基于Biolog表型芯片的立枯丝核菌菌核形成所需代谢底物分析

Biolog phenotypic microarray reveals the metabolic substrates required for sclerotial formation of Rhizoctonia solani

摘要:

目的 分析立枯丝核菌菌核形成所需的代谢底物，旨在了解营养元素和环境因子对其菌核形成的影响规律。方法 基于Biolog表型芯片技术测定663种营养物质、96种渗透压和96种pH环境下立枯丝核菌的菌核形成能力。结果 在供试的营养物质及环境条件中，分别有19/95的碳源、21/95的氮源、16/94的磷硫源、69/94的营养补充物、61/282的肽类氮源、28/96的渗透压环境以及40/96的pH环境可以诱导立枯丝核菌形成菌核；其中，N-乙酰-d-氨基葡萄糖、尿苷3′-单磷酸盐、磷酸胆碱和5种二肽(Arg-Trp、Met-Arg、Pro-Phe、Val-Tyr、Val-Met)，以及10 mmol/L、20 mmol/L硫酸铵(pH 8.0)和pH 4.5+l-脯氨酸3种环境条件可显著促进立枯丝核菌菌核的形成；立枯丝核菌在pH 4.0-10.0范围内均能形成菌核；KEGG分析显示，诱导菌核形成的物质主要参与了代谢途径、ABC转运蛋白、次生代谢物生物合成和d-氨基酸代谢等通路。结论 营养限制和环境胁迫是诱导立枯丝核菌菌核形成的关键因素。在营养受限的条件下，适宜立枯丝核菌形成菌核的物质包括d-山梨醇、d-木糖、N-乙酰-d-半乳糖胺、d-阿拉伯糖和d-松三糖等5种碳源；N-乙酰-d-氨基葡萄糖、腺苷和胸腺嘧啶等3种氮源；尿苷3′-单磷酸盐和磷酸胆碱等2种磷源；吐温-80等1种营养补充物；以及Arg-Trp、Met-Arg、Pro-Phe、Val-Tyr和Val-Met等5种肽类氮源。适宜的渗透压环境为10 mmol/L、20 mmol/L硫酸铵(pH 8.0)，适宜的pH环境为pH 4.0-4.5和pH 9.5-10.0。本研究结果为立枯丝核菌菌核形成机制的研究奠定了基础。

关键词:立枯丝核菌;菌核;物质代谢;表型芯片;KEGG通路

Abstract:

Objective To analyze the metabolic substrates required for the sclerotial formation of Rhizoctonia solani and understand the influence of nutritional elements and environmental factors on this process.Methods Biolog phenotypic microarray was used to study the effects of 663 nutritional substances, 96 osmotic environments, and 96 pH environments on the sclerotial formation of R. solani.Results Among the tested nutritional substances and environmental conditions, 19/95 carbon sources, 21/95 nitrogen sources, 16/94 phosphorus and sulfur sources, 69/94 nutritional supplements, 61/282 peptide nitrogen sources, 28/96 osmotic environments, and 40/96 pH environments induced the sclerotial formation of R. solani. Notably, N-acetyl-d-glucosamine, uridine 3′-monophosphate, phosphoryl choline, and five dipeptides (Arg-Trp, Met-Arg, Pro-Phe, Val-Tyr, and Val-Met), as well as three environmental conditions of 10 mmol/L and 20 mmol/L ammonium sulfate at pH 8.0, and pH 4.5+l-proline, significantly induced the sclerotial formation of R. solani. R. solani formed sclerotia in the environments with a broad range of pH 4.0-10.0. The KEGG analysis indicated that the substances inducing sclerotial formation were primarily involved in metabolic pathways, ABC transporters, secondary metabolite biosynthesis, and d-amino acid metabolism.Conclusion Nutrient limitation and environmental stress are key factors inducing the sclerotial formation of R. solani. Under nutrient-restricted conditions, the suitable substances for inducing sclerotial formation include five carbon sources (d-sorbitol, d-xylose, N-acetyl-d-galactosamine, d-arabinose, and d-melezitose), three nitrogen sources (N-acetyl-d-glucosamine, adenosine, and thymidine), two phosphorus sources (uridine 3′-monophosphate and phosphoryl choline), one nutritional supplement (Tween-80), and five peptide nitrogen sources (Arg-Trp, Met-Arg, Pro-Phe, Val-Tyr, and Val-Met). The suitable osmotic environments were 10 mmol/L and 20 mmol/L ammonium sulfate at pH 8.0, and the suitable pH environments were pH 4.0-4.5 and pH 9.5-10.0. These findings provide a foundation for understanding the sclerotial formation mechanism of R. solani.

Key words:Rhizoctonia solani;sclerotium;material metabolism;phenotypic microarray;KEGG pathway