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作物学报 ›› 2021, Vol. 47 ›› Issue (10): 1913-1926.doi: 10.3724/SP.J.1006.2021.04235

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

叉柱棉sHSP基因家族的鉴定与特征分析

范凯(), 潘鑫峰, 毛志君, 叶方婷, 李兆伟, 林伟伟, 林文雄*()   

  1. 福建农林大学农学院作物遗传育种与综合利用教育部重点实验室, 福建福州 350002
  • 收稿日期:2020-10-29 接受日期:2021-03-19 出版日期:2021-10-12 网络出版日期:2021-03-31
  • 通讯作者: 林文雄
  • 作者简介:E-mail: fankai@fafu.edu.cn
  • 基金资助:
    国家自然科学基金项目(31701470);中国博士后科学基金项目(2017M610388);中国博士后科学基金项目(2018T110637);福建农林大学科技创新专项基金(CXZX2020007A);福建农林大学杰出青年科研人才计划项目(xjq201917)

Identification and analysis of sHSP gene family in Gossypioides kirkii

FAN Kai(), PAN Xin-Feng, MAO Zhi-Jun, YE Fang-Ting, LI Zhao-Wei, LIN Wei-Wei, LIN Wen-Xiong*()   

  1. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, College of Agriculture, Fujian Agriculture and Forestry University, Fuzhou 350002, Fuzhou, China
  • Received:2020-10-29 Accepted:2021-03-19 Published:2021-10-12 Published online:2021-03-31
  • Contact: LIN Wen-Xiong
  • Supported by:
    National Natural Science Foundation of China(31701470);China Postdoctoral Science Foundation(2017M610388);China Postdoctoral Science Foundation(2018T110637);Science and Technology Innovation Special Fund of Fujian Agriculture and Forestry University(CXZX2020007A);Outstanding Youth Scientific Fund of Fujian Agriculture and Forestry University(xjq201917)

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摘要：

植物小分子热激蛋白(small heat shock protein, sHSP)是植物热激蛋白中一类分子量最小的基因家族, 该家族具有保守的α-晶体结构域, 在响应外界环境胁迫过程中具有重要的作用。叉柱棉(Gossypioides kirkii)有关sHSP家族的鉴定与特征分析还未见相关报道。本研究在叉柱棉中共鉴定了39个GksHSP成员, 并且可以将GksHSP成员进一步分为10个亚家族。在GksHSP家族中发现了7个基因复制事件, 并且所有的基因复制事件都是片段复制事件。棉属特有的全基因组复制事件主要导致了GksHSP成员的扩增, 其扩增还与蛋白激酶家族、线粒体载体蛋白家族以及植物生长素响应蛋白家族有关。此外, GksHSP家族可能与ABA和茉莉酸甲酯调控的胁迫响应相关, 并且GksHSP26成员及其在陆地棉中的同源基因可能在胁迫响应中具有关键的作用。本研究的结果可以进一步为今后叉柱棉和棉花的抗逆育种研究提供一定的理论基础。

关键词: 叉柱棉, sHSP, 鉴定, 扩增, 功能

Abstract:

Small heat shock protein (sHSP) has the lowest molecular weight in heat shock protein and sHSP has the highly conserved α-crystallin domain. The sHSP family is very important in response to various stresses, but the identification and analysis of sHSP gene family in Gossypioides kirkii have not been reported. In this study, we identified 39 GksHSPs with 10 subfamilies in Gossypioides kirkii. There were seven gene duplication events in GksHSP family and all of duplicated gene pairs were involved in segmental duplication events. The cotton-specific whole genome duplication event primarily resulted in the GksHSP expansion. The GksHSP expansion was also related to the protein kinase, mitochondrial carrier protein, and auxin responsive protein. Besides, GksHSP members might respond to various stresses by ABA/MeJA-mediated pathways, and GksHSP26 and its corresponding orthologous genes in Gossypium hirsutum were very important in stress responses. These results could provide the theoretical basis of the breeding of stress tolerance in Gossypioides kirkii and cotton.

