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- Title
马尾松 β-蒎烯合酶基因克隆以及对松材线虫侵染的响应.
- Authors
刘 彬; 刘青华; 周志春; 罗 柠; 解懿妮; 陈献志
- Abstract
[Objective] To clone β-pinene synthase gene in Pinus massoniana (PmPinS) and analyze its sequence characteristics and expression pattern after pine wood nematode (PWN) infection in order to provide theoretical basis to probe into the defense mechanism of P. massoniana to pine wood nematode infection. [Method] Based on PCR amplification technology, the full-length coding region sequence of PmPinS was cloned, the homologous amino acid sequence alignment, and the phylogenetic and genetic characteristics were analyzed with bioinformatics method. The expression pattern of PmPinS in resistant and susceptible P. massoniana after inoculation with PWN was analyzed with real-time quantitative PCR. [Result] The full-length coding region sequence of PmPinS was cloned, including a complete open reading frame of 1878 bp, encoding a 625 amino acid, with a molecular weight of 71.95 kD. The secondary structure was α-helix and β-fold, containing the typical terpenoid synthase domains including RR (X)8W, DDXXD and NSE / DTE. Phylogenetic analysis revealed that PmPinS and (-) β-pinene synthase in P. contortus were aggregated into one branch with a sequence similarity of 93.16%. The expression of PmPinS in resistant P. massoniana was higher than the susceptible ones after PWN inoculation, which showed an up-regulation trend within the following 30 days after inoculation. However, the expression of PmPinS in susceptible P. massoniana began to fell rapidly 1 day after inoculation. 2 years after inoculation, the expression level of PmPinS was highly expressed in the stem tissue of resistant P. massoniana. In addition, exogenous application of α-pinene, β-pinene and their mixed solutions to PWN showed that α-pinene and β-pinene could significantly inhibit the survival rate of PWN, especially for the mixed solution with more inhibition. [Conclusion] PmPinS, as a member of the monoterpene synthase family of P. massoniana, may be involved in the synthesis of β-pinene with GPP catalyzed intricately. The PmPinS is highly specific-tissue expressed in the stem of resistant P. massoniana, and its possible catalytic products, α-pinene and β- pinene, have significant inhibitory effects on PWN, which might reveal that PmPinS plays a positive regulatory role in the defensive process to PWN.
- Subjects
PINEWOOD nematode; AMINO acid sequence; NEMATODE infections; SURVIVAL analysis (Biometry); CONIFER wilt; SEQUENCE alignment
- Publication
Forest Research, 2020, Vol 33, Issue 6, p1
- ISSN
1001-1498
- Publication type
Article
- DOI
10.13275/j.cnki.lykxyj.2020.06.001