枯草芽孢杆菌NCD-2对棉花根系分泌物L-脯氨酸响应的转录-蛋白质组学联合分析  

Transcriptome and Proteome Analysis of Bacillus subtilis NCD-2 Response to L-proline from Cotton Root Exudates

作　　者：赵卫松[1] 郭庆港[1] 董丽红[1] 王培培[1] 苏振贺 张晓云[1] 鹿秀云[1] 李社增[1] 马平[1] ZHAO WeiSong;GUO QingGang;DONG LiHong;WANG PeiPei;SU ZhenHe;ZHANG XiaoYun;LU XiuYun;LI SheZeng;MA Ping(Plant Protection Institute of Hebei Academy of Agricultural and Forestry Sciences/IPM Centre of Hebei Province/Key Laboratory of IPM on Crops in Northern Region of North China,Ministry of Agriculture and Rural Affairs,Baoding 071000,Hebei)

机构地区：[1]河北省农林科学院植物保护研究所/河北省农业有害生物综合防治工程技术研究中心/农业农村部华北北部作物有害生物综合治理重点实验室,河北保定071000

出　　处：《中国农业科学》2021年第21期4585-4600,共16页Scientia Agricultura Sinica

基　　金：国家自然科学基金(31801786);国家现代农业产业技术体系(CARS-15-18);河北省农林科学院科学技术研究与发展计划(201820101);国家公益性行业(农业)科研专项(201503109)。

摘　　要：【目的】棉花根系分泌物L-脯氨酸是影响生防菌株定殖的关键因素之一,前期研究发现L-脯氨酸能够提高枯草芽孢杆菌(Bacillus subtilis)NCD-2生物膜形成能力。通过高通量测序技术分析L-脯氨酸调控枯草芽孢杆菌NCD-2生物膜形成和生防潜力相关的基因,为深入认识棉花根系分泌物与生防菌株的分子互作关系打下基础。【方法】以枯草芽孢杆菌NCD-2菌株为供试材料,外源添加浓度为10 mg·mL^(-1)的L-脯氨酸共培养24 h后,分别进行转录组(RNA-seq)和同位素标记相对定量蛋白质组技术(iTRAQ)分析,对所获得的转录组和蛋白质组数据进行生物信息学分析,利用实时荧光定量PCR(RT-qPCR)验证不同代谢通路中部分差异基因的表达情况。【结果】转录组分析发现,L-脯氨酸和NCD-2菌株共培养后,获得1071个差异表达基因(DEG),其中602个基因上调,469个基因下调。GO分析发现在生物过程、细胞组分和分子功能方面分别有49、14和30个功能条目显著富集。KEGG代谢通路主要富集在化合物代谢、鞭毛组装、细菌运动和趋化作用。蛋白质组学分析发现,筛选到211个差异表达蛋白(DEP),其中118个蛋白上调,93个蛋白下调。GO分析发现在生物过程和分子功能方面分别有13和8个功能条目显著富集。KEGG代谢通路主要富集在氨基酸代谢、碳水化合物类代谢、鞭毛组装和ABC转运蛋白。进一步转录-蛋白质组学联合分析发现,在L-脯氨酸作用下,检测到共有的显著差异表达基因(或蛋白)112个,其中38个基因(或蛋白)下调,74个基因(或蛋白)上调。GO功能显著富集主要集中在营养库活性、催化活性、细胞膜、定位、细胞脂质代谢过程、氧化还原过程、sigma因子活性、转运活性、芽孢形成9个方面。KEGG代谢通路主要富集在能量代谢、ABC转运蛋白、抗生素生物合成、鞭毛组装、运动或趋化作用和双组分系统方面。RT-qPCR验证了26个�【Objective】L-proline in root exudates of cotton is one of the key factors affecting the colonization of biocontrol microorganisms.Previous studies showed that L-proline could improve the ability of Bacillus subtilis NCD-2 biofilm formation.The objective of this study is to explore the regulatory genes related to the biofilm formation and biocontrol potential of strain NCD-2 through high-throughput sequencing technology,and to lay a foundation for further understanding the molecular interaction between cotton root exudates and biocontrol strain.【Method】Strain NCD-2 was co-cultured with L-proline at the concentration of 10 mg·mL^(-1)for 24 h,and it was analyzed by transcriptome(RNA-seq)and isotope labeled relative quantitative proteomics(iTRAQ).The expression of some differential genes in different metabolic pathways was verified by RT-qPCR.【Result】Transcriptome analysis showed that 1071 differentially expressed genes(DEGs)were obtained after L-proline and NCD-2 co-culture,of which 602 genes were up-regulated and 469 genes were down-regulated.GO analysis showed that 49,14 and 30 functional items were significantly enriched in biological process,cell component and molecular function,respectively.KEGG pathways were mainly enriched in compound metabolism,flagellum assembly,bacterial motility or chemotaxis.Proteomic analysis showed that a total of 211 differentially expressed proteins(DEPs)were detected compared to the control,of which 118 proteins were up-regulated and 93 proteins were down-regulated.GO analysis showed that 13 and 8 functional items were significantly enriched in biological process and molecular function,respectively.KEGG pathways were mainly enriched in amino acid metabolism,carbohydrate metabolism,flagellum assembly and ABC transporter.Further transcriptional-proteomics analysis revealed that 112 DEGs(or DEPs),including 38 down-regulated genes(or proteins)and 74 up-regulated genes(or proteins),were detected.GO functional items were significantly enriched in nine aspects,including nutrie

关 键 词：L-脯氨酸 枯草芽孢杆菌 转录组 蛋白质组 互作关系 

分 类 号：S562[农业科学—作物学]