大豆孢囊线虫果胶酸裂解酶基因Hg-pel-5的克隆与分析

doi:10.3864/j.issn.0578-1752.2012.05.005

Abstract

Abstract: Abstract:【Objective】The objective of this study is to clone and analyze the pectate lyase genes from soybean cyst nematode (Heterodera glycines), understand the parasitism and pathogenicity of plant parasitic nematodes, and to develop new control strategies for soybean cyst nematode. 【Method】A noval pectate lyase gene Hg-pel-5 was cloned from H. glycines by EST analysis and RACE. The tissue localization and developmental expression of pectate lyase were analysed at different life stages of H. glycines by in situ mRNA hybridization and RT-PCR amplification, and the copy number of the gene was identified by Southern blot. 【Result】 A novel parasitism gene encoding pectate lyase, named Hg-pel-5, was cloned from H. glycines. The full length cDNA of Hg-pel-5 consists of 957 bp, with a 681 bp ORF encoding a 227 amino acid residues. The structure of Hg-pel-5 genomic DNA includes 2 extons and 1 intron. The predicted protein includes a signal peptide of 20 amino acids on the N-terminal and 4 conserved regions characteristic of class Ⅲ pectate lyases of microbes. In situ mRNA hybridization analyses showed that the transcripts of Hg-pel-5 was expressed in subventral oesophageal gland cells of H. glycines. RT-PCR analysis confirmed that Hg-pel-5 transcriptions were mainly expressed at the pre-parasitic and early parasitic second-stage juveniles. Southern blotting confirmed that Hg-pel-5 was of nematode origin and a member of a small multigene family. 【Conclusion】 A novel pectate lyase gene was cloned and analyzed from H. glycines, and these results imply that the Hg-pel-5 might play an important role during the early stages of plant parasitism.

Key words: Heterodera glycines, pectate lyase, gene cloning, in situ hybridization, Southern blot

PENG Huan, PENG De-Liang, HUANG Wen-Kun, HE Wen-Ting, HU Xian-Qi. Molecular Cloning and Analysis of a Novel Pectate Lyase Gene Hg-pel-5 from Soybean Cyst Nematode[J].Scientia Agricultura Sinica, 2012, 45(5): 854-866.

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Molecular Cloning and Analysis of a Novel Pectate Lyase Gene Hg-pel-5 from Soybean Cyst Nematode

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