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作物学报 ›› 2023, Vol. 49 ›› Issue (9): 2446-2461.doi: 10.3724/SP.J.1006.2023.24186

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

比较转录组分析花生种子休眠调控网络

王菲菲1(), 张胜忠1, 胡晓辉1, 崔凤高1, 钟文3, 赵立波4, 张天雨3, 郭进涛5, 于豪谅6, 苗华荣1,*(), 陈静1,*()   

  1. 1山东省花生研究所, 中国山东青岛 266100
    2佐治亚大学蒂芙顿校区园艺系, 美国佐治亚州蒂芙顿 31793
    3山东省种子管理总站, 中国山东济南 250100
    4青岛市农业技术推广中心, 中国山东青岛 266071
    5新郑市和庄农业服务中心, 中国河南新郑 451150
    6烟台枫林食品股份有限公司, 中国山东烟台 264108
  • 收稿日期:2022-08-10 接受日期:2023-02-21 出版日期:2023-09-12 网络出版日期:2023-03-03
  • 通讯作者: *陈静, E-mail: mianbaohua2008@126.com; 苗华荣, E-mail: 1649813628@qq.com
  • 作者简介:王菲菲, E-mail: wangfeifeisj@163.com
  • 基金资助:
    国家自然科学基金青年基金项目(32001584);国家自然科学基金青年基金项目(32201876);山东省自然科学基金面上项目(ZR2022MC045);山东省农业良种工程(2020LZGC001);山东省农业科学院创新工程(CXGC2022A03);山东省农业科学院创新工程(CXGC2022A21);青岛市民生科技计划项目(20-3-4-26-nsh);新疆维吾尔自治区重大科技专项(2022A02008-3)

Comparative transcriptome profiling of dormancy regulatory network in peanut

WANG Fei-Fei1(), ZHANG Sheng-Zhong1, HU Xiao-Hui1, CHU Ye2, CUI Feng-Gao1, ZHONG Wen3, ZHAO Li-Bo4, ZHANG Tian-Yu3, GUO Jin-Tao5, YU Hao-Liang6, MIAO Hua-Rong1,*(), CHEN Jing1,*()   

  1. 1Shandong Peanut Research Institute, Qingdao 266100, Shandong, China
    2Department of Horticulture, University of Georgia Tifton Campus, Tifton 31793, GA, United States
    3Shandong Seed Administration Station, Jinan 250100, Shandong, China
    4Qingdao Agricultural Technology Extension Center, Qingdao 266071, Shandong, China
    5Agricultural Service Center of Hezhuang, Xinzheng, Zhengzhou 451150, Henan, China
    6Yantai Fenglin Foodstuff Co., Ltd, Yantai 264108, Shandong, China
  • Received:2022-08-10 Accepted:2023-02-21 Published:2023-09-12 Published online:2023-03-03
  • Supported by:
    Youth Fund Project of the National Natural Science Foundation of China(32001584);Youth Fund Project of the National Natural Science Foundation of China(32201876);General Project of Shandong Natural Science Foundation(ZR2022MC045);Shandong Province Agriculture Improved Seed Project(2020LZGC001);Innovation Project of Shandong Academy of Agriculture Sciences(CXGC2022A03);Innovation Project of Shandong Academy of Agriculture Sciences(CXGC2022A21);Qingdao People’s Livelihood Science and Technology Project(20-3-4-26-nsh);Major Science and Technology Program of Xinjiang Uygur Autonomous Region(2022A02008-3)

