草履蚧对寄主挥发物感受的研究

发布时间：2018-05-12 05:05

本文选题：草履蚧 + 趋性反应　；参考：《山西大学》2015年硕士论文

【摘要】：草履蚧Drosicha corpulenta (Kuwana)属昆虫纲Insecta,半翅目Hemiptera,蚧总科Coccoidea,绵蚧科Monophlebidae,是一种危害多种林木和果树的植食性害虫,在我国分布广泛。本研究以昆虫寄主选择机制为依据,结合触角感受器的超微结构与功能研究,为草履蚧的生物防治提供理论依据。研究内容包括：(1)利用Y型嗅觉仪和蜜露显色法,观察寄主与非寄主植物的气味对草履蚧行为的影响；(2)运用光学显微镜、扫描电镜和透射电镜技术,研究各龄期草履蚧触角感受器的种类、形态、数量及分布；(3)草履蚧雌成虫触角气味结合蛋白(OBPs)的氨基酸序列分析。结果如下：1、研究了草履蚧2龄若虫和雌成虫分别对12种寄主植物和6种非寄主植物的趋性反应,并采用蜜露显色法测定了4种驱避作用较强的非寄主植物乙醇提取液对草履蚧蜜露排泄量的影响。草履蚧若虫和雌成虫对3种寄主植物柿树、枣树、杨树的气味均表现出显著的正趋性,其中柿树对若虫和雌成虫的引诱率分别达到68.92%和74.68%,诱集效果达到极显著水平；而对非寄主植物花椒、臭椿、薄荷、连翘的挥发物,以及上述4种植物的乙醇提取液表现出显著的负趋性,这些植物的挥发物和提取液对草履蚧具有显著的驱避作用,而榆叶梅植株挥发物仅对雌成虫具有显著驱避作用；拒食作用测定发现,经臭椿和花椒乙醇提取液处理后,草履蚧若虫的蜜露排泄量分别减少了70.57%和70.07%,雌成虫的减少了80.53%和72.42%,减少量均达到极显著水平。2、草履蚧2龄若虫和雌雄成虫触角形态及感受器的数量、分布和超微结构特征如下：草履蚧2龄若虫和雌成虫触角呈短棒状,深棕色,由柄节、梗节、鞭节组成,从2龄若虫发育到雌成虫鞭亚节由4个增加为6个。触角感器共有6种类型9种形态：毛形感器(Ⅰ和Ⅱ)、锥形感器(Ⅰ和Ⅱ)、刺形感器、腔锥形感器、钟形感器、钉形感器(Ⅰ和Ⅱ)。草履蚧雄虫触角为黑色、环生细长感觉毛呈念珠状,由柄节、梗节和8节鞭亚节组成。雄虫触角感器共有7种类型10种形态：毛形感器(Ⅰ、Ⅱ和Ⅲ)、锥形感器(Ⅰ和Ⅱ)、刺形感器(Ⅰ和Ⅱ)、腔锥形感器、钟形感器、钉形感器和周边具橛感器。各类型感器在各龄期触角上的形态、数量和分布不完全相同,毛形、刺形、钉型感受器的长度均随着龄期的增加而增加,其余感器变化不大。绝大部分触角感受器分布在触角的背、腹面和外侧面：毛形感器数量最多,在各龄期各节均有分布,约占80%；钟形感器仅为1个,着生于梗节腹面前缘；若虫和雌成虫具有两种形态的钉型感器,分别分布于触角顶节和第一、二鞭节腹面,雄成虫仅有1种着生于触角顶节；锥形感器雌雄成虫比若虫多1种；若虫和雌成虫的刺形感器仅有1种,主要分布于柄节和梗节,此外,雄成虫还有1种着生在顶节；腔锥感器均为1种,主要散生于梗节和顶节上；周边具橛感器仅存在于雄成虫梗节前缘。超微结构表明：锥形感器表皮壁薄、壁孔多,淋巴腔内有大量神经树突,是一种嗅觉感器；雄成虫的Ⅰ型毛形感器壁较厚,壁上有孔,淋巴腔内有2个神经树突,可能具有嗅觉感受功能；其Ⅱ型毛形感受器壁孔较少,基部淋巴腔内有一管状体,行使嗅觉感受和机械感受的功能；若虫、雌成虫的毛形感受器Ⅰ型与雄成虫的毛形感受器Ⅲ型、剌形感器Ⅱ型,这三种感器特点为表皮壁厚且无孔,淋巴腔内无树突分支,其功能与已知的典型机械感受器一周边具橛感器、钟形感受器一致。雌成虫的刺形感受器与雄虫的刺形感器Ⅰ型表皮壁无孔,内部树突鞘将树突分支包被在内淋巴腔内,可能行使机械感受和化学感受功能；钉型感器壁较薄、壁孔较多,内有大量神经树突,顶部有侧孔,腔锥形感受器具有顶孔,两者具有味觉感受功能。3.提取雌成虫触角RNA,反转录获得cDNA,进行转录组测序分析：筛选得到了17条草履蚧预测的OBP氨基酸序列,有6个保守的半胱氨酸位点,符合昆虫OBP结构特点‘'C1-X15-39-C2-X3-C3-X21-44-C4-X7-12-C5-X8-C6"。通过比对分析,表明其中14条预测序列属于OBPs家族。比对结果表明,草履蚧与同一亚目的蚜科昆虫相比,OBPs同源性为28%-35%,而DcOBP-3与苜蓿盲蝽Adelphocoris lineolatus的OBP4同源性最高,达到了46%
[Abstract]:The Paramecium Drosicha corpulenta (Kuwana) genus Insecta, Hemiptera Hemiptera, cecacroidea Coccoidea, miecacid Monophlebidae, is a kind of phytophagous pest that endanger a variety of trees and fruit trees. It is widely distributed in our country. This study is based on the mechanism of insect host selection, combined with the ultrastructure and function of antennae receptor. The biological control of parameces was provided as a theoretical basis. The contents of the study included: (1) the effects of the odor of host and non host plants on the behavior of Paramecium scale were observed by using the Y olfactory apparatus and the honeydew coloration method. (2) the species, morphology, quantity and score of the antennal sensilla of the Paramecium scale were studied by optical microscopy, scanning electron microscopy and transmission electron microscopy. (3) the amino acid sequence analysis of the antennal odorant binding protein (OBPs) of the female adults of the Paramecium scale. The results are as follows: 1, the taxis reaction of the 12 host plants and 6 non host plants of the 2 instar and female adults of the Paramecium scale, respectively, were studied, and the honeydew color method was used to determine the 4 extracts of the ethanol extract of the non host plants. The scent of the 3 host plant persimmons, jujube trees, and poplar trees showed significant positive taxis. The inducement rate of persimmon to nymphs and female adults was 68.92% and 74.68% respectively, and the effect of the lure reached a very significant level, while the non host plant pepper, Ailanthus, menthol, forsythia forsythia. The volatiles of the 4 plants and the ethanol extracts of these plants showed significant negative taxis. The volatiles and extracts of these plants had significant repellent to the Paramecium. The volatiles of the plants of the plant had a significant repellent to the female adults; the antifeeding test found that the Paramecium was treated by the extract of Ailanthus alon and Zanthoxylum. The nectar excretion of the scale nymphs decreased by 70.57% and 70.07% respectively, the decrease of female adults was 80.53% and 72.42%, and the decrease reached an extremely significant level of.2. The antennae morphology and the number, distribution and ultrastructure of the 2 nymph and