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    白细胞激肽及其受体对美国白蛾取食和生长发育的影响

    Effects of leucokinin neuropeptide and its receptor on feeding and growth of Hyphantria cunea

    • 摘要:
      目的 探究神经肽LK及其受体对美国白蛾生长发育的影响。
      方法 构建美国白蛾基因LK/LKR dsRNA合成的体外原核表达系统,并饲喂美国白蛾幼虫表达dsRNA的菌液,统计其取食量、体重和发育历期。对美国白蛾4龄幼虫注射LK1和LK2短肽,检测72 h后中肠和脂肪体组织中消化酶活性及海藻糖、糖原和葡萄糖含量,并用RT-qPCR技术检测相关基因的相对表达量。
      结果 细菌介导的原核表达系统可有效沉默LKLKR基因,影响美国白蛾4龄幼虫48 h内取食量,但不显著(P > 0.05),并导致体重累计增长率显著提高(P < 0.05)(除L4440-HcLKR处理组幼虫外),发育历期延迟。注射LKs短肽的美国白蛾4龄幼虫取食量和体重累计增长率显著降低,24 h粪便含水量显著升高,但幼虫体内含水量无显著变化;进一步研究显示72 h中肠的α-淀粉酶以及脂肪体中的脂肪酶、α-淀粉酶、胰蛋白酶活性显著增强;中肠的糖原含量显著减少,而脂肪体中的糖原、海藻糖含量显著升高,葡萄糖含量显著降低;中肠和脂肪体中的脂肪酶基因LIP(除脂肪体的LK1组)和α-淀粉酶基因(amy-1和amy-2)表达量显著升高;中肠海藻糖酶基因(Tre1和Tre2)与海藻糖转运蛋白基因Tret1表达量显著升高;脂肪体Tre1(除LK2组)和Tre2基因表达量显著下降,Tret1基因(LK2组)表达量显著升高。
      结论 本研究明确了白细胞激肽参与调节美国白蛾取食、能量代谢和生长发育过程,为研发绿色杀虫剂防控美国白蛾提供理论依据。

       

      Abstract:
      Objective This paper investigates the effects of leucokinin (LK) neuropeptide and its receptor on growth of Hyphantria cunea.
      Method An in vitro prokaryotic expression system was developed for synthesize of LK/LKR dsRNA. The 4th instar larvae of Hyphantria cunea were fed with dsRNA-expressing bacteria. The food intake, body mass and developmental duration were measured and recorded. Subsequently, the 4th instar larvae of Hyphantria cunea were injected with short LK1 and LK2 peptide. Digestive enzyme activities, trehalose, glycogen, and glucose content in the midgut and fat body were detected after 72 h. And the relative expression level of related genes was detected using RT-qPCR technology.
      Result Bacteria-mediated RNAi effectively silenced LK and LKR genes. It affected the food intake of the 4th instar larvae of Hyphantria cunea within 48 h, but it was not significant (P > 0.05). It led to a significant increase in relative growth rate within 48 h, and a delay in developmental duration. It led to an increase in feeding amount (P > 0.05) and a significant increase in relative accumulation growth rate within 48 h, and delayed the development time. After injection of LKs short peptide, the feeding amount and relative accumulation growth rate of 4th instar larvae of Hyphantria cunea were significantly reduced, and the water content of feces was significantly increased at 24 h after injection, but the water content of larvae was not significantly different. Further studies showed the activities of α-amylase in the midgut, and lipase, α-amylase, trypsin, trehalase in the fat body were significantly enhanced at 72 h after injection. The content of glycogen in the midgut decreased significantly. The content of glycogen and trehalose in the fat body increased significantly, while the content of glucose was significantly reduced. The expression level of LIP genes (excluding LK1 group in the fat body), and amy-1 and amy-2 genes significantly increased in the midgut and fat body. The expression level of Tre1, Tre2 and Tret1 genes increased significantly in the midgut. The expression level of Tre1 (excluding LK2 group) and Tre2 decreased, while the expression level of Tret1 (LK2 group) significantly increased in the fat body.
      Conclusion This study reveals that LK is involved in regulating feeding, energy metabolism and growth in Hyphantria cunea, providing a theoretical basis for the development of green insecticides to control Hyphantria cunea.

       

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