基于转录组测序筛选蒙古冰草抗旱相关基因  

Screening Genes Related to Drought Resistance in Agropyron mongolicum Keng Based on Transcriptome

作　　者：樊璐 唐俊 马琳[2] 张志鹏 蒋庆雪 李俄仁措 马巧玲 王学敏[2] 赵威[1] FAN Lu;TANG Jun;MA Lin;ZHANG Zhi-peng;JIANG Qing-xue;LIE Ren-cuo;MA Qiao-ling;WANG Xue-min;ZHAO Wei(College of Agriculture,Henan University of Science and Technology,Luoyang,Henan Province 471000,China;Institute of Animal Sciences of Chinese Academy of Agricultural Sciences,Beijing 100193,China;Qinghai Sanjiang Group Co.,LTD,Xining,Qinghai Province 810100,China)

机构地区：[1]河南科技大学农学院,河南洛阳471000 [2]中国农业科学院北京畜牧兽医研究所,北京100193 [3]青海省三江集团有限责任公司,青海西宁810100

出　　处：《草地学报》2024年第11期3344-3357,共14页Acta Agrestia Sinica

基　　金：内蒙古自治区种业科技创新重大示范工程“揭榜挂帅”项目(2022 JBGS0014);财政部和农业农村部:国家现代农业产业技术体系(CARS-34)资助。

摘　　要：为挖掘蒙古冰草(Agropyron mongolicum Keng)抗旱相关基因并探究蒙古冰草响应干旱胁迫分子机制,本研究以蒙古冰草为材料进行转录组测序。首先将不同生长期及干旱胁迫处理后的盆栽蒙古冰草组织按照根、茎、叶、花(穗)4类分别进行混样并利用三代全长转录组测序技术建立蒙古冰草转录本参考序列数据库,共获得26027条转录本,包含25578个基因,利用BlastX在KEGG,KOG,Nr,Swissprot等数据库中进行比对注释,共有25453个转录本得到注释。其次利用二代转录组测序技术对干旱胁迫处理前后水培蒙古冰草幼苗进行测序,发现干旱胁迫处理2 h,6 h后与处理前相比分别有2669个,3668个差异表达基因。此外,根据转录组及荧光定量PCR验证结果,发现基因ERF053,ERF055,NAC100,bZIP23,ERF5和NAM-B1在干旱处理后表达量较高且均上调可能是抗旱调控关键基因,为后续蒙古冰草抗旱相关候选基因克隆及功能验证奠定了基础。To dig drought-resistant genes and explore the molecular mechanism of Agropyron mongolicum Keng responding to drought stress,transcriptome analysis was carried out in this study using Agropyron mongolicum Keng as the material.In this study,the transcriptome sequencing was performed on Agropyron mongolicum Keng seedlings in pots during different growth periods and after drought stress treatment according to the four categories of roots,stems,leaves and flowers(spikes),and the reference sequence database of Agropyron mongolicum Keng was established by third-generation full-length transcriptome sequencing technology,which obtained a total of 26027 transcripts,containing 25578 genes.BlastX was used to compare the annotations in KEGG,KOG,Nr,Swissprot and other databases,using BlastX,25453 transcripts were annotated.Secondly,the second-generation transcriptome sequencing technology was used to sequence the differentially expressed genes in hydroponically grown Agropyron mongolicum Keng seedlings before and after drought stress treatment,compared with before treatment,2669 and 3668 differentially expressed genes were detected after 2-hour and 6-hour drought stress treatment,respectively.In addition,based on transcriptome analysis and qRT-PCR validation results,the genes ERF053,ERF055,NAC100,bZIP23,ERF5,and NAM-B1 were found to have higher and up-regulated expression after drought treatment,which may be the key genes of drought regulation,and this lays a foundation for the subsequent cloning and functional validation of the drought-related candidate genes in Agropyron mongolicum Keng.

关 键 词：蒙古冰草 干旱 三代全长转录组 二代转录组测序 转录因子 

分 类 号：S543.9[农业科学—作物学]