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作物学报

作物学报 ›› 2021, Vol. 47 ›› Issue (4): 613-625.doi: 10.3724/SP.J.1006.2021.04144

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

花生荚果发育过程中的microRNA鉴定与表达分析

胡冬秀(), 刘浩(), 洪彦彬, 梁炫强, 陈小平*()   

  1. 广东省农作物遗传改良重点实验室 / 国家油料作物改良中心南方分中心 / 广东省农业科学院作物研究所, 广东广州 510640
  • 收稿日期:2020-07-02 接受日期:2020-10-14 出版日期:2021-04-12 网络出版日期:2020-11-18
  • 通讯作者: 陈小平
  • 作者简介:胡冬秀, E-mail: HuDX1017@163.com;|刘浩, E-mail: liuhao@gdaas.cn
  • 基金资助:
    广东省重点研发计划项目(2020B020219003);国家自然科学基金项目(31771841);广东省基础与应用基础研究基金(2020A1515010021)

Identification and expression analysis of microRNA during peanut (Arachis hypogaea L.) pod development

HU Dong-Xiu(), LIU Hao(), HONG Yan-Bin, LIANG Xuan-Qiang, CHEN Xiao-Ping*()   

  1. Guangdong Provincial Key Laboratory of Crop Genetic Improvement / South China Peanut Sub-Center of National Center of Oilseed Crops Improvement / Crops Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, Guangdong, China
  • Received:2020-07-02 Accepted:2020-10-14 Published:2021-04-12 Published online:2020-11-18
  • Contact: CHEN Xiao-Ping
  • Supported by:
    Guangdong Provincial Major Research and Development Project(2020B020219003);National Natural Science Foundation of China(31771841);Guangdong Basic and Applied Basic Research Foundation(2020A1515010021)

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摘要：

花生(Arachis hypogaea L.)是典型“地上开花、地下结果”的作物, 为从转录后调控水平解析此独特的果实发育现象, 本文应用small RNA测序技术研究荚果发育11个时期果壳及种子中的microRNA及其靶基因。通过测序分别获得212个已知的microRNA和112个新microRNA, 其中, 已知microRNA包括197个保守microRNA和15个花生特异microRNA, 新microRNA来自62个新的microRNA前体序列。表达分析发现, 67个microRNA及其靶基因在荚果发育的11个时期存在时空特异性表达, 部分microRNA的表达量积累阶段性调节果壳与种子的发育, 表明microRNA参与了花生荚果暗发育的整个过程。此外, 对28个microRNA与30个靶基因进行荧光定量PCR验证发现, microRNA和靶基因的表达趋势与测序结果基本一致。本研究通过对花生荚果不同发育时期的果壳和种子进行small RNA测序, 鉴定参与调控花生荚果膨大相关的microRNA, 为研究黑暗条件下植物果实发育的调控机制与花生遗传改良奠定理论基础。

关键词: 花生, 荚果发育, small RNA测序, microRNA, 靶基因

Abstract:

“Aerial flower and subterranean fruit” is a distinct feature in peanut (Arachis hypogaea L.). To dissect this character at post-transcription level, small RNA sequencing was performed to identify microRNAs in peanut pod shell and seed during eleven developmental stages. Sequencing analysis identified 212 known microRNAs, including 197 conserved and 15 specific microRNA. In addition, 112 novel microRNAs from 62 novel microRNA precursors were identified. Among the known and new microRNAs, 67 microRNAs and their target genes showed differentially expressed patterns during peanut pod development. Expression trend analysis revealed stage-specific and tissue-specific expression of microRNA and their target genes during pod shell and seed development, implying that microRNAs probably played a role in peanut pod development. To validate expression profiles from small RNA sequencing, quantitative real-time RT-PCR were performed using 28 microRNAs and 30 target genes, revealing consistent expression profiles with sequencing results. The data regarding microRNA and their target genes generated in this study would contribute to understanding the molecular mechanism of plant fruit development under darkness and to crop improvement.

