided an easy way for carrot genetic transformation. To improve the efficiency of antibiotic selection in this method, we report the combined use of an improved green-fluorescent protein, referred to as smGFP, to establish a versatile selection method for carrot callus transformation system. By combining antibiotic selection with the bright fluorescence observed in the callus tissue, we were able to easily identify stable transformants in early stage of the transformation process. In addition to the GFP expression of the callus cells, the transgenic nature of callus cells was confirmed with Southern and Western analysis. We found we can link the simplicity of carrot-callus-cell transformation, early detection of stable transformants with antibiotic selection, visualization of GFP fluorescence, and molecular analysis (Southern and Western) of callus tissue (non-photosynthetic tissue) to provide a more efficient way in identifying stable transformants at early stage of carrot transformation.

  8. In Vitro Callus Induction and Growth of Stevia (Stevia rebaudiana Bertoni M. with Difference Concentrations of PEG (Polyethylene Glycol and Light Conditions

    Directory of Open Access Journals (Sweden)

    Mohamad Ana Syabana

    2017-06-01

    Full Text Available Stevia (Stevia rebaudiana Bert. M. is known as a natural non-caloric sweetener. This plants contain glycoside such steviosida type, mainly on the leave contain sweetness level between 200-300 cane sugar but the calorie is very low. This research was aimed to determine difference effects of PEG (Polyethylene Glycol concentrations and the light conditions on  Stevia (Stevia rebaudiana Bert. M. callus induction  in vitro. This research was conducted from April to June 2016 at Biotechnology Laboratory Faculty of Agriculture, Sultan Ageng Tirtayasa University. This research used a completely randomized design (CRD, which consisted of two factors with three replications. Concentrations of PEG as first factor consisted of four levels (0 mg/L, 5 mg/L, 15 mg/L, and 25 mg/L. Light conditions as second factor consisted of two levels (Dark and Light. The results showed that the concentrations of PEG (Polyethylene Glycol did not significantly effect the time of callus appearance and diameter of callus on 4, 5 and 6 weeks after planting. Dark condition was the best conditions for callus induction of stevia. The texture of callus was compact on all treatments and the callus dominant color produced is golden brown

  9. Morpho-anatomical characterization of embryogenic calluses from immature zygotic embryo of peach palm during somatic embryogenesis =

    Directory of Open Access Journals (Sweden)

    Simone de Alencar Maciel

    2010-04-01

    Full Text Available The objective of this study was to morpho-anatomically characterizenodular embryogenic calluses from zygotic embryos of peach palm during the induction of somatic embryogenesis. Immature zygotic embryos were pre-treated in MS medium added to Picloram and 2,4-D (25 μM and BAP (0, 5, 10 μM. After three months, primary calluses were transferred to MS induction medium added to Picloram and 2,4-D (450 μM. After six months, the embryogenic calluses were then histologically analyzed and cultivated in the maturation medium. The competent tissues of the zygotic embryos differentiated embryogenic calluses under action of both Picloram and 2,4-D auxins (450 μM, where the presence of multi-granular structures were observed. Histological observations showed that in the nodular embryogenic calluses, the outlying parenchymal cells exhibit cellular characteristics of high mitotic activity. Differentiation of tracheal elements exists in embryogenic calluses connecting the callus to the explant. The evaluated cytokinin/auxin interaction influences the development of embryogenic calluses and globular structures.O objetivo deste trabalho foi caracterizar morfoanatomicamente calos nodulares embriogênicos originados de embriões zigóticos de pupunheira durante a indução da embriogênese somática. Embriões zigóticos imaturos de pupunha foram inicialmente pré-tratados em meio de cultura MS, solidificado com 2,5 g L-1 de phytagel® e suplementado com Picloram e 2,4-D na concentração de 25 μM e BAP (0, 5, 10 μM. Após três meses, os calos primários foram transferidos para meio de indução, com Picloram e 2,4-D (450 μM. Após seis meses, os calosnodulares embriogênicos formados foram então analisados histologicamente e repicados para o meio de maturação para a progressão das estruturas multigranulares embriogênicas. Verificou-seque os tecidos competentes dos embriões zigóticos imaturos diferenciaram nódulos embriogênicos pela ação de ambas

  10. Plant germination and production of callus from the yellow hornpoppy (Glaucium flavum): the first stage of micropropagation.

