转双价抗虫基因BmkIT-Chitinase玉米株系的获得

通过超声波辅助花粉介导法,将双价抗虫基因BmkIT-Chitinase分别导入以玉米自交系昌7-2及郑58的花粉为受体的不同基因型的自交系中。本研究共处理玉米雌穗1072穗,获得T0代种子1563粒,经卡那霉素初筛,T1代～T4代PCR及SouthernBlot杂交分子跟踪检测共获得20个转化株系,田间抗虫性鉴定表明共有16个转化株系与对照在抗虫性方面有显著差异,且此抗性随着各代稳定遗传。农艺性状调查结果表明,所获得的转基因玉米株系中大部分材料的农艺性状与对照无显著差异,除了N55材料及N20-1材料。N55材料的穗位高度与对照相比略低6±0.5cm,而穗粒数增加75±5粒。而N20-1材料百粒重增加5±0.5g。因此,转入此双价抗虫基因对玉米农艺性状影响不是很大。经过分子检测、田间抗虫性鉴定及农艺性状调查我们最终选育了9个转双价抗虫基因昌7-2自交系优良株系,6个郑58转双价抗虫基因自交系优良株系。

Acquirement of Transgenic Maize Lines with Binary Insect Resistant BmkIT-Chitinase

The binary insect resistant BmkIT-Chitinase gene was introduced into two different genotypicmaize inbred lines,using the Chang7-2 pollen and Zheng58 pollen as receptor respectively,with the method of sonication assisted pollen mediated transformation.In this study,1 072 corn ears were treated to generate 1 563 transgenic maize seeds of T0 generation.Totally 20 transformed lines came out by continuous validations in the generations from T1 to T4,through the Kanamycin resistance screening,PCR and Southern blotting.Field insect resistant evaluation shows that there are significant differences on insect resistance between 16 transformed lines and their control groups,moreover this insect resistant can be steadily inherited within generations.The investigation of the agronomic traits in the filed demonstrates that there are no any differences on agronomic traits between the most of transformed lines and their control groups,except the Line N55 and Line N20-1.The Height of Ear setting position of Line N55 is 6±0.5 cm lower than that of the control group,but the number of kernels increased 75±5 per ear.The 100-seed weight of Line N20-1 increased 5±0.5 g.Therefore,it could say there is no significant influence on the agronomic traits by transferring this binary insect resistant gene.In this research,9 elite transgenic lines derived from Chang7-2 and 6 elite transgenic lines derived from Zheng58 were acquired,through molecular validation,field insect resistant evaluation and agronomic assessment.