发菜伴生细菌的分离鉴定及其对发菜生理代谢的影响  

Isolation and Identification of Associated Bacteria from Nostoc flagelliforme and Its Effects on Physiological Metabolism of Nostoc flagelliforme

作　　者：韩成凤 林亚慧 刘姝婷 韩培培[1] 贾士儒[1] HAN Chengfeng;LIN Yahui;LIU Shuting;HAN Peipei;JIA Shiru(College of Biotechnology,Tianjin University of Science&Technology,Tianjin 300457,China)

机构地区：[1]天津科技大学生物工程学院,天津300457

出　　处：《天津科技大学学报》2023年第1期1-9,共9页Journal of Tianjin University of Science & Technology

基　　金：国家自然科学基金资助项目(32172191)。

摘　　要：为了探究发菜(Nostoc flagelliforme)伴生菌对其生理代谢的影响,进一步了解发菜的生长体系以及发菜与伴生菌的相互作用,本文采用平板划线法从发菜固体培养基上分离出11株伴生菌,利用高效液相色谱-四极杆飞行时间质谱(LC-qToF-MS)鉴定伴生菌产生的N-酰基高丝氨酸内酯类分子(AHLs)种类.通过质谱图可知微杆菌属(Microbacterium sp.)ABNF-1产生C_(12)-HSL,根瘤菌属(Rhizobium sp.)ABNF-4产生C_(6)-HSL、C10-HSL、C_(12)-HSL,戈登氏菌属(Gordonia sp.)ABNF-9产生C_(6)-HSL、C10-HSL,假单胞菌属(Pseudomonas balearica sp.)ABNF-10产生C10-HSL、C_(12)-HSL.对发菜AHLs产生菌共培养的代谢产物分析表明,主成分分析(PCA-X)和正交偏最小二乘法判别分析(OPLS-DA)的评分都呈现明显的离散现象,表明共培养组的代谢产物发生了变化,发菜的脂肪酸代谢和碳代谢发生了显著变化.通过外源添加AHLs发现,当群体感应信号分子AHLs浓度为5μmol/L以上时,AHLs能够促进荚膜多糖的积累,提高细胞总糖含量,对发菜多糖含量影响显著,这进一步说明AHLs能够影响发菜细胞的生长代谢.In our study we intended to study,the effects of the associated bacteria of Nostoc flagelliforme on its physiological metabolism so as to further understand the growth system of N.flagelliforme and the interaction between N.flagelliforme and associated bacteria.Eleven strains of associated bacteria were isolated from N.flagelliforme solid medium by plate scribing method.The species of N-acylhomoserine lactone-like molecules(AHLs)were identified in associated bacteria by high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(LC-qToF-MS).From the mass spectrum,we found that Microbacterium sp.ABNF-1 produced C_(12)-HSL,Rhizobium sp.ABNF-4 produced C_(6)-HSL,C10-HSL and C_(12)-HSL,Gordonia sp.ABNF-9 produced C_(6)-HSL and C10-HSL,and Pseudomonas balearica sp.ABNF-10 produced C10-HSL and C_(12)-HSL.The analysis of metabolites in the co-culture of N.flagelliforme and AHLs producing bacteria showed that the score of principal component analysis(PCA-X)and orthogonal partial least squares discrimination analysis(OPLS-DA)showed obvious dispersion,indicating that the metabolites in the co-culture group changed,and the fatty acid metabolism and carbon metabolism of N.flagelliforme changed significantly.Through exogenous addition of AHLs,we found that when the concentration of signal molecules exceeded 5μmol/L,it promoted the accumulation of capsular polysaccharide,increased the total sugar content of cells,and significantly affected the polysaccharide yield of N.flagelliforme,which further indicated that AHLs could affect the growth and metabolism of N.flagelliforme cells.

关 键 词：发菜 群体感应信号分子AHLs 代谢产物 发菜多糖 

分 类 号：Q819[生物学—生物工程]