Successful genetic transformation of Chinese cabbage using phosphomannose isomerase as a selection marker.
A mannose selection system was adapted for use in the Agrobacterium-mediated transformation of Chinese cabbage. This system makes use of the pmi gene that encodes phosphomannose isomerase, which converts mannose-6-phosphate to fructose-6-phosphate. Hypocotyl explants from 4-5-day-old seedlings of Chinese cabbage inbred lines were pre-cultured for 2-3 days and then infected with Agrobacterium. Two genes (L: -guluno-gamma-lactone oxidase, GLOase, and jasmonic methyl transferase, JMT) were transformed into Chinese cabbage using the transformation procedure developed in this study. We found that supplementing the media with 7 g l(-1) mannose and 2% sucrose provides the necessary conditions for the selection of transformed plants from nontransformed plants. The transformation rates were 1.4% for GLOase and 3.0% for JMT, respectively. The Southern blot analysis revealed that several independent transformants (T (0)) were obtained from each transgene. Three different inbred lines were transformed, and most of the T (1) plants had normal phenotypes. The transformation method presented here for Chinese cabbage using mannose selection is efficient and reproducible, and it can be useful to introduce a desirable gene(s) into commercially useful inbred lines of Chinese cabbage.
Min
Byung-Whan
BW
Department of Plant Resources, Sangju National University, Sangju, Gyeongbuk, Korea.
Cho
Yi-Nam
YN
Song
Min-Jung
MJ
Noh
Tae-Kyung
TK
Kim
Bong-Kyu
BK
Chae
Won-Ki
WK
Park
Young-Soo
YS
Choi
Yang-Do
YD
Harn
Chee-Hark
CH
D001937:Brassica / Q000235:genetics*; D015870:Gene Expression; D005818:Genetic Engineering; D005819:Genetic Markers / Q000235:genetics; D050544:L-Gulonolactone Oxidase / Q000235:genetics; D008358:Mannose; D008359:Mannose-6-Phosphate Isomerase / Q000032:analysis / Q000235:genetics*; D008780:Methyltransferases / Q000235:genetics; D012231:Rhizobium; D012641:Selection, Genetic; D014170:Transformation, Genetic
D005819:Genetic Markers; D050544:L-Gulonolactone Oxidase; D008780:Methyltransferases; C425043:jasmonic acid carboxyl methyltransferase; D008359:Mannose-6-Phosphate Isomerase; D008358:Mannose