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果胶裂解酶基因在侵染过程中大量表达,调控 和 的形态发育,是该病毒对茶树完全毒力所必需的。

Pectate Lyase Genes Abundantly Expressed During the Infection Regulate Morphological Development of and Is Required for Full Virulence to Tea Plants.

机构信息

College of Tea Science and Tea Culture, Zhejiang A&F University, Hangzhou, Zhejiang, China.

Tea Research Institute, Chinese Academy of Agricultural Sciences/National Center for Tea Improvement/Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture and Rural Affairs of the People's Republic of China, Hangzhou, Zhejiang, China.

出版信息

mSphere. 2023 Feb 21;8(1):e0067722. doi: 10.1128/msphere.00677-22. Epub 2023 Jan 24.

Abstract

Colletotrichum camelliae is the dominant species causing foliar diseases of tea plants (Camellia sinensis) in China. Transcriptome data and reverse transcription-quantitative PCR (qRT-PCR) analysis have demonstrated that the pectate lyase genes in (s) were significantly upregulated during infectious development on tea plants (cv. ). To further evaluate the biological functions of s, we established a polyethylene glycol (PEG)-mediated protoplast transformation system of and generated targeted deletion mutants of seven s. Phenotypic assays showed that the genes contribute to mycelial growth, conidiation, and appressorium development. The polypeptides encoded by each gene contained a predicted N-terminal signal peptide, and a yeast invertase secretion assay suggested that each CcPEL protein could be secreted. Cell death-suppressive activity assays confirmed that all seven CcPELs did not suppress Bax-induced cell death in tobacco leaf cells. However, deletion of significantly reduced necrotic lesions on tea leaves. Taken together, these results indicated that s play essential roles in regulating morphological development, and is required for full virulence in . In this study, we first established a PEG-mediated protoplast transformation system of and used it to investigate the biological functions of seven pectate lyase genes (s) which were abundantly expressed during infection. The results provided insights into the contributions of pectate lyase to mycelial growth, conidial production, appressorium formation, and the pathogenicity of We also confirmed the secretory function of CcPEL proteins and their role in suppressing Bax-induced cell death. Overall, this study provides an effective method for generating gene-deletion transformants in and broadens our understanding of pectate lyase in regulating morphological development and pathogenicity.

摘要

炭疽菌是引起中国茶树(Camellia sinensis)叶部病害的优势种。转录组数据和反转录定量 PCR(qRT-PCR)分析表明,在感染茶树(cv.)的过程中,(s)中的果胶裂解酶基因显著上调。为了进一步评估 s 的生物学功能,我们建立了一种聚乙二醇(PEG)介导的 原生质体转化系统,并生成了 7 个 s 的靶向缺失突变体。表型分析表明,这些基因与菌丝生长、分生孢子形成和附着胞发育有关。每个 基因编码的多肽都含有一个预测的 N 端信号肽,酵母转化酶分泌实验表明每个 CcPEL 蛋白都可以分泌。细胞死亡抑制活性分析证实,所有 7 个 CcPEL 蛋白均不能抑制烟草叶细胞中 Bax 诱导的细胞死亡。然而,缺失 显著减少了茶树叶上的坏死斑。综上所述,这些结果表明 s 在调节形态发育中起重要作用,而 在 中完全毒力需要 。在这项研究中,我们首次建立了一种 PEG 介导的原生质体转化系统,并利用它来研究在感染过程中大量表达的七个果胶裂解酶基因(s)的生物学功能。结果为果胶裂解酶对菌丝生长、分生孢子产生、附着胞形成和 的致病性的贡献提供了新的认识。我们还证实了 CcPEL 蛋白的分泌功能及其在抑制 Bax 诱导的细胞死亡中的作用。总的来说,这项研究为在 中生成基因缺失转化体提供了一种有效的方法,并拓宽了我们对果胶裂解酶在调节形态发育和致病性中的作用的理解。

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