    Science.gov (United States)

    Mohamed, M E; Arafa, A M; Soliman, S S; Eldahmy, S I

    2014-09-01

    The yellow hornpoppy, Glaucium flavum Cr. (Fam. Papaveraceae) is a perennial herb, distributed in the Mediterranean region, including Egypt. The plant contains many benzyl isoquinoline alkaloids from the aporphine type such as glaucine, isoboldine, 1-chelidonine, 1-norchelidonine and 3-O-methylarterenol, making it to display various medicinal activities including antitussive, anticancer, antioxidant, antimicrobial, antiviral, hypoglycemic, analgesic, antipyretic, bronchodilator and anti-inflammatory effects. The plant is now rare and endangered in the Egyptian flora due to urban sprawl. The present study looks into Glaucium flavum seeds' in vitro germination as well as the ability of the explants taken from the growing seedlings to form stable callus lines in order to enable micropropagation as a way to save the rare plant. The study also scans the production of different medicinally valuable alkaloids, particularly glaucine, in produced callus.

  11. Marketing Novel Fruit Products

    NARCIS (Netherlands)

    ’T Riet, Van Jonathan; Onwezen, M.C.; Bartels, Jos; Lans, Van Der I.A.; Kraszewska, Magdalena

    2016-01-01

    The purpose of this study was to compare the influence of four different marketing claims and price information on consumers’ product choices for novel fruits and novel fruit products, using a choice experiment. In total, 1,652 people in Greece (n = 400), the Netherlands (n = 419), Poland (n =

  12. Marketing Novel Fruit Products

    NARCIS (Netherlands)

    ’T Riet, Van Jonathan; Onwezen, M.C.; Bartels, Jos; Lans, Van Der I.A.; Kraszewska, Magdalena

    2016-01-01

    The purpose of this study was to compare the influence of four different marketing claims and price information on consumers’ product choices for novel fruits and novel fruit products, using a choice experiment. In total, 1,652 people in Greece (n = 400), the Netherlands (n = 419), Poland (n = 42

  13. [Effects of arbuscular mycorrhiza fungi (AMF) on the plant growth, fruit yield, and fruit quality of cucumber under salt stress].

    Science.gov (United States)

    Han, Bing; Guo, Shi-Rong; He, Chao-Xing; Yan, Yan; Yu, Xian-Chang

    2012-01-01

    By adopting organic substrate culture, and salt-sensitive cucumber variety 'Jinchun No. 2' was used as test material, this paper studied the effects of inoculating arbuscular mycorrhiza fungi (AMF) on the plant growth, fruit yield, and fruit quality of cucumber under salt stress. AMF-inoculation could effectively promote the plant growth and nutrient uptake, and improve the fruit yield and fruit nutrient quality, compared with ordinary cultivation. Under salt stress, the plant growth was inhibited, and the plant N, P, K, Cu, and Zn contents and K+/Na+ ratio, fruit yield, and fruit soluble protein, total sugar, vitamin C, and nitrate contents decreased, while inoculation with AMF could mitigate the inhibitory effect of salt stress on the plant growth, made the plant N, P, K, Cu, and Zn contents increased by 7.3%, 11.7%, 28.2%, 13.5%, and 9.9%, respectively, and made the plant K+/Na+ ratio, fruit yield, and fruit soluble protein, total sugar, and vitamin C contents have an obvious increase and the fruit nitrate content have a significant decrease. It was suggested that AMF could promote the plant growth and nutrient uptake of cucumber under salt stress, increase the plant salt-tolerance, and improve the fruit yield and its nutrient quality.

  14. Relationship among bone mineral density, collagen composition, and biomechanical properties of callus in the healing of osteoporotic fracture

    Institute of Scientific and Technical Information of China (English)

