Transforming the snowdrop lectin (Galanthus nivalis agglutinin, GNA) gene to soybean by pollen tube pathway technique
2006
Liu Depu, Yuan Ying, Tang Kexuan
Chinese. 采用花粉管通道技术,用雪花莲凝集素基因(Galanthusnivalisagglutinin,GNA)转化吉林省主推品种吉林20号、吉林30号、吉林45号品种大豆。通过接蚜鉴定和PCR鉴定,从所获得的种子苗中筛选出转基因植株。对转基因植株的后代进行分子生物学鉴定:(1)PCR分析,转基因植株97TGR1和97TGR2的T2代表现阳性,第5代表现阳性纯合;97TGR1、97TGR2和98FD1￣98FD20的T3代Westernblotting检测结果证明,GNA基因在蛋白质水平有表达,最高表达量占总可溶性蛋白的0.7%;97TGR1、98TGR2和99JI45TGR2的Southernblotting检测结果显示,GNA基因已插入大豆基因组;(2)遗传学分析,97TRG1的T2代呈孟德尔3:1分离,97TGR2的T3代出现种皮颜色不规则分离。经过抗蚜性鉴定和连续的筛选,获得抗性纯系;(3)抗蚜性鉴定,转基因株的T1、T2世代转基因植株可抑制蚜虫繁殖量50%￣90%;(4)品系鉴定,转基因大豆的抗蚜性达到农学标准抗(R)和高抗(HR)水平;大面积环境释放试验自然感蚜鉴定,转基因系蚜虫发生的高峰比对照延迟,高峰期...
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