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转录组测序分析青稞对啶磺草胺的耐药性

蔡青 翁华

蔡青,翁华. 转录组测序分析青稞对啶磺草胺的耐药性 [J]. 福建农业学报,2024,39(4):492−502 doi: 10.19303/j.issn.1008-0384.2024.04.014
引用本文: 蔡青,翁华. 转录组测序分析青稞对啶磺草胺的耐药性 [J]. 福建农业学报,2024,39(4):492−502 doi: 10.19303/j.issn.1008-0384.2024.04.014
CAI Q, WENG H. Transcriptome Analysis on Pyroxsulam-resistance of Naked Barley [J]. Fujian Journal of Agricultural Sciences,2024,39(4):492−502 doi: 10.19303/j.issn.1008-0384.2024.04.014
Citation: CAI Q, WENG H. Transcriptome Analysis on Pyroxsulam-resistance of Naked Barley [J]. Fujian Journal of Agricultural Sciences,2024,39(4):492−502 doi: 10.19303/j.issn.1008-0384.2024.04.014

转录组测序分析青稞对啶磺草胺的耐药性

doi: 10.19303/j.issn.1008-0384.2024.04.014
基金项目: 青海省科技厅基础研究项目(2023-ZJ-930M)
详细信息
    作者简介:

    蔡青(1999 —),女,硕士研究生,主要从事杂草治理与利用研究,E-mail:2663146708@qq.com

    通讯作者:

    翁华(1979 —),女,硕士,副研究员,主要从事杂草治理与利用研究,E-mail:wenghua_0872@163.com

  • 中图分类号: S512

Transcriptome Analysis on Pyroxsulam-resistance of Naked Barley

  • 摘要:   目的  挖掘青稞代谢啶磺草胺可能涉及的重要基因,明确青稞对啶磺草胺的代谢解毒机制,为啶磺草胺的科学使用和耐受除草剂的青稞品种选育提供理论基础。  方法  以啶磺草胺处理0 、1 、6 d的敏感青稞品种青0160以及耐药青稞品种青0306的叶片为材料,借助高通量测序技术进行转录组测序结果分析。  结果  GO富集分析结果表明,啶磺草胺处理前后差异表达基因均显著富集在光合作用上。KEGG富集分析表明,啶磺草胺处理1 d后,谷胱甘肽代谢途径中抗氧化剂相关基因以及苯丙素和吡啶生物碱生物合成途径中与应激相关基因显著富集;啶磺草胺处理6 d后,与维持细胞功能的一系列生物过程及氨基酸的生物合成和代谢相关的基因显著富集。进一步分析表明,两个品种中的SOD21972、POD55052、CAT15170、DHAR59510、APX42784、GR34873、GSTs849、GSTs36979、GSTs31507、GSTs15106、GSTs20762表达上调,PSⅡ55705Cc31194、Cc17547、Cc17551、CYP12424受到抑制;其中POD55052在敏感青稞中表达量高于耐药青稞,SOD21972、CAT15170、DHAR59510、APX42784、GR34873、GSTs849、GSTs36979、GSTs31507、GSTs15106、GSTs20762、PSⅡ55705、Cc31194、Cc17547、Cc17551、CYP12424在耐药青稞中表达量高于敏感青稞,这些基因可能参与了青稞耐啶磺草胺机制。qRT-PCR相对表达量和转录组测序结果变化趋势基本一致,证明转录组测序结果具有可靠性。  结论  明确了青稞对啶磺草胺的代谢解毒机制,为青稞耐受啶磺草胺提供分子基础,对啶磺草胺安全用药及选育青稞耐除草剂品种有重要意义。
  • 图  1  敏感、耐药青稞对啶磺草胺的响应

    A:处理前;B:处理1 d后;C:处理6 d后。

    Figure  1.  Responses to pyroxsulam of herbicide-sensitive and herbicide-resistant naked barley

    A: before treatment; B: 1 d after treatment; C: 6 d after treatment.

    图  2  基因表达间的相关性矩形分析

    Figure  2.  Rectangular analysis of correlation between gene expressions

    图  3  敏感、耐药青稞对啶磺草胺的差异表达基因

    A:R0 vs S0 火山图;B:R1 vs S1 火山图;C:R6 vs S6 火山图;D:每个比较组合的差异表达基因数。

    Figure  3.  DEGs of herbicide-sensitive and herbicide-resistant naked barley response to pyroxsulam

    A: R0 vs. S0 volcano; B: R1 vs. S1 volcano;C: R6 vs. S6 volcano;D: number of DEGs in each comparison combination.

