沙棘挥发物及其抗虫机制的初步研究
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摘要
沙棘木蠹蛾(Holcocerus hippophaecolus Hua、Chou、Fang et Chen)是近几年在我国内蒙古、辽宁、山西、陕西和宁夏等地沙棘林大面积爆发的钻蛀性害虫,对我国的沙棘产业发展和生态环境构成严重威胁和巨大损失。本文重点是研究沙棘挥发物及对沙棘木蠹蛾趋性行为的影响,为沙棘木蠹蛾的成灾机理提供一定的理论依据。
首先优化了动态顶空采样法采集痕量植物挥发物的实验条件,筛选出美国产食品级聚氯乙烯管(PVC)符合沙棘挥发物的采集；同时初步鉴定了在近自然状态下,沙棘枝叶主要挥发物组分主要有壬烷、辛烷、3-甲基丁醇、戊醛、2-甲基丁醛、2,3-丁二酮、2-壬烯醇、2-苯甲酸叶醇酯。
利用动态顶空采集、热脱附-气质联用(TCT-GC／MS)法分析鉴定了沙棘人工林与天然林中健康和虫害沙棘枝叶和基干部位释放的挥发性物质,共鉴定出29种化合物。结果显示,人工林中戊醛和2-苯甲酸叶醇酯、5-甲基-5-羟基己酸内酯、3-甲基丁醇含量较天然林高；天然林沙棘释放的挥发物以顺式-2-癸醇、二壬烯醇、庚醛、2-甲基丁醛为主。虫害后,沙棘人工林与天然林都会释放桉树脑、萜品醇、2-莰酮萜烯类物质,并且天然林中萜品醇和2-莰酮的相对含量高于人工林,同时醇类、醛类物质虫害后也有增加。
触角电位实验表明,沙棘木蠹蛾对戊醛的反应最大,其次为2,3-丁二酮和三甲基丁醇,对萜品醇反应最小。四种单体化合物对沙棘木蠹蛾的触角电位反应活性在原液的液体石蜡溶液的1／10浓度以下呈正相关关系。
对沙棘木蠹蛾的8种寄主树种进行挥发物的动态顶空采集-热解析气质联用测定,结果表明中国沙棘亚种、橙色、阿亚甘卡两个品种释放的挥发物中,酯类、醛类物质所占的比例较大,相对含量较高的化合物主要有3-甲基戊醛、2-苯乙基异烟酸酯、辛醛、2-乙基己醇、壬醛、2-乙基己基醋酸酯、癸醛、丁基丁酸酯、2-乙基己基安息香酸酯、辛基邻苯二甲酸酯等；丘中和太阳两个沙棘品种释放的挥发物中,以萜烯类所占比例较大,醛类和酯类物质的种类变化不大,但是相对含量明显较橙色和中国的降低,萜类物质主要就是α-蒎烯、水芹烯、三蒈烯。乌兰格木和楚伊两个沙棘品种所释放的挥发物中,醛类和酯类物质总的相对含量较丘中、向阳高而比橙色和中国低,萜类物质释放量低于丘中和向阳；白榆醇类醛类酯类和萜烯类物质各自相对含量接近。
沙棘木蠹蛾成虫对三种沙棘品种(亚种)和白榆的嗅觉行为反应测定结果表明,橙色沙棘、中国沙棘、太阳沙棘和白榆四个树种对沙棘木蠹蛾引诱程度具有一定程度的引诱作用,中国沙棘对沙棘木蠹蛾的引诱作用最强,达到差异极显著水平(p=0.0043),其次为橙色,达到差异显著水平(p=0.047)。
Holcocerus hippophaecolus is a destructive trunk borer of Hippophae rhamnoides L.,in recent years, seabuckthorn carpenter moth H. hippophaecolus broken out in Inner Mongolia Autonomous region,Ningxia Hui Autonomous region,Shanxi,Shaanxi,Liaoning,Hebei and Gansu provinces,and caused a heavy ecological and economic loss.The emphasis of this dissertation is laid on the volatiles of H. rhamnoides and its effect to H. hippophaecolus,which can provide theoretical basis for mechanisms of occurrence of the H. hippophaecolus.
Firstly, the experimental conditions of dynamic headspace air-circulation collection method were optimized,screening the PVC pipe made in USA is suitable to collect trace volatiles; Furthermore,the main volatiles from Hippophae rhamnoides included Nonane、Octane、3-methyl-1-Butanol、Pentanal、2-methyl-Butanal、2,3-Butanedione、2-Hexen-l-ol、2-Hexenyl benzoate in close natural state.
