A simple and efficient Agrobacterium tumefaciens-mediated plant transformation of Brassica rapa ssp. pekinensis.
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.
Department of Bioresources and Food Science, School of Life and Environmental Sciences, Konkuk University, Seoul, 143701, South Korea.
Venkidasamy
V
Baskar
Department of Bioresources and Food Science, School of Life and Environmental Sciences, Konkuk University, Seoul, 143701, South Korea.
Baniekal H
BH
Gangadhar
Department of Bioresources and Food Science, School of Life and Environmental Sciences, Konkuk University, Seoul, 143701, South Korea.
Se Won
SW
Park
Department of Bioresources and Food Science, School of Life and Environmental Sciences, Konkuk University, Seoul, 143701, South Korea. nileshivraj@gmail.com.
Shivraj Hariram
SH
Nile
Acetosyringone; Agrobacterium tumefaciens; Brassica rapa; Cultivation; Transformation