    SHEN Bin; MU Jian-xiong; PEI Fu-xing

    2007-01-01

    Objective: To study the change and relationship among bone mineral density (BMD), collagen composition and biomechanical properties of the callus in the healing process of osteoporotic fracture.Methods: The osteoporotic rat model and fracture model were established through bilateral ovariectomy(OVX) and osteotomy of the middle shaft of the right hind tibiae, respectively. Ninety female SD rats were randomly divided into OVX group and sham group. With the samples of blood and callus, roentgenoraphic and histological observation were performed for the assessment of the healing progress of the fracture, and the serum concentration of TRAP-5b, proportion of type Ⅰ collagen,BMD and biomechanical properties of the callus were measured.Results: The OVX group experienced a significant delay of fracture healing. The mean serum concentration of TRAP-5b of rats in the OVX group was much higher than that in the sham group after the operation (P < 0.05), but the difference at the same time point after fracture was smaller than that before fracture (P < 0.05 ). The BMD of the callus in both groups reached the peak value at the 6 th week after fracture while the proportion of the type Ⅰ collagen and the biomechanical strength reached the peak at the 8th week.Conclusions: The deficiency of estrogen after the ovariectomy could induce the up-regulation of the osteoclasts activities, whereas the potency of further activation after fracture was depressed. Although the synthesis of collagen together with its mineralization determines the biomechanical properties of new bone, the accumulation of collagen could be assessed as an index in the prediction of biomechanical strength of bones independent of the bone mineral deposition.

  15. Nitric oxide increases mitochondrial respiration in a cGMP-dependent manner in the callus from Arabidopsis thaliana.

    Science.gov (United States)

    Wang, Xiaomin; Li, Jisheng; Liu, Jie; He, Wenliang; Bi, Yurong

    2010-11-01

    Nitric oxide (NO) acts as a key molecule in many physiological processes in plants. In this study, the roles of NO in mitochondrial respiration were investigated in the calli from wild-type Arabidopsis and NO associated 1 mutant (Atnoa1) which has a reduced endogenous NO level. Long-term exposure of wild-type Arabidopsis callus to sodium nitroprusside (SNP) increased mitochondrial respiration in both cytochrome and alternative pathways. In Atnoa1 callus, the capacity of both the cytochrome pathway and the alternative pathway was lower than that in wild-type callus. Further study indicated that NO enhanced the transcript abundance of genes encoding mitochondrial respiration-chain proteins as well as the protein expression of the NADH-ubiquinone reductase 75 kDa subunit and the alternative oxidase 1/2 in wild-type and Atnoa1 calli. 2-Phenyl-4,4,5,5-tetremethy-limidazolinone-1-oxyl-3-oxide (PTIO), a NO scavenger, inhibited the effects of NO in both calli. Co-incubation of callus with 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), a guanylate cyclase inhibitor, also abolished NO effects. The membrane-permeable cGMP analog 8Br-cGMP mimicked NO effects. Moreover, the alternative pathway showed a higher sensitivity to the cellular cGMP changes than the cytochrome pathway did in gene transcription, protein expression and O(2) consumption. Taken together, NO could enhance mitochondrial respiration in both cytochrome and alternative pathways in a cGMP-dependent manner in Arabidopsis.

  16. 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

  17. Biosynthesis of Silver Nanoparticles Using Taxus yunnanensis Callus and Their Antibacterial Activity and Cytotoxicity in Human Cancer Cells

    Directory of Open Access Journals (Sweden)

    Qian Hua Xia

    2016-09-01

    Full Text Available Plant constituents could act as chelating/reducing or capping agents for synthesis of silver nanoparticles (AgNPs. The green synthesis of AgNPs has been considered as an environmental friendly and cost-effective alternative to other fabrication methods. The present work described the biosynthesis of AgNPs using callus extracts from Taxus yunnanensis and evaluated their antibacterial activities in vitro and potential cytotoxicity in cancer cells. Callus extracts were able to reduce silver nitrate at 1 mM in 10 min. Transmission electron microscope (TEM indicated the synthesized AgNPs were spherical with the size range from 6.4 to 27.2 nm. X-ray diffraction (XRD confirmed the AgNPs were in the form of nanocrystals. Fourier transform infrared spectroscopy (FTIR suggested phytochemicals in callus extracts were possible reducing and capping agents. The AgNPs exhibited effective inhibitory activity against all tested human pathogen bacteria and the inhibition against Gram-positive bacteria was stronger than that of Gram-negative bacteria. Furthermore, they exhibited stronger cytotoxic activity against human hepatoma SMMC-7721 cells and induced noticeable apoptosis in SMMC-7721 cells, but showed lower cytotoxic against normal human liver cells (HL-7702. Our results suggested that biosynthesized AgNPs could be an alternative measure in the field of antibacterial and anticancer therapeutics.

