本文選題：褐飛虱 + 取食行為��； 參考：《華中農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：褐飛虱(Nilaparvata lugens(St?l))是水稻上一種非常重要的刺吸式口器害蟲(chóng)。作為近年來(lái)褐飛虱田間防治的常用殺蟲(chóng)劑,吡蚜酮一直被認(rèn)為是通過(guò)抑制褐飛虱對(duì)韌皮部的取食使其饑餓而死,而近期研究表明吡蚜酮可能作用于昆蟲(chóng)TRPV通道(Transient receptor potential vaniloid channel,瞬時(shí)感受器電位香草酸受體亞型通道)。本研究通過(guò)RNA干擾的方法沉默褐飛虱TRPV通道兩個(gè)亞型Inactive(Iav)和Nanchung(Nan)的基因,結(jié)果表明褐飛虱TRPV通道與取食行為關(guān)系密切。具體結(jié)果如下:1.克隆褐飛虱Nan和Iav基因,分別與果蠅和豌豆蚜的Nan基因和Iav基因進(jìn)行序列比對(duì)。其中,Iav基因克隆片段與豌豆蚜和果蠅Iav基因相似度分別為83%和85%;Nan基因克隆片段與豌豆蚜和果蠅的相似度分別為85%和83%。進(jìn)一步驗(yàn)證克隆所得片段即分別為褐飛虱Iav和Nan基因片段。2.將褐飛虱分別注射dsIav和dsNan 1 d和3 d后,Iav和Nan基因的表達(dá)量顯著低于注射dsGFP的褐飛虱。Iav干擾后統(tǒng)計(jì)褐飛虱7 d內(nèi)的存活率,結(jié)果表明,第3 d和第4 d存活率顯著低于對(duì)照,而其他天數(shù)與對(duì)照相比無(wú)顯著差異,而Nan基因干擾之后褐飛虱7 d之后存活率僅為26.67%,顯著低于對(duì)照的存活率;RNA干擾后,褐飛虱的足均無(wú)異常情況。3.Iav基因和Nan基因干擾后,通過(guò)EPG監(jiān)測(cè)成蟲(chóng)在水稻植株上的取食,結(jié)果表明,Iav和Nan基因干擾之后褐飛虱韌皮部取食均受到抑制。其中,Iav基因干擾后,口針的刺探次數(shù)和路徑波時(shí)間延長(zhǎng)而韌皮部取食時(shí)間和刺探次數(shù)顯著低于對(duì)照。Nan基因干擾后,與對(duì)照相比木質(zhì)部取食時(shí)間和非刺探時(shí)間延長(zhǎng),而韌皮部的取食時(shí)間和韌皮部刺探次數(shù)降低。4.將RNA干擾之后的褐飛虱通過(guò)EPG監(jiān)測(cè)成蟲(chóng)對(duì)液態(tài)飼料取食,發(fā)現(xiàn)Iav基因和Nan基因干擾后對(duì)蔗糖溶液的取食與對(duì)照相比均無(wú)顯著差異。綜上所述,褐飛虱TRPV兩個(gè)亞型基因的沉默會(huì)抑制褐飛虱對(duì)水稻植株的取食,但不影響對(duì)液態(tài)飼料的取食,因此褐飛虱TRPV不影響褐飛虱的主動(dòng)抽吸能力但是否參與到昆蟲(chóng)-植物的互作則有待進(jìn)一步驗(yàn)證。本研究證明褐飛虱TRPV通道與取食調(diào)控關(guān)系密切,對(duì)褐飛虱的取食機(jī)理、吡蚜酮作用靶標(biāo)的研究以及褐飛虱的防治提供了理論基礎(chǔ)。
[Abstract]:The brown planthopper Nilaparvata lugensus Stleridae is a very important prickly mouthpiece pest in rice. As a commonly used insecticide for the control of brown planthopper in the field in recent years, pipriapyrone has been thought to starve brown planthopper to hunger by inhibiting its intake of phloem. Recent studies have shown that pipriapyrone may act on the transient receptor potential vaniloid channel, transient receptor subtype channel of TRPV channel in insects. In this study, RNA interference was used to silence the genes of two subtypes of TRPV channel of brown planthopper, I. E. Inactivev) and Nanchungus Nan. the results showed that the TRPV channel of BPH was closely related to feeding behavior. The results are as follows: 1. The Nan and Iav genes of brown planthopper were cloned and sequenced with the Nan gene and Iav gene of Drosophila melanogaster and pea aphid respectively. The similarity between the cloned fragment of Iav gene and the Iav gene of the pea aphid and Drosophila was 83% and 85%, respectively, and the similarity between the cloned fragment of Nan gene and the aphid of pea and Drosophila was 85% and 83%, respectively. It was further confirmed that the cloned fragments were Iav and Nan gene fragments of brown planthopper. The expression of Iav and Nan genes in dsIav and dsNan were significantly lower than those in the control group after 1 and 3 days of dsIav and dsNan injection, respectively. The results showed that the survival rate of BPH within 7 days was significantly lower than that of the control group on day 3 and day 4. However, there was no significant difference between the other days and the control, but the survival rate of BPH after Nan gene interference was only 26.67 after 7 days, which was significantly lower than that of the control. 3. There was no abnormality in the foot of BPH after the interference of Iav gene and Nan gene. The feeding of adults on rice plants was monitored by EPG. The results showed that the phloem feeding of brown planthopper was inhibited after the interference of Iav and Nan genes. After the interference of Iav gene, the times of oral needle prying and path wave were prolonged, while the feeding time and times of probing in phloem were significantly lower than those of control group. The feeding time and non-prying time of xylem were significantly longer than those of control group. However, the feeding time of phloem and the times of phloem pricking were decreased by 4. 4. Brown planthopper (BPH) with RNA interference was used to monitor adult feeding on liquid feed by EPG. It was found that there was no significant difference in sucrose solution intake between Iav gene and Nan gene. In conclusion, the silencing of TRPV subtype genes inhibited the feeding of brown planthopper to rice plants, but did not affect the feeding of liquid feed. Therefore, TRPV does not affect the active suction ability of BPH, but whether or not it is involved in the insect-plant interaction needs to be further verified. This study demonstrated that the TRPV channel of brown planthopper is closely related to feeding regulation, which provides a theoretical basis for the study of the feeding mechanism of brown planthopper, the target of pyridoxone action and the control of brown planthopper.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S435.112.3

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