Key words: peanut, pod development, small RNA sequencing, microRNA, target gene

图1

花生荚果发育的表型特征 A: 试验样品信息; B: 花生荚果种子发育的表型特征。P0: 地上果针; P1: 地下果针; P2~P5: 果壳膨大期; P6~P9: 果实充实期; P9~P10: 成熟期。"

图2

花生荚果small RNA测序结果的分析 A: 全部以及特异性small RNA的长度分布和频率; B: small RNA统计分析; C: small RNA的分类。P0: 地上果针; P1: 地下果针; P2~P5: 果壳膨大期; P6~P9: 果实充实期; P9~P10: 成熟期。"

图3

microRNA分析流程"

图4

新的Pre-microRNA二级结构分析"

附表1

花生荚果发育过程中新的和已知的microRNAs鉴定"

编号
ID		 序列
Sequence		 长度
Length(nt)		 编号
ID		 序列
Sequence		 长度
Length(nt)
New microRNAs
ahy-miR-s1-3p UGUAGAAUUUCUGCUCCUUGG 21 ahy-miR-s33-3p UGUUCAUAGCGGUGCUUGCUG 21
ahy-miR-s1-5p UAGGAUGGCAGAUAUUCUUUUCA 23 ahy-miR-s33-5p CAGUGUGGCCGGACUGGAUCAUC 23
ahy-miR-s2-3p UCCCUUUGGAUGUCGUCGUGC 21 ahy-miR-s34-3p AUUGCCUACUGAUUGCAUCCCACA 24
ahy-miR-s2-5p ACUUCUGACAGACGUAGGGAAG 22 ahy-miR-s34-5p AGGGAUGCUAUCCACAGUCGAUAG 24
ahy-miR-s3-3p UUCAUGACAGAGUGCUGGCUC 21 ahy-miR-s35-3p UAACGAGUCGAGCUCGAGCUAGCU 24
ahy-miR-s3-5p AAGGUCUCUGUCUCUGUGGU 20 ahy-miR-s35-5p UUUAGGCUCAAGCUUGGCUCACCA 24
ahy-miR-s4-3p AACCAAAUGAUGAACCAUUCU 21 ahy-miR-s36-3p UCAGUGUUGCCAGGCGCCUGGG 22
ahy-miR-s4-5p AAUGUUGAAGAAUUAGGUUGC 21 ahy-miR-s36-5p CAACGUCUGCCACCACUGAUG 21
ahy-miR-s5-5p CAACAGCACCCUCCACCUGAG 21 ahy-miR-s37-3p UUGCGUUGGAGAUGUUGUCCAAUG 24
ahy-miR-s6-5p AGCGAUUGUUUCUCUUGAUGCAGU 24 ahy-miR-s37-5p AUUGGACAACAUCUCCAACGUAAC 24
ahy-miR-s7-3p GAUGAAUGCAGCUCCCUAUAA 21 ahy-miR-s38-3p UUAAAGUCUGGCCUGACCUAGCCU 24
ahy-miR-s7-5p AUGGGGCUGGAGUUUGGCUC 20 ahy-miR-s38-5p GCUACAGGCCAGGCUUAGACC 21
ahy-miR-s8-3p AGGGGUGAUCUGCCUAGUC 19 ahy-miR-s39-3p UGAAGGAAGUAGUGGCGACCC 21
ahy-miR-s8-5p UGAGGCACGCACUCCAGAUGA 21 ahy-miR-s40-3p UUGAGGACUUGAUAUAUGUCG 21
ahy-miR-s9-3p CGUGCGCGGUCUUGCACUGCU 21 ahy-miR-s41-3p GUUAGUAAUUCGAACCACCCUGGU 24
ahy-miR-s9-5p CAAGUCGUGCACGGCUGGCCU 21 ahy-miR-s41-5p CACCCUGGUUCGAAUUACGCUUAA 24
ahy-miR-s10-5p UUGGAAAUGUAUGGCGAGCCU 21 ahy-miR-s42-3p AGAUAUACUUGUUACGCUUG 20
ahy-miR-s11-3p UUUGAGAUUGGAUAAUUGCCU 21 ahy-miR-s42-5p CGGCAGUGAUGAGGCUCUAACAUG 24
