A simple and efficient Agrobacteriumtumefaciens-mediated plant transformation of Brassica rapa ssp. pekinensis

• Published in: 3 Biotech 2016-03, Vol.6 (1)
• Main Authors: Baskar, Venkidasamy, Gangadhar, Baniekal H., Park, Se Won, Nile, Shivraj Hariram
• Format: Article
• Language: English
• Subjects: Agriculture; Bioinformatics; Biomaterials; Biotechnology; Cancer Research; Chemistry; Chemistry and Materials Science; Original; Original Article; Stem Cells
• ISSN: 2190-572X; 2190-5738
• DOI: 10.1007/s13205-016-0402-1

The present study aims to investigate the numerous factors influencing Agrobacterium tumefaciens -mediated genetic transformation of Chinese cabbage ( Brassica rapa ssp. pekinensis ). Factors affecting transformation efficiency, such as age of explants, Agrobacterium concentration, and effect of acetosyringone, pre-cultivation, infection and co-cultivation time of Agrobacterium were examined. The pre-cultured hypocotyls from young seedlings prior to exposure to Agrobacterium showed higher shoot regeneration. The plant transformation with the modest A. tumefaciens concentrations (0.8 OD) and the 3 days co-cultivation periods increased transformation efficiency. Plant growth hormones [1-naphthyl acetic acid (NAA) and 6-benzyl amino purine (BAP)] were essential for callus and shoot formation. Root formation was effective in half strength MS medium without supplementation of root-inducing hormones. To maintain selection pressure, plant subculture was carried out every 2 weeks with selective antibiotics. The putative transgenic plants were acclimatized in the greenhouse. Polymerase chain reaction was performed to confirm the integration of T-DNA into the genome of transgenic plants. A transformation efficiency of 15 % was obtained. This protocol allows effective transformation and indirect regeneration of Brassica rapa .