摘要
【目的】为了探明胶孢炭疽菌(Colletotrichum gloeosporioides)漆酶基因(lac1)的序列特征,进一步研究胶孢炭疽菌的分子致病机制,【方法】以杧果炭疽病菌为材料,采用同源克隆、SEFA-PCR和RT-PCR技术对lac1基因进行扩增、并对所获得的lacl基因进行了序列特征和预测蛋白保守结构域的分析。【结果】结果表明,该基因DNA和cD-NA全长分别为2 996 bp、1 773 bp,编码区含3个内含子(大小分别为47 bp、50 bp和59 bp),推测编码590个氨基酸,其分子质量约为1174.50 kDa,等电点PI为5.36。系统进化树结果显示该基因编码的氨基酸序列与多种真菌的含铜氧化物酶聚为一类,其中与C.lagenarium laccase(BAB32575.1)的同源性最高,达73%。【结论】lac1基因具备真菌漆酶基因家族的序列特征,推测lac1基因可能参与调控C.gloeosporioides菌丝生长、色素生成和致病性等。
[Objective]The objective of this study was to reveal sequence features of laccase（lacl）, which would be used in further study of pathogenesis of C. gloeosporioides on Mango. [Method]A laccase gene homolog lacl was cloned from C. gloeosporioides on mango by the method of homologous cloning, SEFA- PCR and RT-PCR, and sequence characterization and the conserved domains on predicted protein of lac 1 gene were also analyzed. [Result]The results showed that the complete DNA and eDNA sequence of lacl gene were 2 996 bp and 1 773 bp in size, respectively, with three introns of 47 bp, 50 bp and 59 bp, re- spectively, encoding a putative protein of 590-amino acid with a molecular weight of 1174.50 kDa and isoelectric point of 5.36. Phylogenetic analysis indicated that putative protein of lacl was homologous to C. lagenarium laccase （BAB32575.1） with a similarity of 73%, and was also homologous to copper-containing oxidases in some other fungi at some similarities. [Conclusion]The studies suggested that lacl gene had the sequence characteristics of laeease gene family in fungi,which meant that LA C1 might involve in mycelia growth, pigment production and pathogenicity of C. gloeosporioides.
出处
《果树学报》
CAS
CSCD
北大核心
2013年第2期202-206,共5页
Journal of Fruit Science
基金
公益性行业(农业)科研专项(201203092-2)
环植所基本科研业务费(2010hzsZDZX002)
关键词
杧果
胶孢炭疽菌
漆酶
克隆
序列分析
Mango
Colletotrichum gloeosporioides
Laccase
Cloning
Sequence analysis
