Boualem Khadidja, Waché Yves, Garmyn Dominique, Karbowiak Thomas, Durand Alain, Gervais Patrick, Cavin Jean-François
Laboratoire GPMA, IFR92, ENSBANA, Université de Bourgogne, 1 Esplanade Erasme, F-21000 Dijon, France.
Res Microbiol. 2008 Mar;159(2):110-7. doi: 10.1016/j.resmic.2007.10.004. Epub 2007 Nov 12.
Based on bioinformatic data on model fungi, the rodA and wetA genes encoding, respectively, a RodA hydrophobin protein and the WetA protein involved in conidiation mechanisms, were PCR-cloned and characterized for the first time in Penicillium camemberti. These results, completed by a sequence of the brlA gene (available in GenBank), which encodes a major transcriptional regulator also involved in the conidiation mechanism, were used to compare, by qRT-PCR, the expression of the three genes in liquid and solid cultures in a synthetic medium. While expression of the brlA and wetA genes increased dramatically in both culture conditions after 4 days of growth, expression of the rodA gene increased only with conidiation and in the solid culture, and this expression was correlated with production and secretion of a RodA protein outside the hyphae, which became very hydrophobic. In liquid cultures, no production of RodA occurred in mycelia, which remained hydrophilic, and no conidiation was detected despite formation of swellings at the tips of hyphae. The absence of conidiation in liquid culture correlated with the lack of rodA gene expression, which could be regulated by the medium composition independently of brlA and wetA genes expression.
基于模式真菌的生物信息学数据,首次在卡门柏青霉中通过PCR克隆并鉴定了分别编码RodA疏水蛋白和参与分生孢子形成机制的WetA蛋白的rodA和wetA基因。编码同样参与分生孢子形成机制的主要转录调节因子的brlA基因序列(可在GenBank中获取)完善了这些结果,通过qRT-PCR用于比较这三个基因在合成培养基中的液体和固体培养物中的表达。在生长4天后,brlA和wetA基因的表达在两种培养条件下均显著增加,而rodA基因的表达仅在分生孢子形成时以及固体培养物中增加,并且这种表达与菌丝体外RodA蛋白的产生和分泌相关,该蛋白变得非常疏水。在液体培养物中,菌丝体中未产生RodA,其仍保持亲水性,尽管在菌丝尖端形成了肿胀,但未检测到分生孢子形成。液体培养物中分生孢子形成的缺失与rodA基因表达的缺乏相关,rodA基因表达可由培养基成分独立于brlA和wetA基因表达进行调控。