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林业科学 ›› 2017, Vol. 53 ›› Issue (6): 105-110.doi: 10.11707/j.1001-7488.20170612

• 论文与研究报告 • 上一篇    下一篇

松材线虫CYP450基因与松树蒎烯类物质代谢的相关性

王璇1, 理永霞2, 刘振宇3, 吕全2, 贾秀贞2, 张星耀2   

  1. 1. 中国林业科学研究院林业新技术研究所 北京 100091;
    2. 中国林业科学研究院森林生态环境与保护研究所 国家林业局森林保护学重点实验室 北京 100091;
    3. 山东农业大学 泰安 271018
  • 收稿日期:2015-11-16 修回日期:2016-03-10 出版日期:2017-06-25 发布日期:2017-07-14
  • 通讯作者: 理永霞
  • 基金资助:
    浙江省省院合作林业科技项目(2014SY17);林业公益性行业科研专项经费(201204501)

Relationship between the Cytochrome P450 Gene of Pine Wood Nematode and the Accumulation of Pine Pinene

Wang Xuan1, Li Yongxia2, Liu Zhenyu3, lü Quan2, Jia Xiuzhen2, Zhang Xingyao2   

  1. 1. Research Institute of Forestry New Technology, CAF Beijing 100091;
    2. Key Laboratory of Forest Protection of State Forestry Administration Research Institute of Forest Ecology, Environment and Protection, CAF Beijing 100091;
    3. Shandong Agricultural University Tai'an 271018
  • Received:2015-11-16 Revised:2016-03-10 Online:2017-06-25 Published:2017-07-14

摘要: [目的] 通过比较松材线虫CYP450基因和寄主萜烯类物质代谢的时间特异性,研究松材线虫解毒相关CYP450基因与寄主蒎烯类物质代谢之间的相关性,为该病致病机制的深入研究提供基础。[方法] 松材线虫接种5年生马尾松后,通过实时定量PCR研究松材线虫CYP450基因和寄主萜烯合成酶基因表达模式,同时通过气相色谱法检测寄主松树主要萜烯类物质代谢规律。[结果] 松材线虫接种5年生马尾松后,松树蒎烯合成酶基因分别在第6天和第21天上调表达,而寄主松树体内α-蒎烯和β-蒎烯也具有2个峰值,分别在第9天和第27天时含量最高。松材线虫CYP-33C9基因在第12天和第15天大量表达,CYP-33C4基因在接种后第21天表达量最高。[结论] 松材线虫的入侵引起松树蒎烯合成酶基因上调表达,生物合成大量蒎烯类物质,松材线虫CYP450基因响应松树蒎烯类物质的大量积累而上调表达。因此,根据松材线虫CYP450基因与松树蒎烯类物质代谢在时间上的相关性,推测CYP450基因可能参与了松材线虫蒎烯类物质代谢过程,可能是松材线虫致病相关基因之一。

关键词: 松材线虫, 蒎烯代谢, 细胞色素P450, 致病机制

Abstract: By comparing the expression pattern of cytochrome P450 (CYP450) genes of Pine Wood Nematode (PWN) (Bursaphelenchus xylophilus)and the accumulation of pine terpene, the relationship between the CYP450 genes of PWN and the metabolism of pine pinene was analyzed to provide fundamental information for PWN pathogenic mechanisms study. [Method] Five-years Pinus massoniana were inoculated with PWN. Real-time Quantitative PCR was conducted to detect expression patterns of apin gene of pine trees and CYP450 family genes of PWN. Terpene metabolism in infected P. massoniana was detected by Gas Chromatography. [Result] The expression of pinene synthase genes were dramatically induced at 6 dpi (days post inoculation) and 21 dpi. Both α-pinene and β-pinene were accumulated with two peaks at 9dpi and 27dpi, respectively. Correspondingly, the expression of PWN CYP-33C9 dramatically increased at 12dpi and 15dpi, and the expression of CYP-33C4 were highest at 21dpi. [Conclusion] The pine tree pinene synthase gene was highly induced by PWN inoculation which resulted in sharp accumulation of the α-pinene and β-pinene. In responding to the accumulation of pine terpene, the expression of PWN CYP450 genes dramatically increased. Therefore, according to the temporal correlation between the pinene metabolism and the PWN CYP450 genes expression we inferred that the PWN CYP450 genes might participate in the terpene metabolism and might be one of the pathogenic genes in the PWN pathogenic process.

Key words: pine wood nematode, terpene metabolism, cytochrome P450, pathological mechanisms

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