标题: |
灰葡萄孢分生孢子产生相关基因的克隆及功能分析 |
英文标题: |
Cloning and functional analysis of a gene related to conidiospore formation in Botrytis cinerea |
作者: |
王璇,邢继红,赵斌,韩建民,董金皋 |
出版时间: |
2013-01-01 |
机构: |
河北农业大学 |
关键词: |
灰葡萄孢,T-DNA插入突变体,TAIL-PCR,孢子发育 |
英文关键词: |
Botrytis cinerea,T-DNA insertional mutant,TAIL-PCR,Conidiation |
刊名: |
微生物学通报 |
英文刊名: |
Microbiology China |
ISSN: |
0253-2654 |
卷号: |
003 |
基金: |
^A河北省科技计划项目^B12226507^D1%^A河北农业大学青年科学基金项目^BQJ201235^D2 |
页码: |
533-543 |
分类号: |
S432.4 |
摘要: |
【目的】克隆灰葡萄孢分生孢子产生相关基因,并研究其功能,为进一步研究灰葡萄孢分生孢子产生机理和灰葡萄孢侵染及致病机理奠定基础。【方法】通过筛选灰葡萄孢ATMT突变体库,获得一株不能产生分生孢子的突变菌株BCt78,采用PCR和SouthernBlotting技术,对突变菌株BCt78进行分子鉴定。利用TAIL-PCR技术获得T-DNA插入位点的侧翼序列;将所获得侧翼序列与灰葡萄孢基因组数据库中的已知基因序列进行BLAST分析,推测出T-DNA的插入位点;通过PCR进一步验证T-DNA的插入位点,利用RT-PCR技术确定突变基因;最后对突变菌株的菌落形态、生长速度、胞壁降解酶活力、粗毒素的生物活性、对番茄叶片的致病能力及部分致病相关基因的表达情况进行研究。【结果】TAIL-PCR结果证实T-DNA插入到灰葡萄孢BC1G_12707.1基因的ATG起始密码子区;RT-PCR结果证实突变基因为BC1G_12707.1,该基因DNA全长为135 bp,编码一个44个氨基酸的假定蛋白(Hypothetical protein)。突变菌株在PDA培养基上菌落呈灰白色,生长速度减慢,不能产生分生孢子及菌核;对番茄叶片的致病... |
英文摘要: |
[Objective] To further investigate molecular mechanism of conidiospore formation,infection and pathogenicity of Botrytis cinerea,the gene related to conidiospore formation was cloned and characterized.[Methods] A mutant with no conidiospore,named BCt78,was found by screening T-DNA insertional mutant library of B.cinerea and it was testified by PCR and Southern Blotting techniques.The flanking sequence of T-DNA insertion site was acquired by using TAIL-PCR.The sequence of the T-DNA inserted gene was obtained... |