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Leaf Explants (leaf + explant)

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Life Sciences	88%

Selected Abstracts

Factors affecting adventitious regeneration from in vitro leaf explants of ,Improved French' plum, the most important dried plum cultivar in the USA

ANNALS OF APPLIED BIOLOGY, Issue 1 2010
C. Petri
An adventitious shoot regeneration protocol from in vitro leaves of the most important dried plum cultivar in the USA, ,Improved French', has been established. Factors affecting regeneration were studied in order to optimise regeneration. The proliferation medium in which the shoots, used as the source of leaf explants, were cultured had a strong influence on subsequent regeneration. Shoot regeneration was observed at a mean frequency of 52% when a Murashige-based and Skoog-based shoot culture medium with 3 ,M N6 -benzylaminopurine and 0.25 ,M indole-3-butyric acid (IBA) was employed compared with shoot regeneration frequencies of less than 5% for a Quoirin-based and Lepoivre-based shoot culture medium, with 8.9 ,M N6 -benzylaminopurine and 0.49 ,M IBA. The shoot regeneration medium contained ,-naphthaleneacetic acid at 2.0,6.0 ,M and thidiazuron at 4.5,15.0 ,M. 2,4 Dichlorophenoxy-acetic acid at 9.0 ,M was included in the medium but only for the first 4 days of culture. Shoot regeneration frequencies were positively related to thidiazuron concentration and significantly greater (P < 0.05) for 9,15 ,M thidiazuron than for the media with 4.5 ,M thidiazuron. Leaf explants, incubated in a 16-h-light/8-h-dark photoperiod or in the dark for 1 week followed by exposure to light, showed significantly more organogenic activity (P < 0.01) than was observed for leaves cultured in the dark for 2 or 3 weeks before they were transferred to the light. The utilisation of Bacto agar (0.7%) as the gelling agent increased organogenesis compared with media gelled with TC Agar (0.7%), or an agar,gellan gum blend (AgargelÔ) (0.45%). The addition of the ethylene inhibitor silver thiosulphate at 60,120 ,M also improved organogenesis. When all the studied factors were optimised, a regeneration rate of 65% was achieved. Rooting frequency of regenerated shoots was significantly increased (P < 0.05) by the use of full-strength Murashige and Skoog salts (40%) or 100 mg L,1 phloroglucinol (53%) to the rooting medium. [source]

Characteristics of an isomenthone-rich somaclonal mutant isolated in a geraniol-rich rose-scented geranium accession of Pelargonium graveolens

FLAVOUR AND FRAGRANCE JOURNAL, Issue 5 2001
Ritika Gupta
Abstract A somaclonal essential oil mutant (IRPG) was identified among the regenerants induced in callus cultures initiated with the leaf explants of geranium Pelargonium graveolens cv. Kunti. While the shoot essential oil of the wild-type parent was rich in geraniol, the oil of the IRPG had isomenthone as the major monoterpenoid component. The IRPG oil had about 71% isomenthone, 6% citronellol and 3% geraniol, as compared to the parental variety oil, in which isomenthone, citronellol and geraniol contents were about 8%, 13% and 40%, respectively. Copyright © 2001 John Wiley & Sons, Ltd. [source]

Genome-wide Characterization of Long Terminal Repeat -retrotransposons in Apple Reveals the Differences in Heterogeneity and Copy Number between Ty1 -copia and Ty3 -gypsy Retrotransposons

JOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 9 2008
Hai-Yue Sun
Abstract The conserved domains of reverse transcriptase (RT) genes of Ty1- copia and Ty3- gypsy groups of long terminal repeat (LTR) retrotransposons were isolated from the Malus domestica genome using degenerate oligonucleotide primers. Sequence analysis showed that 45% of Ty1- copia and 63% of Ty3- gypsy RT sequences contained premature stop codons and/or indels disrupting the reading frame. High heterogeneity among RT sequences of both Ty1- copia and Ty3- gypsy group retrotransposons was observed, but Ty3- gypsy group retrotransposons in the apple genome are less heterogeneous than Ty1- copia elements. Retrotransposon copy number was estimated by dot blot hybridizations for Ty1- copia (,5 000) and Ty3- gypsy (,26 000). All elements of the two types of LTR retrotransposons comprise approximately 38% of the M. domestica genome, with the Ty3- gypsy group contribution being higher (33.5%) than the Ty1- copia one (4.6%). Transcription was not detected by reverse transcription-polymerase chain reaction for either Ty1- copia or Ty3- gypsy retrotransposons in the leaves of plants in vitro or in leaf explants cultured on medium supplemented with high concentration benzylaminopurine. This research reveals the differences in heterogeneity and copy number between Ty1- copia and Ty3- gypsy retrotransposons in the apple genome. Ty1- copia retrotransposon has higher heterogeneity than Ty3- gypsy retrotransposon, but the latter has a higher copy number, which implies that Ty3- gypsy retrotransposons may play a more important role in the apple genome evolution. [source]

Development of a strategy for transgenic studies and monitoring of transgene expression in two closely related Moricandia species possessing a C3 or C3,C4 intermediate photosynthetic phenotype

