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Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (12): 2185-2191.doi: 10.3724/SP.J.1006.2012.02185

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Establishment of a Transformation System Using Inbred Line of Yellow-Seeded Brassica napus

LIN Na,LIU Lie-Zhao,YIN Jia-Ming,WANG Rui,CHAI You-Rong,LI Jia-Na*   

  1. College of Agronomy and Biotechnology, Southwest University / Chongqing Rapeseed Engineering & Technology Research Center / Engineering Research Center of South Upland Agriculture, Ministry of Education, Chongqing 400716, China
  • Received:2011-12-01 Revised:2012-07-05 Online:2012-12-12 Published:2012-10-08
  • Contact: 李加纳, E-mail: ljn1950@swu.edu.cn

Abstract:

In this research, we established a transformation system using an inbred line of yellow-seeded Brassica napus. Hypocotyl explants precultured for 5–7 d on Murashige and Skoog medium containing 200 μmol L–1 acetosyringone were cocultured with Agrobacterium tumefaciens strain LBA4404 (pCNR) for 63–69 hours.The plasmid pCNR was constructed by inserting Δ6-fatty acid desaturase gene from Rhizopusstolonifer into plant high-efficient expression vector pCAMBIA2301G. Kanamycin-tolerant shoots were regenerated on shoot induction medium for three months after Agrobacterium inoculation. The average transformation efficiency was about 1.3% under optimal conditions. Results from GUS assay and PCR analysis of transformed plants indicated that the introduced genewas integrated into B. napus genomes.The Southern blot revealed that those transformants carried one or two copies of the goal gene. The fatty acids of the transgenic plant seeds were analyzed by GC, and the γ-linolenic content was 8.2%.

Key words: Brassica napus, Δ6-fatty acid desaturase, Transformation

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