山新杨生长素关键基因响应木霉和链格孢菌诱导的转录组分析

发布时间：2018-05-15 20:25

本文选题：山新杨 + 棘孢木霉　；参考：《东北林业大学》2017年硕士论文

【摘要】：本研究以山新杨(Populus davidiana×P.alba var.pyramidalis)组培苗为试材,在山新杨根际、成熟叶片分别接种棘孢木霉(Trichoderma asperellum)、链格孢菌(Alternaria alternata),构建山新杨与木霉菌、链格孢菌、木霉-链格孢菌互作48 h时茎尖、成熟叶(病原菌侵染)、幼根等12个样品的cDNA文库,在转录组水平分析山新杨生长素合成和信号转导关键基因对木霉菌定殖、链格孢菌侵染的表达调控。主要研究结果为:通过转录组测序共得到107.72 Gb Clean Data,Q30碱基百分比均不小于93.07%。参考毛果杨基因组组装后得到的基因与COG、KOG、GO、KEGG、NR、Swiss-Prot数据库进行比对,共有40151条基因得到注释。以Fold Change≥1且FDR0.05为标准检测差异基因,发现链格孢菌侵染后山新杨叶片(病原菌侵染)差异基因最多;而木霉菌定殖后山新杨茎尖的差异基因最多;而木霉-链格孢菌共同诱导后山新杨幼根中差异基因最多。在生长素合成途径中获得10条关键差异基因,包括YUCCA、ALDH、UGT74B家族基因。对差异基因表达模式分析,发现木霉菌定殖主要会导致山新杨幼根和茎尖中ALDH3I1基因(编码蛋白参与色胺合成生长素途径)上调表达;链格孢菌侵染对山新杨茎尖中YUCCA(编码蛋白参与生长素合成主要途径-吲哚丙酮酸途径)表达影响最大,会使其显著下调;木霉-链格孢菌共同诱导会导致山新杨成熟叶片、幼根中UGT74B(编码蛋白参与吲哚乙醛肟合成生长素途径)基因上调表达。在生长素信号转导途径中获得118条关键差异基因,包括AUX1、ARF、AUX/IAA、SAUR、GH3、E1、E2、SCF复合体(E3)家族基因。对差异基因表达模式分析,发现木霉菌定殖主要影响山新杨茎尖组织中的SAUR、AUX/IAA基因表达变化,链格孢菌侵染主要诱导成熟叶片(病原菌侵染)中SAUR、AUX/IAA、GH3基因表达模式发生变化;木霉-链格孢菌共同诱导会使山新杨幼根中SAUR、AUX/IAA、GH3基因差异表达。此外,不同条件诱导山新杨对泛素介导的AUX/IAA蛋白水解相关基因影响均较小。通过对基因结构及功能分析,共获得33条ARF基因,其中有22条在不同样品中存在差异表达,对差异基因表达模式分析,发现木霉菌定殖主要影响茎尖中ARF基因的表达;链格孢菌侵染对山新杨成熟叶片中ARF基因表达影响最大;木霉-链格孢菌共同诱导对山新杨幼根中ARF基因影响较大,并且会使8条ARF基因下调表达。此外,多数差异表达的ARF基因变化量较小。我们选取4条ARF基因进行荧光定量PCR(RTqPCR)检测,其表达趋势与转录组测序结果基本一致。
[Abstract]:In this study, Populus davidiana x P.alba var.pyramidalis was used as a test material. In the new Yang Genji, mature leaves were inoculated with Trichoderma acanthomiasa (Trichoderma asperellum) and cyclosporin (Alternaria alternata) respectively. The stem apex and mature leaf (pathogenic bacteria) were constructed at 48 h each other. Infection), the cDNA Library of 12 samples, such as young roots, was used to analyze the regulation of the expression of the key genes of new poplar and signal transduction on the colonization of Trichoderma. The main results were: 107.72 Gb Clean Data was obtained by the sequence of transcriptional sequences, and the percentage of Q30 base was not less than that of 93.07%. reference Yang Ji. Compared to the COG, KOG, GO, KEGG, NR, Swiss-Prot database, 40151 genes were annotated. The difference gene was detected with Fold Change > 1 and FDR0.05 as the standard. 10 key differentially genes, including YUCCA, ALDH and UGT74B family genes, were obtained in the pathway of auxin synthesis. The analysis of differential gene expression patterns showed that the colonization of Trichoderma could lead to the ALDH3I1 gene (coded egg) in the young root and the tip of the stem. White ginseng and chromamine synthesis of auxin pathway was up-regulated, and the infection of cyclosporin had the greatest influence on the expression of YUCCA (encoded protein in the main pathway of auxin synthesis, indolo pyruvate pathway) in the tip of new poplar. 118 key differential genes were obtained in the auxin signal transduction pathway, including AUX1, ARF, AUX/IAA, SAUR, GH3, E1, E2, and SCF complex (E3) family genes. The analysis of the differential gene expression patterns showed that Trichoderma colonization mainly affected SAUR, AUX/IAA base in the stem tip tissue of new poplar. The expression patterns of SAUR, AUX/IAA and GH3 were changed mainly in the induction of mature leaves (pathogen infection) by the infection of cyclosporin, and the co induction of Trichoderma Alternaria would make the SAUR, AUX/IAA, and GH3 genes expressed in the young roots of new poplar. In addition, different conditions were used to induce the genes related to the ubiquitin mediated AUX/IAA protein hydrolysis. 33 ARF genes were obtained through the analysis of gene structure and function, of which 22 of them were expressed differently in different samples. The analysis of the differential gene expression patterns showed that the colonization of Trichoderma mainly affected the expression of ARF gene in the stem tip; the infection of cyclosporin had the greatest influence on the expression of ARF gene in the mature leaves of new poplar; The effect of CO induction of mycophenium on the ARF gene in young root of poplar was larger, and the expression of 8 ARF genes was downregulated. In addition, most of the differentially expressed ARF gene changes were small. We selected 4 ARF genes for fluorescence quantitative PCR (RTqPCR) detection, and the expression trend was basically consistent with the sequence of the transfer group.

【学位授予单位】：东北林业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S763.7

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