香蕉枯萎病菌cyp1基因的克隆与序列分析  

Cloning and Sequence Analysis of cyp1 Gene of Fusarium oxysporum f sp.cubense

作　　者：杨腊英[1] 黄小娟[1] 谢玉萍[1] 羊玉花[1] 杨歆璇[1] 方晓东 黄俊生[1] 

机构地区：[1]中国热带农业科学院环境与植物保护研究所海南省热带农业有害生物检测监控重点实验室,海南儋州571737 [2]华大基因研究院,广东深圳518000

出　　处：《热带作物学报》2010年第2期248-252,共5页Chinese Journal of Tropical Crops

基　　金：中央级公益性科研院所基本科研业务费专项(No.2008HZS1J003);国家科技支撑计划项目(No.2007BAD48B01);2009年度公益性行业(农业)科技专项经费项目(No.200903049)资助

摘　　要：为了解亲环素基因(cyp1,其编码亲和蛋白)在尖孢镰刀菌古巴专化型生理小种1号和生理小种4号之间及与其他镰刀菌之间的差异,分析cyp1基因与两生理小种之间的寄主选择性差异的关系,通过比对现有镰刀菌属的亲环素基因序列,设计并合成引物,采用PCR和RT-PCR方法扩增了尖孢镰刀菌古巴专化型生理小种1号和生理小种4号2个生理小种的cyp1基因,并测序进行比较分析,同时对编码蛋白进行了氨基酸序列比对和功能分析。结果表明,2个生理小种cyp1基因全长均为1066bp,开放阅读框均为675bp,编码224个氨基酸,基因序列间差异性为0.8%,编码蛋白间仅存在1个氨基酸的差异;两生理小种的cyp1基因序列与镰刀菌属其他专化型或不同种间存在差异性,与不同属之间其他种间的差异最大达50%,比对蛋白序列发现仅存在0.85%的差异,这进一步证明亲和蛋白在真菌中较保守。从cyp1基因序列及其编码蛋白的氨基酸序列在两生理小种之间的差异性分析结果推测,2个生理小种寄主选择性差异与cyp1基因并无明显对应关系,这为进一步研究cyp1基因功能奠定了基础。In order to know the differences of cyp1 gene between Fusarium oxysporum f sp.cubense（Foc）race 1 and race 4 and other Fusarium,and analyze whether the host selective difference between the races 1 and 4 were related with Cyclophilin A（cyp1）gene,the cyp1 genes of races 1 and 4 were amplified by Polymerase Chain Reaction and Reverse Transcription Polymerase Chain Reaction and sequenced,and the gene sequences and the encoded protein sequences were aligned by using the DNAStar software.The results revealed that the full length of cyp1 from Foc races 1 and 4 were both 1 066 bp,with their open reading frames being 675 bp,encoding 224 amino acids.The sequences of the cyp1 gene of Foc race 1 showed 99.2% similarity to that of Foc race 4 and there was only one amino acid different in the sequences.The cyp1 gene sequences of these two races were different from those of other specialized forms or species of Fusarium or from those of other species of different genera（the highest being 50.9% by alignment）,and the protein blast result showed only 0.85% difference.This proved that the cyp1 protein was conserved in fungi,.It was inferred from the differences of the cyp1 gene sequence and protein sequences from FOC1 and FOC4 isolates that there was no obvious relation between the cyp1 gene and the host selective preference of FOC1 and FOC4.These results would help to know the function of the cyp1 gene from FOC1 and FOC4 in the future researches.

关 键 词：尖孢镰刀菌古巴专化型 生理小种 亲和蛋白基因 寄主选择性 

分 类 号：S432.4[农业科学—植物病理学]