  18. In Vitro Callus Induction and Regeneration Potentiality of Aromatic Rice (Oryza sativa L. Cultivars in Differential Growth Regulators

    Directory of Open Access Journals (Sweden)

    Zahida Yesmin Roly

    2014-06-01

    Full Text Available Aromatic rice (Oryza sativa L. cultivars are strong aromatic rice cultivars which can thrive well in rice fields prone to flood, drought and other soil constraints. The present investigation was undertaken to determine a suitable media compositions for callus induction and regeneration using immature embryo of six aromatic grown rice cultivars of Bangladesh, namely, Chinigura, Kalijira, Radhuni Pagal, Modhumala, Kataribog and Mohonbhog. For callus induction different concentrations and combinations of 2, 4-Dichlorophenoxyaceticacid (2, 4-D along with NAA were evaluated. Maximum callus induction (97.22% was observed in Kalijira when 2 mg/L of 2, 4-D and 0.5 mg/l NAA was used and less Modhumala (66.67% and remaining cultivars showed moderate. For regeneration initially different concentrations and combinations of 6-BenzylAminoPurine (BAP and Indole-3-Acetic Acid (IBA were tested. Maximum regeneration frequency (91.67% was observed Kalijira when the optimum concentrations and combinations of 0.5 mg/l of BAP + 0.1 of mg/l IBA were used. Presently optimized regeneration method holds promise for facilitating the deployment of agronomical important trait through genetic transformation for the improvement of this important food crops.

  19. Regeneration of roots from callus reveals stability of the developmental program for determinate root growth in Sonoran Desert Cactaceae.

    Science.gov (United States)

    Shishkova, Svetlana; García-Mendoza, Edith; Castillo-Díaz, Vicente; Moreno, Norma E; Arellano, Jesús; Dubrovsky, Joseph G

    2007-05-01

    In some Sonoran Desert Cactaceae the primary root has a determinate root growth: the cells of the root apical meristem undergo only a few cell division cycles and then differentiate. The determinate growth of primary roots in Cactaceae was found in plants cultivated under various growth conditions, and could not be reverted by any treatment tested. The mechanisms involved in root meristem maintenance and determinate root growth in plants remain poorly understood. In this study, we have shown that roots regenerated from the callus of two Cactaceae species, Stenocereus gummosus and Ferocactus peninsulae, have a determinate growth pattern, similar to that of the primary root. To demonstrate this, a protocol for root regeneration from callus was established. The determinate growth pattern of roots regenerated from callus suggests that the program of root development is very stable in these species. These findings will permit future analysis of the role of certain Cactaceae genes in the determinate pattern of root growth via the regeneration of transgenic roots from transformed calli.

  20. Short-term effects of carbon dioxide on carnation callus cell respiration. [Dianthus Caryophyllus L. ; Elodea canadensis

    Energy Technology Data Exchange (ETDEWEB)

    Palet, A.; Ribas-Carbo, M.; Argiles, J.M.; Azcon-Bieto, J. (Univ. de Barcelona (Spain))

    1991-06-01

    The addition of potassium bicarbonate to the electrode cuvette immediately stimulated the rate of dark O{sub 2} uptake of photomixotrophic and heterotrophic carnation (Dianthus caryophyllus L.) callus, of Elodea canadensis (Minchx) leaves, and of other plant tissues. This phenomenon occurred at pH values lower than 7.2 to 7.8, and the stimulation depended on the concentration of gaseous CO{sub 2} in the solution. These stimulatory responses lasted several minutes and then decreased, but additional bicarbonate or gaseous CO{sub 2} again stimulated respiration, suggesting a reversible effect. Carbonic anhydrase in the solution increased the stimulatory effect of potassium bicarbonate. The CO{sub 2}/bicarbonate dependent stimulation of respiration did not occur in animal tissues such as rat diaphragm and isolated hepatocytes, and was inhibited by salicylhydroxamic acid in carnation callus cells and E. canadensis leaves. This suggested that the alternative oxidase was engaged during the stimulation in plant tissues. The cytochrome pathway was severely inhibited by CO{sub 2}/bicarbonate either in the absence or in the presence of the uncoupler carbonylcyanide m-chlorophenyl hydrazone. The activity of cytochrome c oxidase of callus tissue homogenates was also inhibited by CO{sub 2}/bicarbonate. The results suggested that high carbon dioxide levels (mainly free CO{sub 2}) partially inhibited the cytochrome pathway (apparently at the oxidase level), and this block in electron transport elicited a large transient engagement of the alternative oxidase when present uninhibited.

