Agrobacterium-mediated transformation of Brassica campestris ssp. Parachinensis with synthetic Bacillus thuringiensis cry1Ab and cry1Ac genes

Y Xiang; W-K R Wong; M C Ma; R S C Wong

doi:10.1007/s002990050007

Agrobacterium-mediated transformation of Brassica campestris ssp. Parachinensis with synthetic Bacillus thuringiensis cry1Ab and cry1Ac genes

Plant Cell Rep. 2000 Jan;19(3):251-256. doi: 10.1007/s002990050007.

Authors

Y Xiang¹, W-K R Wong¹, M C Ma², R S C Wong¹

Affiliations

¹ Department of Biochemistry, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China, , , , , , CN.
² Department of Entomology, University of Maryland, College Park, MD 20742-5141, USA, , , , , , US.

PMID: 30754903
DOI: 10.1007/s002990050007

Abstract

An effective plant regeneration procedure and a gene transfer system via Agrobacterium tumefaciens were developed in Brassica campestris ssp. parachinensis. Hypocotyls from 5-day-old seedlings with 2 days pre-culture were infected with Agrobacterium strain MOG301 harboring a binary vector containing a synthetic Bacillus thuringiensis (B.t.) cry1Ab or cry1Ac gene with full codon-modification. After culture and selection on MS medium supplemented with 4.0 mg/l BAP, 2.0 mg/l NAA, 70 μM AgNO₃ and 50 mg/l kanamycin, a number of kanamycin-resistant plantlets were regenerated. PCR and Southern blotting analysis were used to identify and characterize the transgenic plants with the integrated cry1Ab or cry1Ac gene. Western blotting analysis of the transgenic plants confirmed the expression of insecticidal proteins encoded by cry1Ab or cry1Ac. Subsequent bioassay with larvae of the Diamondback moth, Plutella xylostella, demonstrated that the transgenic plants were resistant to feeding damage.

Keywords: B. campestris ssp.parachinensis; B.t. insecticidal protein gene; Insect resistance; Key words Agrobacterium-mediated transformation; Transgenic plants.