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偏肿革裥菌木屑处理下纤维素酶基因表达

Expression Analysis of Cellulase Genes in Lenzites gibbosaInduced by Sawdust
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摘要 白腐真菌能够降解木质纤维基质的所有成分,为了深入探讨在分子层面其对纤维素降解的机制,检测了白腐真菌偏肿革裥菌(Lenzites gibbosa)在木质和非木质环境下滤纸酶、β-葡萄糖苷酶、内切葡聚糖酶和外切葡聚糖酶等纤维素酶的活性变化。并在转录组测序的基础上,对偏肿革裥菌的各种纤维素酶基因进行筛选,分析表达量,结合功能注释、代谢通路富集与生物信息学分析探讨纤维素酶的具体催化分解功能,从而分析这些纤维素酶基因的表达规律。结果表明:滤纸酶、β-葡萄糖苷酶、内切葡聚糖酶和外切葡聚糖酶活性被木屑诱导效果明显,但并非一直被诱导表达;转录组测序共筛选出46个纤维素酶基因,京都基因与基因组百科全书数据库(KEGG)富集分析发现纤维素酶基因显著富集于淀粉和蔗糖代谢通路(ko00500)和氰基氨基酸代谢通路(ko00460),生物信息学分析发现β-葡萄糖苷酶基因产物中具有较多的非分泌蛋白及跨膜蛋白;β-葡萄糖苷酶基因的表达与其酶活变化关联性较差,其原因:一是对不在淀粉和蔗糖代谢通路中的相应同工酶基因未进行定量分析,二是纤维素酶活性不只受到基因转录水平的控制,且纤维素酶活性与基因转录水平没有直接的关系与关联,主要受到底物和培养条件的制约。本研究为深入了解白腐真菌的木材降解机制提供了纤维素酶基因及其表达的资料。 White rot fungi can degrade all components of lignocellulosic substrates.In order to investigate the mechanism of cellulose degradation at the molecular level,the activity changes of cellulase,such as filter paper enzyme,β-glucosidase,endoglucanase and exoglucanase,were detected in the white rot fungus Lenzites gibbosa in woody and non-woody environments,and with transcriptome sequencing,various cellulase genes of L.gibbosa were screened,and the expression level was analyzed,and the specific catalytic decomposition function of cellulase was explored by combining functional annotations,metabolic pathway enrichment analysis and bioinformatics analysis,and to analyze the regular pattern of these cellulase genes.The results showed that the activities of filter paper,β-glucosidase,endoglucanase and exoglucanase were obviously induced by sawdust,but not always induced and expressed;a total of 46 cellulase genes were screened by transcriptome sequencing.By KEGG enrichment analysis,cellulase genes were significantly enriched in starch and sucrose metabolism pathway(ko00500)and cyanoamino acid metabolism pathway(ko00460).By bioinformatics analysis,β-glucosidase gene products contained more non-secreted proteins and transmembrane proteins.The expressions ofβ-glucosidase genes havepoor correlationwith the changes of their enzyme activities.It is speculated that one is that the corresponding isoenzyme genes that are not in the starch and sucrose metabolic pathway have not been quantitatively analyzed,and the other is that cellulase activities are not only controlled by the levels of gene transcription,and there is no direct relationship and correlation between cellulase activities and the levels of gene transcription.The cellulase activities may be primarily constrained by substrate and culture conditions.This study would provide cellulase genes and their regular information for the in-depth understanding of the wood degradation mechanism of white rot fungi.
作者 谷新治 张俊 池玉杰 Gu Xinzhi;Zhang Jun;Chi Yujie(Northeast Forestry University,Harbin 150040,P.R.China;Xinjiang Ecology and Geography Institute,Chinese Academy of Sciences)
出处 《东北林业大学学报》 CAS CSCD 北大核心 2023年第4期108-114,共7页 Journal of Northeast Forestry University
基金 国家自然科学基金项目(30671700) 黑龙江省自然科学基金项目(C2016006) 东北亚生物多样性研究中心双一流专项-自主创新-科技创新团队项目(411146030416)。
关键词 偏肿革裥菌 转录组 纤维素酶 淀粉和蔗糖代谢通路 Lenzites gibbosa Transcriptome Cellulase Starch and sucrose metabolism pathway
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