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作物学报

作物学报 ›› 2023, Vol. 49 ›› Issue (5): 1410-1425.doi: 10.3724/SP.J.1006.2023.21036

• 研究简报 • 上一篇    下一篇

小麦G2-like转录因子家族基因鉴定与表达模式分析

贾玉库(), 高宏欢(), 冯健超, 郝紫瑞, 王晨阳, 谢迎新, 郭天财, 马冬云()   

  1. 河南农业大学农学院/国家小麦工程技术研究中心/河南省小麦技术创新中心, 河南郑州 450046
  • 收稿日期:2022-05-13 接受日期:2022-07-22 出版日期:2023-05-12 网络出版日期:2022-08-18
  • 通讯作者: *马冬云, E-mail: xmzxmdy@126.com
  • 作者简介:贾玉库, E-mail: 13633969649@163.com;
    高宏欢, E-mail: 2972620298@qq.com第一联系人：**同等贡献
  • 基金资助:
    国家科技支撑计划项目(2015BAD26B00);河南省科技攻关项目(212102110281)

Genome-wide identification and expression analysis of G2-like transcription factors family genes in wheat

JIA Yu-Ku(), GAO Hong-Huan(), FENG Jian-Chao, HAO Zi-Rui, WANG Chen-Yang, XIE Ying-Xin, GUO Tian-Cai, MA Dong-Yun()   

  1. College of Agronomy, Henan Agricultural University/National Engineering Research Center for Wheat/Technology Innovation Center of Henan Wheat, Zhengzhou 450046, Henan, China
  • Received:2022-05-13 Accepted:2022-07-22 Published:2023-05-12 Published online:2022-08-18
  • Contact: *E-mail: xmzxmdy@126.com
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    National Science and Technology Support Program of China(2015BAD26B00);Science and Technology Project of Henan Province(212102110281)

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摘要：

Golden2-Like (G2-like)转录因子, 是属于MYB类转录因子中GARP超家族的成员, 在调节叶绿体发育中起重要作用。本研究利用生物信息学方法对小麦G2-like基因进行了全基因组鉴定, 并对其理化性质、亚细胞定位、启动子顺式作用元件及对非生物胁迫和激素的响应模式进行了分析。从小麦中共鉴定出87个G2-like基因, 不均匀的分布在小麦21条染色体上, 系统发育分析将这些基因分为14个亚族。蛋白质二级结构预测表明, 小麦G2-like基因的氨基酸序列均以α螺旋和随机卷曲为主要结构。启动子顺式作用元件分析表明其上游2 kb区域含有7种(P-box、SpI、LTR、ABRE、MBS、TGA-element和AE-box)与逆境胁迫诱导相关顺式作用元件。其中Ta3AG2-like19含有顺式调控元件结合位点最多, 一共有18个结合位点。qRT-PCR验证发现, Ta3AG2-like19、Ta3AG2-like20、Ta4AG2-like29和Ta6AG2-like52在PEG和盐胁迫下以及GA、IAA和ABA激素诱导下表达量显著上调, 这些基因可能介导了小麦对多种非生物逆境的响应。

关键词: 小麦, G2-like转录因子, 生物信息学, 逆境胁迫, 表达分析

Abstract:

Golden2-like (G2-like) transcription factor, a member of the GARP superfamily of MYB transcription factors, plays an important role in regulating chloroplast development. In this study, genome-wide identification of G2-like genes in wheat was carried out by bioinformatics methods, and their physicochemical properties, subcellular localization, cis-acting elements of promoters, and response patterns to abiotic stresses and hormones were analyzed. A total of 87 G2-like genes were identified from wheat, which distributed in evenly on 21 chromosomes in wheat. Phylogenetic analysis showed that these genes were divided into 14 subfamilies, and fragment replication was the main reason for the expansion of this gene family. The prediction of protein secondary structure revealed that α helix and random curl were the main amino acid sequences of G2-like gene in wheat. Promoter cis-acting elements showed that there were seven cis-acting elements (P-box, SpI, LTR, ABRE, MBS, TGA-Element, and AE-box) in 2-kb region upstream of the promoter. Among them, Ta3AG2-Like19 contained the most cis-regulatory element binding sites with a total of 18 binding sites. The qRT-PCR revealed. that the relative expression levels of Ta3AG2-like19, Ta3AG2-Like20, Ta4AG2-Like29, and Ta6AG2-Like52 were significantly up-regulated under PEG and salt stresses, and induced by GA, IAA, and ABA hormones. These genes may mediate the response of wheat plant to various abiotic stresses.

