不同寄主来源豌豆蚜的寄主适应性研究

发布时间：2018-06-10 05:50

本文选题：豌豆蚜 + 植物次生物质　；参考：《河北大学》2017年硕士论文

【摘要】：昆虫的寄主适应性研究一直以来都是昆虫学和昆虫-植物互作研究的热点,豌豆蚜Acyrthosiphon pisum作为刺吸式口器昆虫的模式种被发现存在多个明显寄主分化的生物型。前期研究表明,豌豆蚜的寄主分化不仅与体内共生菌密切相关,还受到寄主植物免疫应答、昆虫自身代谢机制的影响。许多研究发现不同寄主来源的豌豆蚜在各自原始寄主上展现出更高的种群适合度,但相关机制性研究却一直不清楚。为此,本文以两种生物型豌豆蚜及其各自的原始寄主紫花苜蓿和豌豆为研究对象,从植物次生代谢物质、蚜虫取食行为、解毒代谢方面阐明不同寄主来源豌豆蚜的寄主适应性差异。主要结果有:(1)利用刺探电位仪(Electrical penetration graph,EPG)研究两种生物型豌豆蚜分别在原始寄主植物和非原始寄主上的取食行为。分析发现,豌豆型豌豆蚜在非原始寄主紫花苜蓿上与苜蓿型豌豆蚜相比,刺探波(pd波)个数显著增加、韧皮部被动取食(E2波)持续时间显著缩短。苜蓿型豌豆蚜在非原始寄主豌豆上也有类似现象。进一步调查两种生物型豌豆蚜的种群动态,发现豌豆型豌豆蚜在紫花苜蓿上的种群数量显著少于苜蓿型豌豆蚜,而两种生物型豌豆蚜在豌豆上种群数量无显著差异。因此,苜蓿型豌豆蚜经过一段时间的驯化可以成功定殖于非原始寄主豌豆,而豌豆型豌豆蚜却不能够很好地适应非原始寄主紫花苜蓿。(2)为进一步探索豌豆型豌豆蚜不能适应紫花苜蓿是否与紫花苜蓿的次生代谢物质有关,我们选定了9种代表性的紫花苜蓿叶片次生代谢物质,包括原儿茶酸、绿原酸、香草酸、芦丁、咖啡酸、对香豆素、染料木苷、阿魏酸和染料木黄酮,定量了两种生物型豌豆蚜为害后紫花苜蓿叶片中这些次生代谢物的含量。发现豌豆型豌豆蚜显著上调紫花苜蓿叶片中染料木黄酮的含量,而苜蓿型豌豆蚜显著下调染料木黄酮的含量,其余8种次生物质均未发生显著性变化;(3)利用人工饲料外源添加不同浓度染料木黄酮进一步研究染料木黄酮对两种生物型豌豆蚜的取食效率和存活率的影响,发现添加1ppm和10ppm染料木黄酮会显著提高苜蓿型豌豆蚜的取食效率,且染料木黄酮各浓度对其存活率没有影响;而对于豌豆型豌豆蚜,10ppm染料木黄酮显著提高其取食效率,但是1ppm和10ppm都显著降低其存活率。这些结果表明两种生物型豌豆蚜对染料木黄酮的解毒能力和代谢策略可能存在差异;(4)为了解紫花苜蓿上两种生物型豌豆蚜消化和解毒的差异,我们通过转录组测序的方法分析了两种生物型豌豆蚜取食紫花苜蓿两周后其体内差异性基因表达。发现相比于苜蓿型豌豆蚜,豌豆型豌豆蚜上调了161个基因,下调了37个基因,这些差异表达基因主要包括氨基酸代谢、碳水化合物代谢、能量代谢、脂肪代谢、外源物质的降解和代谢等。上调的基因中有4个是消化酶相关基因,3个是解毒酶相关基因,这为不同生物型豌豆蚜通过解毒酶和消化酶响应不同寄主植物提供了分子证据。以上结果发现,苜蓿型豌豆蚜取食会降低紫花苜蓿中染料木黄酮的含量,因而不需要大量合成解毒酶来削弱染料木黄酮的不利影响,进而可以将更多的营养和能量用于生长和繁殖。豌豆型豌豆蚜取食为害会显著诱导紫花苜蓿中染料木黄酮含量的增加,而染料木黄酮有利于豌豆型豌豆蚜的取食,却显著降低其存活率。这说明染料木黄酮是豌豆型豌豆蚜的取食刺激因子,导致蚜虫摄入过多的次生代谢物,诱导体内消化酶和解毒酶相关基因大量上调,进而不利于自身生长发育和种群适合度。综上所述,本研究以昆植互作和协同进化为主线,从豌豆蚜消化酶和解毒酶相关基因表达、取食行为、种群动态以及植物次生物质代谢等角度,拓展了刺吸式口器昆虫寄主适应性研究,为刺吸式口器害虫的持续性治理和种群分化研究提供了理论依据。
[Abstract]:The study on host adaptability of insects has always been a hot spot in the study of Entomology and insect plant interaction. The Acyrthosiphon Pisum of pea aphid, as a pattern species of spiny mouthed mouthparts, has been found to have many distinct host differentiation types. The influence of host plant immune response and insect's own metabolic mechanism. Many studies have found that pea aphids of different host sources show higher population fitness on their original host, but the related mechanisms are not clear. For this reason, two species of pea aphids and their respective original host alfalfa and peas were found in this paper. The main results were as follows: (1) two species of pea aphids were studied on the original host plant and non primitive host by Electrical penetration graph (EPG). Compared with the alfalfa aphid, the number of spying wave (PD wave) increased significantly and the duration of passive feeding (E2 wave) of phloem was significantly shortened. Alfalfa type pea aphid was also similar to the non original host pea. Further investigation of two species of biogenic pea aphids was found. The population dynamics showed that the number of pea aphids on alfalfa was significantly less than that of alfalfa, but there was no significant difference in the number of two species of pea aphid on pea. Therefore, the alfalfa type pea aphid could be successfully colonized by a period of time domestication, but the pea aphid could not be very good. Well adapted to the non original host alfalfa. (2) in order to further explore the failure of the pea aphid to adapt to the secondary metabolites of alfalfa, we selected 9 representative secondary metabolites of alfalfa leaf, including protocatechuic acid, Lv Yuan acid, vanillin, rutin, caffeic acid, coumarin, dyestuff Glucoside, ferulic acid and genistein were used to quantify the content of these secondary metabolites in the leaves of Alfalfa after two biological types of pea aphids. It was found that pea type pea aphid significantly increased the content of genistein in the leaves of alfalfa, while alfalfa type pea aphid significantly lowered the content of genistein, and the other 8 kinds of biomass were not distributed. 