  1. Production of plant virus inhibitor by Phytolacca americana suspension culture.

    Science.gov (United States)

    Misawa, M; Hayashi, M; Tanaka, H

    1975-09-01

    The inhibitory activity of tobacco mosaic virus (TMV) infection was assayed with the extracts of various callus tissues derived from the intact plants. Phytolacca americana callus was selected as a producer of the virus inhibitor and its cultural conditions in suspension were examined for cell growth and the inhibitor production. A modified liquid medium containing twofold concentrations of all components in that of Murashige and Skoog plus2,4-D (1.0 mg/liter) and sucrose (6%), but without any vitamins and glycine was chosen for production of higher levels of the inhibitor. TMV infections in tobacco, bean, and tomato plants were markedly inhibited by the introduction of the disrupted whole broth of suspension cultured P. americana.

  2. Rhodiola rosea in vitro culture - phytochemical analysis and antioxidant action

    Directory of Open Access Journals (Sweden)

    Mirosława Furmanowa

    2014-01-01

    Full Text Available Various Rhodiola rosea organs and tissues from in vitro culture were studied in two areas: searching for the biological active four phenolic compounds and measurement of antioxidant activity of dry residues of EtOH-extracts soluble in PBS using chemiluminescence method. For phytochemical investigation HPLC method was used. Salidroside was typical for organs of intact plant, rosavin for roots of different origin and shoots of intact plants, triandrin was more typical for tissue cultured in vitro, but it was found in all studied samples. Caffeic acid was detected mainly in green and yellow lines of callus, less in other Rhodiola rosea plant material. The highest antioxidant activity showed the extract from 1,5-years-old callus of green and yellow line, which contained caffeic acid and triandrin. Further observations are in progress.

  3. Androgenesis in anther culture of Lithuanian spring barley cultivars

    OpenAIRE

    Asakavičiūtė, Rita; Pašakinskienė, Izolda

    2006-01-01

    The method of anther culture was used for the production of doubled haploids in Lithuanian spring barley cultivars. Two methods, (i) regeneration from callus (Szarjeko’s method) and (ii) direct regeneration from embryoids (Caredda’s method) were applied to determine the androgenic potential according to the green regenerant yield and other morphogenetic factors. Green double haploid regenerants were obtained in four Lithuanian spring barley cultivars (‘Aura’, ‘Aidas’, ‘Alsa’ and ‘Auksiniai’) ...

  4. Establishment of suspension cell culture of Gymnema sylvestre R.Br.- A threatened anti-diabetic plant

    Directory of Open Access Journals (Sweden)

    Karthic Raju

    2012-04-01

    Full Text Available A cell suspension culture was established from leaf explants of wild Gymnema sylvestre plants collected from Muniyankudisai,Tamilnadu, India. Murashige and Skoog medium supplemented with 9.0 μM l-1 of 2, 4- Dichlorophenoxy acetic acid and 2.1 μM l-1Benzyl adenine produced yellow friable callus with green patches.The cells were subcultured conscientiously twelve times to getconsistent growth of the cells in suspension. From the 10thsubculture onwards callus cells acclimatized to grow in suspensionwith aggregation and reached 168.6 g l-1 fw and 5.16 g l-1 dw of cellbiomass.

  5. Microstructures of external periosteal callus of repaired femoral fracture in children.

    Science.gov (United States)

    Cui, F Z; Wen, H B; Su, X W; Zhu, X D

    1996-01-01

    In order to understand further the mechanism of bone fracture repair, and thus to innovate better operative treatment for bone fracture and to design new implant materials for bone repair, microstructures of external periosteal callus (EPC) of repaired femoral fracture in both children and adults were investigated by using a scanning electron microscope, transmission electron microscopy, and an X-ray microdiffractometer. The repair time after the fractures in children and adults is on average 155 and 370 days, respectively. Collagen fibrils making up children's EPC (CEPC) are underdeveloped and insufficiently mineralized by hydroxyapatite (HA), while those from adults' EPC (AEPC) are similar to normal bone. A lot of particles loaded by brushite (DCPD) minerals were found among the collagen fibrils of CEPC. The main mineral phases in CEPC consist of DCPD and HA, while only HA exists in AEPC. Deposition of DCPD minerals could have compensated for the insufficient mineralization of the collagen fibrils of CEPC, thereby making fractured bone repair more rapidly in children than in adults.