    图  4  差异表达基因的GO富集分析

    A:R0 vs S0的GO富集分析;B:R1 vs S1的GO富集分析;C:R6 vs S6的GO富集分析。

    Figure  4.  GO enrichment analysis on DGEs

    A: R0 vs. S0 GO enrichment analysis; B: R1 vs. S1 GO enrichment analysis; C: R6 vs. S6 GO enrichment analysis.

    图  5  抗氧化系统相关基因表达量分析

    Figure  5.  Gene expression related to antioxidants

    图  6  代谢酶GSTs相关基因表达量分析

    Figure  6.  Expressions of GSTs-related genes of metabolic enzymes

    图  7  光合作用相关基因表达量分析

    Figure  7.  Expressions of photosynthesis-related genes

    图  8  qRT-PCR验证基因表达

    Figure  8.  qRT-PCR verification of gene expression

    表  1  Real-time qRT-PCR分析所用引物序列

    Table  1.   Sequence of primer for qRT-PCR analysis

    基因名称
    Gene name
    正向引物(5'–3′)
    Forward primer(5'–3′)
    反向引物(5'–3′)
    Reverse primer(5'–3′)
    gene_07062 CCAGTCAAGCATCAAAAGCA GCAGAAAGGCCACTGTTTTC
    gene_12424 CAAGGAGAGCGGTGCTAAAC AGCTCCTCTTGCAGCTTCTG
    gene_26867 GACGGAGATGTGCAGAGTGA GTGCTGGTTTCACCACCTTT
    gene_59076 ACCGCAGCATAGCAGAAGAT AAGGGAGAAGCACCAGTTCA
    Actin
    (青稞
    Hulless barley)
    CTATTCAGGCCGTGCTTTCC CCAGCGAGATCCAAACGAAG
    下载: 导出CSV

    表  2  差异表达基因显著富集KEGG途径

    Table  2.   Significantly enriched KEGG-pathway of DGEs

    途径注释
    Description of pathway
    差异表达基因
    DEGs
    途径注释
    Description of pathway
    差异表达基因
    DEGs
    R0 vs S0 R6 vs S6
    碳代谢 Carbon metabolism 139 核糖体 Ribosome 109
    光合生物中的碳固定
    Carbon fixation in photosynthetic organisms
    41 氨基酸生物合成
    Biosynthesis of amino acids
    90
    丙酮酸代谢 Pyruvate metabolism 43 碳代谢 Carbon metabolism 101
    氨基酸生物合成 Biosynthesis of amino acids 113 乙醛酸和二羧酸代谢
    Glyoxylate and dicarboxylate metabolism
    36
    乙醛酸和二羧酸代谢
    Glyoxylate and dicarboxylate metabolism
    42 光合作用-天线蛋白
    Photosynthesis - antenna proteins
    16
    甘氨酸、丝氨酸和苏氨酸代谢
    Glycine, serine and threonine metabolism
    39 光合生物的碳代谢
    Carbon fixation in photosynthetic organisms
    33
    氧化磷酸化 Oxidative phosphorylation 63 光合作用 Photosynthesis 32
    柠檬酸循环(TCA循环)
    Citrate cycle (TCA cycle)
    33 甘氨酸、丝氨酸和苏氨酸代谢
    Glycine, serine and threonine metabolism
    32
    光合作用 Photosynthesis 40 丙氨酸、天冬氨酸和谷氨酸代谢
    Alanine, aspartate and glutamate metabolism
    23
    光合作用-天线蛋白
    Photosynthesis - antenna proteins
    16 精氨酸生物合成 Arginine biosynthesis 17
    丁酸代谢 Butanoate metabolism 14 莨菪烷、哌啶和吡啶生物碱生物合成
    Tropane, piperidine and pyridine alkaloid biosynthesis
    9
    磷酸戊糖途径 Pentose phosphate pathway 31 2-氧代羧酸代谢
    2-Oxocarboxylic acid metabolism
    22
    R1 vs S1 丙酮酸代谢 Pyruvate metabolism 28
    核糖体 Ribosome 50 烟酸和烟酰胺代谢
    Nicotinate and nicotinamide metabolism
    11
    光合作用-天线蛋白
    Photosynthesis -antenna proteins
    9
    下载: 导出CSV
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  • 收稿日期:  2023-08-22
  • 修回日期:  2024-02-09
  • 网络出版日期:  2024-05-08
  • 刊出日期:  2024-04-28

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