3. (3) the effects of genistein on the feeding efficiency and survival rate of two biogenic pea aphids were further studied by adding different concentrations of genistein into the artificial diet. It was found that the feeding efficiency of Alfalfa type pea aphid was significantly increased by adding 1ppm and 10ppm dye, and the concentration of genistein on it was stored. The survival rate was not affected; and for the pea aphid, 10ppm genistein significantly increased its feeding efficiency, but both 1ppm and 10ppm significantly decreased the survival rate. These results showed that the detoxification ability and metabolic strategies of the two biological type pea aphids for genistein might be different. (4) to understand the two species of pea pea on alfalfa. The difference in the digestion and detoxification of aphids, we analyzed the differential gene expression in two biological type pea aphids after two weeks of feeding alfalfa by transcriptional sequencing. It was found that compared to the alfalfa type pea aphid, the pea aphid up regulated 161 genes and lowered 37 genes. These differentially expressed genes mainly included the amino acid generation. Thanks, carbohydrate metabolism, energy metabolism, fat metabolism, the degradation and metabolism of exogenous substances. 4 of the up-regulated genes are digestive enzyme related genes and 3 are detoxification enzymes related genes. This provides molecular evidence for different biotype pea aphids to respond to different host plants through detoxification enzymes and digestive enzymes. The results are found, alfalfa type pea The intake of aphid can reduce the content of genistein in alfalfa, so there is no need for a large number of synthetic detoxification enzymes to weaken the adverse effects of genistein, and then more nutrients and energy can be used for growth and reproduction. Genistein is beneficial to the feeding of pea type pea aphid, but significantly reduces its survival rate. This shows that genistein is a feeding stimulator of pea type pea aphid, which leads to the excessive intake of secondary metabolites from aphids, and induces a large increase in the genes related to digestive enzymes and detoxification enzymes in the body, which is not conducive to the growth and population fitness of the plant. In this study, the study was based on the interplanting and coevolution of Kun planting, from the expression of genes related to digestive enzymes and detoxification enzymes, feeding behavior, population dynamics, and secondary biomass metabolism of the pea aphid, which extended the study on the host adaptability of the spiny mouthpiece insect, and provided the sustainable control and population differentiation of the spiny sucking oral insect pests. The theoretical basis.
【学位授予单位】：河北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S433

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