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作物学报

作物学报 ›› 2024, Vol. 50 ›› Issue (12): 2998-3012.doi: 10.3724/SP.J.1006.2024.42023

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

代谢组和转录组分析籼稻福香占干旱胁迫的分子响应

王颖姮1,2(), 崔丽丽1,2, 蔡秋华1,2, 林强1,2, 吴方喜1,2, 陈飞鹤1,2, 谢鸿光1,2, 朱永生1,2, 陈丽萍1,2, 谢华安1,2, 张建福1,2,*()   

  1. 1福建省农业科学院水稻研究所, 福建福州 350019
    2农业农村部华南杂交水稻种质创新与分子育种重点实验室 / 福州(国家)水稻改良分中心 / 福建省作物分子育种工程实验室 / 福建省水稻分子育种重点实验室 / 福建省作物种质创新与分子育种省部共建国家重点实验室培育基地 / 杂交水稻国家重点实验室华南研究基地 / 水稻国家工程研究中心, 福建福州 350003
  • 收稿日期:2024-05-08 接受日期:2024-08-15 出版日期:2024-12-12 网络出版日期:2024-09-02
  • 通讯作者: *张建福, E-mail: jianfzhang@163.com
  • 作者简介:E-mail: 183016337@qq.com
  • 基金资助:
    福建省自然科学基金(2022J01451);福建省人民政府-中国农业科学院协同创新工程“5511”项目(XTCXGC2021001);福建省财政专项-福建省农业科学院科技创新团队(CXTD2021001)

Metabolome and transcriptome analysis reveal molecular response to drought stress in indica rice Fuxiangzhan

WANG Ying-Heng1,2(), CUI Li-Li1,2, CAI Qiu-Hua1,2, LIN Qiang1,2, WU Fang-Xi1,2, CHEN Fei-He1,2, XIE Hong-Guang1,2, ZHU Yong-Sheng1,2, CHEN Li-Ping1,2, XIE Hua-An1,2, ZHANG Jian-Fu1,2,*()   

  1. 1Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350019, Fujian, China
    2Key Laboratory of Germplasm Innovation and Molecular Breeding of Hybrid Rice for South China, Ministry of Agriculture and Rural Affairs / Fuzhou Branch, National Rice Improvement Center of China / Fujian Engineering Laboratory of Crop Molecular Breeding / Fujian Key Laboratory of Rice Molecular Breeding / Incubator of National Key Laboratory of Fujian Germplasm Innovation and Molecular Breeding Between Fujian and Ministry of Sciences & Technology / Base of South China, National Key Laboratory of Hybrid Rice / National Rice Engineering Laboratory of China, Fuzhou 350003, Fujian, China
  • Received:2024-05-08 Accepted:2024-08-15 Published:2024-12-12 Published online:2024-09-02
  • Contact: *E-mail: jianfzhang@163.com
  • Supported by:
    Natural Science Foundation of Fujian Province(2022J01451);Collaborative Innovation Project “5511” of Fujian Provincial People’s Government and Chinese Academy of Agricultural Sciences(XTCXGC2021001);Science and Technology Innovation Team of Fujian Academy of Agricultural Sciences(CXTD2021001)

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摘要：

干旱是影响农业生产最重要的原因之一。本研究以优质香稻福香占为材料, 对其苗期干旱响应的生理指标、激素代谢物及基因表达调控网络进行分析。干旱处理后, 福香占干旱存活率、抗氧化酶活性高于恢复系明恢63、明恢86和干旱敏感的丽江新团黑谷, 但电导率较低且过氧化物累积较少。IAA、ICA、ABA、cZ和SA等5种激素的代谢物含量上升, tZ、DHZ、GA1、JA等11种激素的代谢物含量下降。6118个差异表达基因(DGEs)中有2615个上调、3503个下调, 涉及了光合作用、能量代谢、转录调控、氧化还原、离子结合等生物学过程和氨基酸、糖、脂肪酸、激素等合成代谢及植物激素信号转导等相关途径。激素代谢组和转录组学分析同时鉴别出植物激素信号转导、玉米素生物合成、类胡萝卜素生物合成和色氨酸代谢4个KEGG代谢通路, 并构建了差异表达基因调控网络。转录因子、抗氧化酶基因、渗透调节等相关的28个干旱应答关键基因在福香占干旱处理后的表达量均上调。福香占受干旱胁迫后, 激素水平发生了变化; 抗逆应答的基因表达量均上调, 包括转录因子、抗氧化系统基因、渗透调节及其他耐旱基因; 抗氧化酶活性相关生理指标发生改变。研究结果有助于进一步挖掘抗旱基因, 服务水稻抗旱育种。

