NaCl处理谷子萌发期种子的转录组学分析  

Transcriptomics Analysis of Na Cl Response in Foxtail Millet(Setaria italica L.) Seeds at Germination Stage

作　　者：潘教文[1] 李臻[1] 王庆国[1] 管延安[2,3] 李小波 戴绍军 丁国华[4] 刘炜[1,3] PAN JiaoWen;LI Zhen;WANG QingGuo;GUAN YanAn;LI XiaoBo;DAI ShaoJun;DING GuoHua;LIU Wei(Biotechnology Research Center,Shandong Academy of Agricultural Sciences/Shandong Provincial Key Laboratory of CropGenetic Improvement,Ecology and Physiology,Jinan 250100;Crop Research Institute,Shandong Academy of AgriculturalSciences/Shandong Engineering Laboratory for Featured Crops,Jinan 250100;College of Life Sciences,Shandong NormalUniversity,Jinan 250014;School of Life Sciences and Technology,Harbin Normal University,Harbin 150025;College of Lifeand Environmental Sciences,Shanghai Normal University/Development Center of Plant Germplasm Resources,Shanghai 200234)

机构地区：[1]山东省农业科学院生物技术研究中心/山东省作物遗传改良与生理生态重点实验室,济南250100 [2]山东省农业科学院作物研究所/山东省特色作物工程实验室,济南250100 [3]山东师范大学生命科学学院,济南250014 [4]哈尔滨师范大学生命科学与技术学院,哈尔滨150025 [5]上海师范大学生命与环境科学学院/植物种质资源开发协同创新中心,上海200234

出　　处：《中国农业科学》2019年第22期3964-3975,共12页Scientia Agricultura Sinica

基　　金：山东省自然科学基金青年基金(ZR2019QC003);现代农业产业技术体系建设专项资金(CARS-06);山东省农业科学院重大科技成果培育计划(2015CGPY10);山东省农业科学院农业科技创新工程(CXGC2018E13);山东省农业科学院青年英才计划(2016-2018)

摘　　要：【目的】谷子具有抗旱、耐贫瘠的特性,逐渐成为研究禾本科作物的模式作物。本研究以前期筛选并获得的谷子耐盐品种和敏感品种为材料,通过RNA-Seq测序筛选鉴定谷子的盐胁迫响应基因,揭示其响应盐害机制,为培育谷子抗盐新种质提供理论依据。【方法】对14个谷子品种进行了萌发期抗盐筛选。谷子不同品种的种子经150 mmol·L-1NaCl处理萌发培养7 d后,分别统计各品种种子的萌发率、根长和芽长,综合各项指标鉴定到豫谷2为耐盐品种、安04为盐敏感品种。以这两个品种盐胁迫前、后萌发期种子为材料,应用RNA-Seq进行转录组测定,分析鉴定盐害下差异表达基因(differentially expression gene,DEG),并对DEG进行GO和KEGG功能富集分析。同时应用qRT-PCR对随机挑选的15个DEG表达模式进行验证,并与RNA-Seq结果进行相关性分析。【结果】耐盐品种豫谷2、盐敏感品种安04种子经盐胁迫处理后,分别鉴定出2786个和4413个DEG;其中,每个品种在NaCl处理前及处理后分别鉴定出1470和3826个DEG。GO和KEGG聚类分析显示,NaCl处理下参与信号转导、抗氧化、有机酸的合成及转运以及次生代谢等相关的DEG在谷子萌发期种子应对盐胁迫响应过程中具有关键性的作用,DEG主要集中在胁迫响应、离子的跨膜转运、氧化还原、次生代谢及有机酸、多胺、苯丙烷的合成等过程。qRT-PCR对15个DEG表达模式的检测结果与RNA-seq结果相关系数为0.8817。其中编码离子通道的基因(HKT)、过氧化物酶基因(POD)、黄烷酮-3-羟化酶基因(FL3H)、MYB转录因子基因等在豫谷2中表达量较高,显示其在谷子萌发期种子响应盐胁迫中可能发挥一定作用。【结论】谷子耐盐品种及盐敏感品种的种子对盐胁迫的响应程度具有一定的差异性,且品种对盐害的耐受能力主要与基因对胁迫的响应程度相关,而与DEG的数量相关性不大。[Objective] Foxtail millet (Setaria italica L.) remarkably adapts to adverse ecologies, including drought, barren regions and high soil salinity. These characteristics promote it as a very promising model crop for exploring basic biology processes of plants. Unveil of mechanisms of foxtail millet under salinity is of immense significance. [Method] In this research, 14 millet varieties at germination stage were used for resistance screening under 150 mmol L -1 NaCl. The germination rate, root and bud length of each variety were measured after been treated for 7 d, and the salt tolerance variety Yugu 2 and salt susceptible variety An 04 were obtained. These two varieties before and after salt treatment were used for transcriptomes analysis by RNA-seq sequencing. The functional and pathway enrichments of differentially expressed genes (DEGs) were also performed by GO and KEGG analysis. Fifteen DEGs were randomly selected for further qRT-PCR analysis to verify the RNA-seq data. [Result] Totally 2 786 and 4 413 DEGs were identified in Yugu 2 and An 04, and there were 1 470 and 3 826 DEGs within each cultivar before and after salt treatment, respectively. GO and KEGG enrichment analysis suggested that DEGs were mainly clustered into signaling, antioxidant system, organic acid biosynthesis and transport, and secondary metabolism, and the DEGs participated in response to stress, ion transmembrane transport, redox homeostasis, secondary metabolism, organic acid, polyamine and phenylpropanoid biosynthetic process. The qRT-PCR results showed high correlation coefficient of 0.8817 with the data of RNA-seq. Especially the genes such as cation transporter (HKT8), peroxidase (POD), flavanone 3-dioxygenase (FL3H) and MYB transcription factors, which displayed higher vary degrees in Yugu2 under salinity may play key roles in salt response process of foxtail millet. [Conclusion]There was difference in the response degree of salt tolerance variety and salt sensitive variety of millet under salinity. Moreover, the resistance trait f

关 键 词：谷子 盐胁迫 萌发期种子 转录组 分子机制 

分 类 号：S51[农业科学—作物学]