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转录组和代谢组联合分析茶树对茶饼病病原菌的防御反应

姚玉仙 张明泽 舒烨 罗福丹 全昌彬 刘荣 向志攀 梅鑫

姚玉仙,张明泽,舒烨,等. 转录组和代谢组联合分析茶树对茶饼病病原菌的防御反应 [J]. 福建农业学报,2024,39(X):1−16
引用本文: 姚玉仙,张明泽,舒烨,等. 转录组和代谢组联合分析茶树对茶饼病病原菌的防御反应 [J]. 福建农业学报,2024,39(X):1−16
YAO Y X, ZHANG M Z, SHU Y, et al. Transcriptome and Metabolome Analysis of Tea Leaves in Response to Tea Blister Blight [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−16
Citation: YAO Y X, ZHANG M Z, SHU Y, et al. Transcriptome and Metabolome Analysis of Tea Leaves in Response to Tea Blister Blight [J]. Fujian Journal of Agricultural Sciences,2024,39(X):1−16

转录组和代谢组联合分析茶树对茶饼病病原菌的防御反应

基金项目: 贵州省基础研究(自然科学)项目(黔科合基础〔2020〕1Y120);贵州省教育厅自然科学基金项目(黔教合KY字〔2019〕211);贵州省教育厅贵州省高等学校茶树特征性成分研究重点实验室项目(黔教技〔2023〕027号);贵州省大学生创新创业训练计划项目(S202210670021)
详细信息
    作者简介:

    姚玉仙(1988 —),女,硕士,讲师,主要从事植物分子生物学研究,E-mail:yaoyuxian0520@126.com

    通讯作者:

    张明泽(1986 —),男,博士,副教授,主要从事植物分子遗传学研究,E-mail: zhangmingze20@163.com

  • 中图分类号: S435.711

Transcriptome and Metabolome Analysis of Tea Leaves in Response to Tea Blister Blight

  • 摘要:   目的  探明茶树对茶饼病菌的抗性分子机制,挖掘抗病相关基因,为茶树抗性育种提供依据。  方法  通过转录组测序和代谢组分析,比较茶树健康叶片(CK)和感染茶饼病的叶片(TB)中的差异表达基因(DEGs)和差异代谢物(DAMs)。  结果  转录组数据显示,样品CK和TB之间共有1009个 DEGs;GO 富集分析表明,差异基因参与了细胞壁代谢及调控几丁质酶活性、氧化还原酶活性和木葡聚糖:木葡基转移酶活性;KEGG 代谢途经分析表明,DEGs显著富集在“类黄酮生物合成”“苯丙素生物合成”“氨基糖和核苷糖代谢”“甘油酯代谢”和“芪类,二芳基庚烷和姜酚生物合成”途径;DEGs 中包含47个转录因子,分属21个转录因子家族,主要包括bHLH、SBP、AP2/ERF-AP2和MYB等,这些转录因子可能是茶树抵御茶饼病侵染过程中重要的调控基因。利用广泛靶向代谢组学技术分析,共发现353个DAMs,DAMs主要富集于“类黄酮生物合成”“赖氨酸生物合成”和“丙氨酸、天门冬氨酸和谷氨酸代谢”途径。利用转录组联合代谢组分析发现,显著共同富集的途径是“类黄酮生物合成”“苯丙素生物合成”和“芪类,二芳基庚烷和姜酚生物合成”;筛选了与苯丙素类及类黄酮生物合成途径相关的20个DEGs和15个DAMs,其中,CSS00117414CL)、CSS0002940DFR)、CSS0015968DFR)和CSS0010687ANS)等DEGs在感病叶中上调表达,根皮素、根皮苷、4-羟基苯乙烯、对香豆酰喹啉酸、二氢杨梅素、表没食子儿茶素和芍药素-3-O葡萄糖苷等DAMs在感病叶中积累。  结论  “苯丙素类生物合成”和“类黄酮生物合成”等代谢途径中的 DEGs在茶树响应茶饼病侵染中发挥重要作用,根皮素、根皮苷以及表没食子儿茶素等DAMs可能是茶树抵御茶饼病侵染的重要次生代谢产物。
  • 图  1  样本相关性热图及各样本间的主成分分析

    A. 转录组样本相关性热图;B. 转录组各样本间的主成分分析;C. 代谢组样本相关性热图;D. 代谢组各样本间的主成分分析。

    Figure  1.  Heatmap of the Pearson correlation between samples and principal component analysis plots of samples

    A. Heatmap of the Pearson correlation between samples in transcriptome; B. Principal component analysis plots of samples in transcriptome; C. Heatmap of the Pearson correlation between samples in metabolome; D. Principal component analysis plots of samples in metabolome.

    图  2  样品CK和TB中的差异表达基因

    Figure  2.  Differentially expressed genes in CK vs. TB

    图  3  差异表达基因的GO富集分析

    A. 生物学过程;B. 细胞组分;C. 分子功能。

    Figure  3.  GO enriched analysis of differentially expressed genes

    A. Biological process; B. Cellular component; C. Molecular function.

    图  4  差异表达基因的KEGG富集分析

    A. KEGG代谢通路分类统计;B. KEGG前20条显著性富集代谢通路。

    Figure  4.  KEGG enrichment analysis of differentially expressed genes

    A. Classification statistical diagram of KEGG metabolic pathway; B. The top 20 metabolic pathways of KEGG were significantly enriched.

