Agrobacterium-mediated transformation of cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis)

F-L Zhang; Y Takahata; M Watanabe; J-B Xu

doi:10.1007/s002990050775

Agrobacterium-mediated transformation of cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis)

Plant Cell Rep. 2000 May;19(6):569-575. doi: 10.1007/s002990050775.

Authors

F-L Zhang¹, Y Takahata¹, M Watanabe¹, J-B Xu²

Affiliations

¹ Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan e-mail: ytakahata@iwate-u.ac.jp, Fax. +81-19-6216177, , , , , , JP.
² Beijing Vegetable Research Center, Beijing 100089, China, , , , , , CN.

PMID: 30754819
DOI: 10.1007/s002990050775

Abstract

A procedure for producing transgenic Chinese cabbage plants by inoculating cotyledonary explants with Agrobacterium tumefaciens strain EHA101 carrying a binary vector pIG121Hm, which contains kanamycin-resistance and hygromycin-resistance genes and the GUS reporter gene, is described. Infection was most effective (highest infection frequency) when explants were infected with Agrobacterium for 15 min and co-cultivated for 3 days in co-cultivation medium at pH 5.2 supplemented with 10 mg/l acetosyringone. Transgenic plants of all three cultivars used were obtained with frequencies of 1.6-2.7% when the explants were regenerated in shoot regeneration medium solidified with 1.6% agar. A histochemical GUS assay and PCR and Southern blot analyses confirmed that transformation had occurred. Genetic analysis of T1 progeny showed that the transgenes were inherited in a Mendelian fashion.

Keywords: Agrobacterium tumefaciens; Cotyledonary explant; Genetic transformation; Key words Chinese cabbage (Brassica campestris L. ssp. pekinensis).