Key words: Gossypioides kirkii, sHSP, identification, expansion, function

图1

拟南芥和叉柱棉中sHSP成员的系统发育分析"

表1

本研究中GksHSP成员的鉴定以及结构分析"

名称
Name		 位点名称
Locus name		 亚家族
Subfamily		 蛋白质长度
Protein length (aa)		 分子量
Molecular weight (kD)		 理论pI
Theoretical pI		 α-螺旋
α-helix		 延伸链
Extended strand		 β-折叠
β-turn		 无规则卷曲
Random coil
GksHSP01 Kirkii_Version3_Juiced.00g028900 CP 144 16.32340 4.95 23 30 6 85
GksHSP02 Kirkii_Version3_Juiced.00g047410 CI 159 18.32872 5.98 28 29 11 91
GksHSP03 Kirkii_Version3_Juiced.00g056780 ER 187 21.44579 6.77 64 31 8 84
GksHSP04 Kirkii_Version3_Juiced.00g059950 CI 153 17.69095 5.35 34 28 7 84
GksHSP05 Kirkii_Version3_Juiced.00g085690 CII 159 18.01872 6.64 52 34 8 65
GksHSP06 Kirkii_Version3_Juiced.00g085700 CII 156 17.66154 6.62 53 38 11 54
GksHSP07 Kirkii_Version3_Juiced.00g091700 CI 156 17.91837 5.79 38 30 10 78
GksHSP08 Kirkii_Version3_Juiced.00g091710 CI 156 17.80026 6.18 36 29 9 82
GksHSP09 Kirkii_Version3_Juiced.00g092160 CI 133 14.79902 9.00 40 22 5 66
GksHSP10 Kirkii_Version3_Juiced.00g096000 MTII 246 27.95450 7.80 82 46 13 105
GksHSP11 Kirkii_Version3_Juiced.00g097580 CIII 102 11.37751 4.37 35 16 5 46
GksHSP12 Kirkii_Version3_Juiced.00g101120 CIV 139 15.76492 5.34 28 29 10 72
GksHSP13 Kirkii_Version3_Juiced.00g120690 CI 159 18.25460 6.18 20 34 10 95
GksHSP14 Kirkii_Version3_Juiced.00g120700 CI 159 18.19460 6.18 28 31 10 90
GksHSP15 Kirkii_Version3_Juiced.00g120710 CI 159 18.27965 6.64 29 32 8 90
GksHSP16 Kirkii_Version3_Juiced.00g134580 CIV 139 15.85984 4.61 30 34 6 69
GksHSP17 Kirkii_Version3_Juiced.00g136170 CII 159 18.18400 6.19 49 31 7 72
GksHSP18 Kirkii_Version3_Juiced.00g136180 CII 153 17.05883 8.59 53 40 9 51
GksHSP19 Kirkii_Version3_Juiced.00g184450 CI 156 17.82725 5.77 32 31 10 83
GksHSP20 Kirkii_Version3_Juiced.00g184460 CI 160 18.36475 6.18 39 33 8 80
GksHSP21 Kirkii_Version3_Juiced.00g184750 CI 135 14.96507 5.91 38 33 4 60
GksHSP22 Kirkii_Version3_Juiced.00g188220 CI 143 16.28479 9.22 25 28 7 83
GksHSP23 Kirkii_Version3_Juiced.00g193540 PX 165 18.81639 5.98 18 40 8 99
GksHSP24 Kirkii_Version3_Juiced.00g194110 CII 158 17.75257 5.97 51 34 11 62
GksHSP25 Kirkii_Version3_Juiced.00g209640 CP 232 25.77139 6.84 39 43 8 142
GksHSP26 Kirkii_Version3_Juiced.00g215570 CI 159 18.38593 5.83 30 30 10 89
GksHSP27 Kirkii_Version3_Juiced.00g230720 SINGLE 140 16.28668 5.39 28 26 10 76
GksHSP28 Kirkii_Version3_Juiced.00g244800 CI 163 18.52218 8.83 33 36 10 84
GksHSP29 Kirkii_Version3_Juiced.00g246020 CI 156 17.87539 6.76 27 39 10 80
GksHSP30 Kirkii_Version3_Juiced.00g256930 CI 144 16.53776 5.58 26 34 8 76
GksHSP31 Kirkii_Version3_Juiced.00g267060 CV 166 19.59569 6.33 57 39 17 53
GksHSP32 Kirkii_Version3_Juiced.00g273810 CI 93 10.50390 4.61 11 32 9 41
GksHSP33 Kirkii_Version3_Juiced.00g301400 PX 141 15.98226 6.92 24 36 10 71
GksHSP34 Kirkii_Version3_Juiced.00g331020 CP 230 25.96835 5.86 40 38 9 143
GksHSP35 Kirkii_Version3_Juiced.00g336540 MTI/CP 181 20.93121 6.23 61 35 10 75
GksHSP36 Kirkii_Version3_Juiced.00g336590 MTI/CP 327 36.83181 5.49 103 48 22 154
GksHSP37 Kirkii_Version3_Juiced.00g343620 CI 149 17.03446 7.83 30 35 9 75
GksHSP38 Kirkii_Version3_Juiced.00g375140 ER 184 20.66178 5.76 57 33 8 86
GksHSP39 Kirkii_Version3_Juiced.00g377690 CV 196 22.48230 5.45 44 40 17 95