  6. Temporomandibular joint ankylosis caused by chondroid hyperplasia from the callus of condylar neck fracture.

    Science.gov (United States)

    Kim, Soung Min; Park, Jung Min; Kim, Ji Hyuck; Kwon, Kwang Jun; Park, Young Wook; Lee, Jong Ho; Lee, Sang Shin; Lee, Suk Keun

    2009-01-01

    A patient who complained of difficulty in opening his mouth after condylar neck fracture 1 year ago presented typical features of temporomandibular joint ankylosis in clinical and radiologic examinations. To demonstrate a possible pathogenesis of temporomandibular joint ankylosis after condylar neck fracture, the fractured condylar portion removed was examined by histologic and immunohistochemical stainings. Interpositional gap arthroplasty was performed by removing the inferomesially displaced fractured condyle, and reconstruction with subcutaneous dermis to the previous vertical height was performed immediately. The fractured condylar portion was almost intact with slight erosion of the condylar cartilage. In the hematoxylin and eosin and Masson trichrome stainings, an extensive chondroid hyperplasia with abundant hyaline cartilage was shown in the removed condylar portion. There were also hyperplastic features of the synovial membrane, which were abnormally distributed throughout the chondroid tissues. In the immunohistochemical stainings of proliferating cell nuclear antigen (PCNA) and bone morphogenetic protein (BMP)-2 and BMP-4, the chondroid tissues were conspicuously hyperplastic and strongly positive for BMP-4 but sparse for BMP-2. From these results, we think that the hyperplastic chondroid tissue was derived from the callus of the primary fractured site of the condylar neck and propose that the chondroid tissue could proliferate continuously because of synovial tissue support from around the temporomandibular joint, resulting in temporomandibular joint ankylosis. This pathogenesis is quite different from those of other diaphyseal fracture of long bones.

  7. Consumer acceptance of novel fruits and fruit products

    NARCIS (Netherlands)

    Bakker, T.; Benninga, J.; Rakowska, J.; Bartels, J.

    2010-01-01

    The task of the Deliverable 1.3.7 Report on case studies of fruit innovations is to provide information on consumers' acceptance of innovative fruit and fruit products selected for case studies in Deliverable 1.3.2 List of selected fruit innovations, and to validate findings from previous stages of

  8. Caribbean Fruit Fly (Diptera: Tephritidae) and Small Fruit in Florida

    Science.gov (United States)

    Tephritid fruit flies are among the most important pests of fruits and vegetables worldwide. The Caribbean fruit fly, Anastrepha suspensa (Loew), is a tephritid pest that became established in Florida following introduction in 1965. Populations of this fruit fly also occur in Puerto Rico and Cuba, ...

  9. Induction of a photomixotrophic plant cell culture of Helianthus annuus and optimization of culture conditions for improved α-tocopherol production.

    Science.gov (United States)

    Geipel, Katja; Song, Xue; Socher, Maria Lisa; Kümmritz, Sibylle; Püschel, Joachim; Bley, Thomas; Ludwig-Müller, Jutta; Steingroewer, Juliane

    2014-03-01

    Tocopherols, collectively known as vitamin E, are lipophilic antioxidants, which are synthesized only by photosynthetic organisms. Due to their enormous potential to protect cells from oxidative damage, tocopherols are used, e.g., as nutraceuticals and additives in pharmaceuticals. The most biologically active form of vitamin E is α-tocopherol. Most tocopherols are currently produced via chemical synthesis. Nevertheless, this always results in a racemic mixture of different and less effective stereoisomers because the natural isomer has the highest biological activity. Therefore, tocopherols synthesized in natural sources are preferred for medical purposes. The annual sunflower (Helianthus annuus L.) is a well-known source for α-tocopherol. Within the presented work, sunflower callus and suspension cultures were established growing under photomixotrophic conditions to enhance α-tocopherol yield. The most efficient callus induction was achieved with sunflower stems cultivated on solid Murashige and Skoog medium supplemented with 30 g l(-1) sucrose, 0.5 mg l(-1) of the auxin 1-naphthalene acetic acid, and 0.5 mg l(-1) of the cytokinin 6-benzylaminopurine. Photomixotrophic sunflower suspension cultures were induced by transferring previously established callus into liquid medium. The effects of light intensity, sugar concentration, and culture age on growth rate and α-tocopherol synthesis rate were characterized. A considerable increase (max. 230%) of α-tocopherol production in the cells was obtained within the photomixotrophic cell culture compared to a heterotrophic cell culture. These results will be useful for improving α-tocopherol yields of plant in vitro cultures.