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摘要：

种子休眠性是花生重要且复杂的农艺性状, 对花生(Arachis hypogaea L.)的产量和品质影响巨大。为深入揭示花生种子休眠维持和解除的分子调控网络, 本研究以强休眠品种花育52号(HY52)和弱休眠突变株系M23、M67为试材, 种子吸胀处理(0 h、12 h、24 h)后测定其激素ABA和GA含量并进行转录组测序。吸胀12 h时M23和M67中GA含量显著高于HY52, ABA含量和ABA/GA比值则低于HY52。测序共得到31,373个差异表达基因(DEGs), 其中ABA和GA生物合成和信号转导相关的基因在种子休眠维持和解除过程中发生显著变化, 挖掘到50个ABA相关基因、8个GA相关基因、49个乙烯相关基因和13个生长素相关基因。此外, 还鉴定到许多参与碳水化合物和脂质代谢、氨基酸代谢途径相关的DEG, 挖掘到糖代谢相关基因5个、脂质代谢相关基因4个; 昼夜节律调控也可能参与花生种子休眠解除。这表明, 花生种子休眠维持和解除的调控是一个复杂网络, 植物激素平衡调控可能只是其中一个重要部分。

关键词: 花生, 休眠维持, 休眠解除, 转录组, 植物激素, 氨基酸代谢

Abstract:

Seed dormancy is an important and complex agronomic trait affecting yield and quality of peanut (Arachis hypogaea L.). Seed dormancy and germination was reported to be regulated by the balance between abscisic acid (ABA) and gibberellic acid (GA). In this study, transcriptomic sequencing was performed with Huayu 52 (HY52), a peanut cultivar with strong dormancy, and two EMS mutant lines from HY52 with a weak level of dormancy. Seeds from these three lines were imbibed for 0, 12, and 24 h before tissue harvesting and RNA seq analysis. GA content of M23 and M67 was significantly higher than HY52 at 12 h after imbibition, however, the ABA content and ABA/GA ratio were lower than HY52. A total of 31,374 differentially expressed genes (DEGs) including biosynthesis and signal transduction related genes of plant hormones such as ABA and GA were discovered. We identified 50 genes related to ABA, 8 genes related to GA, 49 genes related to ethylene, and 13 genes related to auxin. Expression profiles of ABA and GA related genes was consistent with the higher GA and lower ABA content in the mutants compared with HY52 after 12 h and 24 h imbibition. In addition, many DEGs involved in carbohydrate and lipid metabolism, amino acid metabolism, and glutathione metabolism pathway were also identified. There were 5 carbohydrate metabolism related genes (GPT) and 4 lipid metabolism related genes. In addition, differentially regulated circadian rhythm pathways were found to involve in the process of peanut seed dormancy release. These results suggested that the regulation of dormancy maintenance and release was more complicated than phytohormone balance.

Key words: peanut, dormancy maintenance, dormancy release, transcriptome, plant hormone, amino acid metabolism

图1

HY52、M23和M67的休眠性差异 A: HY52、M23和M67的萌发率, B~D: 吸胀12 h和24 h的GA含量、ABA含量和ABA/GA。GA: 赤霉素; ABA: 脱落酸; HY52: 花育52号。星号表示通过单因素方差分析与HY52相比有显著性(*P<0.05, **P<0.01, ***P<0.001)。"

表1

参试样品的转录组数据质量"

