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【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】 To investigate the molecular mechanism of lac1 gene in Colletotrichum gloeosporioides and further study on molecular mechanism of Colletotrichum gloeosporioides. 【Method】 Using Colletotrichum gloeosporioides as the material, The lac1 gene was amplified by cloning, SEFA-PCR and RT-PCR. The lacl gene was sequenced and the predicted protein conserved domains were analyzed. 【Result】 The results showed that the full-length cDNA and cD-NA of the gene were 2 996 bp and 1 773 bp, respectively. The coding region contained 3 introns (47 bp, 50 bp and 59 bp, respectively) Amino acids, the molecular mass of about 1174.50 kDa, isoelectric point PI of 5.36. The phylogenetic tree showed that the amino acid sequence encoded by this gene was clustered with copper-containing oxidases of various fungi, of which the highest homology was 73% with C. lagenarium laccase (BAB32575.1). 【Conclusion】 The lac1 gene has the sequence characteristics of the fungal laccase gene family. It is speculated that the lac1 gene may be involved in the regulation of C. gloeosporioides mycelial growth, pigment production and pathogenicity.