  10. Occurrence of iridoid glycosides in in vitro cultures and intact plants of Scrophularia nodosa L.

    Science.gov (United States)

    Sesterhenn, Katja; Distl, Melanie; Wink, Michael

    2007-03-01

    Shoot, root, and callus cultures of Scrophularia nodosa L. (Scrophulariaceae) were established and cultivated in vitro. Iridoid glycosides, such as harpagoside, aucubin, and catalpol were identified by LC-ESI-MS and their contents determined by HPLC. For comparison intact plants of S. nodosa were analysed. In shoot cultures slightly lower amounts of detectable iridoid glycosides (4.36% dry weight) were determined than in the field grown plants (4.88%). Concentration of harpagoside was highest in leaves of field plants (1.05%) and in flowers of in vitro plantlets (1.10%). For aucubin the highest amount was found in the leaves of in vitro plantlets (1.67%) whereas the levels of aucubin in the leaves of field plants were remarkably lower. Catalpol was produced as a trace compound in intact plants and shoot cultures. Callus and root cultures were apparently not able to synthesise iridoid glycosides.

  11. Variation in bioactive principles of Artemisia amygdalina Decne. in wild and tissue culture regenerants.

    Science.gov (United States)

    Rasool, Rafia; Ganai, Bashir Ahmad; Akbar, Seema; Kamili, Azra Nahaid; Dar, Muhammad Younus; Masood, Akbar

    2013-05-01

    Wild and tissue culture raised regenerants of Artemisia amygdalina, a critically endangered and endemic plant of Kashmir and North West Frontier Provinces of Pakistan were screened for the amount of bioactive principles and in particular antimalarial compound artemesinin. Phytochemical screening of extracts revealed the presence of terpenes, alkaloids, phenolics, tannins (polyphenolics), cardiac glycosides and steroids in wild (aerial, inflorescence) and tissue culture regenerants (in vitro grown plant, callus and green house acclimatized plants). HPLC of Artemisia amygdalina revealed the presence of artemesinin in petroleum ether extracts of wild aerial part, tissue culture raised plant and green house acclimatized plants. Acetonitrile and water in 70:30 ratios at flow rate of 1ml/min was standardised as mobile phase. Retention time for standard chromatogram was 6.7. Wild inflorescences and callus does not produce artemesinin. This is the first report of phytochemical screening and artemesinin estimation of wild and tissue culture raised regenerants of Artemisia amygdalina.

  12. Evaluation of limonoid production in suspension cell culture of Citrus sinensis

    Directory of Open Access Journals (Sweden)

    Elisângela Fumagali Gerolino

    2015-10-01

    Full Text Available ABSTRACTThe use of cell and plant tissue culture techniques to produce economically important active metabolites has been growing. Among these substances are total limonoid aglycones, which are produced by "pera" orange (Citrus sinensis (L. Osbeck, Rutaceae and have received considerable attention because of their anticancer actions. The main objective of the present study was to analyze and compare the levels of limonoid aglycones in seeds, callus cultures (originating from seeds, callus cultures (originating from hypocotyls, cell suspensions from hypocotyls cells, and cell suspensions from cotyledons. The cell cultures or C. sinensis were obtained by inoculating two strains of callus in MS medium supplemented with 2.0 µM 2,4-dichlorophenoxyacetic acid, 7.0 µM benzyl aminopurine, and 3% (w/v sucrose in the dark. The highest concentrations of limonoid aglycone that were obtained were observed in cotyledon cell lines (240 mg/100 g dry weight that were produced on day 21 of culture and hypocotyl cell lines on day 7 (210 mg/100 g dry weight. Explants of different origins under the same culture conditions had different limonoid aglycone content. The present results may suggest strategies for enhancing the productivity of biologically important limonoid aglycones and investigating the complex pathways of these secondary metabolites in plant tissue cultures.

