【作者】 杨立军；

【导师】 刘惠霞；

【作者基本信息】 西北农林科技大学 ， 农业昆虫与害虫防治， 2011， 博士

【摘要】 枣镰翅小卷蛾(Ancylis sativa Liu)是枣树(Zizyphus jujuba Mill.)的重要害虫,隶属鳞翅目(Lepidoptera)卷蛾科(Tortricidae),以幼虫取食枣树芽、叶、花和果实,近年来在陕西和山西枣区危害严重。本文以寄主挥发物在植物与植食性昆虫关系中的作用为切入点,利用固相微萃取(SPME)和气相-质谱联用仪(GC-MS)技术,昆虫触角电位技术(EAG)、昆虫行为测定等研究方法,研究了木枣挥发物对枣镰翅小卷蛾寄主选择的影响,探讨了枣镰翅小卷蛾寄主选择机制,筛选出枣镰翅小卷蛾成虫引诱剂和驱避剂组分,为有效控制枣镰翅小卷蛾的危害提供依据。主要结论如下:利用固相微萃取(SPME)和GC-MS对木枣枝条和不同处理枣吊的挥发性组分鉴定表明:木枣枝条的挥发物组分有9种,枣吊的挥发物组分有24种。不同处理之间,在挥发物组分及比例上都有差异,枣吊虫害后,释放了虫害特异性挥发物—乙酸-4-己烯-1-醇酯。行为测试结果显示:枣吊受人工机械损伤后,对枣镰翅小卷蛾具有驱避作用,而健康和虫害枣吊具有明显的吸引作用。说明木枣不同部位及其损伤后,释放的挥发物成分和浓度都会发生改变,这种变化造成了枣镰翅小卷蛾的不同行为反应。而且,虫害后枣吊释放的挥发物,更加感虫。以木枣和酸枣为材料,在枣树萌芽期,利用固相微萃取(SPME)和GC-MS,分析比较了木枣和酸枣嫩叶的挥发性成分。结果显示,两者的挥发性成分没有差异,但相对含量稍有不同。枣镰翅小卷蛾成虫对木枣和酸枣的EAG反应测定显示,枣镰翅小卷蛾成虫对两种寄主都有强烈的刺激反应,且同一寄主内,雌虫的反应值极显著的高于雄虫。Y-型嗅觉仪测定表明,枣镰翅小卷蛾对木枣具有强烈的趋向反应,而对酸枣的趋向不明显,且雌虫的趋向反应高于雄虫。室内产卵结果显示,木枣上的产卵量显著高于酸枣,与行为测试结果一致,且枣吊上的产卵量极显著的高于枝条。这些结果表明,枣镰翅小卷蛾在选择寄主时,雌虫起主要作用,且对木枣更偏好。枣镰翅小卷蛾室内产卵行为研究结果表明,枣镰翅小卷蛾第一代的产卵量最高;在木枣的不同发育期中,幼叶期产卵量最高。Y-型嗅觉仪结果显示,枣镰翅小卷蛾越冬代和第一代成虫对木枣有较强的趋向反应;木枣的不同发育期中,花期及花期以前,枣镰翅小卷蛾对木枣都有较强的趋向反应,其它时期枣镰翅小卷蛾的趋向不明显。说明枣镰翅小卷蛾的种群动态变化与寄主植物的生长发育有着密切的相关性。利用扫描电镜研究了枣镰翅小卷蛾触角感受器。枣镰翅小卷蛾触角为线型,有柄节、梗节和49～52鞭节组成,触角着生在两个复眼之间。研究发现,共有10种类型的触角感器,分别是:毛形感器、刺形感器(Ⅰ型和Ⅱ型)、腔锥感器、鳞形感器、栓锥感器、氏鬃毛和耳形感器(Ⅰ型、Ⅱ型和Ⅲ型)。雌雄之间感器有性二型现象,如刺形感器在雄虫上只有Ⅰ型。雌雄相比,雄虫触角上毛形感器的数量要比雌虫多,而腔锥感器的数量,雌虫要多于雄虫。毛形感器、耳形感器和腔锥感器在寄主选择和定位中起着重要的作用。枣镰翅小卷蛾的EAG测试结果表明,枣镰翅小卷蛾雌虫、雄虫和交配雌虫的EAG反应差异显著;在所测定的18种化合物中,枣镰翅小卷蛾对乙酸正丁酯的EAG反应值最大,对反-2-己烯醇、乙酸叶醇酯、反-3-己烯醇和反-2-己烯醛也有很强的EAG反应。18种化合物6个浓度梯度的EAG反应结果显示,大部分单体的EAG反应都是随浓度的升高而呈上升趋势,与浓度的变化基本一致。枣镰翅小卷蛾行为测试结果显示,在100μg/μL浓度下,枣镰翅小卷蛾对反-2-己烯醇、乙酸叶醇酯、反-3-己烯醇、乙酸正丁酯、反-2-己烯醛和α-水芹烯均呈现负趋向,说明100μg/μL超出了枣镰翅小卷蛾的反应浓度范围。10μg/μL浓度是枣镰翅小卷蛾对乙酸正丁酯、罗勒烯和反-2-己烯醇反应的最佳浓度。1μg/μL浓度是枣镰翅小卷蛾对反-3-己烯醇反应的最佳浓度,也是对大多数化合物刺激反应的最低浓度。这些结果表明,在枣镰翅小卷蛾寄主选择中,乙酸正丁酯、罗勒烯、反-2-己烯醇、3-蒈烯和反-3-己烯醇起着重要的作用。更多还原