PHYSIOLOGIA PLANTARUM, Issue 1 2003
Vera Thole
In order to establish a model system for comparative studies of C3 and C3,C4 intermediate photosynthesis, the development of efficient transformation systems and the monitoring of transgene behaviour and stability were carried out in two closely related Moricandia species (Brassicaceae): the C3,C4 photosynthetic intermediate species M. arvensis and the C3 species M. moricandioides. In this study the green fluorescent protein (gfp) reporter gene was used as a vital marker gene while the use of the , -glucuronidase (gusA) gene was based on the highly sensitive detection of its activity. For Agrobacterium -mediated transformation of leaf explants, a cauliflower mosaic virus 35S promoter-driven, modified version of gfp, the mgfp5-ER gene and the gusA gene, respectively, were introduced into the new dual binary transformation vector system pGreen/pSoup (Hellens et al. 2000, Plant Mol Bio 42: 819,832). GFP5 produced bright-green fluorescence in transformed tissues that was distinctly detected 5,12 days following transformation in developing calli of the two species. Visual screening, combined with antibiotic selection, enabled early and easy identification of transformation events and contributed to improvements in the transformation strategies. Transgene integration studies demonstrated that mgfp5-ER was inserted with low copy number in the M. arvensis plant lines and the transgene was transmitted in a Mendelian fashion to T1 and T2 progenies. GFP5 expression levels in a population of 100 independent primary transformed M. arvensis plant lines (T0) showed great variation between transformation events (coefficient of variation of 108%). The mgfp5-ER or gusA reporter genes were expressed in 90,95% of the kanamycin-resistant M. arvensis plant lines and in up to 98% of the independent M. moricandioides plant lines. [source]

In vitro selection and plant regeneration of copper-tolerant plants from leaf explants of Nicotiana tabacum L. cv. ,Xanthi'

PLANT BREEDING, Issue 4 2007
G. R. Rout
Abstract Copper tolerance of Nicotiana tabacum L. var. Xanthi in vitro was achieved through plant regeneration from leaf explants on Murashige and Skoog's (MS) medium supplemented with 0.5 mg/l BA, 0.1,0.25 mg/l IAA and 60 ,m Cu. Tolerant organogenic calli showed more vigorous growth in medium containing 60 ,m Cu than the non-tolerant calli. Standard growth parameters such as fresh and dry weight of organogenic callus, growth tolerance index (GTI), enzyme activity (peroxidase and catalase) and copper accumulation were used as indicators of copper tolerance. The activities of peroxidase and catalase as well as estimation of protein, total amino acid and chlorophyll were greater in tolerant calli than non-tolerant ones. The GTI in the 4 weeks after the beginning of treatments yielded significant differences among the tolerant and non-tolerant organogenic callus cultures. The accumulation of copper in the tolerant calli increased significantly with an increase in copper concentration in the medium. Shoot bud regeneration was achieved in both tolerant and non-tolerant organogenic calli on MS medium containing 0.5 mg/l BA and 0.1 mg/l IAA. The tolerant regenerated shoots were rooted on half-strength basal MS medium with 60 ,m Cu for selection of tolerant clones. This study may help in the selection and characterization of Cu-tolerant lines of N. tabacum cv. ,Xanthi' for building conservation strategies and also for phytoremediation programmes. [source]

Factors affecting adventitious regeneration from in vitro leaf explants of ,Improved French' plum, the most important dried plum cultivar in the USA

Alkaloid production in Vernonia cinerea: Callus, cell suspension and root cultures

BIOTECHNOLOGY JOURNAL, Issue 8 2007
Priti Maheshwari
Abstract Fast-growing callus, cell suspension and root cultures of Vernonia cinerea, a medicinal plant, were analyzed for the presence of alkaloids. Callus and root cultures were established from young leaf explants in Murashige and Skoog (MS) basal media supplemented with combinations of auxins and cytokinins, whereas cell suspension cultures were established from callus cultures. Maximum biomass of callus, cell suspension and root cultures were obtained in the medium supplemented with 1 mg/L ,-naphthaleneacetic acid (NAA) and 5 mg/L benzylaminopurine (BA), 1.0 mg/L NAA and 0.1 mg/L BA and 1.5 mg/L NAA, respectively. The 5-week-old callus cultures resulted in maximum biomass and alkaloid contents (750 ,g/g). Cell suspension growth and alkaloid contents were maximal in 20-day-old cultures and alkaloid contents were 1.15 mg/g. A 0.2-g sample of root tissue regenerated in semi-solid medium upon transfer to liquid MS medium containing 1.5 mg/L NAA regenerated a maximum increase in biomass of 6.3-fold over a period of 5 weeks. The highest root growth and alkaloid contents of 2 mg/g dry weight were obtained in 5-week-old cultures. Maximum alkaloid contents were obtained in root cultures in vitro compared to all others including the alkaloid content of in vivo obtained with aerial parts and roots (800 ,g/g and 1.2 mg/g dry weight, respectively) of V. cinerea. [source]