利用HS-SPME-GC/MS法分析横坑切梢小蠹危害细叶云南松诱导抗性挥发物质

Analysis of Induced Resistant Volatile Compounds in Pinus yunnanensis var.tenuifolia Damaged by Tomicus minor(Coleoptera:Scolytidae)by HS-SPME-GC/MS Method

【目的】掌握横坑切梢小蠹危害与细叶云南松应激响应规律,探讨其与抗性有关的代谢挥发物关系,以期提出有效的防治措施。【方法】以广西乐业县雅长林场不同危害程度细叶云南松嫩梢为对象,利用顶空固相微萃取-气相色谱/质谱法研究细叶云南松诱导抗性挥发物质。SPME萃取优化条件为:在75℃固相萃取仪上平衡15 min,用75μm萃取头(DVB/CAR/PDMS)萃取15 min。【结果】重度危害检测出最多挥发组分,主要为萜烯类、醇类。细叶云南松主要代谢挥发产物与其受危害程度密切相关,随着受害程度的增加,α-蒎烯、3-蒈烯、β-石竹烯、β-毕澄茄烯的含量随之增加,β-蒎烯含量逐渐减少,且仅对照含有少量长叶烯成分。重度危害类型中β-蒎烯含量最低仅为对照的47.85%,而3-蒈烯升高幅度最大,高于对照62.83%。单萜类及倍半萜类含氧衍生物总量随受害程度增加而增加,表明含氧化合物增多其抗虫性减弱。【结论】3-蒈烯、β-蒎烯、β-榄香烯、β-石竹烯和β-毕澄茄烯含量差异是导致细叶云南松诱导横坑切梢小蠹虫抗性的主要原因。顶空固相微萃取-气相色谱-质谱联用技术适用于松针挥发性成分的快速分析,并为细叶云南松小蠹诱导抗性物质研究提供科学依据。

【Objective】This study aims to understand the harmfulness of bark beetle,Tomicus minor,and the stress response of Pinus yunnanensis var.tenuifolia,and explore the relationship between the response and metabolic volatiles related to resistance so as to put forward effective control measures.【Method】The induced volatile substances of the pine were studied by headspace solid phase microextraction-gas chromatography/mass spectrometry with the shoots in different damage degrees in Yachang Forest Farm,Leye County,Guangxi.The optimum conditions of SPME were as follows:equilibrium for 15 min on solid phase extraction instrument at 75℃and extraction for 15 min with 75 micron extraction head(DVB/CAR/PDMS).【Result】The most volatile components were detected in severely damaged shoots,mainly terpenes and alcohols.The main metabolic volatile products of the pine were closely related to its damage degree.With the increase of the damage degree,the contents ofα-pinene,3-pinene,β-caryophyllene andβ-picengene increased,while the content ofβ-pinene decreased gradually,and only a small amount of longifolene was found in the control.In severely damaged shoots,β-pinene content was the lowest,only 47.85%of the control,while 3-pinene increased the most,higher than 62.83%of the control.The total amount of monoterpenoids and sesquiterpenoids oxygen-containing derivatives increased with the degree of damage,indicating that the insect resistance decreased with the increase of oxygen-containing compounds.【Conclusion】The differences in the contents of 3-carene,β-pinene,β-elemene,β-caryophyllene andβ-bichenylsolanee are the main reasons for inducing resistance of the pine to the bark beetle in transverse pits.Headspace solid phase microextraction(HS-SPME)coupled with gas chromatography-mass spectrometry(GC-MS)is suitable for the rapid analysis of volatile components in the pine needles,and this study provides a scientific basis for the study of induced resistance substances of Tomicus minor.