An improved method for Agrobacterium-mediated genetic transformation from cotyledon explants of Brassica juncea
SCIE
SCOPUS
- Title
- An improved method for Agrobacterium-mediated genetic transformation from cotyledon explants of Brassica juncea
- Authors
- Bhuiyan, MSU; Min, SR; Jeong, WJ; Sultana, S; Choi, KS; Lim, YP; Song, WY; Lee, Y; Liu, JR
- Date Issued
- 2011-01
- Publisher
- JAPANESE SOC PLANT CELL & MOLECULAR BIOL
- Abstract
- An efficient Agrobacterium-mediated genetic transformation method was established for Brassica juncea by investigating several factors responsible for successful gene transfer. Four-day-old cotyledon explants from in vitro grown seedlings were co-cultivated with Agrobacterium strain GV3101 harboring the binary vector EnPCAMBIA1302-YCF1, which contained the hygromycin phosphotransferase (HPT) gene as a selectable marker and the yeast cadmium factor 1 (YCF1) gene. Two days co-cultivation period on shoot induction medium (MS medium supplemented with 0.1 mg l(-1) alpha-naphthaleneacetic acid, 1.0 mg l(-1) 6-benzyladenine, and 2.0 mg l(-1) silver nitrate) containing 20 mg l(-1) acetosyringone and five days delaying exposure of explants to selective agent enhanced transformation efficiency significantly. A three-step selection strategy was developed to select hygromycin resistant shoots. Hygromycin-resistant shoots were subsequently rooted on root induction medium. Rooted plantlets were transferred to pot-soil, hardened, and grown in a greenhouse until maturity. Using the optimized transformation procedure, transformation efficiency reached at 16.2% in this study. Southern blot analysis was performed to confirm that transgenes (HPT and YCF1) were stably integrated into the plant genome. All transgenic plants showed single-copy of transgene integration in the host genome. Segregation analysis of T-1 progeny showed that the transgenes were stably integrated and transmitted to the progeny in a Mendelian fashion.
- Keywords
- Cotyledon explants; co-cultivation; genetic transformation; hygromycin; Indian mustard; INDIAN MUSTARD; SHOOT REGENERATION; TRANSGENIC PLANTS; TOLERANCE; L.; TUMEFACIENS; LEAF; ACCUMULATION; ANTIBIOTICS; CADMIUM
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/17503
- DOI
- 10.5511/PLANTBIOTECHNOLOGY.10.0921A
- ISSN
- 1342-4580
- Article Type
- Article
- Citation
- PLANT BIOTECHNOLOGY, vol. 28, no. 1, page. 17 - 23, 2011-01
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- There are no files associated with this item.
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