卵寄生真菌虫草棒束孢几丁质酶基因<em>IFCHI1</em>的克隆与分析

doi:10.7668/hbnxb.2015.02.015

摘要/Abstract

摘要： 为研究卵寄生真菌在寄生并破坏卵的过程中几丁质酶的作用,本研究从烟草南方根结线虫卵上分离得到虫草棒束孢HNE3,采用RT-PCR及RACE技术,首次克隆得到一个几丁质酶基因 IFCHI1 。该基因DNA序列全长1 417 bp,具有1个内含子,长度为61 bp,包含1个1 185 bp的开放阅读框(ORF),编码1个由394个氨基酸组成的蛋白。通过在线软件分析,该蛋白理论分子量为44.1 kDa,等电点为4.88。通过GenBank比对,几丁质酶IFCHI1属于第18家族糖基水解酶,比对分析不同来源真菌几丁质酶,发现该几丁质酶具有典型的催化区保守序列SIGGW和FDGIDIDWE及结合区保守序列GTWEQGVYDY和GLGGAMWWESS。同源性比对发现,该几丁质酶同昆虫寄生真菌几丁质酶同源关系近。结果表明,从虫草棒束孢HNE3克隆得到几丁质酶基因 IFCHI1,为进一步明确该菌产生的几丁质酶对南方根结线虫卵壳降解及降低孵化率的作用机理提供理论依据。

关键词: 虫草棒束孢, 南方根结线虫卵, 克隆, 几丁质酶基因, 序列分析

Abstract: In order to explore the function of chitinase in parasitizing and destroying egg, a nematode egg-paraastic fungus Isaria farinosa HNE3 was isolated from Meloidogyne incognita infecting tobacco in Henan Province.By using RT-PCR and RACE, we had cloned a chitinase gene IFCHI1 for the first time.The full-length DNA of IFCHI1 was 1 417 bp, containing a intron with the length of 61 bp, ORF was 1 185 bp and it encoded 394 amino acids.Analyzing chitinase IFCHI1 by online biological software deduced that its molecular mass and isoelecftric point were 44.1 kDa and 4.88, especially.By compared with other chitinase from different fungus in GenBank, IFCHI1 belongs to glycosyl hydrolase family 18 chitinase.We found two prevalent conserved catalytic domains with the sequence of SIGGW and FDGIDIDWE and two binding domains with the sequence of GTWEQGVYDY and GLGGAMWWESS.Comparison of chitinase IFCHI1 with other chitinases from entomopathogenic fungi revealed that enzymes were highly similar.It was concluded that the chitinase coded by gene IFCHI1 provided theoretical basis for damaging on eggshell structure and inhibiting on egg-hatching of M.incognita.

Key words: Isaria farinosa, Meloidogyne incognita, Cloning, Chitinase gene, Sequence analysis

中图分类号:

王凯, 赵洋, 朱先婷, 才秀华, 武侠. 卵寄生真菌虫草棒束孢几丁质酶基因IFCHI1的克隆与分析[J]. 华北农学报, 2015, 30(2): 78-86. doi: 10.7668/hbnxb.2015.02.015.

WANG Kai, ZHAO Yang, ZHU Xian-ting, CAI Xiu-hua, WU Xia. Cloning and Analysis of Chitinase Gene IFCHI1 from Egg-parasitic Fungus Isaria farinosa[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(2): 78-86. doi: 10.7668/hbnxb.2015.02.015.

http://www.hbnxb.net/CN/Y2015/V30/I2/78

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卵寄生真菌虫草棒束孢几丁质酶基因IFCHI1的克隆与分析

Cloning and Analysis of Chitinase Gene IFCHI1 from Egg-parasitic Fungus Isaria farinosa

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卵寄生真菌虫草棒束孢几丁质酶基因IFCHI1的克隆与分析

Cloning and Analysis of Chitinase Gene IFCHI1 from Egg-parasitic Fungus Isaria farinosa

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