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草业学报 ›› 2024, Vol. 33 ›› Issue (6): 126-144.DOI: 10.11686/cyxb2023251

• 研究论文 • 上一篇    

野生稻CBF/DREB1转录因子的鉴定与生物信息学分析

霍晨敏1(), 袁敏3, 张宝文2, 王瑞菊2()   

  1. 1.河北经贸大学生物科学与工程学院,河北 石家庄 050061
    2.分子细胞生物学教育部重点实验室,河北省细胞生物学基础学科研究中心,河北省细胞信号与环境适应协同创新中心,河北省分子细胞生物学重点实验室,河北师范大学生命科学学院,河北 石家庄 050024
    3.华北理工大学生命科学学院,河北 唐山 063210
  • 收稿日期:2023-07-19 修回日期:2023-09-28 出版日期:2024-06-20 发布日期:2024-03-20
  • 通讯作者: 王瑞菊
  • 作者简介:E-mail: ruijuw@hebtu.edu.cn
    霍晨敏(1978-),女,河北井陉人,副教授,博士。E-mail: huochenmin@163.com
  • 基金资助:
    河北省自然科学基金优秀青年科学基金项目(C2021205011);河北省重点研发计划项目(20326513D);河北经贸大学校内科研基金项目(2019ZD05);河北省教育厅科学研究项目(BJ2021029)

Genome-wide identification and bioinformatics analysis of CBF/DREB1 transcription factors in wild rice

Chen-min HUO1(), Min YUAN3, Bao-wen ZHANG2, Rui-ju WANG2()   

  1. 1.College of Bioscience and Engineering,Hebei University of Economics and Business,Shijiazhuang 050061,China
    2.Ministry of Education Key Laboratory of Molecular and Cellular Biology,Hebei Research Center of the Basic Discipline of Cell Biology,Hebei Collaboration Innovation Center for Cell Signaling and Environmental Adaptation,Hebei Key Laboratory of Molecular and Cellular Biology,College of Life Sciences,Hebei Normal University,Shijiazhuang 050024,China
    3.College of Life Sciences,North China University of Science and Technology,Tangshan 063210,China
  • Received:2023-07-19 Revised:2023-09-28 Online:2024-06-20 Published:2024-03-20
  • Contact: Rui-ju WANG

摘要:

低温冷害是限制栽培稻地域分布和产量的重要因素之一,严重影响我国的粮食安全。以CBF/DREB1为核心的C-重复结合因子-冷调节基因(CBF-COR)途径是水稻适应低温的重要信号传导途径。基于稻属9种植物的全基因测序结果和隐马尔可夫模型(HMM)搜索,共鉴定出71个CBF/DREB1基因序列。所有基因均无内含子,由单个外显子组成。大部分DREB1s呈弱酸性,所有DREB1s亲水性平均值<0。进化树分析进一步将其分为3个组:DREB1A/1B/1HDREB1C/1E/1F/1G和DREB1D/1I/1J,各组成员除AP2保守结构域和侧翼序列外,其他保守基序组成差别较大。适应性进化分析粳亚种和其他水稻的直系同源基因对,发现OsDREB1A/ObDREB1AOsDREB1D/OnDREB1DOsDREB1I/OsIDREB1IOsDREB1J/OsIDREB1J四对基因非同义替换/同义替换(Ka/Ks)值均>1,受到正选择压力。旁系同源基因之间启动子顺式元件种类和数量差异较大,直系同源基因的启动子顺式元件种类和数量很相似。对日本晴、93-11和东乡野生稻7 d幼苗4 ℃ 冷处理2 h的实时荧光定量PCR分析发现早期冷响应基因可分为激活型(DREB1A/1B/1C/1E/1F/1G/1H)和抑制型(DREB1D/1I/1J)。在日本晴、93-11和东乡野生稻冷激活型基因中,DREB1B/1G/1H均是冷处理后响应最快的一级冷响应因子,且启动子均含有至少一个拷贝的CAMTA转录因子结合元件CM2(CCGCGT)。OrDREB1B/1C/1E/1F/1G/1H在东乡野生稻4 ℃冷处理4 h后的表达量均高于栽培稻同源基因的表达量。9种稻属植物的CBF/DREB1基因亚家族的分子进化分析结果为低温抗性基因的挖掘和利用提供了前期基础。

关键词: 野生稻, CBF/DREB1, 保守基序, 选择压力, 启动子分析

Abstract:

Low temperature and cold damage are important factors limiting the regional distribution and yield of cultivated rice, and this seriously affects food security in China. The C-repeat binding factor cold-regulated gene (CBF-COR) pathway with CBF/DREB1 transcription factors as the core component is an important signal transduction pathway involved in the cold acclimation of rice. A total of 71 members of the CBF/DREB1 gene subfamily were identified based on whole genome sequencing results and hidden markov model (HMM) searches of nine species in the rice (Oryza) genus. All genes were intron-free and consisted of a single exon. Most of them encoded weakly acidic proteins. All putative DREB1 proteins had a grand average of hydropathicity (GRAVY) of <0. A phylogenetic analysis divided into three groups: Group I (DREB1A/1B/1H), Group Ⅱ(DREB1C/1E/1F/1G), and Group Ⅲ (DREB1D/1I/1J). The motif composition differed among the groups, although all of the putative proteins contained the characteristic AP2 domain and flanking sequence. An adaptive evolutionary analysis of japonica subspecies and other rice orthologuous gene pairs showed that the nonsynonymous nucleotide substitution rate/synonymous nucleotide substitution rate (Ka/Ks) values for the gene pairs OsDREB1A/ObDREB1AOsDREB1D/OnDREB1D,OsDREB1I/OsIDREB1I, and OsDREB1J/OsIDREB1J were greater than 1, indicating that they were under positive selection pressure. The types and numbers of promoter elements were similar in orthologs, but differed between paralogs. Real-time quantitative PCR analyses were conducted for 7-day-old seedlings of Nipponbare, 93-11 and Dongxiang wild rice after 2 hours of cold treatment at 4 ℃. The results showed that, during the early cold response, some DREB genes were activated (DREB1A/1B/1C/1E/1F/1G/1H) and others were inhibited (DREB1D/1I/1J). Among the cold-activated genes in Nipponbare, 93-11, and Dongxiang wild rice, DREB1B/1G/1H were the first and the fastest to respond to cold. All of these three genes had at least one copy of the CAMTA transcription factor binding element CM2 (CCGCGT) in their promoter regions. The transcript levels of OrDREB1B/1C/1E/1F/1G/1H in Dongxiang wild ricewere higher than those of their homologs in cultivated rice after 4 hours of cold treatment at 4℃. The results of this molecular evolutionary study on the CBF/DREB1 gene subfamily in nine species in the Oryza genus provide a preliminary basis for the mining and utilization of low-temperature resistance genes.

Key words: wild rice, CBF/DREB1, motif, selective pressure, promoter analysis