图2

GksHSP成员的系统发育(左)、保守基序(中)以及基因结构(右)分析"

表2

通过MEME程序识别有关GksHSP成员的基序信息"

基序ID
Motif ID		 基序序列
Motif sequence		 基序长度
Motif length (aa)
MEME-1 RIDWKETPEAHVFKADVPGLKKEEVKVEVEDDRVLQISGERNVEKEDKNDT 51
MEME-2 NAKMDQIKASMEBGVLTVTVP 21
MEME-3 ERSSGKFMRRFRLPE 15
MEME-4 MAMIPSFFGNRRSSI 15
MEME-5 FDPFSLDVWDPFKDF 15
MEME-6 HVFKADLPGLKKEEVKVEVED 21
MEME-7 KLEVKKPDVKAIEIS 15
MEME-8 SLSTRSPETSAFVNA 15
MEME-9 MLDIPDETEKSPNAPSRAYVRDAKAMAAT 29
MEME-10 MEFPFPSDQHSPLYHYLLPSPPLFSNQLLPENHLNWTQTP 40
MEME-11 QQREGKKKDWRSCNWWEYGYVRRLELPZDADWRKIEAFLSNDVVLEIRIPRN 52
MEME-12 MDFRIMGFDSPLLHT 15
MEME-13 WTNRSYSSYBTSLQLPD 17
MEME-14 VLVIKGERKEE 11
MEME-15 QERAVEKRPKRLAMDVSPFGLLDPLSPMRSMRQMLDTMDRIFEDAMIFPGSNRRQGG 57
MEME-16 DWKETPEA 8
MEME-17 PFSVSFPSKNPCNSRLSVVRAZAAGDNNKDTSVDVHVNKDN 41
MEME-18 GSFYIDPADVPDRVEVLARAA 21
MEME-19 NIQIHVEKGKIMEIFGQ 17
MEME-20 RAPWDIKDGEH 11

图3

GksHSP成员在叉柱棉12条染色体上的分布"

表3

叉柱棉中复制的sHSP成员的Ka和Ks分析"

复制基因1
Duplicated gene 1		 复制基因2
Duplicated gene 2		 亚家族
Subfamily		 非同义
替换率
Ka		 同义替换率
Ks		 非同义替换率/同义替换率
Ka/Ks		 纯化选择
Purifying selection		 复制类型
Duplicated type
GksHSP05 GksHSP17 CII 0.063959 0.809673 0.078994 是Yes 片段复制事件
Segmental
GksHSP06 GksHSP18 CII 0.090699 0.742610 0.122136 是Yes 片段复制事件Segmental
GksHSP07 GksHSP20 CI 0.045458 0.629437 0.072220 是Yes 片段复制事件Segmental
GksHSP09 GksHSP21 CI 0.159061 0.715073 0.222441 是Yes 片段复制事件Segmental
GksHSP06 GksHSP24 CII 0.113536 1.039721 0.109199 是Yes 片段复制事件Segmental
GksHSP18 GksHSP24 CII 0.103983 0.730373 0.142370 是Yes 片段复制事件Segmental
GksHSP25 GksHSP34 CP 0.125830 0.606857 0.207347 是Yes 片段复制事件Segmental

图4

GksHSP成员的基因复制事件分析 A: 复制的GksHSP成员分析, 与GksHSP家族有关的复制框使用不同的颜色在染色体上展示。B: 复制事件在不同亚家族的分布。C: 复制的GksHSP同义替换率分布。"

图5

与GksHSP家族有关的复制框分析 A~E: 与GksHSP家族有关的复制框1 (A)、复制框2 (B)、复制框3 (C)、复制框4 (D)和复制框5 (E)的微共线分析。F: 与GksHSP家族有关的5个复制框的基因注释分布。G~I: 与GksHSP家族有关的5个复制框的Ka(G)、Ks(H)和Ka/Ks(I)分析。*表示在P < 0.05水平上差异显著。"

图6

GksHSP成员的启动子区中与胁迫响应相关的顺式调控元件分析 GksHSP亚家族用不用的颜色和字母表示。每一个颜色的方框代表一种顺式调控元件。"

图7

在陆地棉中GksHSP26同源基因的识别和表达分析 A: GksHSP26与其在陆地棉中的同源基因Gh_D08G181200和Gh_A08G183800的氨基酸序列分析。B~C: 在高温、干旱和盐胁迫下Gh_D08G181200 (B)和Gh_A08G183800 (C)的表达分析。"

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