Key words: wheat, G2-like transcription factors, bioinformatics, adversity stress, the relative expression level

表1

部分G2-like基因荧光定量所用引物"

基因ID Gene ID 上游引物Forward primer (5°−3°) 下游引物Reverse primer (5°−3°)
TraesCS1D02G305200.2 GACACCCGAGTTACATCAGCG CCTATGGTGTTCTTGTTTGTCCC
TraesCS2A02G441300.1 CGGGCTTGTCGGAGAATGGAAC TCCTGACTTCGTACACGGTAGAGC
TraesCS3A02G361800.1 GAAGTACCGCCTCTACCTCAAG CTGGTAGGAAACGTACGGATAC
TraesCS3A02G526600.1 GGCAACCGGGACAACAACTCT CCCTGCATCCGCTTGACGT
TraesCS4A02G130000.1 TTCCAGGAGAGTCAGGGCTTA CCTCATAATGGTTTTTGGGGT
TraesCS4B02G172900.1 AATCCAATACGGAGAAGGTGC CAAGGGGCTCAGAAGACTCG
TraesCS4B02G174600.1 AGCGACGGCTACATGAACAACTG GTGTCTCGTGTGCCTTCTCCAAC
TraesCS4D02G175000.1 CGGTCCTGCTGTGACATCTGATG TGTGACGCAGCCGAATTGGAAG
TraesCS4D02G176500.1 GCCAGTCAGCCACCACCTTAATAC TGCACCTCCAGTTGTTCATGTAGC
TraesCS5A02G178500.1 TAAGGACAATGGGTGTAAAGGG TGAGCATCAAGAAGATAGGACC
TraesCS5D02G183000.1 AGATGGGTCCTACCTTCTTGATGC AGGCGAACTGCTCTGAGACGAT
TraesCS6A02G109700.1 GAACACCAATAGCCGCCTCTACG ACGCCCAGTCTTGATTCCTGAAAC
TraesCS6A02G385500.1 GCAAGTACCTGGAGATGGTGATCG GACTGCGTGTTTGAATGCTGGAAC
TraesCS7B02G315800.1 TCCATTGTCTCATGTCTCGCA AGGCAGAGCACCCATTCGC
TraesCS7D02G334100.1 AACGGCAACACTGGCAGAAC TCTTCGTCTTCCTCGCCTCTGC

表2

小麦G2-like转录因子家族成员基本信息"

