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

F. L. Zhang, Y. Takahata, M. Watanabe, J. B. Xu

Research output: Contribution to journalArticlepeer-review

39 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)569-575
Number of pages7
JournalPlant Cell Reports
Volume19
Issue number6
DOIs
Publication statusPublished - 2000 May
Externally publishedYes

Keywords

  • Agrobacterium tumefaciens
  • Chinese cabbage (Brassica campestris L.ssp. pekinensis)
  • Cotyledonary explant
  • Genetic transformation

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

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