转新型抗虫基因Vip3A棉花新种质的创制  

Development of Cotton Germplasm Introduced with Novel Insect Resistant Gene Vip3A

作　　者：张安红 王志安 肖娟丽 刘圆 罗晓丽 ZHANG Anhong;WANG Zhi'an;XIAO Juanli;LIU Yuan;LUO Xiaoli(Institute of Cotton,Shanxi Agricultural University(Shanxi Academy of Agricultural Sciences),Yuncheng,Shanxi 044000,China;Shanxi Key Laboratory of Cotton Germplasm Resources Utilization and Molecular Design Breeding,Yuncheng,Shanxi 044000,China)

机构地区：[1]山西农业大学(山西省农业科学院)棉花研究所,山西运城044000 [2]棉花种质资源利用与分子设计育种山西省重点实验室,山西运城044000

出　　处：《西北植物学报》2020年第8期1287-1293,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：转基因生物新品种培育重大专项(2018ZX08009-01B);国家重点研发计划(2018YFD0100301)。

摘　　要：该研究构建植物表达载体pBin438-Vip3A,通过农杆菌介导法转化棉花品种‘冀合713’,将新型抗虫基因Vip3A导入到棉花植株,创制对棉铃虫抗性的转基因棉花新种质。结果表明:(1)PCR检测Vip3A基因已经导入到棉花基因组中且能够稳定遗传。(2)室内抗虫性鉴定表明,与对照相比转基因植株对棉铃虫的抗性显著提高,并获得2株高抗和3株抗棉铃虫的转Vip3A基因株系。(3)Southern blotting结果显示,转基因株系BV01为单拷贝。(4)Elisa检测表明,外源Vip3A基因在BV01的根、茎、叶、花、种子中都有表达,在叶片中的Vip3A蛋白表达为苗期>蕾期>花期>铃期>吐絮期。该研究创制了新型抗虫转基因棉花材料,为培育棉花新型抗虫品种提供了种质资源。The plant expression vector pBin438-Vip3A was constructed and transformed into cotton variety“Jihe 713”by agrobacterium-mediated transformation.A new insect-resistant gene,Vip3A,was introduced into cotton plants to create new transgenic cotton germplasm resistant to cotton bollworm.The results showed that:(1)Vip3A gene has been transformed into the cotton genome and can be inherited stably.(2)In laboratory,two transgenic lines exhibited high resistance rate to bollworm,and three transgenic lines did resistance rate compared to the control.(3)Based on Southern blotting analysis,the transgenic line BV01 contained a single copy of insertion.(4)According to the results of Elisa,the foreign Vip3A gene was expressed in root,stem,leaf,flower and seed of BV01.Moreover,there were significant differences in Vip3A protein contents of the functional leaves among different stages,in order from high to low:seedling stage,bud stage,anthesis stage,boll stage and bolling stage.In this study,we successfully introduced the Vip3A gene into cotton plants,which possessed high resistance to cotton bollworm,laying a solid foundation for breeding new cotton cultivars with high resistance to cotton bollworm.

关 键 词：抗虫性 VIP3A基因 棉花 棉铃虫 

分 类 号：Q785[生物学—分子生物学] Q786