    图  5  茶树感染茶饼病菌后转录因子变化情况

    A. 转录因子类别及数量统计;B. 转录因子表达热图。

    Figure  5.  Changes of transcription factors in tea plants infected with E. vexans

    A. Classification and quantity statistics of transcription factors; B. The expression heatmap of transcription factors.

    图  6  CK vs. TB的差异代谢物火山图

    Figure  6.  Differential metabolite volcano map of CK vs. TB

    图  7  差异代谢物KEGG通路富集分析

    Figure  7.  Enriched KEGG pathways of differentially metabolites

    图  8  前20个差异代谢物差异倍数柱状图

    Figure  8.  Histogram of fold change for the first 20 differential metabolites

    图  9  转录-代谢KEGG共有富集通路气泡图

    A. Venn 图;B. 代谢通路分析。圆点代表DEGs, 三角形代表DAMs。-lg (P value) 越大,富集可信度越高。

    Figure  9.  Bubble diagram of enriched pathways shared by transcriptome and metabolome through KEGG analysis

    A. Venn diagram; B. Metabolic pathway analysis. Triangles are DEGs and dots are DAMs. The bigger the -lg (P value) is, the more reliable the enrichment is.

    图  10  苯丙素类和类黄酮生物合成途径的关键DEGs和DAMs

    流程图中红色方框表示TB中的上调代谢物,绿色方框表示TB中的下调代谢物。

    Figure  10.  The key DEGs and DAMs of the phenylpropanoid and flavonoid biosynthesis pathway

    The red boxes in the flow chart represent up-regulated metabolites in TB, and the green boxes represent down-regulated metabolites in TB.

    图  11  部分差异表达基因的qRT-PCR验证分析

    “*” 和“**”分别代表CK与TB相比差异显著(P<0.05)和差异极显著(P<0.01)。

    Figure  11.  qRT-PCR verification analysis of partial differentially expressed genes

    "*" and "**" represent significant differences (P<0.05) and extremely significant differences (P<0.01) between CK and TB, respectively.

    表  1  qRT-PCR基因及引物

    Table  1.   Genes and primers for qRT-PCR

    基因
    Gene
    正向引物
    Forward primer (5'-3')
    反向引物
    Reverse primer (5'-3')
    CSS0007745 CCAAGAGTGTGGAAGGGTATG CAGAGGGAGGCCAAATCTTATG
    CSS0013831 CATTGCTTCTTGGCCCTACTA TGACAGTGCCATCACCATAAA
    CSS0038049 GATGCCATGGCTTATGGATTTG GACCTATGCGGTAGACAGTTTC
    CSS0008562 AGTGCCTCATCAACCATCAC AGGAAGAAGAAGAGGAGGTAGAG
    CSS0042430 GAGACTCAGAGGACTCGAAAGA CAGACTCGGACGCCTTTATG
    CSS0005594 AATATGGGTGCCGGGTATTG TCTTCTCCTCCGATGGTACTT
    CsEF1KA280301.1 TTGGACAAGCTCAAGGCTGAACG ATGGCCAGGAGCATCAATGACAGT
    下载: 导出CSV

    表  2  样本测序数据的质量控制分析

    Table  2.   Analysis of quality control for transcriptome sequencing data

    样本
    Sample
    过滤序列
    Clean reads
    过滤碱基
    Clean bases/G
    Q20/% Q30/% GC含量
    GC Content /%
    匹配序列(比对效率/%)
    Mapped reads (Efficiency/%)
    CK1 40107608 6.00 98.87 96.51 45.31 35374239 (88.20)
    CK2 40805448 6.10 98.72 96.33 45.11 35890315 (87.95)
    CK3 40738514 6.10 98.48 95.89 44.84 35837746 (87.97)
    TB1 41033002 6.14 98.77 96.24 45.42 36088623 (87.95)
    TB2 40766338 6.10 98.87 96.54 45.25 35886380 (88.03)
    TB3 41144588 6.16 98.71 96.28 45.21 36124713 (87.80)
    总计 Total 244595498 36.59
    下载: 导出CSV

    表  3  显著差异的黄酮类物质

    Table  3.   Flavonoids with significant differences

    编号
    code
    代谢物名称
    Compound
    差异倍数
    Fold_change
    P
    P-value
    变量重要性投影
    VIP
    差异类型
    Type
    NEG_q122 二氢杨梅素 Dihydromyricetin 1.10 0.00 1.33 上调 Up
    NEG_q260 对香豆酰奎宁酸 P-Coumaroyl quinic Acid 1.22 0.00 1.38
    NEG_q264 根皮素 Phloretin 1.56 0.02 1.27
    NEG_q265 根皮苷 Phlorizin 1.29 0.04 1.28
    POS_q141 (-)-表没食子儿茶素 Epigallocatechin 1.05 0.02 1.29
    NEG_q79 芹菜素 Apigenin 0.65 0.01 1.37 下调 Down
    NEG_q94 儿茶素没食子酸酯 Catechin Gallate 0.95 0.03 1.30
    NEG_q160 没食子儿茶素没食子酸酯 Gallocatechin Gallate 0.95 0.02 1.30
    NEG_q145 表儿茶素没食子酸酯 (-)-Epicatechin Gallate 0.73 0.01 1.27
    NEG_q92 儿茶素 Catechin 0.65 0.00 1.38
    NEG_q98 绿原酸 Chlorogenic Acid 0.68 0.00 1.36
    NEG_q120 二氢山奈酚 Dihydrokaempferol 0.57 0.00 1.35
    NEG_q158 (+)-没食子儿茶素 Gallocatechin 0.91 0.01 1.32
    NEG_q240 柚皮素查耳酮 Naringenin Chalcone 0.66 0.00 1.38
    NEG_q241 柚皮苷 Naringin 0.76 0.00 1.32
    NEG_q307 花旗松素 Taxifolin 0.80 0.00 1.35
    下载: 导出CSV
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  • 收稿日期:  2024-05-14
  • 修回日期:  2024-07-18
  • 网络出版日期:  2024-11-11

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