ahy-miR-s12-3p UUGGAAAAGGUCGGUAGAAAG 21 ahy-miR-s43-3p UGGUAAGAGUUUGAUUAAGUA 21
ahy-miR-s12-5p UUCACCGACUCUGUUCAGACU 21 ahy-miR-s43-5p CUUAAUCAAACUCUUAUCAUC 21
ahy-miR-s13-5p UUCAGAGGCUUGUUUUAUCUC 21 ahy-miR-s44-3p UACUGAAAUCAGAUCAUCCGAUUA 24
ahy-miR-s14-3p GCAAACAGGUUCGGUGAAAAC 21 ahy-miR-s44-5p AUCGGAUGGUCCGAUUUGUGUACC 24
ahy-miR-s14-5p UUUACCAGAUCCUGUUUGUUC 21 ahy-miR-s45-3p UCUAACCGAACCGAACCGAAC 21
ahy-miR-s15-3p UUGUCUCUAAGUGGUGGUUCCA 22 ahy-miR-s45-5p UCGGUUCGGUUUUCGGUUCUGACC 24
ahy-miR-s15-5p GAACUGUUCUUCUUAGUUGCGCCU 24 ahy-miR-s46-3p UGAAGAUUGUUGGGUAGUGUU 21
ahy-miR-s16-3p CAGCUGUUUGCGGACAAGUCUGGC 24 ahy-miR-s46-5p AACAUGGUGAAAGGUUUUCAUU 22
ahy-miR-s16-5p AGACUGUGCGCCGAUAUGCCCGUG 24 ahy-miR-s47-3p UUGAUAUGGUGUAUUCUGCAGCCU 24
ahy-miR-s17-5p UUUGGCAGCUCAAUCACUUGG 21 ahy-miR-s47-5p GUUGUUCAGGUGUAUUAUAUCAGA 24
ahy-miR-s18-3p CACUUCUUAUGCUUGUUGGAG 21 ahy-miR-s48-3p AUUUUUAAUCGAUUGUUUUGUGAU 24
ahy-miR-s18-5p CGCAUGGAUGGAGAAUCUCGCU 22 ahy-miR-s48-5p GACCAAUCGAUUGAUUUUUG 20
ahy-miR-s19-5p UCUUGAAAGCUGAGCAUUGGC 21 ahy-miR-s49-3p CCAACGUUGGAGCAAAAGUUAGA 23
ahy-miR-s20-3p ACCGGACUUGCGCCCCAGUCGUA 23 ahy-miR-s49-5p AACUUUUUGGCUAACGUUGGCGCC 24
ahy-miR-s20-5p UCACAGGCAGCAUAUCAGGUUG 22 ahy-miR-s50-3p UCCAAGAUGUGGCAUAUCAGUUG 23
ahy-miR-s21-5p ACGAAACUGUGUGAAGUAAGGCCU 24 ahy-miR-s50-5p UUUCAUAUGCCGUUCAGAGAGU 22
ahy-miR-s22-3p UUAAAUCAGUCUGUGGUACCC 21 ahy-miR-s51-3p UUCUCGGAUUAAUCAGGCUCAGCC 24
ahy-miR-s22-5p CCGCGGAUUGAUUUUUACCAGA 22 ahy-miR-s51-5p CUGCCUGUUUAGUUCGCGGACU 22
ahy-miR-s23-3p UUGAUUGAGUGACUGUAUAGCU 22 ahy-miR-s52-5p UCAGUGUUUGCUCGCCUCUUC 21
ahy-miR-s23-5p CUAUGAGUUACUCAAUCAAAC 21 ahy-miR-s53-3p CUGGAGGCCUUUGAAGGAGAC 21
ahy-miR-s24-3p UGGCUUGGUCUGACGUGACUCAGC 24 ahy-miR-s53-5p UCCCUCAAAGGCUUCCAGUA 20
ahy-miR-s24-5p AUGAGUCGAGUCGUGAGCCAAGCU 24 ahy-miR-s54-3p AACCGACACGUGAGCUCAUGGCCA 24
ahy-miR-s25-3p UUUUCCUCUUAAUGGAUCCUCUGU 24 ahy-miR-s54-5p UUAGCUAUCGGAUGUGUCGAGUCU 24
