农杆菌介导多年生黑麦草转化体系的建立  

Establishment of an Agrobacterium Mediated Transformation System for Lolium perenne L.

作　　者：王奇丽[1] 何近刚[2] 陈彦龙[2] 张小芸[2] 吴金霞[2] 

机构地区：[1]山西农业大学,山西太谷030801 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《中国农业科技导报》2009年第2期119-123,共5页Journal of Agricultural Science and Technology

基　　金：国家973计划项目(2007CB108900)资助

摘　　要：以胚性愈伤组织系,作为转化受体,利用农杆菌介导的遗传转化方法,将报告基因gus和抗除草剂基因bar成功转入多年生黑麦草。共计获得52株T0代抗性植株。利用PCR方法对转基因植株的bar基因进行扩增,确定阳性株系27株。对阳性植株叶片进行GUS染色,有15株叶片呈现较强的蓝色。对再生植株和野生型植株叶片喷施5‰的除草剂Basta溶液,GUS阳性的植株全部表现为Basta抗性。以上结果表明,以黑麦草成熟胚胚性愈伤组织为受体,通过优化农杆菌介导的黑麦草遗传转化体系,初步建立了多年生黑麦草的转基因平台,这对黑麦草的遗传改良具有重要的意义。The vector pDM805 which harbors the selection gene bar and reporter gene gus was transformed into embry- ogenic callus lines of ryegrass. In total, 52 individual regenerated plants were obtained from bialaphos resistant callus lines. Among them, 27 were confirmed by bar PCR analysis; 15 of the 27 PCR positive plants showed strong blue color with GUS staining. The results suggested that through optimizing callus induction and plant regeneration conditions, an efficient Agrobacterium mediated transformation protocol was established in ryegrass, which paves the way for its genetic improvement through genetic engineering in the near future.

关 键 词：多年生黑麦草 农杆菌介导转化 GUS染色 PCR检测 Basta筛选 

分 类 号：Q786[生物学—分子生物学]