Dissecting the Biology of the Fungal Wheat Pathogen Zymoseptoria tritici: A Laboratory Workflow

Current Protocols in Microbiology Pub Date : 2020-11-11 DOI:10.1002/cpmc.128

Wagner C. Fagundes, Janine Haueisen, Eva H. Stukenbrock

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引用次数: 6

Abstract

The fungus Zymoseptoria tritici is one of the most devastating pathogens of wheat. Aside from its importance as a disease-causing agent, this species has emerged as a powerful model system for evolutionary genetic studies of crop-infecting fungal pathogens. Z. tritici exhibits exceptionally high levels of genetic and phenotypic diversity as well as morphological plasticity, which can make experimental studies and comparability of results obtained in different laboratories, e.g., from infection assays, challenging. Therefore, standardized experimental methods are crucial for research on Z. tritici biology and the interaction of this fungus with its wheat host. Here, we describe a suite of well-tested and optimized protocols ranging from isolation of Z. tritici field specimens to analyses of virulence assays under controlled conditions. Several biological and technical aspects of working with Z. tritici under laboratory conditions are considered and carefully described in each protocol. © 2020 The Authors.

Basic Protocol 1: Purification of Z. tritici field isolates from leaf material

Basic Protocol 2: Molecular identification of Z. tritici isolates

Support Protocol 1: Rapid extraction of Z. tritici genomic DNA

Support Protocol 2: Extraction of high-quality Z. tritici genomic DNA

Basic Protocol 3: In vitro culture and long-term storage of Z. tritici isolates

Basic Protocol 4: Analysis of Z. tritici virulence in wheat

Support Protocol 3: Preparation of Z. tritici inoculum

Abstract Image

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小麦真菌病原菌酵母酵母的生物学剖析:实验室工作流程

小麦酵母菌是小麦最具破坏性的病原菌之一。除了作为致病因子的重要性外，该物种已成为作物感染真菌病原体进化遗传研究的强大模型系统。小麦小麦表现出极高的遗传和表型多样性以及形态可塑性，这使得实验研究和不同实验室(例如感染分析)获得的结果的可比性具有挑战性。因此，标准化的实验方法对于研究小麦小麦黑穗病菌的生物学特性及其与小麦寄主的相互作用至关重要。在这里，我们描述了一套经过良好测试和优化的方案，范围从田间样品的分离到受控条件下的毒力分析。在实验室条件下处理麦曲霉的几个生物学和技术方面都被考虑并仔细描述在每个方案中。©2020作者。基本方案1:从叶片材料中分离麦兹黑麦田间分离物的纯化基本方案2:分离麦兹黑麦分子鉴定支持方案1:快速提取麦兹黑麦基因组dna支持方案2:提取高质量的麦兹黑麦基因组dna基础方案3:分离麦兹黑麦离体培养和长期储存基本方案4:小麦中麦兹黑麦毒力分析支持方案3:制备麦兹黑麦接种物

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Current Protocols in Microbiology Immunology and Microbiology-Parasitology

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期刊介绍： Current Protocols in Microbiology provides detailed, step-by-step instructions for analyzing bacteria, animal and plant viruses, fungi, protozoans and other microbes. It offers updated coverage of emerging technologies and concepts, such as biofilms, quorum sensing and quantitative PCR, as well as proteomic and genomic methods. It is the first comprehensive source of high-quality microbiology protocols that reflects and incorporates the new mandates and capabilities of this robust and rapidly evolving discipline.