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作 者:叶岚 刘辉[1,2] 李珊珊 向文胜 张艳艳 YE Lan;LIU Hui;LI Shanshan;XIANG Wensheng;ZHANG Yanyan(State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China;College of Life Science,Northeast Agricultural University,Harbin 150030,Heilongjiang,China)
机构地区:[1]中国农业科学院植物保护研究所、植物病虫害生物学国家重点实验室,北京100193 [2]东北农业大学生命科学学院,黑龙江哈尔滨150030
出 处:《微生物学报》2023年第2期717-730,共14页Acta Microbiologica Sinica
基 金:国家自然科学基金(31872936)。
摘 要:【目的】研究蓝灰链霉菌中Aco类群感效应信号分子合酶基因scy A1缺失对细胞生理代谢和调控网络造成的广泛性扰动,揭示Scy A1对细胞生命活动的全局性调控作用。【方法】通过测定细胞干重确定scy A1缺失对液体发酵条件下细胞生长的影响。采用RNA-Seq比较分析探究蓝灰链霉菌scy A1突变株和野生型NMWT1发酵培养3d和6d全基因组范围内的显著差异表达基因。【结果】scy A1缺失不影响液体发酵条件下细胞生物量的积累。比较转录组分析显示scy A1缺失后,糖酵解和三羧酸循环途径基因表达表现出双向显著差异变化趋势;L-半乳糖形成UDP-葡萄糖途径、戊糖磷酸途径、氨基酸(L-缬氨酸、L-异亮氨酸和L-色氨酸)合成途径和嘌呤核苷酸降解途径相关基因均表现出显著上调趋势。众多次级代谢生物合成基因簇、保守转录调控因子和菌丝体结构性蛋白组分编码基因表达显著下调,而少数则表达水平显著上调。【结论】Scy A1广泛影响了菌株的初级代谢、次级代谢、保守调控因子和菌丝体结构相关基因的表达。总之,本研究丰富了我们对Aco类群感效应信号分子合酶功能的认知。[Objective]To investigate the effects of deletion of an Aco-like autoregulator synthase gene,scyA1,on the physiological metabolism and regulatory network of cells,and thus to reveal the global roles of Scy A1 in cellular activities.[Methods]Dry cell weight was measured to determine the effect of scyA1 deletion on the growth of cells in liquid fermentation.Comparative RNA-Seq analysis between the scyA1 mutant and the wild type NMWT1 was performed to explore the genome-wide differentially expressed genes after 3 and 6 days of fermentation.[Results]The deletion of scyA1 did not affect the biomass of cells in liquid fermentation.After the deletion of scyA1,the expression of the genes involved in glycolysis and tricarboxylic acid cycle showed significant bidirectional differences,and the expression levels of the genes involved in the pathway from L-galactose to UDP-glucose,the pentose phosphate pathway,amino acid(L-valine,L-isoleucine,and L-tryptophan)biosynthesis,and purine nucleotide degradation were significantly upregulated.Moreover,the expression levels of a variety of gene clusters for the biosynthesis of secondary metabolites and the genes encoding conserved transcriptional regulators and mycelium structural proteins were significantly downregulated,while those of a few were significantly upregulated.[Conclusion]Scy A1 affected primary metabolism,secondary metabolism,and expression of conserved regulatory factors and mycelium structure-related genes of the test strain.The results of this study demonstrated new details of the global function of the Aco-like autoregulator synthase in Streptomyces.
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