【摘要】：褐飛虱是一種典型的殺蟲(chóng)劑誘導(dǎo)的再猖獗型水稻重大害蟲(chóng)。目前,有關(guān)藥劑誘導(dǎo)褐飛虱再猖獗機(jī)制方面研究?jī)H局限于藥劑對(duì)雌蟲(chóng)生殖的刺激效應(yīng),忽視了雄蟲(chóng)在再猖獗中的作用。本論文將兩性交配的褐飛虱雌雄作為一個(gè)整體,采用RNAi飼喂法探討雄蟲(chóng)生殖相關(guān)基因NlSPATA5和NlPHF7經(jīng)交配對(duì)雌蟲(chóng)生殖及種群增長(zhǎng)的影響,進(jìn)一步明確褐飛虱雄蟲(chóng)精子形成基因NlSPATA5和精巢發(fā)育基因NlPHF7在褐飛虱再猖獗中的作用,從雄蟲(chóng)生殖效應(yīng)角度來(lái)揭示褐飛虱再猖獗的另一新機(jī)制。1.沉默褐飛虱雄蟲(chóng)NlSPATA5基因?qū)Υ葡x(chóng)生殖及其子代種群的影響NlSPATA5基因組織特異性表達(dá)分析,結(jié)果表明NlSPATA5基因僅在雄蟲(chóng)的內(nèi)生殖器和脂肪體中特異性表達(dá)。采用dsRNA飼喂法對(duì)NlSPATA5基因沉默率分析,結(jié)果表明NlSPATA5基因在不同發(fā)育時(shí)期(羽化后1、3、5、7d)基因表達(dá)量下降了約41.0%～67.3%;dsNlSPATA5處理后雄蟲(chóng)附腺蛋白質(zhì)含量與對(duì)照相比(Control和dsGFP)下降了約29.7%,并且導(dǎo)致輸精管發(fā)育畸形。此外,dsNlSPATA5處理經(jīng)交配對(duì)雌蟲(chóng)生殖力也產(chǎn)生一定影響,與對(duì)照相比,產(chǎn)卵量下降32.9%,產(chǎn)卵前期延長(zhǎng)31.3%,以及子代種群數(shù)量減少53.7%和卵孵化率下降16.9%,但是對(duì)產(chǎn)卵歷期和子代性別比沒(méi)有顯著影響。沉默NlSPATA5基因表達(dá)導(dǎo)致卵巢發(fā)育延遲,脂肪體和卵巢內(nèi)蛋白質(zhì)含量分別下降了 27.9%和49.6%,以及Nlvg表達(dá)水平下降了 34.4%。另外,不管在羽化后2天(2DPE)或羽化后3天(3DPE),脂肪體內(nèi)類酵母共生菌含量與對(duì)照相比均顯著下降。然而,無(wú)論是雌蟲(chóng)或雄蟲(chóng),dsNlSPATA5處理對(duì)褐飛虱成蟲(chóng)壽命和體重均沒(méi)有顯著性影響。2.沉默褐飛虱雄蟲(chóng)NlPHF7基因?qū)Υ葡x(chóng)生殖及其子代種群的影響NlPHF7基因組織特異性表達(dá)分析,結(jié)果表明NlPHF7基因僅在雄蟲(chóng)的內(nèi)生殖器和脂肪體中特異性表達(dá)。采用dsRNA飼喂法對(duì)NlPHF7基因沉默率分析,結(jié)果表明NlPHF7基因在不同發(fā)育時(shí)期(羽化后1、3、5、7d)基因表達(dá)量下降了約44.3%～60.2%;dsNlPHF7處理后雄蟲(chóng)附腺蛋白質(zhì)和精氨酸含量于對(duì)照相比分別下降了約16.6%和37.6%,雄成蟲(chóng)體重也下降了約21.3%,并且導(dǎo)致輸精管和精巢發(fā)育畸形。此外,dsNlPHF7處理經(jīng)交配對(duì)雌蟲(chóng)的生殖力也產(chǎn)生一定影響,與對(duì)照相比,產(chǎn)卵量下降了 40.9%,子代種群數(shù)量減少了 66.1%和卵孵化率下降了 18.0%,但對(duì)產(chǎn)卵前期、產(chǎn)卵歷期和子代性別比均沒(méi)有顯著性影響。沉默NlPHF7基因?qū)е侣殉舶l(fā)育延遲,雌成蟲(chóng)體重下降了 14.5%,卵巢內(nèi)蛋白質(zhì)含量下降了40.9%,以及Nlvg表達(dá)量也下降了 53.2%;然而,dsNlPHF7處理經(jīng)交配對(duì)褐飛虱雌蟲(chóng)脂肪體內(nèi)蛋白質(zhì)含量無(wú)顯著性影響;不管在羽化后2天(2DPE)或3天(3DPE),脂肪體內(nèi)類酵母共生菌含量與對(duì)照相比均顯著性下降。然而,無(wú)論是雌蟲(chóng)或雄蟲(chóng),dsNlPHF7處理對(duì)褐飛虱成蟲(chóng)壽命均沒(méi)有顯著性影響。
[Abstract]:Brown planthopper is a typical insecticide-induced recurrent rice pest. At present, the research on the mechanism of reactivity induced by insecticides is limited to the stimulative effect of insecticides on female reproduction, and neglects the role of males in the reproduction of brown planthopper. In this paper, the male and female sex of hermaphroditic brown planthopper (BPH) was studied by RNAi feeding method to study the effect of mating of NlSPATA5 and NlPHF7 genes on female reproduction and population growth. The roles of male spermatogenesis gene NlSPATA5 and spermatozoa development gene NlPHF7 in the reoccurrence of brown planthopper were further clarified, and another new mechanism of brown planthopper reoccurrence was revealed from the angle of male reproductive effect. 