褐飞虱TRPV通道两个亚型基因与取食行为关系

发布时间：2018-06-03 13:03

本文选题：褐飞虱 + 取食行为　；参考：《华中农业大学》2017年硕士论文

【摘要】：褐飞虱(Nilaparvata lugens(St?l))是水稻上一种非常重要的刺吸式口器害虫。作为近年来褐飞虱田间防治的常用杀虫剂,吡蚜酮一直被认为是通过抑制褐飞虱对韧皮部的取食使其饥饿而死,而近期研究表明吡蚜酮可能作用于昆虫TRPV通道(Transient receptor potential vaniloid channel,瞬时感受器电位香草酸受体亚型通道)。本研究通过RNA干扰的方法沉默褐飞虱TRPV通道两个亚型Inactive(Iav)和Nanchung(Nan)的基因,结果表明褐飞虱TRPV通道与取食行为关系密切。具体结果如下:1.克隆褐飞虱Nan和Iav基因,分别与果蝇和豌豆蚜的Nan基因和Iav基因进行序列比对。其中,Iav基因克隆片段与豌豆蚜和果蝇Iav基因相似度分别为83%和85%;Nan基因克隆片段与豌豆蚜和果蝇的相似度分别为85%和83%。进一步验证克隆所得片段即分别为褐飞虱Iav和Nan基因片段。2.将褐飞虱分别注射dsIav和dsNan 1 d和3 d后,Iav和Nan基因的表达量显著低于注射dsGFP的褐飞虱。Iav干扰后统计褐飞虱7 d内的存活率,结果表明,第3 d和第4 d存活率显著低于对照,而其他天数与对照相比无显著差异,而Nan基因干扰之后褐飞虱7 d之后存活率仅为26.67%,显著低于对照的存活率;RNA干扰后,褐飞虱的足均无异常情况。3.Iav基因和Nan基因干扰后,通过EPG监测成虫在水稻植株上的取食,结果表明,Iav和Nan基因干扰之后褐飞虱韧皮部取食均受到抑制。其中,Iav基因干扰后,口针的刺探次数和路径波时间延长而韧皮部取食时间和刺探次数显著低于对照。Nan基因干扰后,与对照相比木质部取食时间和非刺探时间延长,而韧皮部的取食时间和韧皮部刺探次数降低。4.将RNA干扰之后的褐飞虱通过EPG监测成虫对液态饲料取食,发现Iav基因和Nan基因干扰后对蔗糖溶液的取食与对照相比均无显著差异。综上所述,褐飞虱TRPV两个亚型基因的沉默会抑制褐飞虱对水稻植株的取食,但不影响对液态饲料的取食,因此褐飞虱TRPV不影响褐飞虱的主动抽吸能力但是否参与到昆虫-植物的互作则有待进一步验证。本研究证明褐飞虱TRPV通道与取食调控关系密切,对褐飞虱的取食机理、吡蚜酮作用靶标的研究以及褐飞虱的防治提供了理论基础。
[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.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S435.112.3

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