[1]梁曼,邓晟,张昕,等.大丽轮枝菌菌核型菌株T-DNA插入突变体库的构建及微菌核发育异常突变体的筛选[J].江苏农业学报,2015,(03):520-525.[doi:10.3969/j.issn.1000-4440.2015.03.009]
　LIANG Man,DENG Sheng,ZHANG Xin,et al.Construction of T-DNA inserted transformation library of sclerotium type Verticillium dahliae strain and screening of mutants with abnormal microsclerotia development[J].,2015,(03):520-525.[doi:10.3969/j.issn.1000-4440.2015.03.009]
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大丽轮枝菌菌核型菌株T-DNA插入突变体库的构建及微菌核发育异常突变体的筛选()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年03期
页码:
520-525
栏目:
植物保护
出版日期:
2015-06-30

文章信息/Info

Title:
Construction of T-DNA inserted transformation library of sclerotium type Verticillium dahliae strain and screening of mutants with abnormal microsclerotia development
作者:
梁曼12; 邓晟2; 张昕2; 林玲2
（1.南京师范大学生命科学学院，江苏南京210046；2.江苏省农业科学院植物保护研究所，江苏南京210014）
Author(s):
LIANG Man12; DENG Sheng2; ZHANG Xin2; LIN Ling2
（1.College of Life Science, Nanjing Normal University, Nanjing 210046, China;2.Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China）
关键词:
农杆菌介导的转化; 大丽轮枝菌; 突变体库; 微菌核
Keywords:
Agrobacterium-mediated transformation; Verticillium dahliae; mutant library; microsclerotia
分类号:
Q343.5
DOI:
10.3969/j.issn.1000-4440.2015.03.009
文献标志码:
A
摘要:
为阐明大丽轮枝菌微菌核形成发育的分子机理，利用农杆菌介导的遗传转化方法，建立了包含2 000个转化子的大丽轮枝菌菌核型菌株V08DF1的T-DNA插入突变体库，并在查氏、查氏-根或PDA培养基上培养，观察各转化子的菌落形态。从中筛选出130个菌落特征与野生型菌株V08DF1有明显差异的突变体菌株，其中14个突变体菌株为微菌核发育受阻。对这14 个微菌核发育异常的突变体菌株进行T-DNA 插入的PCR 验证，均能扩增到潮霉素抗性基因，Southern 杂交显示其中7 个为单拷贝插入，5个为双拷贝插入，2个为三拷贝插入，表明T-DNA 已经成功整合到这些突变株的基因组DNA 中。
Abstract:
To illustrate the molecular mechanism underlying microsclerotial development in Verticillium dahliae, agrobacterium tumefaciens-mediated transformation (ATMT) was applied for insertional mutagenesis of V. dahliae conidia. A T-DNA insertion mutant library of a sclerotium-type strain V08DF1 including 2 000 transformants were constructed. All of the 2 000 transformants cultured on two kinds of medium plates (Czapek-Dox medium plate and Czapek-Dox plus cotton root extract medium plate, or Czapek-Dox medium plate and PDA medium plate) were observed for colony morphology. 130 mutants with significant difference in colony morphology from V08DF1 were selected, among which 14 showed abnormal microsclerotia development. Southern blot analysis indicated that T-DNAs were inserted randomly into the V. dahliae genome and that seven mutants out of baove 14 were single-copy T-DNA insertion, five were two-copy T-DNA insertion, and two were three-copy T-DNA insertion. 

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相似文献/References:

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备注/Memo

备注/Memo:
收稿日期：2015-01-04 基金项目：国家自然科学基金项目（31201462）；国家科技支撑计划项目（2012BAD19B05）；国家现代农业技术体系棉花产业技术体系（CARS-18-16）；江苏省农业科技自主创新基金项目[CX(13)5022] 作者简介：梁曼（1992-），女，安徽六安人，硕士研究生，从事大丽轮枝菌微菌核发育分子机理研究。（Tel）025-84390769；（E-mail）liangman19921116@sina.com 通讯作者：林玲，（E-mail）linling@jaas.ac.cn
更新日期/Last Update: 2015-06-30