抗病基因PR1在大豆中的遗传转化与多抗材料的培育  

Genetic Transformation of Disease Resistance Gene PR1 in Soybean and Cultivation of Multi-resistant Materials

作　　者：曲硕 焦耀磊 付加禹 崔慧敏 蔡奇 王丕武[1] Qu Shuo;Jiao Yaolei;Fu Jiayu;Cui Huimin;Cai Qi;Wang Piwu(Center for Plant Biotechnology,Jilin Agricultural University,Changchun,130118)

机构地区：[1]吉林农业大学,生物技术中心,长春130118

出　　处：《分子植物育种》2023年第1期174-184,共11页Molecular Plant Breeding

基　　金：国家转基因生物新品种培育重大专项(2016ZX08004-004)资助

摘　　要：单一抗病、抗虫品种在农业生产中已经不能满足需要,本研究通过基因工程技术构建抗病基因pCAMBIA-3301-PR1表达载体,通过花粉管通道法转化外源抗病基因PR1到含有CryAB-13-1转基因高世代受体大豆中,得到兼具抗病抗虫新材料。对T_(2),转基因植株进行常规PCR检测,SouthernBlot杂交,Real TimePCR检测,室内、室外抗虫鉴定,室内抗病鉴定。结果表明,在T_(2)代常规PCR检测中,阳性植株共15株。SouthernBlot杂交结果显示,PR1与CryAB-13-1基因均以单拷贝子的形式整合在大豆中。Real Time PCR表明PR1和CryAB-13-1基因在不同组织中表达量有差异,PR1基因在植物根、茎、叶中表达量平均分别为2.88、1.48、5.86,CryAB-13-1基因在根、茎、叶中表达量平均分别为3.03、1.32、7.11。室外抗虫鉴定阳性植株CryAB-13-1基因的植株虫食率为3.47%,抗虫级别上属于R-抗,抗虫级别从感虫到抗虫级别。室内采用圆盘分隔法进行抗虫鉴定,转化植株芽长有明显增长。室内抗病鉴定未转化植株平均死亡率高达58.34%,转化PR1基因植株平均死亡率为15.38%,抗病能力明显提高。因此得到兼具抗病抗虫新材料。A single drought-resistant and insect-resistant variety can no longer meet the needs in agricultural production. In this study, an expression vector for the drought-resistant gene pCAMBIA-3301-PR1 was constructed through genetic engineering technology, and the disease-resistant gene PR1 was transformed into the CryAB-13-1transgenic recipient soybean through the pollen tube method. In the process, new materials with both disease resistance and insect resistance were obtained. T2transgenic plants were test by conventional PCR detection, Southern Blot hybridization, Real Time PCR detection, indoor and outdoor pest resistance identification, indoor disease resistance identification. The results showed that there were 15 positive plants in the routine PCR test of T2generation. The results of Southern Blot showed that both PR1 and CryAB-13-1 genes were integrated in soybeans as single copies. Real Time PCR showed that the expression levels of PR1 and CryAB-13-1 genes were different in different tissues. The average expression levels of PR1 gene in plant roots, stems, and leaves were 2.88, 1.48, 5.86, respectively, and CryAB-13-1 gene were 3.03, 1.32, and 7.11 respectively. The insectivorous rate of the CryAB-13-1gene of the plants with positive insect resistance identification in the outdoor was 3.47%, which is R-resistant, and the insect resistance level ranges from susceptible to insect resistant. The disc inoculation method was used indoors for pest resistance identification, and the bud length of the transformed plants changed significantly. The average mortality rate of untransformed plants in indoor disease resistance identification was as high as 58.34%, and the average mortality rate of plants transformed with PR1 gene was 15.38%. The disease resistance ability was significantly improved. Therefore, a new material with resistance to diseases and insects was obtained.

关 键 词：大豆 PR1基因 CryAB-13-1基因 抗虫 抗病 

分 类 号：S565.1[农业科学—作物学]