RICE SCIENCE

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Breeding and Identification of Insect-Resistant Rice by Transferring Two Insecticidal Genes, sbk and sck

  1. 1Institute of Food Crops, Jiangsu Academy of Agricultural Sciences/Jiangsu High Quality Rice R&D Center, Nanjing 210014, Chin; 2College of Agriculture, Nanjing Agricultural University, Nanjing 210095, China
  • Online:2013-01-28 Published:2013-01-06
  • Contact: LU Chuan-gen
  • Supported by:

    This research was financially supported by the Natural Science Foundation of Jiangsu Province, China (Grant No. BK2008348) and the China National Science & Technology Major Project for Breeding New Plant Varieties of Genetically Modified Organisms (GMOs) (Grant No. 2008ZX08001-004).

Abstract:

The plasmid of pCDMARUBA-Hyg, which contained two insect-resistance genes, sbk (modified from Cry1A(c)) and sck (modified from CpTI), was transformed into an Agrobacterium EHA105 for infection of the calli of a super japonica rice Nanjing 45. Primarily, using polymerase chain reaction (PCR) detection with the primers of sbk and sck genes, 42 positive transgenic plants that were marker-free and contained the two target genes were selected from 97 regenerated plants. Results of southern-blotting indicated that 23, 11, 5, 2 and 1 plants had one, two, three, four and five copies of the transformed genes, respectively. Analysis of reverse transcription PCR (RT-PCR) and Bt gene testing paper showed that 28 T3 generation plants derived from four transgenic plants having a single copy were insect-resistant. Feeding experiment with rice stem borer revealed that the insect resistance was greatly increased with the larva mortality ranging from 94% to 100%. In addition, among the transgenic plants, three T3 transgenic plants possessed some desirable characteristics for breeding and production, such as plant height, seed-setting rate, 1000-grain weight and larva mortality. The mechanism of insect resistance of Bt gene and its application in rice transgenic research were also briefly discussed.

Key words: Oryza sativa , sbk gene , sck gene , insect resistance , transgene , breeding