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林业科学 ›› 2022, Vol. 58 ›› Issue (2): 159-170.doi: 10.11707/j.1001-7488.20220216

• 研究论文 • 上一篇    下一篇

千年桐根部黄酮类化合物生物合成对枯萎病菌侵染的响应

王嘉1,2,梁晓洁1,高暝1,吴立文1,赵耘霄1,汪阳东1,黄世清3,张永志3,傅火勇3,陈益存1,*   

  1. 1. 中国林业科学研究院亚热带林业研究所 杭州 311400
    2. 南京林业大学 南京 210037
    3. 安吉县龙山林场 湖州 313306
  • 收稿日期:2020-12-21 出版日期:2022-02-25 发布日期:2022-04-26
  • 通讯作者: 陈益存
  • 基金资助:
    国家科技基础资源调查专项“工业油料树种种质资源调查收集”课题(2019FY100803)

Response of Flavonoids Biosynthesis in Roots of Vernicia montana to Fusarium Wilt Infection

Jia Wang1,2,Xiaojie Liang1,Ming Gao1,Liwen Wu1,Yunxiao Zhao1,Yangdong Wang1,Shiqing Huang3,Yongzhi Zhang3,Huoyong Fu3,Yicun Chen1,*   

  1. 1. Research Institute of Subtropical Forestry, CAF Hangzhou 311400
    2. Nanjing Forestry University Nanjing 210037
    3. Anji Longshan Forest Farm Huzhou 313306
  • Received:2020-12-21 Online:2022-02-25 Published:2022-04-26
  • Contact: Yicun Chen

摘要:

目的: 油桐枯萎病是一种土传的根部真菌病害,严重危害油桐主栽品种三年桐,极大地限制了其规模化栽培。而同属的千年桐具有抗枯萎病能力,其根部防御作用机制的研究可以为抗枯萎病防治和抗性育种提供思路。方法: 利用乙酸乙酯萃取法获得三年桐和千年桐根部提取物;基于高效液相色谱和串联质谱检测病原菌侵染后三年桐和千年桐根部代谢物成分;利用Illumina HiSeqTM2000、检测病原菌侵染过程中三年桐和千年桐根部基因表达变化规律和通路变化,并利用实时定量PCR试验验证基因表达规律;利用R软件包等生物信息学方法进行数据分析。结果: 1) 与三年桐相比,千年桐根部提取物对油桐枯萎病菌的生长有明显的抑制作用;2) 病原菌侵染后千年桐根部产生的芒柄花苷、橙皮苷等异黄酮和黄烷酮化合物是三年桐的1 000倍以上;3) 病原菌侵染后千年桐根部负责黄酮类化合物生物合成的上游关键通路“苯丙烷类生物合成途径”显著富集;4) 苯丙烷类生物合成途径中有4个中心基因,包括4-香豆酸CoA连接酶、β-D-木糖苷酶、β-葡萄糖苷酶和过氧化物酶N1,通过实时定量PCR试验验证在病原菌侵染早期上调表达,且与其他1 625基因具有极高的相关关系,激活苯丙烷类生物合成途径从而产生黄酮类化合物抵御病原菌入侵。结论: 通过代谢组和转录组联合分析,揭示了千年桐根部苯丙烷类生物合成途径在枯萎病病原菌入侵后发生响应,产生芒柄花苷、橙皮苷等黄酮类化合物,从而抵御病原菌入侵。

关键词: 油桐, 枯萎病, 黄酮类化合物, 苯丙烷类生物合成途径, 中心基因

Abstract:

Objective: Objective: The tung oil tree, Vernicia fordii, is suffered from the soil-borne Fusarium oxysporum f. sp. fordii, Fof-1. However its sister species, Vernicia montana, shows high resistance to the pathogen. This study aims to investigate the resistance mechanism of V.montana, so as to provide ideas for resistance breeding of V. fordii. Method: The ethyl acetate extraction method was used to extract the root metabolites of V.montana and V. fordii after infection. The ultra performance liquid chromatography and tandem mass spectrometry were used to detect the root metabolites. The Illumina HiSeqTM2000 was used to detect the changes of gene expression mode and the pathway in the roots of V.montana and V. fordii during the infection by the Fusarium wilt pathogen. The gene expression was further validated by qRT-PCR method. The data were analyzed using R soft package and other bioinformatics methods. Result: 1) Comparing with that of V. fordii, the root extract of V. Montana had an obvious inhibition effect on the growth of Fof-1. 2) After being infected, the isoflavone and flavanone, such as formononetin 7-O-glucoside (Ononin) and hesperetin 7-rutinoside (hesperidin), in the roots of V. montana were 1 000 folds higher than those in the roots of V. fordii. 3) The phenylpropanoid biosynthesis pathway, the upstream key pathway responsible for flavonoid biosynthesis, was significantly enriched in the roots of resistant V. montana upon Fof-1 infection. 4) There were four hub genes, including 4-coumarate: CoA ligase (4CL), β-D-xylosidase, β-glucosidase, and peroxidase N1 with high connection with the other 1625 genes, which were all up-expressed in the early stage of Fof-1 infection in the phenylpropanoid biosynthesis pathway. Conclusion: Based on the metabonomic and transcriptomic analyses, it is revealed that the phenylpropanoid biosynthesis pathway actively responds to Fof-1 infection in the roots of V. montna, and the oflavone and flavanone, such as on onin and hesperidin, are correspondingly produced for the resistance to Fusarium pathogen Fof-1 in the roots of V. montana.

Key words: tung oil trees, Fusarium wilt disease, flavonoids, phenylpropanoid biosynthesis, hub genes

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