小黄鳃金龟寄主植物的挥发物鉴定及其电生理活性研究

发布时间：2018-03-09 23:08

本文选题：金龟　切入点：小黄鳃金龟　出处：《长江大学》2015年硕士论文　论文类型：学位论文

【摘要】：金龟是鞘翅目昆虫中非常重要的一个类群,其幼虫称为蛴螬,是种类最多、分布最广、为害最重的地下害虫。在我国重要的蛴螬种类有30余种,南北方均有发生,且常是多种蛴螬混合发生。其中,小黄鳃金龟(Melubohusju veseensbrenske)是重要的园林植物害虫,主要为害草坪、各种花卉、灌木及乔木。地下害虫金龟在求偶、聚集、觅食等活动中常常是通过借助化学气味物质传递信息的。研究这些化学气味物质和寄主植物挥发物的化学本质,可以为解释金龟在求偶、聚集、觅食等活动现象的本质,并为研究金龟引诱剂提供理论依据。本试验以小黄鳃金龟为研究对象,根据小黄鳃金龟在盛发期群集为害寄主植物的特点,调查了整个发生期里7种不同寄主植物上小黄鳃金龟的数量及其在丁香Syringa Linn,上的发生规律；并利用顶空法收集了7种植物的挥发物,采用现代化学分析的方法如气相色谱(GC)、气相色谱-质谱联用(GC-MS)和电生理的方法如触角电位仪(EAG)、气相色谱-触角电位联用(GC-EAD)等技术鉴定了寄主植物对小黄鳃金龟有电生理活性的挥发物；并挑选了7种挥发物标准样品做EAG分析。根据小黄鳃金龟在7种不同寄主植物上的分布数量,确定了小黄鳃金龟的优势寄主为丁香,次要寄主为榆树Ulmus pumila L.和山楂Crataegus pinnatifida Bunge第三寄主植物金银木Lonicera maackii (Rupr.) Maxim、桃树Amygdalus persica L和梨树Pyrus sorotina及非寄主植物为银杏Ginkgo biloba L.。通过丁香植物上小黄鳃金龟发生规律可知,小黄鳃金龟的年发生时间为6月上旬到7月中下旬,盛发期在6月下旬。从小黄鳃金龟对7种植物挥发物的EAG分析中可知,小黄鳃金龟雌雄成虫对丁香植物挥发物的EAG反应最大,分别为2.02±0.079 mV和2.03±0.069mV,与其它植物挥发物的电生理反应差异达到显著水平。对银杏植物挥发物的EAG反应最小,分别为0.26土0.043 mV和0.37±0.071 mV,与其它植物挥发物的电生理反应差异达到显著水平。采用GC-EAD和GC-MS技术,共鉴定出15种植物挥发物对小黄鳃金龟有电生理活性。这些挥发物分别为E-3-己烯醛或1-戊烯-3-醇,E-罗勒烯,Z-罗勒烯,p-伞花烃,E-3-己烯乙酸酯或E-2-戊烯-1-醇,1-己醇,苯甲醚,E-3-己烯醇,Z-3-己烯醇,Z-2-己烯醇或Z-2-己烯乙酸酯或壬醛,氧化芳樟醇,庚醇,芳樟醇,葵醛,乙酸苄酯。丁香、榆树和山楂挥发物中对小黄鳃金龟有活性的化合物比较多,金银木、银杏中有活性的挥发物比较少。丁香、榆树和山楂中化合物基本上类似,金银木和银杏中挥发物也基本类似。在金银木、银杏挥发物样品中有活性的化合物几乎都出现在丁香、山楂和榆树挥发物样品中。在用小黄鳃金龟雌雄成虫做EAG实验时,测试了连接不同触角鳃瓣对结果的影响,发现4种连接方式之间几乎没有显著性差异。因此,在做鳃金龟EAG实验时,可以选择一种简单的连接触角鳃瓣的方式即可。在小黄鳃金龟对7种挥发物标准样品的EAG分析实验中可知,小黄鳃金龟雌雄成虫对苯甲醚标准样品的EAG反应差异显著,在0.001 μg、0.01μg和O.1雌三种浓度下,雌虫的EAG反应分别为0.189±0.021 mV、0.225±0.038 mV和0.577±0.260 mV,雄虫的EAG反应分别为0.527±0.055 mV、0.887±0.107 mV和2.348±0.266 mV。随着挥发物样品剂量的增加,EAG反应值也随之增加。在小黄鳃金龟雄虫对7种挥发物标准样品的EAG实验中,1μg的顺-3-己烯醇样品的EAG反应为1.243±0.162 mV,10μg的顺-3-己烯醇样品的EAG反应为1.771±0.159 mV, EAG反应变化增加了1.4倍,而100μg的顺-3-己烯醇样品的EAG反应为6.149±0.467 mV, EAG反应变化增加了3.5倍。小黄鳃金龟在不同寄主植物上的分布数量与小黄鳃金龟对植物挥发物的EAG反应结果相一致,在丁香上金龟的数量最多,同时对丁香挥发物的EAG反应也最大,说明小黄鳃金龟在寻找寄主时是通过感受植物释放的挥发物中的组分来定位寄主植物的。对小黄鳃金龟有活性的挥发物都是常见的绿叶挥发物,但每种寄主植物释放的挥发物种类不同,说明小黄鳃金龟在寄主定位时是通过感受某些植物挥发物的特定组合,而不是感知某一个特定的植物挥发物。
[Abstract]:Rich is a very important species of Coleoptera insects whose larvae, called grubs, is the most species, the most widely distributed underground pests in China. The most important grub species are more than 30 species, both north and South have occurred, and is often a mixture of white grubs. Among them, Miss Daisy Sai Jingui (Melubohusju veseensbrenske) is an important pest of garden plants, mainly on the lawn, all kinds of flowers, shrubs and trees. The underground pest beetles in courtship, aggregation, often foraging activities is through the use of chemical odors passing information. These chemical substances and odors of host plant volatile chemical nature of volatiles, can explain the beetle in courtship, aggregation, foraging activities such as nature, and provide theoretical basis for the study of beetle attractant. In this experiment, the Yellow beetle as the research object, according to the Yellow beetle in the peak period of the host cluster damage The characteristics of the plant, investigated 7 different host plants on the Yellow beetle of the whole period and number of Linn in the occurrence of the clove Syringa; and a collection of 7 kinds of plant volatiles by headspace method, using modern methods such as gas chromatography chemical analysis (GC), gas chromatography mass spectrometry combined with (GC-MS) and electrophysiological methods such as electroantennography (EAG), gas chromatography electroantennogram (GC-EAD) combined with techniques such as identification of host plants on the electrophysiological activity of yellow beetle volatiles; and selected 7 kinds of volatiles of standard sample for EAG analysis. According to the distribution of the number of yellow melolonthidae in 7 different host plants, the Yellow beetle is the predominant host