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α-甘露糖苷酶SSc4基因过表达质粒构建与油菜转化

Over-expression Plasmid Construction of α-mannosidase Gene SSc4 and Rapeseed Transformation
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摘要 菌核病是由核盘菌引起的一种世界性的真菌病害,对我国油菜生产危害尤为严重。但是现在已知的所有植物资源中都找不到对核盘菌完全具有抗性的基因资源。本研究采用"以毒攻毒"的途径,将核盘菌自身参与致病的一个α-甘露糖苷酶SSc4基因转到甘蓝型油菜中,以期获得对能对核盘菌产生抗性的油菜种质新资源。本研究克隆核盘菌的SSc4基因,构建了其过表达质粒,并通过农杆菌介导转化油菜,实验共筛选到159株抗性苗,其中获得了27株转基因油菜,转化率约为1%。对其中的9株转基因苗进一步作RT-PCR检测,显示有6株转基因植株中SSc4基因高表达,实现了设计目标。 Sclerotinia caused by Sclerotinia sclerotiorum is a worldwide fungal diseases, and causes a huge loss on rapeseed yield in China. However, gene resource that entirely resistant to the disease are not found in any known plant resources. Therefore, in order to obtain new germplasm resources of rapeseed that are resistant to Sclerotinia sclerotiorum, we adopted "combat poison with poison"strategy and transferred an alpha-mannosidase SSc4 gene from Sclerotinia itself into Brassica napus Xiangyou 15. In this research, we cloned SSc4 gene, constructed its over-expression plasmid, and transformed into Brassica napus Xiangyou 15 through Agrobacterium tumefaciens Gv3101. 159 resistant seedlings were gained, 27 of those are transgenic plants through PCR identification, which means transformation rate is about 1%. The RT-PCR results showed that the SSc4 was highly expressed in 6transgenic plants selected from 9 transgenic plants, which suggested the design objectives were achieved.
作者 魏然 何雯 刘春林 Wei Ran He Wen Liu Chunlin(Agricultural college, Htman Agricultural Univesity, Changsha, 410128)
出处 《分子植物育种》 CAS CSCD 北大核心 2017年第1期155-159,共5页 Molecular Plant Breeding
基金 国家支撑计划项目(2013BAD01B03-10)资助
关键词 核盘菌 α-甘露糖苷酶基因 过表达质粒 油菜转化 Sclerotinia sclerotiorum α-mannosidase gene Over-expression plasmid Rapeseed transformation
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