样品名称
Sample name		 原始读长
Raw reads		 有效读长
Clean reads		 有效碱基
Clean bases (Gb)		 有效读长的比例
Valid ratio (reads) (%)		 基因组上的比对率
Mapped ratio (%)		 ≥Q30 (%) GC含量
GC content (%)
HY52_0_1 47,336,764 46,882,472 7.03 99.04 92.15 94.54 46.00
HY52_0_2 40,239,562 39,850,56 5.98 99.03 92.52 96.56 45.50
HY52_0_3 42,126,764 41,757,916 6.26 99.12 92.29 94.37 47.00
M23_0_1 53,524,396 47,595,296 7.14 88.92 84.53 90.44 48.50
M23_0_2 54,811,458 53,791,824 8.07 98.14 91.70 96.29 48.00
M23_0_3 58,964,636 58,493,438 8.77 99.20 92.87 96.57 45.50
M67_0_1 44,573,54 44,010,372 6.60 98.74 92.02 95.24 47.00
M67_0_2 43,794,954 43,380,290 6.51 99.05 91.63 92.85 45.50
M67_0_3 43,654,578 43,168,590 6.48 98.89 92.05 93.96 46.00
HY52_12_1 43,666,144 43,006,610 6.45 98.49 92.15 95.24 49.00
HY52_12_2 40,974,886 40,618,232 6.09 99.13 92.86 93.45 46.00
HY52_12_3 48,252,204 47,774,688 7.17 99.01 92.33 94.51 45.50
M23_12_1 44,263,360 43,728,20 6.56 98.79 92.87 94.23 47.00
M23_12_2 44,207,630 43,791,190 6.57 99.06 92.59 94.40 46.00
M23_12_3 57,707,210 57,167,752 8.58 99.07 93.87 96.48 46.50
M67_12_1 47,514,726 46,870,288 7.03 98.64 92.41 95.52 47.00
M67_12_2 47,016,098 46,486,226 6.97 98.87 92.85 94.35 45.00
M67_12_3 41,492,948 41,145,894 6.17 99.16 92.88 93.62 45.00
HY52_24_1 51,701,800 50,178,368 7.53 97.05 93.48 96.37 47.50
HY52_24_2 39,391,916 39,016,270 5.85 99.05 92.14 93.33 46.50
HY52_24_3 48,847,594 48,142,426 7.22 98.56 93.28 96.44 47.00
M23_24_1 42,599,196 41,768,316 6.27 98.05 92.71 94.42 46.00
M23_24_2 63,130,436 62,578,018 9.39 99.12 93.84 95.84 45.00
M23_24_3 60,429,598 59,883,810 8.98 99.10 94.47 96.12 46.50
M67_24_1 44,066,802 43,632,302 6.54 99.01 94.07 95.96 47.50
M67_24_2 41,088,448 40,772,706 6.12 99.23 94.22 96.06 45.50
M67_24_3 42,584,300 42,214,250 6.33 99.13 93.06 92.79 45.00

表2

花育52号全长转录组的功能注释"

数据库
Database		 总基因簇
Total_unigene		 KOG数据库
KOG database		 KEGG数据库
KEGG database		 NR数据库
NR database		 SwissProt数据库
SwissProt
database		 GO数据库
GO database		 注释基因总数
Overall_
annotated
Gene_Number 47,697 25,840 19,192 43,623 33,161 19,468 43,930
Annotation_Ratio — 54.18% 40.24% 91.46% 69.52% 40.82% 92.10%

图2

不同处理差异表达基因比较及韦恩图 HY52: 花育52号; DEGs: 差异表达基因。"

图3

HY52、M23和M67的差异基因表达谱 HY52 (A)和M67 (C)的表达谱聚成2个组, 即上调和双向表达谱, M23 (B)则聚成3个组, 即上调、下调和双向表达谱。表达谱编号标记在左上角, 每个表达谱对应的P值标记在左下角, 括号中为每个表达谱中差异基因的数量。HY52: 花育52号。"

图4

花生种子休眠保持过程中差异基因GO富集分析 HY52: 花育52号。"

图5

花生种子休眠保持过程中差异基因KEGG富集分析 HY52: 花育52号。"

图6

花生种子休眠解除过程中差异基因GO富集分析"

图7

花生种子休眠解除过程中差异基因KEGG富集分析"

图8

花生种子吸胀过程中与激素合成、代谢和信号转导相关的差异基因表达热图 A: ABA、GA、乙烯和生长素的生物合成、代谢和信号转导示意图; B: 与植物激素相关的差异表达基因热图。HY52: 花育52号。"

图9

花生种子吸胀过程中与碳水化合物、脂质合成、代谢相关的差异基因表达热图 A: 脂质代谢相关差异表达基因热图; B: 碳水化合物合成和代谢相关差异表达基因热图。HY52: 花育52号。"

图10

HY52_12 vs M67_12 (A) and HY52_24 vs M67_24 (B)比较组前20个GO条目中与植物激素和胁迫相关的差异表达基因热图分析 HY52: 花育52号。"

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