摘要
尖孢镰刀菌在与寄主的相互作用中分泌几个特定的富含半胱氨酸的小分子量蛋白进入木质部中启动致病力,被称为SIX(secreted in xylem)蛋白,为明确其在不同寄主中的作用,本研究比较分析了几种尖孢镰刀菌专化型中SIX1、SIX4、SIX6、SIX8同源基因序列。根据已完成的尖孢镰刀菌古巴专化型1号(Foc1)与4号生理小种(Foc4)全基因组测序序列信息及相关SIX基因序列设计引物,应用PCR方法扩增分析56株尖孢镰刀菌古巴专化型与18株其它专化型及非致病型尖孢镰刀菌菌与其它种或属共21株菌株中的SIX1、SIX4、SIX6、SIX8基因。结果表明:设计的SIX1、SIX4、SIX6、SIX8基因引物均不能从非致病性尖孢镰刀菌与其它镰刀属种或其它属的菌株DNA中扩增出目的条带;SIX1基因的2个引物均能从供试的Foc菌株DNA中扩增出目的条带,同时可从部分其它专化型菌株DNA中扩增出目的条带;SIX4基因引物仅能从供试的尖孢镰刀菌番茄专化型与部分甘蓝专化型菌株DNA中扩增出目的条带;SIX6引物仅能从供试Foc1、Foc2、Foc4菌株DNA中扩增出目的条带;SIX8基因引物能从所有供试的致病尖孢镰刀菌中DNA扩增出目的条带。研究发现的SIX6基因序列提供了快速鉴定尖孢镰刀菌古巴专化型的检测方法,同时为深入研究尖孢镰刀菌各个专化型中SIX基因的功能奠定基础。
During infection of the host, Fusarium oxysporum secretes several unique proteins, called 'secreted in xylem'(SIX) proteins, into the xylem sap. For understanding the roles of SIX proteins in diverse hosts, the sequence of SIX1, SIX4, SIX6, SIX8 gene were cloned and analyzed. Based on the genomic sequences of F. oxysporum f. sp.cubense race 1(Foc1) and race 4(Foc4) and the gene sequences in the Gen Bank, the SIX genes' primers were designed.Then using a PCR and sequencing approach, homologs of F. oxysporum f. sp. lycopersici(Fol) SIX1, SIX4, SIX6, and SIX8 in the genome of 56 isolates of F. oxysporum f. sp. cubense and 18 isolates of other eight F. oxysporum formae speciales and two non-pathogen F. oxysporum and total 19 isolates of F. monilifarme and F. solani and some other genus pathogens of tropical crops were identified. SIX1, SIX4, SIX6, and SIX8 genes were not detected in all tested two non-pathogen F. oxysporum and total 19 isolates of F. monilifarme and F. solani and some other genus pathogens of tropical crops. SIX1 gene were detected in all tested isolates of Foc by using the two primers, and SIX1 gene with Foc4-six1-f/Foc4-six1-r primers or Foc4-324six1-f/Foc4-six1-r primers were also detected in some other different F. oxysporum formae speciales. SIX4 gene with the synthesized primers was only detected in Fol and some isolates of F. oxysporum f. sp. conglutinans. SIX6 gene with the designed and synthesized primers was only detected in all tested isolates of Foc. SIX8 gene with the designed and synthesized primers was detected in all tested pathogenici isolates of F. oxysporum formae speciales. A specific diagnostic for Foc based on Foc SIX6 is described and the results provide a foundation for further studying the SIX genes' function of F. oxysporum formae speciales.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2015年第8期1739-1746,共8页
Genomics and Applied Biology
基金
国家自然科学基金项目(31371900)资助
关键词
尖孢镰刀菌专化型
效应基因
香蕉枯萎病菌
分子鉴定
Fusarium oxysporum formae speciales
Effector genes
F.oxysporum f.sp.cubense
Molecular diagnostic
