南方农业学报2012，Vol.43Issue(2)：141-145,5.DOI:10.3969/j:issn.2095-1191.2012.02.141
Bt抗虫水稻转育后代两种筛选方法的比较
Comparison of two screening methods for detecting insect-resistant offspring of Bt-transgenic rice
摘要
Abstract
[Objective]Two screening techniques, viz., marker-assisted selection and herbicide resistance screening were compared to develop an efficient and accurate selection method for detecting insect-resistant offsprings of Bt-transgenic rice. [Methodjln the first method, two DNA markers linked to CrylC* and Cry2A* genes were respectively used to screen the genomic DNA extracted from seedlings of the transgenic offspring plants. In the second method, adult plant leaves were sprayed with 300 mg/L herbicide Bastar and the withered and yellow leaves were counted to calculate the rate of positive plants harboring target genes. Moreover, the Bt protein content in the resistant plants was quantified using ELISA in order to verify the accuracy of two screening methods. [Results ]The consistency rate of two screening methods was 98.2% in screening of 169 individual plants generated from 4 combinations by using TIC-19 as donor parent. In screening of 141 individual plants generated from 3 combinations by using T2A-1 as donor parent, the consistency rate of two methods was 98.6%. The ELISA assay results showed that the Bt protein in 144 individually selected plants (from positive plants), using both of the two methods, had corresponding expression and was increased or decreased substantially compared to donor parents T1C-19 and T2A-21. [Conclusion)Both the marker-assisted selection and herbicide resistance screening techniques were highly accurate in selecting insect-resistance offspring of Bt-transgenic rice. The former one can be used in lab at any stage but the cost was relatively higher, while the latter one is cost-effective and convenient, however, easy to be affected by environmental factors. In general, this study provides useful information for breeding Bt-transgenic rice.关键词
Bt抗虫水稻/分子标记辅助选择/除草剂抗性筛选/Cry1C*/Cry2A*Key words
Bt transgenic rice/ marker-assisted selection/ herbicide resistance screening/ CrylC */ Cry2A *分类
农业科技引用本文复制引用
李孝琼,郭嗣斌,韦宇,李杰民,刘开强,高国庆..Bt抗虫水稻转育后代两种筛选方法的比较[J].南方农业学报,2012,43(2):141-145,5.基金项目
国家863计划重点项目(2010AA101803) （2010AA101803）
国家转基因生物新品种培育重大专项项目(2011ZX08001-001) （2011ZX08001-001）
广西科学研究与技术开发计划项目(桂科攻10100005-8) （桂科攻10100005-8）
广西农业科学院科技发展基金项目(201003Z) （201003Z）