关键词: 福香占, 抗旱, 生理特性, 激素代谢组, 调控网络

Abstract:

Drought is one of the most significant factors affecting agricultural production. In this study, the physiological indexes, hormone metabolites, and gene expression regulation network of drought response were analyzed for the high grain quality fragrant rice Fuxiangzhan. After drought stress, Fuxiangzhan exhibited higher drought survival rates and antioxidant enzyme activity compared to restorer lines Minghui63, Minghui86, and the drought-sensitive line Lijiangxintuanheigu, while showing lower membrane iron leakage and less peroxide accumulation. Five hormone metabolites (IAA, ICA, ABA, cZ, and SA) increased, whereas 11 hormone metabolites, including tZ, DHZ, GA1, and JA, decreased. A total of 6118 differentially expressed genes (DEGs) were identified, including 2615 up-regulated and 3503 down-regulated genes, which are involved in biological processes such as photosynthesis, energy metabolism, transcriptional regulation, REDOX, and ion binding, as well as molecular functions related to amino acids, sugars, fatty acids, hormones, and other anabolic metabolism and plant hormone signal transduction. KEGG pathways involving plant hormone signal transduction, zein biosynthesis, carotenoid biosynthesis, and tryptophan metabolism were identified from hormone metabolites and transcriptome analysis. Differentially expressed gene regulatory networks of the four pathways were constructed. The expression levels of 28 drought response genes related to transcription factors, antioxidant enzymes, and osmotic regulation were all up-regulated after drought stress in Fuxiangzhan. Our conclusion is that the hormone levels in Fuxiangzhan change after drought stress. The expression of anti-stress genes, including transcription factors, antioxidant system genes, osmoregulation, and other drought tolerance genes, were up-regulated. These changes lead to alterations in the activity of antioxidant enzymes and other physiological indexes. These results are helpful for further exploration of drought-resistant genes and serve in rice drought resistance breeding.

Key words: Fuxiangzhan, drought resistance, physiological properties, hormone metabolome, regulatory network

附表1

qRT-PCR引物序列"

基因登录号
Gene ID		 基因符号
Gene symbol		 上游引物
Forward primer (5′-3′)		 下游引物
Reverse primer (5′-3′)
LOC_Os01g63770 OsAUX1 TGATGGTCGTCATGGCTTATAA GAGGAATCGCCGATTACAATTG
LOC_Os03g08850 OsAFB5 CTGATAAATAGCCAGCCTGAGA GACTCCAGCTATCTTACCTTCC
LOC_Os01g53880 OsIAA6 ACCTGATCTTAACTCCGTGTTT TTCTACACTCAACTTAGGCTGG
LOC_Os04g56850 OsARF11 CAGCCTGTCATTGATTCGATTT TACTGTTTTGCTCCGAAGTACT
LOC_Os01g69920 OsHk3 GTTTCATGGACATACAGATGCC ATAGCCATCCATTTCGCTTTTC
LOC_Os07g25710 OsPHR2 TCTTCAGTTTCAAGCAATGAGC CCAGCTGCAATAGAACTGTTTT
LOC_Os05g38290 OsPP2C49 CGTGATCCGCCATTATTATTGG ATCGCAGGAGTAAATTAGGAGG
LOC_Os05g49730 OsPP2C51 CGAAGTTGTAGTAGCCGGAG CTTCACCGACCGGATTCTCTTC
LOC_Os01g64970 OsSAPK4 GTTTGTGTGCAGCTATTGTGTA CGCCAAACAGAAGCAAATTTAC
LOC_Os07g25590 OsTDC2 ATTGTTTGCCCTTCGAGTATTG TCAAACGACCTAACCCACTAAA
LOC_Os11g43900 OsTCTP CGTTCAAGGAGCTATTGATGTG GTCACAAACTGCTTCTTGTCAA
LOC_Os07g25540 OsYUCCA6 CTCAAGGGAAGTGACTTCTTCA TTGTGAAGCCAACAGAGTAGAG
LOC_Os01g07500 OsTAR2 CTTCTGCAACTTCACCAAGGAG AATCCTCCACATCCTCCCTATC
LOC_Os04g31040 OsVDE GGTGAAACAGGACTTGATGAAC TTACTTGTTGCCTTGTACTTGC
LOC_Os07g05940 OsNCED4 GCGACAAGCTTAGCTCAAATTA GCTAAACTATTTCAACTCCCTAGC
LOC_Os02g47510 OsNCED1 AGCACCAATGATACAAACCAAC TGTGTGCACTTATACTACGTGT
LOC_Os12g42280 OsNCED5 TTTGCTTTGCTTGTACAGACAG CACTGCAACTATCCCTATCACT
LOC_Os06g39880 OsCYP734A4 GCTAGCTAGGAAAAGACAGGAA GTACCACTAGTCTGTTAGCGAG

