本文選題：豌豆蚜 + 植物次生物質(zhì)　； 參考：《河北大學(xué)》2017年碩士論文

【摘要】：昆蟲的寄主適應(yīng)性研究一直以來都是昆蟲學(xué)和昆蟲-植物互作研究的熱點,豌豆蚜Acyrthosiphon pisum作為刺吸式口器昆蟲的模式種被發(fā)現(xiàn)存在多個明顯寄主分化的生物型。前期研究表明,豌豆蚜的寄主分化不僅與體內(nèi)共生菌密切相關(guān),還受到寄主植物免疫應(yīng)答、昆蟲自身代謝機(jī)制的影響。許多研究發(fā)現(xiàn)不同寄主來源的豌豆蚜在各自原始寄主上展現(xiàn)出更高的種群適合度,但相關(guān)機(jī)制性研究卻一直不清楚。為此,本文以兩種生物型豌豆蚜及其各自的原始寄主紫花苜蓿和豌豆為研究對象,從植物次生代謝物質(zhì)、蚜蟲取食行為、解毒代謝方面闡明不同寄主來源豌豆蚜的寄主適應(yīng)性差異。主要結(jié)果有:(1)利用刺探電位儀(Electrical penetration graph,EPG)研究兩種生物型豌豆蚜分別在原始寄主植物和非原始寄主上的取食行為。分析發(fā)現(xiàn),豌豆型豌豆蚜在非原始寄主紫花苜蓿上與苜蓿型豌豆蚜相比,刺探波(pd波)個數(shù)顯著增加、韌皮部被動取食(E2波)持續(xù)時間顯著縮短。苜蓿型豌豆蚜在非原始寄主豌豆上也有類似現(xiàn)象。進(jìn)一步調(diào)查兩種生物型豌豆蚜的種群動態(tài),發(fā)現(xiàn)豌豆型豌豆蚜在紫花苜蓿上的種群數(shù)量顯著少于苜蓿型豌豆蚜,而兩種生物型豌豆蚜在豌豆上種群數(shù)量無顯著差異。因此,苜蓿型豌豆蚜經(jīng)過一段時間的馴化可以成功定殖于非原始寄主豌豆,而豌豆型豌豆蚜卻不能夠很好地適應(yīng)非原始寄主紫花苜蓿。(2)為進(jìn)一步探索豌豆型豌豆蚜不能適應(yīng)紫花苜蓿是否與紫花苜蓿的次生代謝物質(zhì)有關(guān),我們選定了9種代表性的紫花苜蓿葉片次生代謝物質(zhì),包括原兒茶酸、綠原酸、香草酸、蘆丁、咖啡酸、對香豆素、染料木苷、阿魏酸和染料木黃酮,定量了兩種生物型豌豆蚜為害后紫花苜蓿葉片中這些次生代謝物的含量。發(fā)現(xiàn)豌豆型豌豆蚜顯著上調(diào)紫花苜蓿葉片中染料木黃酮的含量,而苜蓿型豌豆蚜顯著下調(diào)染料木黃酮的含量,其余8種次生物質(zhì)均未發(fā)生顯著性變化;(3)利用人工飼料外源添加不同濃度染料木黃酮進(jìn)一步研究染料木黃酮對兩種生物型豌豆蚜的取食效率和存活率的影響,發(fā)現(xiàn)添加1ppm和10ppm染料木黃酮會顯著提高苜蓿型豌豆蚜的取食效率,且染料木黃酮各濃度對其存活率沒有影響;而對于豌豆型豌豆蚜,10ppm染料木黃酮顯著提高其取食效率,但是1ppm和10ppm都顯著降低其存活率。這些結(jié)果表明兩種生物型豌豆蚜對染料木黃酮的解毒能力和代謝策略可能存在差異;(4)為了解紫花苜蓿上兩種生物型豌豆蚜消化和解毒的差異,我們通過轉(zhuǎn)錄組測序的方法分析了兩種生物型豌豆蚜取食紫花苜蓿兩周后其體內(nèi)差異性基因表達(dá)。發(fā)現(xiàn)相比于苜蓿型豌豆蚜,豌豆型豌豆蚜上調(diào)了161個基因,下調(diào)了37個基因,這些差異表達(dá)基因主要包括氨基酸代謝、碳水化合物代謝、能量代謝、脂肪代謝、外源物質(zhì)的降解和代謝等。上調(diào)的基因中有4個是消化酶相關(guān)基因,3個是解毒酶相關(guān)基因,這為不同生物型豌豆蚜通過解毒酶和消化酶響應(yīng)不同寄主植物提供了分子證據(jù)。以上結(jié)果發(fā)現(xiàn),苜蓿型豌豆蚜取食會降低紫花苜蓿中染料木黃酮的含量,因而不需要大量合成解毒酶來削弱染料木黃酮的不利影響,進(jìn)而可以將更多的營養(yǎng)和能量用于生長和繁殖。豌豆型豌豆蚜取食為害會顯著誘導(dǎo)紫花苜蓿中染料木黃酮含量的增加,而染料木黃酮有利于豌豆型豌豆蚜的取食,卻顯著降低其存活率。這說明染料木黃酮是豌豆型豌豆蚜的取食刺激因子,導(dǎo)致蚜蟲攝入過多的次生代謝物,誘導(dǎo)體內(nèi)消化酶和解毒酶相關(guān)基因大量上調(diào),進(jìn)而不利于自身生長發(fā)育和種群適合度。綜上所述,本研究以昆植互作和協(xié)同進(jìn)化為主線,從豌豆蚜消化酶和解毒酶相關(guān)基因表達(dá)、取食行為、種群動態(tài)以及植物次生物質(zhì)代謝等角度,拓展了刺吸式口器昆蟲寄主適應(yīng)性研究,為刺吸式口器害蟲的持續(xù)性治理和種群分化研究提供了理論依據(jù)。
[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.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S433

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本文編號：2002186