基因名
Gene name		 登录号
Accession number		 氨基酸长度
Length of amino acids		 分子量
Molecular weight (kD)		 等电点
Isoelectric point		 不稳定
系数
Instability coefficient		 疏水性指数
Hydrophobicity index		 α螺旋
α-helix
(%)		 延伸链
Extending chain
(%)		 β折叠
β-folding
(%)		 无规则
卷曲
Random crimp (%)		 亚细胞
定位
Subcellular localization
Ta1AG2-like1 TraesCS1A02G202500.1 353 38.40 8.82 41.82 0.724 28.33 9.07 10.20 52.41 N
Ta1AG2-like2 TraesCS1A02G258400.1 312 31.91 6.59 53.71 0.459 27.56 5.77 6.41 60.26 N
Ta1AG2-like3 TraesCS1A02G305500.2 407 45.60 6.35 39.59 0.776 37.59 9.09 2.46 50.86 N
Ta1BG2-like4 TraesCS1B02G216600.1 352 38.35 8.73 45.09 0.693 23.86 10.23 7.95 57.95 N
Ta1BG2-like5 TraesCS1B02G268900.1 308 31.59 6.59 52.98 0.447 27.27 7.79 7.79 57.14 N
Ta1BG2-like6 TraesCS1B02G316200.2 407 45.83 6.58 39.24 0.755 34.89 8.11 5.16 51.84 N
Ta1DG2-like7 TraesCS1D02G205800.1 352 38.33 8.78 45.10 0.749 26.14 9.66 8.81 55.40 N
Ta1DG2-like8 TraesCS1D02G257700.1 309 31.51 6.57 53.38 0.441 27.18 7.12 7.44 58.25 N
Ta1DG2-like9 TraesCS1D02G305200.2 407 45.62 6.47 39.89 0.761 33.66 9.09 3.19 54.05 N
Ta2AG2-like10 TraesCS2A02G100600.1 355 38.90 7.70 58.76 0.686 42.82 12.11 5.63 39.44 N
Ta2AG2-like11 TraesCS2A02G441300.1 279 30.63 6.19 35.67 0.589 45.88 9.68 4.30 40.14 M
Ta2AG2-like12 TraesCS2A02G488200.1 452 49.66 6.11 67.78 0.637 29.20 11.28 3.10 56.42 N
Ta2BG2-like13 TraesCS2B02G117800.1 357 39.07 7.26 57.22 0.681 42.02 10.36 3.92 43.70 N
Ta2BG2-like14 TraesCS2B02G515800.1 455 49.59 6.25 67.20 0.559 29.67 11.43 3.96 54.95 N
Ta2DG2-like15 TraesCS2D02G100100.1 357 38.95 7.71 58.72 0.644 49.30 9.24 3.92 37.54 N
Ta2DG2-like16 TraesCS2D02G488500.1 452 49.56 6.10 69.81 0.610 30.31 12.39 3.98 53.32 N
Ta3AG2-like17 TraesCS3A02G105500.2 430 46.95 5.11 54.42 0.578 29.77 6.05 3.02 61.16 P
Ta3AG2-like18 TraesCS3A02G161000.1 509 54.84 5.69 58.88 0.501 25.34 5.70 2.75 66.21 N
Ta3AG2-like19 TraesCS3A02G361800.1 225 24.23 5.85 62.38 0.568 26.67 7.11 6.22 60.00 N
Ta3AG2-like20 TraesCS3A02G526600.1 285 29.70 7.37 46.77 0.540 29.82 10.18 8.77 51.23 N
Ta3BG2-like21 TraesCS3B02G124100.1 432 46.98 5.12 57.01 0.579 26.85 3.70 3.24 66.20 P
Ta3BG2-like22 TraesCS3B02G191600.1 503 54.28 5.79 56.66 0.490 27.83 5.17 2.19 64.81 N
Ta3BG2-like23 TraesCS3B02G394200.1 225 24.31 5.84 59.07 0.566 29.33 5.33 5.78 59.56 N
Ta3BG2-like24 TraesCS3B02G594300.1 287 29.91 6.81 50.21 0.506 25.09 11.50 8.01 55.40 N
Ta3DG2-like25 TraesCS3D02G107800.1 432 47.11 5.07 53.12 0.618 30.09 6.25 3.01 60.65 P
Ta3DG2-like26 TraesCS3D02G168200.1 377 40.53 4.84 55.49 0.415 33.16 10.34 4.77 51.72 N
Ta3DG2-like27 TraesCS3D02G355500.1 227 24.43 5.84 61.74 0.541 25.55 6.17 4.85 63.44 N
Ta3DG2-like28 TraesCS3D02G531900.1 281 29.27 6.83 43.21 0.514 32.03 9.25 7.47 51.25 N
Ta4AG2-like29 TraesCS4A02G130000.1 354 39.68 8.28 57.73 0.711 36.44 8.47 3.39 51.69 N
Ta4AG2-like30 TraesCS4A02G131700.1 486 53.39 6.26 60.73 0.773 30.25 4.94 2.88 61.93 P
Ta4AG2-like31 TraesCS4A02G180900.1 341 37.76 7.02 48.84 0.709 27.86 13.49 4.99 53.67 N
Ta4BG2-like32 TraesCS4B02G172900.1 437 48.08 5.99 62.97 0.806 27.69 5.26 2.06 64.99 P
Ta4BG2-like33 TraesCS4B02G174600.1 355 39.82 7.06 58.02 0.701 39.44 9.30 3.38 47.89 N
Ta4DG2-like34 TraesCS4D02G071300.2 224 24.03 9.33 45.49 0.559 35.71 10.71 3.12 50.45 N
Ta4DG2-like35 TraesCS4D02G131500.1 341 37.91 8.54 44.89 0.728 29.62 15.25 5.28 49.85 N
Ta4DG2-like36 TraesCS4D02G175000.1 451 49.30 5.89 60.76 0.792 28.82 4.43 1.55 65.19 P
Ta4DG2-like37 TraesCS4D02G176500.1 354 39.73 8.57 57.50 0.703 41.81 7.06 3.67 47.46 N
Ta5AG2-like38 TraesCS5A02G153000.1 326 35.77 7.32 53.69 0.678 33.13 13.50 6.75 46.63 N
Ta5AG2-like39 TraesCS5A02G178500.1 255 27.69 8.71 53.52 0.500 40.00 11.76 4.71 43.53 N
Ta5AG2-like40 TraesCS5A02G178600.1 286 30.98 5.76 53.63 0.836 33.22 6.29 4.90 55.59 P
Ta5BG2-like41 TraesCS5B02G151800.1 337 37.06 7.35 46.54 0.677 31.45 12.46 5.93 50.15 N
Ta5BG2-like42 TraesCS5B02G176200.1 255 27.72 8.71 53.85 0.510 40.39 10.59 5.10 43.92 N