ahy-miR-s25-5p AGAGAGGAUCCUUAUCUGGGAACU 24 ahy-miR-s55-3p UUUGAAACAGAAUCAGUUGGGAUC 24
ahy-miR-s26-3p UCCUUGGACCUUGGGUGUGAU 21 ahy-miR-s55-5p UUGGUGACUGAUUUGUGAAUCAAA 24
ahy-miR-s26-5p AGGACCCAACUCUCUUGAAGGAGA 24 ahy-miR-s56-3p UUCUGCGCGGUCGCGUCGCUGACG 24
ahy-miR-s27-3p UGAGGGGGUUGAGGGUGCUGGGGU 24 ahy-miR-s56-5p UGAGCCAUGAGACCGCGUCACUGC 24
ahy-miR-s27-5p UUCAGGAUCUCUAUUACUGGC 21 ahy-miR-s57-5p UUGAGUACCGUCGGAUUUAUCAUC 24
ahy-miR-s28-3p GUUACUGAUUUACUGGUUCACG 22 ahy-miR-s58-3p ACUUUAGUGGCUGUUCGCUCACUU 24
ahy-miR-s28-5p AGAACCGGACCGGUCAAUAAACCA 24 ahy-miR-s58-5p AUAGAGAGUAGGCCAGUAGAAGCC 24
ahy-miR-s29-3p UUCUCCCUUGGUAGUGGCGAAGC 23 ahy-miR-s59-3p UUGUUGUGGCGCCAACGUUUGCCU 24
ahy-miR-s29-5p CAUCGCCAACUCCAAGGAAG 20 ahy-miR-s59-5p CAAACGUUGGCGCAAGCUUUUGCU 24
ahy-miR-s30-3p GGAAGAUUGUUGGUUAGUGUU 21 ahy-miR-s60-3p AGGAGCUCUGCUGUGUCUUGAUGG 24
ahy-miR-s30-5p GACUAAUGACAUUCAACCUC 20 ahy-miR-s60-5p UUCCACGGCAUGACUCUCUAAACC 24
ahy-miR-s31-3p AAUUUAGACAAUUCAUCCGAU 21 ahy-miR-s61-3p ACAGUGAGGUUUGUAAGAAAAAGC 24
ahy-miR-s31-5p AAAUCGGAUGAAUUGUCUAAAUUC 24 ahy-miR-s61-5p UUUUUCUGUUUAAUUUUGUCU 21
ahy-miR-s32-3p AAUAAAACAAGUUUUGACUGU 21 ahy-miR-s62-3p UCAAACGAGGAAAGGCUUAUGG 22
ahy-miR-s32-5p AGCUCAAAUUUGCCUUAUUUA 21 ahy-miR-s62-5p AUCUAGCAGCACCUUAGGAUGGCA 24
Known and Conserved microRNAs
ahy-miR156b-5p UUGACAGAAGAUAGAGAGCAC 21 ahy-miR168d UCGCUUGGUGCAGGUCGGGAC 21
ahy-miR156b-3p GCUCUCUAAGCUUCUGUCAUC 21 ahy-miR169f CAGCCAAGGAUGACUUGCCGG 21
ahy-miR156c UUGACAGAAGAGAGAGAGCAC 21 ahy-miR169h UGAGCCAAGGAUGGCUUGCCG 21
ahy-miR156d UGACAGAAGAAAGUGAGCAC 20 ahy-miR169l CAGCCAAGAAUGACUUGCCGG 21
ahy-miR156e UGACAGAAGAGAGUGAGCACA 21 ahy-miR169p UGAGCCAAGGAUGACUUGCCG 21
ahy-miR156f UGACAGAAGAGAAUGAGCAC 20 ahy-miR169r UAGCCAAGGAUGACUUGCCU 20
ahy-miR156g UGACAGAAGAGAGGGAGCAC 20 ahy-miR169s AAGCCAAGGAUGACUUGCCGG 21
ahy-miR156h UGACAGAAGAAAGAGAGCAC 20 ahy-miR171a GGAUAUUGGUACGGUUCAAUC 21
ahy-miR156i ACAGAAGAUAGAGAGCACAG 20 ahy-miR171b UUGAGCCGUGCCAAUAUCACG 21
