褐飞虱雄虫生殖基因NlSPATA5和NlPHF7调控雌虫生殖及种群增长的研究

发布时间：2019-01-01 18:52

【摘要】：褐飞虱是一种典型的杀虫剂诱导的再猖獗型水稻重大害虫。目前,有关药剂诱导褐飞虱再猖獗机制方面研究仅局限于药剂对雌虫生殖的刺激效应,忽视了雄虫在再猖獗中的作用。本论文将两性交配的褐飞虱雌雄作为一个整体,采用RNAi饲喂法探讨雄虫生殖相关基因NlSPATA5和NlPHF7经交配对雌虫生殖及种群增长的影响,进一步明确褐飞虱雄虫精子形成基因NlSPATA5和精巢发育基因NlPHF7在褐飞虱再猖獗中的作用,从雄虫生殖效应角度来揭示褐飞虱再猖獗的另一新机制。1.沉默褐飞虱雄虫NlSPATA5基因对雌虫生殖及其子代种群的影响NlSPATA5基因组织特异性表达分析,结果表明NlSPATA5基因仅在雄虫的内生殖器和脂肪体中特异性表达。采用dsRNA饲喂法对NlSPATA5基因沉默率分析,结果表明NlSPATA5基因在不同发育时期(羽化后1、3、5、7d)基因表达量下降了约41.0%～67.3%;dsNlSPATA5处理后雄虫附腺蛋白质含量与对照相比(Control和dsGFP)下降了约29.7%,并且导致输精管发育畸形。此外,dsNlSPATA5处理经交配对雌虫生殖力也产生一定影响,与对照相比,产卵量下降32.9%,产卵前期延长31.3%,以及子代种群数量减少53.7%和卵孵化率下降16.9%,但是对产卵历期和子代性别比没有显著影响。沉默NlSPATA5基因表达导致卵巢发育延迟,脂肪体和卵巢内蛋白质含量分别下降了 27.9%和49.6%,以及Nlvg表达水平下降了 34.4%。另外,不管在羽化后2天(2DPE)或羽化后3天(3DPE),脂肪体内类酵母共生菌含量与对照相比均显著下降。然而,无论是雌虫或雄虫,dsNlSPATA5处理对褐飞虱成虫寿命和体重均没有显著性影响。2.沉默褐飞虱雄虫NlPHF7基因对雌虫生殖及其子代种群的影响NlPHF7基因组织特异性表达分析,结果表明NlPHF7基因仅在雄虫的内生殖器和脂肪体中特异性表达。采用dsRNA饲喂法对NlPHF7基因沉默率分析,结果表明NlPHF7基因在不同发育时期(羽化后1、3、5、7d)基因表达量下降了约44.3%～60.2%;dsNlPHF7处理后雄虫附腺蛋白质和精氨酸含量于对照相比分别下降了约16.6%和37.6%,雄成虫体重也下降了约21.3%,并且导致输精管和精巢发育畸形。此外,dsNlPHF7处理经交配对雌虫的生殖力也产生一定影响,与对照相比,产卵量下降了 40.9%,子代种群数量减少了 66.1%和卵孵化率下降了 18.0%,但对产卵前期、产卵历期和子代性别比均没有显著性影响。沉默NlPHF7基因导致卵巢发育延迟,雌成虫体重下降了 14.5%,卵巢内蛋白质含量下降了40.9%,以及Nlvg表达量也下降了 53.2%;然而,dsNlPHF7处理经交配对褐飞虱雌虫脂肪体内蛋白质含量无显著性影响;不管在羽化后2天(2DPE)或3天(3DPE),脂肪体内类酵母共生菌含量与对照相比均显著性下降。然而,无论是雌虫或雄虫,dsNlPHF7处理对褐飞虱成虫寿命均没有显著性影响。
[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.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S435.112.3
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本文编号：2397977