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作物学报 ›› 2022, Vol. 48 ›› Issue (3): 644-655.doi: 10.3724/SP.J.1006.2022.02089

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

水稻延伸因子复合体家族基因鉴定及非生物胁迫诱导表达模式分析

巫燕飞(), 胡琴, 周棋, 杜雪竹, 盛锋*()   

  1. 湖北大学生命科学学院/省部共建生物催化与酶工程国家重点实验室, 湖北武汉 430062
  • 收稿日期:2020-12-21 接受日期:2021-07-12 出版日期:2022-03-12 网络出版日期:2021-08-09
  • 通讯作者: 盛锋
  • 作者简介:E-mail: Byronwyf@163.com
  • 基金资助:
    湖北省技术创新专项重大项目(2018ABA080);湖北省技术创新专项重大项目(2020BBB052);湖北省自然科学基金重点项目(2019CFA027);湖北省自然科学基金项目(2020CFB261)

Genome-wide identification and expression analysis of Elongator complex family genes in response to abiotic stresses in rice

WU Yan-Fei(), HU Qin, ZHOU Qi, DU Xue-Zhu, SHENG Feng*()   

  1. State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, Wuhan 430062, Hubei, China
  • Received:2020-12-21 Accepted:2021-07-12 Published:2022-03-12 Published online:2021-08-09
  • Contact: SHENG Feng
  • Supported by:
    Major Program of Technological Innovation of Hubei Province(2018ABA080);Major Program of Technological Innovation of Hubei Province(2020BBB052);Key Program of Natural Science Foundation of Hubei Province(2019CFA027);Natural Science Foundation of Hubei Province(2020CFB261)

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摘要：

延伸因子复合体(Elongator, ELP)是一类在真核细胞转录中起延伸RNA聚合酶II作用的蛋白质复合物, 这类复合体在植物生长发育以及生物和非生物胁迫响应等方面均发挥着重要作用。本研究利用生物信息学的方法对水稻(Oryza sativa) ELP基因家族(OsELPs)成员进行鉴定, 并进一步对其理化性质、亚细胞定位、染色体定位、启动子顺式作用元件及在非生物胁迫下的表达模式进行分析。结果表明: OsELPs基因家族含有6个成员、随机分布在水稻5条染色体上, 这6个基因编码含250~1344个氨基酸残基的蛋白质, 其分子量介于27.97~148.99 kD, 等电点介于5.01~8.63。系统进化分析将来源于水稻(Oryza sativa)、拟南芥(Arabidopsis thaliana)、酵母(Saccharomyces cerevisiae)和人类(Homo sapiens)的ELPs蛋白分为4个亚组(Group I~Group IV), 其中第I亚组包含OsELP1, 第II亚族包含OsELP2和OsELP5, 第III亚组包含OsELP4, 而第IV亚组包含OsELP3和OsELP6。OsELPs的启动子区域中存在多种顺式作用元件, 主要响应光、植物激素、干旱、低温、防御和逆境等信号。在PEG、低温、高盐以及脱水等4种非生物胁迫下, 6个OsELPs基因均有不同程度的差异表达, 其中OsELP6在4种非生物胁迫下均显著诱导上调表达, 该基因可能介导水稻对多种非生物逆境的综合抗性。

关键词: 水稻(Oryza sativa), 延伸因子复合体, 生物信息学, 非生物逆境胁迫

Abstract:

Elongator complex (ELP) is a class of protein complex that elongates RNA polymerase II in eukaryotic transcription, which plays an important role in plant growth and development, and resistance to biotic and abiotic stresses. In this study, we identified the ELP family genes and explored the physical and chemical properties, subcellular location, chromosome location, promoter cis-acting elements prediction, and expression patterns under abiotic stresses in rice (Oryza sativa) ELP family genes by bioinformatics methods. A total of six OsELPs members were preliminarily identified, which randomly distributed on five chromosomes and encoded protein containing 250 to 1344 amino acid in rice. Their molecular weight and isoelectric point were 27.97-148.99 kD and 5.01-8.63, respectively. Phylogenetic analysis showed that the ELPs proteins from rice (Oryza sativa), Arabidopsis (Arabidopsis thaliana), yeast (Saccharomyces cerevisiae), and human (Homo sapiens) could be divided into four groups (Group I-Group IV). And subgroup I contained OsELP1, subgroup II contained OsELP2 and OsELP5, subgroup III contained OsELP4, subgroup IV contained OsELP3 and OsELP6. There were a variety of cis-acting elements in the promoter regions of OsELPs, which mainly responded to light, phytohormone, drought, low temperature, defense, and stress stimulant signals. The induced expression patterns confirmed that all OsELPs genes were differentially expressed with different degrees under various abiotic stresses including PEG, low temperature, salt, and dehydration. Among them, OsELP6 was significantly up-regulated under the four kinds of abiotic stresses, which may mediate the comprehensive resistance to various abiotic stresses in rice.

Key words: rice (Oryza sativa), Elongator complex, bioinformatics, abiotic stresses

表1

用于qRT-PCR分析的OsELPs基因和内参基因的引物列表"

引物名称
Primer name		 序列
Sequence (5°-3°)		 长度
Length (bp)
qELP1-F CATCTGTACTGATGGCAATTCG 22
qELP1-R ACGGGCTAAAGTAGTCAGTATG 22
qELP2-F GGAGCGTTCAAGATTCATCTTC 22
qELP2-R AATGTTCCAGAGCTCGATCATT 22
qELP3-F TATCAGAAATGGCACCTCCCTT 22
qELP3-R GCCTCTTCAACATTAGCAGAAG 22
qELP4-F CAGGACCTATGAAGGAACCC 20
qELP4-R CTATCCTCAAAGCCTCGTCA 20
qELP5-F GTGATTCTGATGATGAGCAACC 22
qELP5-R CAAGTACGATCAAGGCATTCTG 22
qELP6-F CGTAAGATGGGCTGTAACCTAT 22
qELP6-R TATCAGAAATGGCACCTCCCTT 22
qActin-F CTATCCTCAAAGCCTCGTCA 20
qActin-R TACCACCACTGAGAACGATGT 21

表2

水稻中的ELP基因家族的基本信息"

基因名称
Gene name		 RAP-DB ID
Gene ID		 基因座
LOC loucs		 氨基酸数
AA size (aa)		 分子量
Molecular weight
(kD)		 等电点
Isoelectric point		 亚细胞定位
Subcellular
localization
OsELP1 Os07g0563700 LOC_Os07g37640 1344 148.99 5.47 PMa, Nuclb/Mbb
OsELP2 Os08g0493900 LOC_Os08g38570 850 92.67 6.69 Extraa/PMa, Mbb
OsELP3 Os04g0484900 LOC_Os04g40840 574 63.78 8.63 M.t.ma, Nuclb
OsELP4 Os06g0639600 LOC_Os06g43280 378 27.46 7.96 Nucla, C.t.sb
OsELP5 Os03g0201700 LOC_Os03g10460 385 42.52 5.01 Nucla, Cytob
OsELP6 Os03g0284000 LOC_Os03g17560 250 27.97 6.23 Cytoa, Cytob

图1

水稻、拟南芥、人类、酵母ELP蛋白的系统发育树 在本研究中水稻ELP蛋白以黑色圆形显示, 拟南芥ELP蛋白以绿色正方形显示, 人类ELP蛋白以青色菱形显示, 酵母ELP蛋白以蓝色三角形显示。比例尺代表每个同义位点0.2个氨基酸替代。"

图2

水稻OsELPs基因结构"

图3

水稻OsELPs蛋白质保守结构域分析"

图4

预测水稻OsELPs启动子中顺式调控元件"

图5

PEG、低温、高盐处理下OsELPs基因qRT-PCR表达情况"

图6

脱水处理下OsELPs基因qRT-PCR表达情况"

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