Use of cDNA microarrays to monitor transcriptional responses of the chestnut blight fungus Cryphonectria parasitica to infection by virulence-attenuating hypoviruses
Use of cDNA microarrays to monitor transcriptional responses of the chestnut blight fungus Cryphonectria parasitica to infection by virulence-attenuating hypoviruses
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DOI：
10.1128/ec.2.6.1253-1265.2003
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发表时间：
2003-12-01
期刊：
影响因子：
--
通讯作者：
Nuss, DL
中科院分区：
文献类型：
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作者：
Allen, TD;Dawe, AL;Nuss, DL
Hypoviruses是栗疫病菌Cryphonectria parasitica的一个细胞质复制RNA病毒家族。该真菌病毒家族的成员持续改变毒力（低毒力）和相关的真菌发育过程，包括无性和有性孢子形成。为了更好地了解这些变化的分子基础，我们开发了一个C。寄生虫cDNA微阵列监测全球低病毒感染的转录反应。在这份报告中，一个斑点DNA微阵列代表约2,200 C。寄生虫基因的表达被用于监测原型低病毒CHV 1-EP 713感染后转录谱的变化。由于CHV 1-EP 713感染，295个克隆（2，200个独特cDNA的13.4%）的转录物丰度被鉴定为改变的-132个上调，163个下调。相比之下，只有不到20个特定的C。先前通过北方分析和mRNA差异显示鉴定了寄生虫基因对低病毒感染有反应。一个93%的验证率之间实现实时逆转录PCR结果和微阵列预测。差异表达的基因代表了广泛的生物学功能，包括应激反应、碳代谢和转录调控。这些发现与12.7 kbp的低病毒RNA感染导致C.寄生虫转录组微阵列研究的潜在影响，目前和未来的努力，以建立在细胞基因表达和表型的低病毒介导的变化之间的联系进行了讨论。
Hypoviruses are a family of cytoplasmically replicating RNA viruses of the chestnut blight fungus Cryphonectria parasitica. Members of this mycovirus family persistently alter virulence (hypovirulence) and related fungal developmental processes, including asexual and sexual sporulation. In order to gain a better understanding of the molecular basis for these changes, we have developed a C. parasitica cDNA microarray to monitor global transcriptional responses to hypovirus infection. In this report, a spotted DNA microarray representing approximately 2,200 C. parasitica genes was used to monitor changes in the transcriptional profile after infection by the prototypic hypovirus CHV1-EP713. Altered transcript abundance was identified for 295 clones (13.4% of the 2,200 unique cDNAs) as a result of CHV1-EP713 infection-132 up-regulated and 163 down-regulated. In comparison, less than 20 specific C. parasitica genes were previously identified by Northern analysis and mRNA differential display as being responsive to hypovirus infection. A 93% validation rate was achieved between real-time reverse transcription-PCR results and microarray predictions. Differentially expressed genes represented a broad spectrum of biological functions, including stress responses, carbon metabolism, and transcriptional regulation. These findings are consistent with the view that infection by a 12.7-kbp hypovirus RNA results in a persistent reprogramming of a significant portion of the C. parasitica transcriptome. The potential impact of microarray studies on current and future efforts to establish links between hypovirus-mediated changes in cellular gene expression and phenotypes is discussed.