Co-Innovation Center for Sustainable Forestry in Southern China, College of Biology and the Environment, Nanjing Forestry University, Nanjing, China.
Noda Institute for Scientific Research, Noda, Japan.
Appl Environ Microbiol. 2019 Apr 4;85(8). doi: 10.1128/AEM.00089-19. Print 2019 Apr 15.
Basic-region helix-loop-helix (bHLH) proteins are a superfamily of transcription factors that are often involved in the control of growth and differentiation. Recently, it was reported that the bHLH transcription factor DevR is involved in both asexual and sexual development in and regulates the conidial melanin production in In this study, we identified and characterized an gene that showed high similarity with of and (Ao). In the Ao-disrupted strain, growth was delayed and the number of conidia was decreased on Czapek-Dox (CD) minimal agar plates, but the conidiation was partially recovered by adding 0.6 M KCl. Simultaneously, the overexpression of Ao was induced and resulted in extremely poor growth when the carbon source changed from glucose to polysaccharide (dextrin) in the CD agar plate. Scanning electron microscopy (SEM) indicated that the overexpression of Ao resulted in extremely thin aberrant hyphal morphology. Conversely, the deletion of Ao resulted in thicker hyphae and in more resistance to Congo red relative to the control strain. Quantitative reverse transcriptase PCR (RT-PCR) further indicated that AoDevR significantly affects chitin and starch metabolism, and importantly, the overexpression of Ao inhibited the expression of genes related to starch degradation. A yeast one-hybrid assay suggested that the DevR protein possibly interacted with the promoter of , which encodes a transcription factor involved in amylase production. Importantly, AoDevR is involved in polysaccharide metabolism and affects the growth of the strain. is an industrially important filamentous fungus; therefore, a clear understanding of its polysaccharide metabolism and utilization is very important for its industrial utilization. In this study, we revealed that the basic-region helix-loop-helix (bHLH) transcription factor AoDevR is importantly involved in chitin and starch metabolism in The overexpression of Ao strongly suppressed the expression of amylase-related genes. The results of a yeast one-hybrid assay suggested that the DevR protein potentially interacts with the promoter of , which encodes a transcription factor involved in amylase production and starch utilization. This study provides new insight for further revealing the regulation mechanism of amylase production in .
基本区螺旋-环-螺旋(bHLH)蛋白是转录因子的超家族,通常参与控制生长和分化。最近有报道称,bHLH 转录因子 DevR 参与 和 的无性和有性发育,并调节 的分生孢子黑色素产生。在本研究中,我们鉴定并表征了一个 基因,该基因与 和 (Ao)的 具有高度相似性。在 Ao 敲除菌株中,在 Czapek-Dox (CD)最小琼脂平板上的生长延迟,分生孢子数量减少,但添加 0.6 M KCl 可部分恢复分生孢子形成。同时,Ao 的过表达被诱导,当碳源从葡萄糖变为多糖(糊精)时,在 CD 琼脂平板上导致生长极差。扫描电子显微镜(SEM)表明,Ao 的过表达导致菌丝形态极其异常,非常细。相反,与对照菌株相比,Ao 的缺失导致菌丝较厚,对刚果红的抗性更强。定量逆转录 PCR (RT-PCR)进一步表明,AoDevR 显著影响几丁质和淀粉代谢,重要的是,Ao 的过表达抑制了与淀粉降解相关的基因的表达。酵母单杂交试验表明,DevR 蛋白可能与编码参与淀粉酶产生的转录因子的 基因的启动子相互作用。重要的是,AoDevR 参与多糖代谢并影响 菌株的生长。 是一种具有工业重要性的丝状真菌;因此,对其多糖代谢和利用的清晰理解对其工业利用非常重要。在本研究中,我们揭示了基本区螺旋-环-螺旋(bHLH)转录因子 AoDevR 重要参与 的几丁质和淀粉代谢。Ao 的过表达强烈抑制了与淀粉酶相关的基因的表达。酵母单杂交试验的结果表明,DevR 蛋白可能与编码参与淀粉酶产生和淀粉利用的转录因子的 基因的启动子相互作用。本研究为进一步揭示 在淀粉酶产生中的调控机制提供了新的见解。