Acta Agrestia Sinica ›› 2024, Vol. 32 ›› Issue (11): 3344-3357.DOI: 10.11733/j.issn.1007-0435.2024.11.002

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

FAN Lu1,2, TANG Jun2, MA Lin2, ZHANG Zhi-peng2, JIANG Qing-xue2, LIE Ren-cuo3, MA Qiao-ling3, WANG Xue-min2, ZHAO Wei1   

  1. 1. College of Agriculture, Henan University of Science and Technology, Luoyang, Henan Province 471000, China;
    2. Institute of Animal Sciences of Chinese Academy of Agricultural Sciences, Beijing 100193, China;
    3. Qinghai Sanjiang Group Co., LTD, Xining, Qinghai Province 810100, China
  • Received:2024-03-20 Revised:2024-04-29 Published:2024-11-29

基于转录组测序筛选蒙古冰草抗旱相关基因

樊璐1,2, 唐俊2, 马琳2, 张志鹏2, 蒋庆雪2, 李俄仁措3, 马巧玲3, 王学敏2, 赵威1   

  1. 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
  • 通讯作者: 王学敏,E-mail:zhaowei@haust.edu.cn;赵威,E-mail:wangxuemin@caas.cn
  • 作者简介:樊璐(1997-),女,汉族,内蒙古包头人,硕士研究生,主要从事牧草种质资源创新与利用及植物抗逆基因挖掘研究,E-mail:f1915213249@163.com
  • 基金资助:
    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

Abstract: 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 26 027 transcripts,containing 25 578 genes. BlastX was used to compare the annotations in KEGG,KOG,Nr,Swissprot and other databases,using BlastX,25 453 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.

Key words: Agropyron mongolicum Keng, Drought, Third-generation full-length transcriptome sequencing, Second-generation transcriptome sequencing, Transcription factor

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26 027条转录本,包含25 578个基因,利用BlastX在KEGG,KOG,Nr,Swissprot等数据库中进行比对注释,共有25 453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ERF053ERF055NAC100bZIP23ERF5NAM-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。

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

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