【Abstract】 Ancylis sativa Liu (Lepidoptera: Tortricidae), is one of the most important pests of Chinese jujube tree (Ziziphus jujube Mill).In recent years, it caused serious damage to Chinese jujube tree in Shaanxi and Shanxi province, and sometimes even cause destruction. In this paper we focused on the plant volatiles interact with the insect. Solid phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS), the electroantennogram recording technique (EAG), insect behavior method and other research methods were used to confirm the mechanism of host selection of A. sativa and host volatiles effect. Whilemean, finding the attraction and aversion composition of volatiles to A. sativa is intended to provide scientific and theoretical basis for the effective control on the damage by A. sativa. The main results showed as follows:The analysis of the volatiles composition of branches and shoots in different physiological status from Ziziphus jujuba Mill.cv.Muzao by solid phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) showed that the volatile compounds comprised mainly a range of terpenes and esters, secondly alcohol. The volatiles composition of branch of Z. jujuba Mill.cv.Muzao consisted of 9 compounds, and shoots 24 compounds. The healthy, artificial and infested damage shoots are detected difference in composition and ratio of volatiles. 4-hexen-1-ol, acetate is an herbivore-induced plant volatile which is emitted rapidly after leaf damaged and may attracts natural enemies. The results of olfactory experiment indicated that the artificial damage shoot showed significant repellency against moth of A. sativa while the shoots whatever healthy and infested damage showed attraction.The volatile compounds of tender leaves from Z. jujuba Mill.cv.Muzao and Ziziphus spinosa were identified in the germination stage by SPME and GC-MS, the results showed that Z. jujuba Mill.cv.Muzao and Z. spinosa were no significantdifferencese; however, the relative content was slightly different. EAG response of A. sativa on Z. jujuba Mill.cv.Muzao and Z. spinosa showed that both of the host plants had strong response in millivolts, and the absolute response in millivolts of the female antenna is general substantially higher than the male on the same host. Oflactory responses showed that A. sativa were strongly attracted by Z. jujuba Mill.cv.Muzao, and the females were attracted more strongly than the males, but they were not attraction by Z. spinosa. The test of insect oviposition showed that more eggs were obtained on the Z. jujuba Mill.cv.Muzao than on the Z. spinosa in the laboratory. Our results showed that the preference of A. sativa was Z. jujuba Mill.cv.Muzao. Furthermore, plant volatiles played a great role in oviposition, as in the A. sativa selection for plant host.The experiment under controlled laboratory conditions showed that oviposition and behavioral responses of A. sativa were different under plant volatile stimulus or not. Compared with other generations, first generation laid most eggs. In different developing period of Z. jujuba Mill.cv.Muzao, A. sativa laid most eggs in young leaf stage. Oflactory responses showed that overwintering and first generations of A. sativa made stronger selection responses, whilemean, selection responses of A. sativa were also stronger in the flower stage or before flower stage.The external morphologies of the antennal sensilla on the antennae of both female and male adults of A. sativa were observed using scanning electron microscopy. The antennae of both sexs of A. sativa were filiform in shape, consisting of the scape, pedicel and 49-52 segments of flagellomeres, and located between the compound eyes. Sensilla trichodea, two types of sensilla chaetica, sensilla coeloclnica, sensilla squamiformia, sensilla styloconica, b?hm bristles and three types of sensilla auricillica were observed on the female and male antennae. Two type of sensilla chaetica occurred only on the female antenna. The numbers of sensilla trichodea on the male were significantly greater than those of the female. However, the numbers of sensilla coeloclnica of the female were significantly more than those of the male.EAG responses of A. sativa to host volatile compounds were recorded. The results were significantly different among Virgin female, male and mate female in EAG responses. Five of the 18 compounds can elicit strong electrophysiological response, and butyl acetate had highest in millivolts, secondly was Trans-2-hexen-1-ol, Trans-3-hexen-1-ol, Cis-3-hexenyl acetate and Trans-2-hexen-1-al to A. sativa. EAG responses of the most of volatiles from low to high concentration to A. sativa showed an upward trend. Oflactory responses showed that trans-2-hexen-1-ol, trans-3-hexen-1-ol, cis-3-hexenyl acetate, trans-2-hexen-1-al, butyl acetate andα-Phellandrene were all significant repellency in host selection of A. sativa at the highest concentration (100μg/μL). But butyl acetate, ocimene and trans-2-hexen-1-ol were the most significant attractive to A. sativa at the 10μg/μL concentration. 1μg/μL concentration was perfect in the lowest concentration of the most compounds in behavioral responses of A. sativa. There were no significant differences on treatment and control among the lower three concentrations (0.1μg/μL, 0.01μg/μL, 0.001μg/μL). Thus, our results showed that trans-2-hexen-1-ol, trans-3-hexen-1-ol, cis-3-hexenyl acetate, ocimene, butyl acetate played important roles in host recognition in A. sativa.更多还原