砧木及抗性马尾松松脂组分对松材线虫的响应

Response of Different Rootstocks and Resistant Resin Components of Pinus massoniana to Pine Wood Nematode

[目的]通过对马尾松接种松材线虫,分析接种前后松脂化学组分的变化,为马尾松抗松材线虫病的研究提供理论基础。[方法]以浙江省临海市5年生马尾松无性系为研究对象,提取接种前和接种松材线虫1、7、15 d的不同砧木(马尾松砧木和湿地松砧木)、不同抗性(高抗和易感)马尾松的松脂组分,分析各松脂组分含量及动态变化;利用松脂内所得有效萜类对松材线虫实施外源处理,测定松材线虫存活率。[结果]从不同砧木(马尾松砧木和湿地松砧木)、不同抗性(高抗和易感)马尾松中检出19种主要化学组分,其中,α-蒎烯、β-蒎烯、水芹烯、海松酸、山达海松酸、长叶松酸/左旋海松酸、去氢枞酸、枞酸和新枞酸含量较高。接种松材线虫1 d时马尾松砧木中β-月桂烯含量显著高于湿地松砧木(p<0.05);接种松材线虫7 d与15 d时上述组分含量在不同砧木间无显著差异。不同抗性马尾松中α-蒎烯、β-蒎烯、β-月桂烯、柠檬烯、龙脑、长叶烯、α-石竹烯、反式-β-金合欢烯和新枞酸含量在接种松材线虫后变化规律不同,接种松材线虫1 d时高抗马尾松中α-蒎烯和β-蒎烯含量显著高于易感马尾松;接种松材线虫7 d时高抗马尾松中α-蒎烯、β-蒎烯、柠檬烯和长叶烯含量显著高于易感马尾松(p<0.05);接种松材线虫15 d时高抗马尾松中α-蒎烯、β-蒎烯、柠檬烯和长叶烯含量显著高于易感马尾松(p<0.05)。对上述萜类组分进行梯度浓度松材线虫外源试验:以150 mg·g^(−1)浓度α-蒎烯溶液处理0.5 h时,松材线虫存活率达20%;不同浓度β-蒎烯溶液对松材线虫抑制作用相同,1.20 mg·g^(−1)浓度处理0.5 h时松材线虫存活率达50%左右;以10 mg·g^(−1)浓度柠檬烯溶液处理0.5 h时,松材线虫存活率趋于零;以10 mg·g^(−1)浓度长叶烯溶液处理0.5 h时,松材线虫存活率达18%。[结论]不同砧木对马尾松抗性无明显作用;接种松材线虫后,α-蒎烯、β-蒎烯、柠檬烯和长叶烯含量存在显著差异,且均为高抗马尾松显著高于易感马尾松。α-蒎烯、β-蒎烯、柠檬烯和长叶烯低浓度下抑制松材线虫活性,与马尾松抗性有关。

[Objective]By inoculating Pinus massoniana Lamb.with pine wood nematode,the changes of pine resin components before and after inoculation were analyzed,so as to provide a theoretical basis for the study of resistance of P.massoniana to pine wood nematode disease.[Method]The experimental material were 5-year-old clones of P.massoniana in Linhai City,Zhejiang Province.We extracted the pine resin components of P.massoniana rootstocks and P.elliottii rootstocks,highly resistant and susceptible P.massoniana inoculated with pine wood nematode for 1,7 and 15 days and also analyzed the content and dynamic changes of each pine resin component.The effective terpenes were used to treat pine wood nematode.Then we analyzed the survival rate of pine wood nematodes.[Result]The results showed that 19 oleoresin terpenoids were Identified.The abundant terpenoids wereα-pinene,β-pinene,carylene,pimaric acid,sandaracopimaric acid,palustric acid/levopimaric acid,dehydroabietic acid,neoabietic acid and abietic acid.In the rootstock of P.massoniana,the content ofβ-myrcene was significantly higher than that of P.elliottii rootstock 1day after inoculating with pine wood nematode(p<0.05);the contents of the above components had no significant difference among different rootstocks after inoculating with pine wood nematode 7 days and 15 days.The contents ofα-pinene,β-pinene,β-laurene,limonene,borneol,longifolene,α-caryophyllene,trans-β-farnesene and neoabietic acid in different resistant P.massoniana changed differently after inoculation with pine wood nematode.In the high resistance P.massoniana,the contents ofα-pinene andβ-pinene were significantly higher than those of susceptible P.massoniana 1 day after inoculating with pine wood nematode(p<0.05);In the high resistance P.massoniana,the content ofα-pinene,β-pinene,limonene and longifolene were significantly higher than those of susceptible P.massoniana 7 days after inoculating with pine wood nematode(p<0.05);The situation was the same when pine wood nematode was inoculated for 15 days as when pine wood nematode was inoculated for 7 days.The result of gradient concentration test of the above terpenoids showed that:when the concentration ofα-pinene was 150 mg·g^(−1) for 0.5 h,the survival rate of pine wood nematode was 20%.Different concentrations of β-pinene solution had the same inhibitory effect on pine wood nematode,and the survival rate reached about 50%when treated for 0.5h.When limonene solution concentration was 10mg·g^(−1) for 0.5 h,the survival rate of inhibited pine wood nematode was almost zero.When the concentration of longifolene was 10mg·g^(−1) for 0.5 h,the survival rate of pine wood nematode was 18%.[Conclusion]There are no obvious relationships between different rootstocks and resistance to P.massoniana.There are significant differences inα-pinene,β-pinene,limonene and longifolene contents between high and susceptible P.massoniana inoculated with pine wood nematode,and the content of high resistance P.massoniana was significantly higher than those of susceptible P.massoniana.At low concentrationsα-pinene,β-pinene,limonene and longifolene inhibit the activity of pine wood nematodes,which shows the four terpenoids are related to resistance of P.massoniana.