RNA-seq analyses of gene expression in the microsclerotia of Verticillium dahliae.
RNA-seq analyses of gene expression in the microsclerotia of Verticillium dahliae.
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DOI：
10.1186/1471-2164-14-607
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发表时间：
2013-09-09
期刊：
影响因子：
4.4
通讯作者：
Klosterman SJ
中科院分区：
文献类型：
--
作者：
Duressa D;Anchieta A;Chen D;Klimes A;Garcia-Pedrajas MD;Dobinson KF;Klosterman SJ
土传真菌大丽轮枝菌（Verticillium dahliae）可引起植物的黄萎病。黄萎病难以控制，因为大丽轮枝菌能够作为黑化微菌核在土壤中持续10至15年，使得用于疾病控制的作物轮作策略无效。大丽轮枝菌的微菌核越冬并萌发产生感染性菌丝，引起原发感染。因此，微菌核的形成、维持和萌发是大丽轮枝菌病害循环中至关重要的过程。为了进一步阐明有助于大丽轮枝菌中微菌核生物发生和黑色素合成的分子过程，从大丽轮枝菌菌株VdLs.17（平均值= 5223万个读数）和不产生微菌核的培养物（NoMS，平均值= 5058万个读数）的10天龄产生微菌核（MS）的培养物制备三个重复的RNA-seq文库。对这些文库的差异基因表达的分析揭示了超过200个差异表达的基因，包括黑素生成相关基因四羟基萘还原酶（增加344倍）和scytalone酶（增加231倍）的上调，以及位于菌株VdLs.17的48.8酶黑色素生物合成基因簇中的其他基因。在MS文库中鉴定为差异表达的基因中有近50%编码假设蛋白质。在促进或阻止微菌核发育的生长条件下，使用微阵列方法用来自大丽轮枝菌菌株Dvd-T5和来自无微菌核vdh 1菌株的RNA进行大丽轮枝菌中基因表达的另外的比较分析。使用RT-qPCR或北方杂交证实通过RNA-seq或微阵列分析观察到的所选基因的差异表达。总的来说，从这些调查中获得的数据提供了额外的洞察力的基因表达和分子过程中发生的MS生物发生和成熟的大丽轮枝菌。因此，鉴定的基因产物可能代表通过预防生存结构发育进行疾病控制的新靶点。
The soilborne fungus, Verticillium dahliae, causes Verticillium wilt disease in plants. Verticillium wilt is difficult to control since V. dahliae is capable of persisting in the soil for 10 to 15 years as melanized microsclerotia, rendering crop rotation strategies for disease control ineffective. Microsclerotia of V. dahliae overwinter and germinate to produce infectious hyphae that give rise to primary infections. Consequently, microsclerotia formation, maintenance, and germination are critically important processes in the disease cycle of V. dahliae. To shed additional light on the molecular processes that contribute to microsclerotia biogenesis and melanin synthesis in V. dahliae, three replicate RNA-seq libraries were prepared from 10 day-old microsclerotia (MS)-producing cultures of V. dahliae, strain VdLs.17 (average = 52.23 million reads), and those not producing microsclerotia (NoMS, average = 50.58 million reads). Analyses of these libraries for differential gene expression revealed over 200 differentially expressed genes, including up-regulation of melanogenesis-associated genes tetrahydroxynaphthalene reductase (344-fold increase) and scytalone dehydratase (231-fold increase), and additional genes located in a 48.8 kilobase melanin biosynthetic gene cluster of strain VdLs.17. Nearly 50% of the genes identified as differentially expressed in the MS library encode hypothetical proteins. Additional comparative analyses of gene expression in V. dahliae, under growth conditions that promote or preclude microsclerotial development, were conducted using a microarray approach with RNA derived from V. dahliae strain Dvd-T5, and from the amicrosclerotial vdh1 strain. Differential expression of selected genes observed by RNA-seq or microarray analysis was confirmed using RT-qPCR or Northern hybridizations. Collectively, the data acquired from these investigations provide additional insight into gene expression and molecular processes that occur during MS biogenesis and maturation in V. dahliae. The identified gene products could therefore potentially represent new targets for disease control through prevention of survival structure development.