In order to research the cause of that it is more seriously harmed by H. hippophaecolus in the planted forest of H. rhamnoides than in the natural forest,the headspace volatiles emitted by healthy and verminous leaves and trunk of H. rhamnoides in planted forest and natural forest were examined by adsorption-thermodesorption and GC-MS identification.Twenty nine compounds were yielded.The analysis revealed that the plants in the planted forest released more pentanal、E-2-Hexenyl 2-benzoate、5-Methyl-5-hydroxyhexanoic acid lactone、3-Methyl-1-butanol,while the plants in the natural forest released more Trans-2-decen-ol、2-nonen-1-ol、2-Methyl-butanal、Heptanal. Lots of terpenes were released and the releasing of aldehydes and alcohols were also enhanced when those trees were damaged by the H. hippophaecolus.
The EAG results showed that,H.hippophaecolus is most sensitive to pentanal,following are 2, 3-Butanedione and 3-methyl-l-butanol.The potential reactivities of H.hippophaecolus by different compounds are positive correlation in the decimus concentration.
The volatiles from 8 kinds of host trees of H.hippophaecolus were collected by using dynamic headspace air-circulation collection method and identified by TCT-GC-MS.The result shows that in the volatiles of the H.rhamnoides subsp sinensis、chenggse and ayaganka.The main compounds are 3-Methyl-Pentanal、2-Phenylethyl-isonicotinic acid ester、Octanal、2-Ethyl-l-hexanol、Nonanal、Acetic acid、2-Ethylhexyl、decanal、Butanoic acid,butyl ester、Benzoic acid,2-ethylhexyl ester、DI-N-octyl phthalate; The proportion of terpenes is larger in the volatiles from qiuzhong taiyang, while the relative content of esters and aldehydes is lower than H.rhamnoides subsp sinensis、chenggse and ayaganka,but not in species.The main terpenes are Alpha-pinene、Beta-phellandrene、3-carene. The volatiles from wumu and chuyi, the relative contents of esters and aldehydes are larger than the relative contents from qiuzhong and xiangyang,but lower than chengse and H.rhamnoides subsp sinensis; The relative content of terpenes are lower than qiuzhong and xiangyang.The relative contents of Esters、Aldehydes、Terpenes from Ulmus pumila L.are close to each other.
The behavior responses of H.hippophaecolus to four host trees were measured by the "Y" olfactometer.The result showed that the volatiles from H.rhamnoides subsp sinensis was better than others in attracting H.hippophaecolus, secondly was chengse,the analysis of variance showed that the difference reached at the 5% significant level.
首先优化了动态顶空采样法采集痕量植物挥发物的实验条件,筛选出美国产食品级聚氯乙烯管(PVC)符合沙棘挥发物的采集；同时初步鉴定了在近自然状态下,沙棘枝叶主要挥发物组分主要有壬烷、辛烷、3-甲基丁醇、戊醛、2-甲基丁醛、2,3-丁二酮、2-壬烯醇、2-苯甲酸叶醇酯。
利用动态顶空采集、热脱附-气质联用(TCT-GC／MS)法分析鉴定了沙棘人工林与天然林中健康和虫害沙棘枝叶和基干部位释放的挥发性物质,共鉴定出29种化合物。结果显示,人工林中戊醛和2-苯甲酸叶醇酯、5-甲基-5-羟基己酸内酯、3-甲基丁醇含量较天然林高；天然林沙棘释放的挥发物以顺式-2-癸醇、二壬烯醇、庚醛、2-甲基丁醛为主。虫害后,沙棘人工林与天然林都会释放桉树脑、萜品醇、2-莰酮萜烯类物质,并且天然林中萜品醇和2-莰酮的相对含量高于人工林,同时醇类、醛类物质虫害后也有增加。
触角电位实验表明,沙棘木蠹蛾对戊醛的反应最大,其次为2,3-丁二酮和三甲基丁醇,对萜品醇反应最小。四种单体化合物对沙棘木蠹蛾的触角电位反应活性在原液的液体石蜡溶液的1／10浓度以下呈正相关关系。
对沙棘木蠹蛾的8种寄主树种进行挥发物的动态顶空采集-热解析气质联用测定,结果表明中国沙棘亚种、橙色、阿亚甘卡两个品种释放的挥发物中,酯类、醛类物质所占的比例较大,相对含量较高的化合物主要有3-甲基戊醛、2-苯乙基异烟酸酯、辛醛、2-乙基己醇、壬醛、2-乙基己基醋酸酯、癸醛、丁基丁酸酯、2-乙基己基安息香酸酯、辛基邻苯二甲酸酯等；丘中和太阳两个沙棘品种释放的挥发物中,以萜烯类所占比例较大,醛类和酯类物质的种类变化不大,但是相对含量明显较橙色和中国的降低,萜类物质主要就是α-蒎烯、水芹烯、三蒈烯。乌兰格木和楚伊两个沙棘品种所释放的挥发物中,醛类和酯类物质总的相对含量较丘中、向阳高而比橙色和中国低,萜类物质释放量低于丘中和向阳；白榆醇类醛类酯类和萜烯类物质各自相对含量接近。
沙棘木蠹蛾成虫对三种沙棘品种(亚种)和白榆的嗅觉行为反应测定结果表明,橙色沙棘、中国沙棘、太阳沙棘和白榆四个树种对沙棘木蠹蛾引诱程度具有一定程度的引诱作用,中国沙棘对沙棘木蠹蛾的引诱作用最强,达到差异极显著水平(p=0.0043),其次为橙色,达到差异显著水平(p=0.047)。