ahy-miR156j UGACAGAAGAGGGUGAGCAC 20 ahy-miR171c UAUUGGUGCGGUUCAAUGAGA 21
ahy-miR156k UGUGCUCACUCUCUUCUGUCA 21 ahy-miR171d UUGAGCCGUGCCAAUAUCACU 21
ahy-miR156l UGUCAGAAGAGAGUGAGCAC 20 ahy-miR171e UGAGCCGUGCCAAUAUCACAU 21
ahy-miR156m CGACAGAAGAGAGUGAGCAC 20 ahy-miR171f UGAUUGAGCCGUGCCAAUAUC 21
ahy-miR156n UGACAGAGGAGAGUGAGCAC 20 ahy-miR171g CGAUGUUGGUGAGGUUCAAUC 21
ahy-miR156o UGACAGAAGAGAGCGAGCAC 20 ahy-miR171h UGAUUGAGCCGCGUCAAUAUC 21
ahy-miR156q UGACAGAAGAGAGUGAGCACU 21 ahy-miR171i CGAGCCGAAUCAAUAUCACUC 21
ahy-miR156s CUGACAGAAGAUAGAGAGCAC 21 ahy-miR171j UAUUGGCCUGGUUCACUCAGA 21
ahy-miR157b GCUCUCUAAGCUUCUGUCAUCA 22 ahy-miR171k UUGAGCCGCGCCAAUAUCACA 21
ahy-miR157d UGACAGAAGAUAGAGAGCAC 20 ahy-miR171l UUGAGCCGCGCCAAUAUCACU 21
ahy-miR159a UUUGGAUUGAAGGGAGCUCUG 21 ahy-miR171n UUGAGCCGUGCCAAUAUCACA 21
ahy-miR159b UUUGGAUUGAAGGGAGCUCUU 21 ahy-miR171p UUGAGCCGCGUCAAUAUCUUA 21
ahy-miR159c UUUGGAUUGAAGGGAGCUCCU 21 ahy-miR171t UUGAGCCGCGUCAAUAUCUCA 21
ahy-miR159d AGCUGCUUAGCUAUGGAUCCC 21 ahy-miR172a AGAAUCUUGAUGAUGCUGCAU 21
ahy-miR159e UUUGGAUUGAAAGGAGCUCUU 21 ahy-miR172b GGAGCAUCAUCAAGAUUCACA 21
ahy-miR159f CUUGGAUUGAAGGGAGCUCUA 21 ahy-miR172c GUAGCAUCAUCAAGAUUCACA 21
ahy-miR159g UUGGAUUGAAGGGAGCUCCA 20 ahy-miR172d AGAAUCUUGAUGAUGCUGCAG 21
ahy-miR159h CUUGGAUUGAAGGGAGCUCU 20 ahy-miR172k UGAAUCUUGAUGAUGCUGCAU 21
ahy-miR159i UUUGGACUGAAGGGAGCUCUA 21 ahy-miR390a-5p AAGCUCAGGAGGGAUAGCGCC 21
ahy-miR159j AUUGGAGUGAAGGGAGCUCCA 21 ahy-miR390b CGCUAUCCAUCCUGAGUUUCA 21
ahy-miR159k UUUGGUUUGAAGGGAGCUCUA 21 ahy-miR390c CGCUAUCCAUCCUGAGUUUC 20
ahy-miR159l AUUGGAUUGAAGGGAGCUCCU 21 ahy-miR390d AAGCUCAGGAGGGAUAGCACC 21
ahy-miR159m AUUGGAUUGAAGGGAGCUCCA 21 ahy-miR390e AAGCUCAGGAGGGAUAGCGUC 21
ahy-miR159n CUUGGAUUGAAGGGAGCUCCC 21 ahy-miR390f GAGCUCAGGAGGGAUAGCGCC 21
ahy-miR159 UUUGGAUUGAAGGGAGCUCUA 21 ahy-miR390g CGCUAUCCAUCCUGAGUUCCA 21
ahy-miR319a UUGGACUGAAGGGAGCUCCC 20 ahy-miR393a UCCAAAGGGAUCGCAUUGAUCC 22
