农杆菌介导烟草抗真菌β-1,3-葡聚糖酶基因转化洋桔梗  

Agrobacterium-mediated transformation of tobacco β-1,3-glucanase gene into Eustoma grandiflorum(Raf.) Shinn.

作　　者：付晓佳 张倩[1] 李琳[1] 乔宁宁[1] 王转梅[1] 陈崇顺[1] 

机构地区：[1]南京师范大学生命科学学院/江苏省生物多样性与生物技术重点实验室,江苏南京210023

出　　处：《江苏农业学报》2016年第1期44-50,共7页Jiangsu Journal of Agricultural Sciences

基　　金：江苏高校优势学科建设工程资助项目

摘　　要：为获得洋桔梗抗真菌病害新品种,本研究将烟草抗真菌β-1,3-葡聚糖酶基因(Glu)与质粒p BI121重组,构建了植物表达载体p BI121-Glu,利用农杆菌介导法转化洋桔梗品种Double Mariachi Pink叶块,共获得21个卡那霉素抗性株系。对抗性株系进行PCR扩增,获得10个阳性株系。选取其中生长良好的5个株系进行RT-PCR检测,鉴定出3个株系(EG6、EG11和EG17)为转基因正常转录株系。EG6、EG11和EG17株系及非转基因对照株系的β-1,3-葡聚糖酶的平均活力分别为0.301 U、0.243 U、0.401 U及0.236 U,其中EG6和EG17株系的平均活力显著高于非转基因对照株系。洋桔梗离体叶片的抗灰霉病鉴定显示,EG17病斑面积(0.432 cm2)显著小于非转基因对照株系(2.156 cm2)。此外,EG17对灰霉病的抗性还表现为发病延迟、病斑面积扩大速度减慢等。In order to enhance the resistance to fungi diseases,an antifungal β-1,3-glucanase gene from tobacco was transferred into Double Mariachi Pink lisianthus [Eustoma grandiflorum（ Raf.） Shinn.] through Agrobacterium-mediated transformation by constructing the plant expression vector of p BI121-Glu. 21 kanamycin-resistant lines were obtained,and10 lines carrying the target gene were detected by PCR. RT-PCR analysis confirmed that the β-1,3-glucanase gene was expressed in three lines,EG6,EG11 and EG17,which showed average β-1,3-glucanase activities of 0. 301 U,0. 243 U and0. 401 U,higher than that of non-transformed line（ 0. 236 U）. Compared with non-transformed plants,resistance to Botrytis cinerea in the transgenic line EG17 was improved as smaller lesion area,slower lesion expansion speed and delayed onset of B.cinerea.

关 键 词：洋桔梗 Β-1 3-葡聚糖酶基因 农杆菌介导法 灰霉病抗性 

分 类 号：Q789[生物学—分子生物学]