Ta5BG2-like43 TraesCS5B02G176300.1 286 31.02 5.75 54.74 0.856 29.72 4.55 3.15 62.59 P
Ta5DG2-like44 TraesCS5D02G158300.1 324 35.73 8.68 55.56 0.710 33.95 12.96 5.86 47.22 N
Ta5DG2-like45 TraesCS5D02G183000.1 255 27.73 8.71 53.52 0.509 36.47 10.59 4.31 48.63 N
Ta5DG2-like46 TraesCS5D02G183100.1 286 31.02 5.75 53.39 0.853 31.82 5.59 2.45 60.14 P
Ta6AG2-like47 TraesCS6A02G108800.1 415 45.89 6.18 61.00 0.640 28.19 8.67 2.65 60.48 N
Ta6AG2-like48 TraesCS6A02G109700.1 477 52.25 8.92 51.15 0.699 28.51 11.11 5.24 55.14 N
Ta6AG2-like49 TraesCS6A02G155400.1 362 39.28 8.30 45.73 0.540 39.23 11.33 5.25 44.20 N
Ta6AG2-like50 TraesCS6A02G254700.1 303 33.19 9.44 63.28 0.558 39.93 6.60 6.93 46.53 N
Ta6AG2-like51 TraesCS6A02G269000.1 276 30.36 8.92 29.87 0.561 45.65 10.51 5.07 38.77 N
Ta6AG2-like52 TraesCS6A02G385500.1 406 45.10 5.98 46.92 0.645 30.54 9.36 2.96 57.14 N
Ta6BG2-like53 TraesCS6B02G137300.1 409 45.35 7.23 57.66 0.652 33.25 6.85 1.96 57.95 N
Ta6BG2-like54 TraesCS6B02G138200.1 528 58.32 7.88 54.92 0.705 31.25 8.71 4.36 55.68 N
Ta6BG2-like55 TraesCS6B02G183500.1 362 39.29 7.78 48.07 0.536 41.44 9.39 4.42 44.75 N
Ta6BG2-like56 TraesCS6B02G270800.1 318 34.61 9.27 53.80 0.524 38.68 6.92 4.09 50.31 M
Ta6BG2-like57 TraesCS6B02G296300.1 280 30.91 8.72 28.89 0.585 46.07 11.43 4.64 37.86 N
Ta6BG2-like58 TraesCS6B02G394700.1 534 58.47 5.51 41.81 0.376 33.71 9.55 6.18 50.56 C
Ta6BG2-like59 TraesCS6B02G424600.1 431 48.08 6.09 39.85 0.637 32.02 9.51 2.09 56.38 N
Ta6DG2-like60 TraesCS6D02G097000.1 386 42.63 6.28 52.91 0.682 31.35 8.81 1.30 58.55 N
Ta6DG2-like61 TraesCS6D02G098200.1 475 51.73 8.92 50.50 0.648 36.42 9.26 4.63 49.68 N
Ta6DG2-like62 TraesCS6D02G145300.1 363 39.35 8.30 48.93 0.528 36.09 10.74 3.86 49.31 N
Ta6DG2-like63 TraesCS6D02G236000.1 304 33.43 9.65 59.00 0.578 40.79 7.89 3.29 48.03 N
Ta6DG2-like64 TraesCS6D02G245900.1 277 30.36 8.72 28.00 0.542 47.65 10.11 3.97 38.27 N
Ta6DG2-like65 TraesCS6D02G370300.1 407 45.39 5.90 40.98 0.635 32.92 6.63 2.70 57.74 N
Ta7AG2-like66 TraesCS7A02G339800.1 461 49.17 5.30 65.82 0.319 31.45 8.46 3.25 56.83 N
Ta7AG2-like67 TraesCS7A02G345200.1 512 55.89 9.36 52.35 0.785 33.40 6.84 5.47 54.30 N
Ta7AG2-like68 TraesCS7A02G381900.1 339 36.57 5.69 46.54 0.594 28.91 7.96 5.01 58.11 N
Ta7AG2-like69 TraesCS7A02G415800.1 454 49.57 5.71 65.91 0.706 28.63 7.93 1.32 62.11 N
Ta7AG2-like70 TraesCS7A02G469900.1 249 27.44 6.76 61.15 0.816 31.73 11.65 6.02 50.60 N
Ta7AG2-like71 TraesCS7A02G470100.1 252 27.84 7.09 71.64 0.840 28.57 11.11 6.75 53.57 M
Ta7AG2-like72 TraesCS7A02G470200.1 246 27.56 8.39 71.73 0.725 30.89 10.57 5.28 53.25 N
Ta7AG2-like73 TraesCS7A02G483100.1 348 37.47 6.83 53.15 0.457 39.66 7.18 3.16 50.00 N
Ta7BG2-like74 TraesCS7B02G251400.1 459 49.29 5.24 64.16 0.381 33.99 7.19 2.61 56.21 N
Ta7BG2-like75 TraesCS7B02G315800.1 454 49.53 5.71 67.55 0.719 29.07 7.05 1.54 62.33 N
Ta7BG2-like76 TraesCS7B02G372000.1 250 27.76 7.77 64.83 0.853 28.80 10.40 6.00 54.80 N
Ta7BG2-like77 TraesCS7B02G372100.1 240 27.00 9.12 64.75 0.945 25.00 13.33 7.50 54.17 N
Ta7BG2-like78 TraesCS7B02G372200.1 245 27.17 7.75 72.37 0.736 33.06 6.53 5.71 54.69 N
Ta7BG2-like79 TraesCS7B02G385700.1 348 37.41 7.17 53.37 0.449 39.08 7.18 3.74 50.00 N
Ta7DG2-like80 TraesCS7D02G334100.1 517 55.27 8.40 46.25 0.750 33.27 7.54 4.84 54.35 N
Ta7DG2-like81 TraesCS7D02G347500.1 458 48.96 5.51 65.44 0.369 29.69 6.33 2.84 61.14 N
Ta7DG2-like82 TraesCS7D02G409100.1 454 49.40 5.78 64.55 0.667 28.63 7.27 1.32 62.78 N
Ta7DG2-like83 TraesCS7D02G457300.1 250 27.67 8.33 64.87 0.858 27.20 8.80 6.00 58.00 N
Ta7DG2-like84 TraesCS7D02G457400.1 251 27.67 7.16 73.87 0.881 28.29 7.17 6.77 57.77 N
Ta7DG2-like85 TraesCS7D02G457500.1 246 27.74 9.21 67.18 0.798 32.93 9.35 6.91 50.81 N
Ta7DG2-like86 TraesCS7D02G457600.1 247 27.27 6.50 72.08 0.678 35.22 8.50 4.86 51.42 N
Ta7DG2-like87 TraesCS7D02G470000.1 348 37.40 7.17 52.82 0.449 41.38 6.90 4.89 46.84 N

