热带作物学报2023，Vol.44Issue(12)：2461-2468,8.DOI:10.3969/j.issn.1000-2561.2023.12.011
橡胶树红根病病原菌木聚糖酶编码基因GpTR1774的克隆与表达分析
Cloning and Expression Analysis of Xylanase GpTR1774 Gene from Ganoderma pseudoferreum
摘要
Abstract
Cell wall degrading enzymes are important pathogenic factors of many pathogenic fungi,among which xy-lanase,as the most important hemicellulase,plays an important role in the process of host cell wall degrading by fila-mentous fungi to achieve host infection.In this study,RT-PCR was used to clone the coding region of xylanase GpTR1774 gene of Ganoderma pseudoferreum strain HD3 and analyze its bioinformatics.The root of Hevea tissue cul-tured seedlings Reyan 73397 were infected with HD3,meanwhile infection and destruction process of root cells were observed by electron microscopy.The gene expression of GpTR1774 was determined by qRT-PCR.The results showed that the total length of GpTR1774 cDNA was 780 bp,encoding 259 amino acids,among which the most abundant amino acid was alanine(Ala),accounting for 15.1%.The molecular weight of GpTR1774 protein was 28.12 kDa,fat coeffi-cient was 81.93,and the isoelectric point was 9.07.It was a hydrophilic protein with 15 phosphorylation sites,no signal and peptide transmembrane domain,and was located in the cell solute.The main components of the secondary structure α-helix and random curling are the main components of the secondary structure of GpTR1774 protein,accounting for 38.10%and 41.31%of the amino acid sequence,respectively.Phylogenetic analysis showed that GpTR1774 gene had the highest similarity with xylanase gene of Ganoderma boninense,reaching 87.5%.qRT-PCR showed that the overall gene expression trend of xylanase GpTR1774 was firstly increased and then decreased.The expression level of GPTR1774 increased significantly on the 3rd and 4th day after infection,and reached the highest level on the 4th day,about 16 times of the initial level.The results of this study indicate that the xylanase GpTR1774 gene was likely to be involved in the pathogenesis of G.psedoferreum,providing reference for the pathogenesis analysis and green prevention and control of G.psedoferreum.关键词
热研73397/红根病病原菌/GpTR1774基因/侵染过程/qRT-PCRKey words
Hevea brasiliensis Reyan 73397/Ganoderma pseudoferreum(Wakef)v.Over.et Steinm/GpTR1774/infec-tion process/qRT-PCR分类
林学引用本文复制引用
伏雪,涂敏,蔡海滨,张红骥,于德才,曾霞..橡胶树红根病病原菌木聚糖酶编码基因GpTR1774的克隆与表达分析[J].热带作物学报,2023,44(12):2461-2468,8.基金项目
海南省重点研发计划项目(No.ZDYF2021XDNY291) （No.ZDYF2021XDNY291）
海南省优秀人才团队项目(No.20210203). （No.20210203）