图1

干旱胁迫处理对4份水稻品种的影响 A: 干旱胁迫前和复水后植株形态; B: 存活率; C: 电导率; D: 超氧化物歧化酶; E: 过氧化物酶; F: 过氧化氢酶; G: 丙二醛; H: 过氧化氢含量; I: NBT染色。FXZ: 福香占; MH63: 明恢63; MH86: 明恢86; LTH: 丽江新团黑谷; CK: 对照; DS: 干旱处理; 数据均为平均值±标准差(n = 3), 不同小写字母表示在0.05概率水平差异显著。"

图2

福香占干旱胁迫处理和正常条件下的激素代谢物分析 A: OPLS-DA得分图(x轴代表组间差异分量, y轴代表组内差异分量, 图下方标注了模型的参数); B: 差异代谢物火山图(火山图中每个点代表1个代谢物, x轴代表该组对比各物质的倍数变化取log2的值, y轴表示t检验的P值取log10的值); C: 差异代谢聚类热图(x轴为各样本, y轴为层次聚类后的代谢物Z-score标准化后的定量值, 上方颜色条标注区分了不同组别); D: 差异代谢物KEGG富集网络图(淡黄色节点为通路, 与之相连的小节点是注释到该通路的具体代谢物, 颜色的深浅表示差异倍数取log2的值, 该图显示5个通路); CK: 对照; DS: 干旱处理。"

图3

福香占干旱胁迫处理和正常条件下的转录组分析 A: 相关性分析热图; B: 基因火山图; C: 差异表达基因数量; D: 差异基因GO富集(纵坐标表示GO条目, 横坐标表示q值取-log10的值, 数字表示条目内富集基因的数量); E: 差异基因KEGG富集(纵坐标代表显著富集的KEGG通路名称, 横坐标代表富集基因的比率, 圆点大小代表通路中富集的基因数, 圆点颜色代表q值); CK: 对照; DS: 干旱处理。"

图4

激素代谢组和转录组相关性分析 A: 激素代谢物和基因表达相关性分析的九象限图; B: 激素代谢组和转录组差异代谢通路韦恩图(列出4个共有的代谢通路); CK: 对照; DS:干旱处理。"

图5

福香占干旱胁迫后激素合成及信号转导调控 热图中红色表示基因表达上调, 紫色表示下调。小圆圈表示代谢物, 红色表示上调, 绿色下调; 方框表示基因产物, 红色代表上调, 绿色代表下调, 蓝色代表既有上调又有下调。CK: 对照; DS: 干旱处理; TAM: 色胺; IAN: 吲哚-3-乙腈; IAAld: 吲哚乙醛; IPyA: 吲哚丙酮酸; IAOX : 吲哚乙醛污; IAM: 吲哚乙酰胺; Trp: 色氨酸。"

图6

qRT-PCR验证差异表达基因 黑色柱子表示RNA-seq FPKM值, 红色线条表示qRT-PCR相对表达量; 左边纵坐标轴为RNA-seq FPKM值, 右边纵坐标轴为qRT-PCR相对表达量; CK: 对照, DS: 干旱处理; 数据均为平均值±标准差(n = 4), 不同小写字母表示在0.05概率水平差异显著。"

图7

福香占和丽江新团黑谷干旱处理前后激素相关代谢通路基因表达变化 FXZ: 福香占; LTH: 丽江新团黑谷; CK: 对照; DS: 干旱处理; 数据均为平均值±标准差(n = 4), 不同小写字母表示在0.05概率水平差异显著。"

图8

干旱胁迫下的耐旱应答关键基因表达水平值热图 CK: 对照; DS: 干旱处理; log2(FC)为差异倍数取log2的值。"

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