图1

小麦G2-like基因的系统发育关系、保守蛋白基序结构和基因结构 A: 利用MEGA7基于小麦G2-like蛋白全长序列构建系统发育树。B: 小麦G2-like蛋白的基序组成。不同颜色表示motif 1~motif 10; 蛋白质的长度可以用底部的刻度来估计。C: 小麦G2-like基因的外显子-内含子结构。绿色方框表示未翻译的5-和3-区域;黄色框表示外显子; 黑线表示内含子。"

图2

小麦G2-like基因染色体定位 标度代表兆碱基(Mb)。"

图3

小麦G2-like基因的染色体位置及共线性关系 所有的共线区域和基因由灰线连接, G2-like基因的片段重复基因对用红线标出。"

图4

小麦G2-like基因启动子区域与抗逆相关的7种顺式作用元件 顺式作用元件用不同的彩色方框表示。"

图5

不同时期籽粒、叶片、根和穗中87个G2-like基因表达谱热图 S: 幼苗期; V: 期; R: 生殖期。红色或绿色代表每个样品中每个转录本的较高或较低的相对丰度。"

图6

小麦G2-like基因在热、干旱以及热和干旱共同胁迫下的表达模式 D1、D6:干旱处理1、6 h; H1、H6: 高温处理1、6 h; D+H1、D+H6: 在高温和干旱胁迫下处理时间为1、6 h。红色和绿色分别代表每个样品中每个转录本的较高和较低的相对丰度。"

图7

定量实时PCR分析14个G2-like基因对NaCl、PEG处理的响应 *表示处理间存在显著差异(P <0.05)。"

图8

定量实时PCR分析14个G2-like基因对ABA、GA和IAA处理的响应 *表示处理间存在显著差异(P < 0.05)。"

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