[Preliminary studies on tissue culture and agrobacterium-medicated transformation of Brassica campestris ssp. chinensis].
The hypocotyls and cotyledons of the asepetic seedling of Brassica campestris ssp. chinensis L cv. Pudongaijiecai) were used as explants for tissue culture. Adventitious buds were differentiated on modified MS medium supplemented with TDZ 1-2 mg/L, NAA 0.2-1 mg/L and AgNO3 7.5 mg/L. The percentage of explants which formed buds of cotyledons was about 56%, and that of hypocotyls was about 37%. When the regenerated explants were transferred onto MS medium with 2 i.p. 5 mg/L and NAA 0.1 mg/L for two weeks, whole plantlets were obtained by culturing the regenerated shoots on 1/2 MS medium with NAA 0.1 mg/L. Agrobacterium tumefaciens strain (LBA 4404/PBI 121) carrying the GUS gene and Npt II gene was used for transformation. After 2 days of coculture, the hypocotyls and cotyledons were transferred onto regenerated medium containing CP 300 mg/L for bud formation. After 4-5 weeks, the differentiated buds were transferred onto selection medium with CP 200 mg/L and Km 10 mg/L for 1 month, then the green shoots were transferred onto the rooting medium containing Cef 100 mg/L and Km 20 mg/L. 4-5 weeks later, plantlets with Km resistance were obtained and some of them showed higher enzymatic activities of beta-glucuronidase than control ones.
Wang
L J
LJ
Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shanghai 200032.
Ni
D A
DA
Wang
G Y
GY
Xia
Z A
ZA
Xu
Z K
ZK
D001937:Brassica / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development*; D018548:Cotyledon / Q000187:drug effects / Q000254:growth & development; D046508:Culture Techniques / Q000379:methods; D018546:Hypocotyl / Q000187:drug effects / Q000254:growth & development; D010671:Phenylurea Compounds / Q000494:pharmacology; D030821:Plants, Genetically Modified; D013830:Thiadiazoles; D014170:Transformation, Genetic
D010671:Phenylurea Compounds; D013830:Thiadiazoles; C016785:thidiazuron