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

作　　者：ZHANG Qi-jun LI Cong LIU Shao-kui LAI Dong QI Qing-ming LU Chuan-gen 

机构地区：[1]Institute of Food Crops,Jiangsu Academy of Agricultural Sciences/Jiangsu High Quality Rice R&D Center [2]College of Agriculture,Nanjing Agricultural University

出　　处：《Rice science》2013年第1期19-24,共6页水稻科学（英文版）

基　　金：supported by the Natural Science Foundation of Jiangsu Province, China (Grant No. BK2008348);the China National Science & Technology Major Project for Breeding New Plant Varieties of Genetically Modified Organisms (GMOs) (Grant No. 2008ZX08001-004)

摘　　要：The plasmid of pCDMARUBA-Hyg, which contained two insect-resistance genes, sbk （modified from CrylA（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 .qene and its application in rice transgenic research were also briefly discussed.The plasmid of pCDMARUBA-Hyg, which contained two insect-resistance genes, sbk （modified from CrylA（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 .qene and its application in rice transgenic research were also briefly discussed.

关 键 词：Oryza sativa sbk gene sck gene insect resistance TRANSGENE BREEDING 

分 类 号：S511[农业科学—作物学]