  13. Social attraction mediated by fruit flies' microbiome.

    Science.gov (United States)

    Venu, Isvarya; Durisko, Zachary; Xu, Jianping; Dukas, Reuven

    2014-04-15

    Larval and adult fruit flies are attracted to volatiles emanating from food substrates that have been occupied by larvae. We tested whether such volatiles are emitted by the larval gut bacteria by conducting tests under bacteria-free (axenic) conditions. We also tested attraction to two bacteria species, Lactobacillus brevis, which we cultured from larvae in our lab, and L. plantarum, a common constituent of fruit flies' microbiome in other laboratory populations and in wild fruit flies. Neither larvae nor adults showed attraction to axenic food that had been occupied by axenic larvae, but both showed the previously reported attraction to standard food that had been occupied by larvae with an intact microbiome. Larvae also showed significant attraction to volatiles from axenic food and larvae to which we added only either L. brevis or L. plantarum, and volatiles from L. brevis reared on its optimal growth medium. Controlled learning experiments indicated that larvae experienced with both standard and axenic used food do not perceive either as superior, while focal larvae experienced with simulated used food, which contains burrows, perceive it as superior to unused food. Our results suggest that flies rely on microbiome-derived volatiles for long-distance attraction to suitable food patches. Under natural settings, fruits often contain harmful fungi and bacteria, and both L. brevis and L. plantarum produce compounds that suppress the growth of some antagonistic fungi and bacteria. The larval microbiome volatiles may therefore lead prospective fruit flies towards substrates with a hospitable microbial environment.

  14. FRUIT NUTRITIVE FACTOR NETWORK

    Institute of Scientific and Technical Information of China (English)

    Yanqing QU; Yumei JIANG; Daren HE

    2009-01-01

    As an research example of the widely existing cooperation-competition systems, the authors present an empirical investigation on a fruit nutritive factor network. It is described by a node-weighted bipartite graph. The fruit nutritive factors are defined as the nodes, and two nodes are connected by an edge if at least one fruit contains these two nutritive factors. The fruits are defined as the collaboration acts. The node-weight Writ, which signifies the "importance degree" of each actor node, is defined as the content of a nutritive factor in a fruit. The empirical investigation results show some unique features.The node-weight distributions take so-called "shifted power law" function forms, but the act-weight distribution takes a normal form. The degree and act-degree distributions show impulsive-spectrum like forms. These observations may be helpful for the study of fruits. The network description method proposed in this article may be universal for a kind of cooperation-competition systems.

  15. 湖北麦冬花药愈伤组织诱导及再生植株的获得%Callus induction and plant regeneration of Liriope spicata var. prolifera anther

    Institute of Scientific and Technical Information of China (English)

    周群; 周健丘; 王小刚; 王菁菁; 李敬文; 陈家春

    2009-01-01

    Objective: To study the technique of the callus induction from anther and plant regeneration of medicinal plants Liriope spicata var. prolifera. Method: Callus was induced from anther of L. spicata var. prolifera on a MS medium supplemented with different hormones. The squash methods combined with a microscope were used to analyze chromosomes of regenerated plantlets.regenerated plantlets originated from somatic cells. At the same time, the effects of pretreatment of low temperature at 4 ℃ on the callus induction were studied and discussed. Conclusion: This paper sets up the method of tissue culture of anther somatic-cells and intermediate propagation of L. spicata var. prolifera.%目的:探讨药用植物湖北麦冬花药愈伤组织诱导和植株再生条件.方法:以湖北麦冬花药为外植体,采用MS培养基,附加不同的植物激素进行实验.常规压片法结合显微镜进行再生植株染色体的计数分析.结果:MS+2,4-D 1.0 mg·L~(-1)+KT 2.0 mg·L~(-1)诱导愈伤组织效果最好,愈伤组织诱导率可达41.07%.MS+6-BA 1.5~2.0 mg·L~(-1)+NAA0.1~0.3mg·L~(-1)适于不定芽的诱导,不定芽转入附加NAA 0.1