female and male adult insect of Paramecium scale, distribution and ultrastructure were as follows: the 2 age nymphs of Paramecium scale and female adult antennae were short bar like, dark brown, and dark brown. Crank, skeleton, and flagellum, from 2 old nymph to female adult flagellate from 4 to 6. There are 6 types of antennae in 9 forms: hair sensilla (I and II), cone sensilla (I and II), conical sense organ, cone sense organ, bell shaped sense organ (I and II). The male insect tentacles of Paramecium scale are black, and long and slender sensory hair is present. The rosary, consisting of the knots, the skeleton and the 8 sections of the flagellum. The male tentacle has 7 types and 10 forms: the hair sensilla (I, II and III), the cone sensilla (I and II), the prickly sensilla (I and II), the cone sense organ, the bell shaped sense organ, the peg sense organ and the peripheral peg. The shape, quantity and distribution of the various types of sensilla on the antennae of each age. Not exactly the same, the length of hair shape, prickle shape and nail receptor increased with the age, and the other sensilla changed little. Most of the antennae receptors were distributed on the back of the antennae, in the abdomen and in the outer side: the number of hair sensilla was the most, about 80% in all ages, and only 1 of the bell shaped sensilla, which was born in front of the abdomen. The nymph and female adult have two types of nail sensilla, which are distributed in the antennae and the first, second flagellum, and the male adult is only 1 species born in the antennae; the conical sensilla is 1 more than that of the nymph; there are only 1 spiny receptors of the nymph and the female adult, mainly in the knots and ganglia, and in addition, there are 1 species of male adult worms. There are 1 kinds of cones, which are mainly scattered on the ganglion and the top of the ganglion. The peripheral peg only exists in the front of the male adult worm. The ultrastructure shows that the skin wall of the cone is thin, the wall holes are many, the lymphatic cavity has a large number of nerve dendrites, and it is a sense organ. The wall of the type I sense organ of the male adult is thicker, and the wall has holes and drenching. There are 2 nerve dendrites in the intraluminal cavity, which may have the function of olfactory sensation, and there are fewer wall holes in the type II hair receptor and a tubular body in the base lymph cavity, which exercises the function of olfactory sensation and mechanical feeling; the nymph, the hair receptor type I of the female adult and the male adult's hair receptor type III, the type II type, are characterized by the characteristics of the three sensilla. The epidermis is thick and nonporous, and the lymphatic cavity has no dendritic branches, and its function is consistent with the known typical mechanoreceptor with a peripheral peg and bell receptor. The spiny receptor of the female adult and the stinging sense organ of the male have no pores, and the internal dendrite sheath covers the dendrite branch into the endolymphatic cavity, which may exercise mechanical feeling and chemistry. The wall of the nail sensilla is thinner and more wall hole, there are a large number of nerve dendrites with a large number of nerve dendrites, the top has a lateral hole, and the conical receptor of the cavity has the top hole. Both have the taste sensory function.3. extract the female adult antennae RNA, reverse transcription cDNA, and carry out the transcriptional sequence analysis: the OBP amino acid sequence predicted by 17 Paramecium scale is screened, and there are 6 guarantees. The homocysteine site conformed to the OBP structure of the insect ''C1-X15-39-C2-X3-C3-X21-44-C4-X7-12-C5-X8-C6'. Through comparison and analysis, 14 of the predicted sequences belonged to the OBPs family. Compared with the same subfamily aphid, the OBPs homology of the Paramecium was 28%-35%, and DcOBP-3 and the alfalfa Adelphocoris lineol. The OBP4 homology of atus is the highest, reaching 46%

【学位授予单位】：山西大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433

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