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Development of Agrobacterium-mediated transformation technology for mature seed-derived callus tissues of indica rice cultivar IR64.
- Source :
-
GM Crops & Food . Apr-Jun2012, Vol. 3 Issue 2, p123-128. 6p. - Publication Year :
- 2012
-
Abstract
- Indica rice cultivar IR64 is most recalcitrant to regenerate, which affects the transformation efficiency especially when mature seed-derived callus tissues are used as explants. Therefore, a simple, rapid and improved genetic transformation protocol has been developed for the indica rice cultivar IR64 using Agrobacterium-mediated genetic transformation. With different hormonal combination tested, the maximum callus induction was observed on MS medium supplemented with 2.5 mg/l 2,4-D and 0.15 mg/l BAP from the scutellum explants. Three weeks old scutellum derived callus explants were immersed in Agrobacterium suspension (strain LBA4404, OD600 = 1.0) and co-cultured at 26 ± 2°C in dark for 2 d. The maximum transformation efficiency (12%) was achieved with infection of callus explants for 20 min along with use of 150 &mgr;m acetosyringone. The maximum plant regeneration was observed on MS medium supplemented with 3 mg/l BAP, 1 mg/l Kinetin and 0.5 mg/l NAA. The maximum root induction was observed on MS medium along with 10 g/l glucose and 20 g/l sucrose. The integration of the transgene in T1 transgenic plants was confirmed by polymerase chain reaction and Southern blot analyses. The copy number of transgenes has been found to vary from 1 to 2 in transgenic plants. By using this improved method we have successfully raised transgenic rice plants within 3 mo from seed inoculation to plant regeneration. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 21645698
- Volume :
- 3
- Issue :
- 2
- Database :
- Academic Search Index
- Journal :
- GM Crops & Food
- Publication Type :
- Academic Journal
- Accession number :
- 77410978
- Full Text :
- https://doi.org/10.4161/gmcr.20032