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转录因子 NsdD 调控草酸青霉中参与植物生物质降解酶、分生孢子形成和色素生物合成的基因表达。

Transcription Factor NsdD Regulates the Expression of Genes Involved in Plant Biomass-Degrading Enzymes, Conidiation, and Pigment Biosynthesis in Penicillium oxalicum.

机构信息

State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Science and Technology, Guangxi University, Nanning, Guangxi, China.

State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Life Science and Technology, Guangxi University, Nanning, Guangxi, China

出版信息

Appl Environ Microbiol. 2018 Aug 31;84(18). doi: 10.1128/AEM.01039-18. Print 2018 Sep 15.

Abstract

Soil fungi produce a wide range of chemical compounds and enzymes with potential for applications in medicine and biotechnology. Cellular processes in soil fungi are highly dependent on the regulation under environmentally induced stress, but most of the underlying mechanisms remain unclear. Previous work identified a key GATA-type transcription factor, NsdD (PoxNsdD; also called POX08415), that regulates the expression of cellulase and xylanase genes in PoxNsdD shares 57 to 64% identity with the key activator NsdD, involved in asexual development in In the present study, the regulatory roles of PoxNsdD in were further explored. Comparative transcriptomic profiling revealed that PoxNsdD regulates major genes involved in starch, cellulose, and hemicellulose degradation, as well as conidiation and pigment biosynthesis. Subsequent experiments confirmed that a Δ strain lost 43.9 to 78.8% of starch-digesting enzyme activity when grown on soluble corn starch, and it produced 54.9 to 146.0% more conidia than the Δ parental strain. During cultivation, Δ cultures changed color, from pale orange to brick red, while the Δ cultures remained bluish white. Real-time quantitative reverse transcription-PCR showed that dynamically regulated the expression of a glucoamylase gene (/), an α-amylase gene (/), and a regulatory gene (/), as well as a polyketide synthase gene (//) for yellow pigment biosynthesis and a conidiation-regulated gene (/). Moreover, binding experiments showed that PoxNsdD bound the promoter regions of the above-described genes. This work provides novel insights into the regulatory mechanisms of fungal cellular processes and may assist in genetic engineering of for potential industrial and medical applications. Most filamentous fungi produce a vast number of extracellular enzymes that are used commercially for biorefineries of plant biomass to produce biofuels and value-added chemicals, which might promote the transition to a more environmentally friendly economy. The expression of these extracellular enzyme genes is tightly controlled at the transcriptional level, which limits their yields. Hitherto our understanding of the regulation of expression of plant biomass-degrading enzyme genes in filamentous fungi has been rather limited. In the present study, regulatory roles of a key regulator, PoxNsdD, were further explored in the soil fungus , contributing to the understanding of gene regulation in filamentous fungi and revealing the biotechnological potential of via genetic engineering.

摘要

土壤真菌产生广泛的化合物和酶,具有在医学和生物技术应用的潜力。土壤真菌的细胞过程高度依赖于环境诱导应激下的调节,但大多数潜在机制仍不清楚。先前的工作确定了一种关键的 GATA 型转录因子 NsdD(PoxNsdD;也称为 POX08415),它调节纤维素酶和木聚糖酶基因在 PoxNsdD 中的表达与关键激活剂 NsdD 有 57%至 64%的同一性,参与无性发育在本研究中,进一步探讨了 PoxNsdD 在中的调控作用。比较转录组分析显示,PoxNsdD 调节参与淀粉、纤维素和半纤维素降解以及产孢和色素生物合成的主要基因。随后的实验证实,Δ 株在可溶玉米淀粉上生长时,丧失了 43.9%至 78.8%的淀粉消化酶活性,比 Δ 亲本株产生了 54.9%至 146.0%更多的分生孢子。在培养过程中,Δ 培养物的颜色从浅橙色变为砖红色,而 Δ 培养物保持蓝白色。实时定量逆转录-PCR 显示,动态调节葡糖淀粉酶基因 (/),α-淀粉酶基因 (/), 调节基因 (/), 以及用于黄色色素生物合成的聚酮合酶基因 (//) 和产孢调节基因 (/)的表达。此外,结合实验表明,PoxNsdD 结合了上述基因的启动子区域。这项工作为真菌细胞过程的调控机制提供了新的见解,并可能有助于对 进行遗传工程改造,以用于潜在的工业和医学应用。大多数丝状真菌产生大量的细胞外酶,这些酶在植物生物质的生物炼制中被商业用于生产生物燃料和增值化学品,这可能有助于向更环保的经济转型。这些细胞外酶基因的表达在转录水平上受到严格控制,限制了它们的产量。迄今为止,我们对丝状真菌中植物生物质降解酶基因表达的调控的理解相当有限。在本研究中,进一步探索了关键调节剂 PoxNsdD 在土壤真菌中的调控作用,有助于理解丝状真菌中的基因调控,并通过遗传工程揭示 的生物技术潜力。

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