Suppression subtractive hybridization and comparative expression analysis to identify developmentally regulated genes in filamentous fungi
Suppression subtractive hybridization and comparative expression analysis to identify developmentally regulated genes in filamentous fungi
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DOI：
10.1002/jobm.201200223
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发表时间：
2013-09-01
影响因子：
3.1
通讯作者：
Nowrousian, Minou
中科院分区：
文献类型：
--
作者：
Gesing, Stefan;Schindler, Daniel;Nowrousian, Minou
子囊菌区分四种主要的形态类型的子实体（apothelium，perithelium，pseudothelium和cleistothelium），这些子实体来自祖先的子实体。因此，子实体分化很可能是由一组共同的核心基因控制的。鉴定这些基因的一种方法是寻找具有进化保守表达模式的基因。利用抑制性消减杂交（SSH），我们选择了差异表达的转录本在Pyronema confluens（Pezizales）通过比较两个cDNA文库的性和营养发展，分别。通过定量真实的时间PCR验证从两个文库中选择的基因的表达模式。几个相应的同源基因的表达被认为是保守的两个成员的Sordariales（Sordariamacrospora和Neurosporacrassa），一个衍生的组子囊菌，这是唯一的远亲的Pezizales。用N.差异调节基因的crassa直向同源物揭示了基因NCU 05079在子实体发育期间的功能作用，该基因编码推定的MFS肽转运蛋白。这些数据表明保守的基因表达模式和相应的基因在子实体发育过程中的功能作用，这些基因是进一步功能分析的候选人。
Ascomycetes differentiate four major morphological types of fruiting bodies (apothecia, perithecia, pseudothecia and cleistothecia) that are derived from an ancestral fruiting body. Thus, fruiting body differentiation is most likely controlled by a set of common core genes. One way to identify such genes is to search for genes with evolutionary conserved expression patterns. Using suppression subtractive hybridization (SSH), we selected differentially expressed transcripts in Pyronema confluens (Pezizales) by comparing two cDNA libraries specific for sexual and for vegetative development, respectively. The expression patterns of selected genes from both libraries were verified by quantitative real time PCR. Expression of several corresponding homologous genes was found to be conserved in two members of the Sordariales (Sordaria macrospora and Neurospora crassa), a derived group of ascomycetes that is only distantly related to the Pezizales. Knockout studies with N. crassa orthologues of differentially regulated genes revealed a functional role during fruiting body development for the gene NCU05079, encoding a putative MFS peptide transporter. These data indicate conserved gene expression patterns and a functional role of the corresponding genes during fruiting body development; such genes are candidates of choice for further functional analysis.