ahy-miR319b UUGGACUGAAGGGAGCUCCCU 21 ahy-miR393b UCCAAAGGGAUCGCAUUGAUCU 22
ahy-miR319c UUUGGACUGAAGGGAGCUCCU 21 ahy-miR393c UCCAAAGGGAUCGCAUUGAUC 21
ahy-miR319d UUGGACUGAAGGGAGCUCCU 20 ahy-miR393d AUCAUGCUAUCCCUUUGGAUU 21
ahy-miR319e CUUGGACUGAAGGGAGCUCCC 21 ahy-miR393e AUCAUGCUAUCUCUUUGGAUU 21
ahy-miR319f UUGGACUGAAGGGGCCUCUU 20 ahy-miR393k UUCCAAAGGGAUCGCAUUGAUC 22
ahy-miR319g UUGGACUGAAGGGAGCUCCCA 21 ahy-miR394 UUGGCAUUCUGUCCACCUCC 20
ahy-miR319h UUGGACUGAAGGGUGCUCCCU 21 ahy-miR394c UUGGCAUUCUGUCCACCUCCAU 22
ahy-miR319i UUGGACUGAAGGGGAGCUCCUUC 23 ahy-miR396a UUCCACAGCUUUCUUGAACUA 21
ahy-miR319j CUUGGACUGAAGGGAGCUCCU 21 ahy-miR396b UUCCACAGCUUUCUUGAACUG 21
ahy-miR319k UUGGGCUGAAGGGAGCUCCC 20 ahy-miR396c UUCCACAGCUUUCUUGAACUU 21
ahy-miR319l UUGGACUGAAGGGAGCUCCUUC 22 ahy-miR396d UUCCACGGCUUUCUUGAACUU 21
ahy-miR160a GCGUAUGAGGAGCCAAGCAUA 21 ahy-miR396e UUCCACAGCUUUCUUGAACUGU 22
ahy-miR160b UGCCUGGCUCCCUGGAUGCCA 21 ahy-miR396f UUCCACGGCUUUCUUGAACUG 21
ahy-miR160c UGCCUGGCUCCCUGUAUGCCA 21 ahy-miR396g UCUUCCACAGCUUUCUUGAAC 21
ahy-miR160d UGCCUGGCUCCCUGCAUGCCA 21 ahy-miR396h UUCCACAGCUUUCUUGAACAG 21
ahy-miR160f UGCCUGGCUCCCUGUAUGCCG 21 ahy-miR396i GUUCAAUAAAGCUGUGGGAAG 21
ahy-miR160i CGCCUGGCUCCCUGUAUGCCA 21 ahy-miR396j GUUCAAUAAAGCUGUGGGAAA 21
ahy-MIR160-5p UGCCUGGCUCCCUGAAUGCCA 21 ahy-miR396k GCUCAAGAAAGCUGUGGGAGA 21
ahy-MIR160-3p GCAUGAAGGGAGUCACGCAGG 21 ahy-miR396l AAGAAAGCUGUGGGAGAAUAUGGC 24
ahy-miR162a UCGAUAAACCUCUGCAUCCAG 21 ahy-miR397a UCAUUGAGUGCAGCGUUGAUG 21
ahy-miR162b GGAGGCAGCGGUUCAUCGAUC 21 ahy-miR397b UCAUUGAGUGCAGCGUUGAUGU 22
ahy-miR162c UCGAUAAACCUCUGCAUCCGG 21 ahy-miR397c CCAUUGAGUGCAGCGUUGAUG 21
ahy-miR162d UCGAUAAGCCUCUGCAUCCAG 21 ahy-miR398a UGUGUUCUCAGGUCGCCCCUG 21
ahy-miR162e UGGAGGCAGCGGUUCAUCGAUC 22 ahy-miR398b UGUGUUCUCAGGUCACCCCUU 21
ahy-miR164a UGGAGAAGCAGGGCACGUGCA 21 ahy-miR398c UGUGUUCUCAGGUCGCCCCCG 21
ahy-miR164b UGGAGAAGCAGGGUACGUGCA 21 ahy-miR398d UGUGUUCUCAGGUCACCCCUG 21
