摘要
为了阐明烟草赤星病（tobaccobrown-spotdisease）病原真菌长柄链格孢（Alternarialongipes）对二甲酰亚胺类杀菌剂抗性的分子机制，以不同抗性水平的长柄链格孢菌株为实验材料，通过基因钓鱼（GeneFishing）技术进行基因差异表达分析；并利用基因敲除技术对已克隆到的AlCyP1基因进行功能分析．结果表明：一个亲环蛋白基因AlCyP1的表达量在不同抗性水平的长柄链格孢中存在明显差异；应用DNA步移（DNAwalking）方法克隆得到AlCyP1基因的DNA序列，其开放阅读框中存在2个翻译起始密码子，可以编码产生2种不同长度的多肽，长、短多肽产物分别具有222和188个氨基酸，其中短多肽起始于长多肽的第35个密码子；另外，氨基酸序列同源性分析发现，AlCyP1和其他真菌的亲环蛋白具有很高的同源性，达50．0％～61.3％；利用基因敲除技术对AlCyP1基因进行功能分析发现，该基因通过表达水平的改变参与了长柄链格孢对渗透胁迫的适应调节过程．
In order to clarify the molecular mechanism of tobacco brown-spot disease pathogenic fungi Alternaria longipes resistance to dicarboximide fungicides （DCFs）, GeneFishing technology was conducted to analyse gene differential expression among A. longipes strains with different DCFs- resistant level, and the biological functions of the cloned AlCyP1 gene were analyzed by gene disruption. The results revealed that significant expression difference of a cyclophilin gene - AlCyP1 existed in different DCFs-resistant level strains. Using DNA walking method, the DNA sequence of AlCyP1 gene was cloned. Two translation initiation codons existed within the open reading frame of AlCyP1 gene, which coded two different length polypeptide products. The long and short polypeptide products contained 222 and 188 amino acids, respectively, in which the short one was initiated at codon 35 of the long polypeptide product. In addition, the sequence homology analysis of amino acids showed AlCyP1 sharing high homology （50.0%-61.3% ） with cyclophilins of other fungi. Finally, biological functions of the AlCyP1 gene were analyzed by gene disruption, and the results showed that AlCyP1 gene involved in A. longipes osmotic stress adaptation process dependent of its expression level change.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2011年第2期133-141,共9页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
国家重点基础研究发展计划资助项目(2007CB411600)
云南省烟草公司科技资助项目(04A19)
云南省中青年学术和技术带头人后备人才资助项目(2009CI052)
云南省应用基础研究自筹经费资助项目(2010ZC056)