Holcocerus hippophaecolus is a destructive trunk borer of Hippophae rhamnoides L.,in recent years, seabuckthorn carpenter moth H. hippophaecolus broken out in Inner Mongolia Autonomous region,Ningxia Hui Autonomous region,Shanxi,Shaanxi,Liaoning,Hebei and Gansu provinces,and caused a heavy ecological and economic loss.The emphasis of this dissertation is laid on the volatiles of H. rhamnoides and its effect to H. hippophaecolus,which can provide theoretical basis for mechanisms of occurrence of the H. hippophaecolus.
Firstly, the experimental conditions of dynamic headspace air-circulation collection method were optimized,screening the PVC pipe made in USA is suitable to collect trace volatiles; Furthermore,the main volatiles from Hippophae rhamnoides included Nonane、Octane、3-methyl-1-Butanol、Pentanal、2-methyl-Butanal、2,3-Butanedione、2-Hexen-l-ol、2-Hexenyl benzoate in close natural state.
In order to research the cause of that it is more seriously harmed by H. hippophaecolus in the planted forest of H. rhamnoides than in the natural forest,the headspace volatiles emitted by healthy and verminous leaves and trunk of H. rhamnoides in planted forest and natural forest were examined by adsorption-thermodesorption and GC-MS identification.Twenty nine compounds were yielded.The analysis revealed that the plants in the planted forest released more pentanal、E-2-Hexenyl 2-benzoate、5-Methyl-5-hydroxyhexanoic acid lactone、3-Methyl-1-butanol,while the plants in the natural forest released more Trans-2-decen-ol、2-nonen-1-ol、2-Methyl-butanal、Heptanal. Lots of terpenes were released and the releasing of aldehydes and alcohols were also enhanced when those trees were damaged by the H. hippophaecolus.
The EAG results showed that,H.hippophaecolus is most sensitive to pentanal,following are 2, 3-Butanedione and 3-methyl-l-butanol.The potential reactivities of H.hippophaecolus by different compounds are positive correlation in the decimus concentration.
The volatiles from 8 kinds of host trees of H.hippophaecolus were collected by using dynamic headspace air-circulation collection method and identified by TCT-GC-MS.The result shows that in the volatiles of the H.rhamnoides subsp sinensis、chenggse and ayaganka.The main compounds are 3-Methyl-Pentanal、2-Phenylethyl-isonicotinic acid ester、Octanal、2-Ethyl-l-hexanol、Nonanal、Acetic acid、2-Ethylhexyl、decanal、Butanoic acid,butyl ester、Benzoic acid,2-ethylhexyl ester、DI-N-octyl phthalate; The proportion of terpenes is larger in the volatiles from qiuzhong taiyang, while the relative content of esters and aldehydes is lower than H.rhamnoides subsp sinensis、chenggse and ayaganka,but not in species.The main terpenes are Alpha-pinene、Beta-phellandrene、3-carene. The volatiles from wumu and chuyi, the relative contents of esters and aldehydes are larger than the relative contents from qiuzhong and xiangyang,but lower than chengse and H.rhamnoides subsp sinensis; The relative content of terpenes are lower than qiuzhong and xiangyang.The relative contents of Esters、Aldehydes、Terpenes from Ulmus pumila L.are close to each other.
The behavior responses of H.hippophaecolus to four host trees were measured by the "Y" olfactometer.The result showed that the volatiles from H.rhamnoides subsp sinensis was better than others in attracting H.hippophaecolus, secondly was chengse,the analysis of variance showed that the difference reached at the 5% significant level.
引文
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