Constructing Haustorium-Specific cDNA Libraries from Rust Fungi
Constructing Haustorium-Specific cDNA Libraries from Rust Fungi
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DOI：
10.1007/978-1-61737-998-7_8
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发表时间：
2011-01-01
期刊：
影响因子：
--
通讯作者：
Dodds, Peter
中科院分区：
文献类型：
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作者：
Catanzariti, Ann-Maree;Mago, Rohit;Dodds, Peter
吸器是活体营养型植物病原菌的显著特征。几个高度分化的病原体类已经独立地进化吸器，这表明它们代表了在活植物组织内生长的有效适应。尽管它们在活体营养中具有明显的重要性，但由于活体营养病原体与其宿主的密切联系以及无法在体外产生吸器，因此它们一直难以研究。通过吸器分离技术的发展，这些缺点在锈菌的研究中得到了克服。凝集素伴刀豆球蛋白A（ConA）与锈菌吸器的强结合允许这些结构通过ConA-Sepharose macrobead柱上的亲和层析从受感染的植物组织中纯化。分离过程导致大量产量的完整吸器，保留其细胞质内容物，使它们适合实验。从分离的锈菌吸器的cDNA文库的构建和随后的序列分析提供了显着的深入了解吸器功能在分子水平上，揭示了重要的作用，在营养获取和致病性效应蛋白的传递。本章描述了锈菌吸器特异性cDNA文库的产生。
The haustorium is a distinguishing feature of biotrophic plant pathogens. Several highly diverged pathogen classes have independently evolved haustoria, suggesting that they represent an effective adaptation for growing within living plant tissue. Despite their clear importance in biotrophy, they have been difficult to study due to the close association of biotrophic pathogens with their host and the inability to produce haustoria in vitro. These drawbacks have been circumvented in the study of rust fungi by the development of a haustoria isolation technique. The strong binding of the lectin concanavalin A (ConA) to rust haustoria allows these structures to be purified from infected plant tissue by affinity chromatography on a ConA-Sepharose macrobead column. The isolation process results in substantial yields of intact haustoria that retain their cytoplasmic contents, making them amenable to experimentation. The construction of cDNA libraries from isolated rust haustoria and their subsequent sequence analysis have provided significant insight into haustoria function at a molecular level, revealing important roles in nutrient acquisition and the delivery of pathogenicity effector proteins. The generation of a rust haustorium-specific cDNA library is described in this chapter.