农杆菌介导Cry1 Ab/Ac抗虫转基因玉米植株的获得

转基因技术的迅速发展有效地打破了物种间的遗传壁垒,从而加快了优良基因定向聚合的进程,然而遗传转化率低及转基因安全问题成为制约其进一步发展的限制因素。基于此,本研究以玉米自交系H99为材料,将Ac/Ds双元表达载体(包含抗虫Cry1 Ab/Ac基因和gfp报告基因等)通过农杆菌介导法转入由110粒玉米幼胚诱导出的愈伤组织中,获得28株抗性转基因植株。经PCR及RT-PCR的验证结果显示,其中8株为含有Cry1 Ab/Ac目的基因且该基因有效表达的阳性植株,转化效率达7%。通过农杆菌介导法所获得的玉米转基因植株为今后剔除抗生素筛选标记,从而获得安全的玉米抗虫新种质提供了遗传材料。

Agrobacterium-mediated Cry1 Ab/Acgene transformation in maize

The rapid development of transgenic technologies has enabled researchers to transfer agronomical desirable genes going beyond species borders.However,the drawback of the transgenic efficiency and selectable markers consisting of gene encoding antibiotics or herbicides resistance remains,which potentially poses an environmental health impact.Here,a high throughput method to develop transgenic maize without the contaminations of any selectable marker genes was reported.An Ac(Activator)/Ds(Dissociation) binary vector harboring the Cry1Ab/Ac gene and green fluorescence protein driven by a maize ubiquitin gene promoter was introduced into 110 maize embryo callus through Agrobacterium-mediated transformation.Ac/Ds transposon system promotes the enclosed target genes translocation,and GFP markers make it possible to visually detect the transgenic callus.Based upon this method,more than 8 putative independent transgenic events were identified by target gene specific PCR detection and further confirmed for alien gene expression using RT-PCR.The frequency of transformation reached 7%.The reported method is useful for improving the quantity and quality of transgenic maize lines and their application in cultivar improvement.