1. Effects of silencing NlSPATA5 gene of male brown planthopper on reproduction and offspring of female planthopper the specific expression of NlSPATA5 gene in tissues was analyzed. The results showed that NlSPATA5 gene was expressed only in the male genitalia and fat body. The silencing rate of NlSPATA5 gene was analyzed by dsRNA feeding method. The results showed that the expression of NlSPATA5 gene decreased by 41.0% and 67.3% at different developmental stages (1D after Eclosion). Compared with the control group (Control and dsGFP), the protein content of accessory gland of male worms decreased by 29.7% after dsNlSPATA5 treatment, and resulted in deformity of vas deferens. In addition, the fecundity of female worms was also affected by mating with dsNlSPATA5. Compared with the control, the oviposition decreased by 32.9m, the preoviposition was prolonged by 31.3%, the number of offspring decreased 53.7% and the hatchability of eggs decreased 16.9%. However, there was no significant effect on the oviposition duration and the sex ratio of offspring. Silencing NlSPATA5 gene expression resulted in delayed ovarian development, decreased protein content in fat body and ovary by 27.9% and 49.6%, and decreased Nlvg expression level by 34.4g%. In addition, the content of yeast symbiotic bacteria in adipose body was significantly lower than that in control group, no matter 2 days after Eclosion (2DPE) or 3 days after Eclosion (3DPE). However, dsNlSPATA5 treatment had no significant effect on adult life and body weight of brown planthopper, whether female or male. Effects of silencing NlPHF7 gene of male brown planthopper on reproduction and offspring of female planthopper the specific expression of NlPHF7 gene in tissues was analyzed. The results showed that NlPHF7 gene was expressed only in the male genitalia and fat body. The silencing rate of NlPHF7 gene was analyzed by dsRNA feeding method. The results showed that the expression of NlPHF7 gene decreased about 44.30.2in different developmental stages (1D after Eclosion). After dsNlPHF7 treatment, the contents of protein and arginine in accessory gland of male worm decreased by 16.6% and 37.6%, respectively, and the weight of male adult decreased by 21.3%, which resulted in deformity of vas deferens and spermatozoa. In addition, the fecundity of female was also affected by mating with dsNlPHF7. Compared with the control, oviposition decreased by 40.9%, the number of offspring decreased by 66.1% and the hatchability of eggs decreased by 18.0%, but in the early stage of oviposition, the number of eggs was decreased by 66.1%, and the hatching rate of eggs was decreased by 18.0%. There was no significant effect on the oviposition period and the sex ratio of offspring. The silencing of NlPHF7 gene resulted in delayed ovarian development, the weight of female adult decreased 14.5%, the protein content of ovary decreased 40.9%, and the expression of Nlvg decreased 53.2%. However, after mating with dsNlPHF7, there was no significant effect on protein content in fat of female brown planthopper, but the content of yeast symbiotic bacteria in fat body was significantly lower than that in control group, no matter 2 days after emergence (2DPE) or 3 days (3DPE). However, dsNlPHF7 treatment had no significant effect on adult life of brown planthopper, whether female or male.
【學(xué)位授予單位】：揚(yáng)州大學(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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本文編號(hào)：2397977