clove, secondary host for pumila L. and Crataegus Ulmus Yushu Hawthorn pinnatifida Bunge third host plant Lonicera (Rupr.) maackii and Maxim, peach Amygdalu S persica L and Pyrus sorotina of pear and non host plants for Ginkgo biloba L.. By Ginkgo plant yellow beetle clove occurrence that occurred in early June to mid to late 7 yellow beetle of the year, the peak period in late June. From the Yellow beetle to 7 plant volatiles in the volatile EAG analysis the Yellow beetle, male and female adults of lilac plant were tested by EAG technique, were 2.02 + 0.079 and 2.03 + mV, 0.069mV, and other plant essential differences in electrophysiological responses to volatiles reached significant level. The EAG reaction of Ginkgo plant volatiles is the smallest, respectively 0.26 and 0.37 + 0.043 mV + 0.071 mV. With other plant essential electrophysiological responses to volatiles differences reached significant level. Using GC-EAD and GC-MS technology, we identified 15 plant volatiles have electrophysiological activity to the Yellow beetle. These volatiles were E-3- hexenal or 1- Pentene alcohol -3-, E- ocimene, Z- ocimene, p- p-cymene, E-3- hexenyl acetate or E-2- -1- 1- pentene alcohol, cyclohexanol, benzyl ether, E-3- cis-3-hexenol, Z-3- cis-3-hexenol, cis-3-hexenol Z-2- or Z-2- hexenyl acetate or nonanal, linalool oxide, heptanol, linalool. Sunflower aldehyde, benzyl acetate, clove, elm and volatile compounds in Hawthorn volatiles on the activity of the Yellow beetle more, honeysuckle, volatiles of Ginkgo biloba in activity less. Clove, elm and hawthorn compound basically similar, honeysuckle and ginkgo essential is basically similar. Volatiles in the gold and silver wood of ginkgo the volatile compounds active compounds in almost all samples appeared in clove, hawthorn and elm volatiles in the sample. The Yellow beetle of male and female adults do EAG experiments, connecting the different influence on the results of lamellibranch antenna test, was found between 4 types of connection almost no significant difference. Therefore, doing Beetle EAG experiments, then can choose a simple connection way. In the Yellow tentacles of lamellibranch beetle of 7 kinds of volatiles of standard sample EAG analysis experiment shows that the difference of EAG reaction yellow beetle of male and female adults of anisole standard samples were 0.001 g, 0.01 g and O.1 female three concentrations, respectively, EAG reaction was 0.189 female + 0.021 mV, 0.225 mV + 0.038 and 0.577 + 0.260 mV, respectively. EAG reaction was 0.527 + 0.055 mV male, 0.887 + 0.107 and 2.348 + 0.266 mV mV. with the increase of volatile sample dosage, reaction EAG value also increased. In the Yellow gill the male beetles on 7 kinds of volatiles of standard samples in the EAG experiment, 1 g CIS EAG reaction -3- cis-3-hexenol sample was 1.243 + 0.162 mV, 10 g CIS EAG reaction -3- cis-3-hexenol sample was 1.771 + 0.159 mV, EAG reaction increased 1.4 times, and 100 g Shun -3- sample cis-3-hexenol The EAG reaction was 6.149 + 0.467 mV, EAG reaction increased 3.5 times. The Yellow beetle abundance distribution in different host plants with yellow beetle EAG responses to plant volatiles are consistent, the number of beetles in the lilac on most, while the clove volatiles of EAG reaction is the highest, that the Yellow beetle is to locate the host plant through the plant release of the volatile components in the host finding. The Yellow beetle has volatile activity are common green leaf volatiles, but each kind of host plant volatiles released from different species, that yellow beetle in host location is through feel some specific combinations of plant volatiles, rather than the perception of a specific plant volatiles.

【学位授予单位】：长江大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433.5

【引证文献】