摘要
利用Blast及TCDB数据库对桉树焦枯病菌(Calonectria pseudoreteaudii)的Cpsit1基因进行鉴定;再利用SMART数据库和Prot Param、TMHMM、PHD、Pro Comp 9.0在线分析工具对Cpsit1基因编码蛋白的理化性质、跨膜螺旋、蛋白质二级结构和亚细胞定位进行预测分析;最后采用qRT-PCR方法对CpSit1基因在焦枯病菌侵染桉树过程中的表达情况进行分析.结果表明:CpSit1基因长度为1 780 bp,其编码的蛋白序列共有592个氨基酸残基组成,并将其鉴定为铁载体-铁:H+同向转运蛋白.其保守结构域为MFS_1;共含有14个跨膜螺旋;在二级结构中α螺旋占45.10%,延伸链占26.01%,无规则卷曲占28.89%.通过qRT-PCR相对定量的方法分析焦枯病菌侵染桉树24、48和72 h后CpSit1基因的表达情况,结果显示,CpSit1基因在这3个时段均发生上调表达,但24 h的表达量明显大于48和72 h.说明桉树焦枯病菌CpSit1基因在病原菌侵染寄主的过程中通过调控铁载体-铁化合物的转运来完成铁元素的摄入,协助其在寄主中的定植.
Cpsii1 was identified by Blast and TCDB database,and then the physicochemical property,transmembrane helix, sec-ondary, structure and subcellular localization of Cp5ii1 were analyzed and predicted by SMART database, ProtParam,TMHMM , PHD and ProComp 9.0 online serice. Finally expression of Cp5ii1 gene was detected by qRT-PCR within 24,48 and 72 h Ca/oneciria pseuoreieauii infection of Eucalyptus. Results showed that Cpsit1 was identified as proton-coupled symporter for siderophore, with CpSit1 gene being 1 780 bp and encoding 592 amino acids. And the conserved domain of Cp〉Sii1 is MFS1. a-helix accounted for 45.10% of the secondary structure,with extension chain and random coil reaching 26.01% and 28.89%, respectively. QRT - PCR results revealed that expressionsof CpSit1 gene were all upregulated within 24,48 and 72hC. pseudoreteaudii infection,and the expression level in 24 h was significantly higher than the other 2 stages. It proved that Cp5it1 gene could mediate the iron metabolism pathway of C. pseudoreteaudii by regulating siderophore and assist C. pseudoreteaudii in the process of colonization.
出处
《福建农林大学学报（自然科学版）》
CSCD
北大核心
2017年第4期418-422,共5页
Journal of Fujian Agriculture and Forestry University:Natural Science Edition
基金
福建省财政厅资助项目(K81150002
K81139238
K8112004A)
关键词
桉树
焦枯病
铁载体转运蛋白
生物信息学分析
表达分析
Ucalyptus
Calonectria pseudoreteaudii
siderophore-iron transporter
bioinformatics
gene expression
