Transgenic indica rice plants harboring a synthetic cry2A* gene of Bacillus thuringiensis exhibit enhanced resistance against lepidopteran rice pests
Transgenic indica rice plants harboring a synthetic cry2A* gene of Bacillus thuringiensis exhibit enhanced resistance against lepidopteran rice pests
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DOI：
10.1007/s00122-005-0062-8
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发表时间：
2005-11-01
影响因子：
5.4
通讯作者：
Zhang, QF
中科院分区：
文献类型：
--
作者：
Chen, H;Tang, W;Zhang, QF
利用农杆菌介导法将人工合成的cry 2A * 基因导入籼稻恢复系明恢63。共获得102个独立的转化体。经PCR检测，其中71个转化子为cry 2A * 阳性植株。通过田间鉴定和Southern杂交分析，筛选出4个单拷贝插入的高抗品系（2A-1、2A-2、2A-3和2A-4）。所有4个转基因株系的种子在含有Basta的1/2 MS培养基上萌发时均表现出孟德尔分离。根据T-2代的发芽率（100%）选择纯合的转基因植株。在纯合转基因品系、其衍生的杂交种和其回交后代中测定Cry 2A * 蛋白浓度。四个纯合转基因株系的Cry 2A * 蛋白浓度范围为9.65 - 12.11 μ g/g叶片鲜重。杂种和回交后代的变异很小。在实验室和田间进行了昆虫生物测定。4个转基因株系均对鳞翅目害虫表现出显著的抗性。这些转cry 2A * 基因株系可用于生产抗虫杂交种，并可作为开发双毒Bt水稻的抗性来源。
A novel synthetic cry2A* gene was introduced into the elite indica rice restorer line Minghui 63 by Agrobacterium-mediated transformation. A total of 102 independent transformants were obtained. Among them, 71 transformants were positive cry2A* plants according to PCR analysis. Four highly insect-resistant lines with single-copy insertion (designated as 2A-1, 2A-2, 2A-3, and 2A-4) were selected based on field assessment and Southern blot analysis in the T-1 generation. All four transgenic lines showed Mendelian segregation by seed germination on 1/2 MS medium containing Basta. Homozygous transgenic plants were selected according to germination ratio (100%) in the T-2 generation. Cry2A* protein concentrations were determined in homozygous transgenic lines, their derived hybrids, and their backcross offspring. The Cry2A* protein concentrations of four homozygous transgenic lines ranged from 9.65 to 12.11 mu g/g of leaf fresh weight. There was little variation in the hybrids and backcross offspring. Insect bioassays were conducted in both the laboratory and field. All four transgenic lines were significantly resistant to lepidopteran rice pests. These cry2A* transgenic lines can be used to produce insect-resistant hybrids and serve as a resistant source for the development of two-toxin Bt rice.