ahy-miR164c UGGAGAAGCAGGGCACGUGCG 21 ahy-miR399a CGCCAAAGGAGAGUUGCCCUG 21
ahy-miR164d UGGAGAAGCAGGGCACGUGCU 21 ahy-miR399b UGCCAAAGGAGAGUUGCCCUA 21
ahy-miR164e UGGAGAAGCAGGGCACGUGAA 21 ahy-miR399c UGCCAAAGGAGAUUUGCCCUG 21
ahy-miR164h UGGAGAAGCAGGGCACGUGUG 21 ahy-miR399d UGCCAAAGGAGAGCUGCCCUG 21
ahy-miR166a-5p GGAAUGUUGUCUGGCUCGAGG 21 ahy-miR399e GGGCUUCUCUUUCUUGGCAGG 21
ahy-miR166a-3p UCGGACCAGGCUUCAUUCCCC 21 ahy-miR399f UGCCAAAGGAGAUUUGCCCGG 21
ahy-miR166b UCGGACCAGGCUUCAUUCCCCC 22 ahy-miR399g CGGGGCAAAUCUCCUUUGGCA 21
ahy-miR166c UCGGACCAGGCUUCAUUCCCGU 22 ahy-miR399j UGCCAAAGGAGAGUUGCCCUG 21
ahy-miR166d UCGGACCAGGCUUCAUUCCCU 21 ahy-miR399r UGCCAAAGAAGAUUUGCCCCG 21
ahy-miR166e CUCGGACCAGGCUUCAUUCCC 21 ahy-miR408-3p AUGCACUGCCUCUUCCCUGGC 21
ahy-miR166f UCGGACCAGGCUUCAUCCCCC 21 ahy-miR408a UGCACUGCCUCUUCCCUGGCU 21
ahy-miR166g UCGGACCAGGCUUCAUUCCC 20 ahy-miR408b ACUGGGAACAGGCAGAGCAUGA 22
ahy-miR166h-3p UCUCGGACCAGGCUUCAUUCC 21 ahy-miR408c UGCACUGCCUCUUCCCUGGCUG 22
ahy-miR166i UCGGACCAGGCUUCAUUCUC 20 ahy-miR482c UUUCCAAUUCCACCCAUUCCUA 22
ahy-miR166j-3p UCGGACCAGGCUUCAUUCCCG 21 ahy-miR482d-3p UCUUCCCUACACCUCCCAUACC 22
ahy-miR166j-5p GGAAUGUUGUUUGGCUCGAGG 21 ahy-miR482d-5p UAUGGGGGGAUUGGGAAGGAAU 22
ahy-miR166k UCGAACCAGGCUUCAUUCCCC 21 ahy-miR482e UAUGGGGGGAUUGGGAAGGAA 21
ahy-miR166l UCCGGACCAGGCUUCAUUCCC 21 ahy-miR530 UGCAUUUGCACCUGCACUUUA 21
ahy-miR166m-5p GGAAUGUUGGCUGGCUCGAGG 21 ahy-miR828 UCUUGCUCAAAUGAGUAUUCCA 22
ahy-miR166m-3p UCGGACCAGGCUUCAUUCCUC 21 ahy-miR858 UCUCGUUGUCUGUUCGACCUU 21
ahy-miR166n UCGGACCAGGCUUCAAUCCCU 21 ahy-miR894 CGUUUCACGUCGGGUUCACC 20
ahy-miR166o UCGGACCAGGCUUCAUUCCUU 21 ahy-miR1310 AGGCAUCGGGGGCGCAACGCCC 22
ahy-miR166p UCGGACCAGGCUUCAUUCCUA 21 ahy-miR1507 CCUCGUUCCAUACAUCAUCUAG 22
ahy-miR166q UCGGACCAGGCUUCAUUCCCUU 22 ahy-miR1511 AACCAGGCUCUGAUACCAUG 20
ahy-miR166r UCGGAUCAGGCUUCAUUCCUC 21 ahy-miR1515 UCAUUUUUGCGUGCAAUGAUCC 22
ahy-miR166u UCUCGGACCAGGCUUCAUUC 20 ahy-miR2111 UAAUCUGCAUCCUGAGGUUUA 21
ahy-miR167a AGAUCAUCUGGCAGUUUCACC 21 ahy-miR2118a UUGCCGAUUCCACCCAUUCCUA 22
ahy-miR167b UGAAGCUGCCAGCAUGAUCUA 21 ahy-miR2118b UUACCGAUUCCACCCAUUCCUA 22
ahy-miR167c UGAAGCUGCCAGCAUGAUCUGG 22 ahy-miR3711 AGGCCCUCCUUCUAGCGCCA 20
ahy-miR167d UGAAGCUGCCAGCAUGAUCUGA 22 ahy-miR4995 AGGCAGUGGCUUGGUUAAGGG 21
ahy-miR167e UGAAGCUGCCAGCAUGAUCUC 21 ahy-miR5139 AAACCUGGCUCUGAUACCA 19
ahy-miR167f UGAAGCUGCCAGCAUGAUCUG 21 ahy-miR5141a UUAUCUGUCAGUCGCGUCGGGUCU 24
ahy-miR167j UGAAGCUGCCAGCAUGAUCUUA 22 ahy-miR5141b AGACCCGACGCGACUGACAGAUAA 24
ahy-miR167-5p UGAAGCUGCCAGCAUGAUCUU 21 ahy-miR5538 ACUGAACUCAAUCACUUGCUGC 22
ahy-miR167-3p AGAUCAUGUGGCAGUUUCACC 21 ahy-miR6173a AGCCGUAAACGAUGGAUACU 20
ahy-miR168a UCGCUUGGUGCAGGUCGGGAA 21 ahy-miR6173b AGUAUCCAUCGUUUACGGCU 20
ahy-miR168b UCGCUUGGUGCAGGUCGAGAA 21 ahy-miR6478 CCGACCUUAGCUCAGUUGGUG 21
ahy-miR168c CCCGCCUUGCAUCAACUGAAU 21
Known and Peanut-Specific
ahy-miR3508 UAGAGGGUCCCCAUGUUCUCA 21 ahy-miR3514-3p UCACCGUUAAUACAGAAUCCUU 22
ahy-miR3509-5p AUACUUGAGAGCCGUUAGAUGA 22 ahy-miR3515 AAUGUAGAAAAUGAACGGUAU 21
ahy-miR3510 UUAUACCAUCUUGCGAGACUGA 22 ahy-miR3516 GCUGGGUGAUAUUGACAGAAG 21
ahy-miR3511-3p UGUUACUAUGGCAUCUGGUAA 21 ahy-miR3517 CUGACCACUGUGAUCCCGGAA 21
ahy-miR3511-5p GCCAGGGCCAUGAAUGCAGA 20 ahy-miR3518 UGACCUUUGGGGAUAUUCGUG 21
ahy-miR3512 CGCAAAUGAUGACAAAUAGA 20 ahy-miR3519 UCAAUCAAUGACAGCAUUUCA 21
ahy-miR3513-5p UUAAUUUCUGAGUUUGUCAUC 21 ahy-miR3520-3p AAGGGAGACGUUUGAAUUAUC 21
ahy-miR3513-3p UUGAUAAGAUAGAAAUUGUAU 21

图5

microRNA在花生荚果发育过程中的表达模式分析 P0: 地上果针; P1: 地下果针; P2~P5: 果壳膨大期; P6~P9: 果实充实期; P9~P10: 成熟期。"

图6

花生荚果发育过程中microRNA的趋势分析 P0: 地上果针; P1: 地下果针; P2~P5: 果壳膨大期; P6~P9: 果实充实期; P9~P10: 成熟期。"

图7

靶基因在花生荚果发育过程中的表达模式分析 P0: 地上果针; P1: 地下果针; P2~P5: 果壳膨大期; P6~P9: 果实充实期; P9~P10: 成熟期。"

图8

花生荚果发育过程中靶基因的表达趋势分析 P0:地上果针;P1:地下果针; P2~P5:果壳膨大期; P6~P9:果实充实期; P9~P10:成熟期。"

图9

转录组数据的qRT-PCR验证 P0: 地上果针; P1: 地下果针; P2~P5: 果壳膨大期; P6~P9: 果实充实期; P9~P10: 成熟期。"

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