昆虫学报 ›› 2021, Vol. 64 ›› Issue (12): 1377-1387.doi: 10.16380/j.kcxb.2021.12.003

• 研究论文 • 上一篇    下一篇

营养对褐飞虱生长和繁殖信号通路的影响

康奎1, 蔡永进2, 张道伟1, 龚俊1, 张文庆2,*   

  1. (1. 遵义师范学院生物与农业科技学院, 贵州省赤水河流域动物资源保护与应用研究重点实验室, 贵州遵义 563002; 2. 中山大学生命科学学院, 有害生物控制与资源利用国家重点实验室, 广州 510275)
  • 出版日期:2021-12-20 发布日期:2021-11-26

Effects of nutrition on the growth and reproductive signaling pathways in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)

KANG Kui1, CAI Yong-Jin2, ZHANG Dao-Wei1, GONG Jun1, ZHANG Wen-Qing2,*   

  1.  (1. Laboratory of Regional Characteristic for Conservation and Utilization of Animal Resource in Chishui River Basin, College of Biology and Agriculture, Zunyi Normal University, Zunyi, Guizhou 563002, China; 2. State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-sen University, Guangzhou, 510275, China)
  • Online:2021-12-20 Published:2021-11-26

摘要:

【目的】昆虫感知外界营养状态,在体内营养信号通过许多信号通路进行传递,进而调控昆虫的生长和繁殖。本研究旨在为营养调控褐飞虱Nilaparvata lugens生长和繁殖的分子机制作出初步探索。【方法】在不同营养条件[100%浓度饲料(全纯人工饲料D-97, 对照)、50%浓度人工饲料和25%浓度人工饲料]下饲喂褐飞虱3龄第2天若虫至成虫,统计体重、发育历期、存活率和每卵巢成熟卵量;利用荧光定量PCR检测羽化后第2天褐飞虱成虫不同组织(卵巢、脂肪体和其他组织)中IIS, TOR以及AMPK信号通路相关基因(InR1, InR2, AKT, FoxO, TOR, S6K, 4EBP, AMPKα,AMPKβAMPKγ)的表达量;利用Western blot方法检测羽化后第6天褐飞虱成虫卵巢和其他组织中Akt, FoxO和AMPK蛋白磷酸化水平;同时检测羽化后第6天褐飞虱成虫不同组织中的ROS水平以及不同龄期褐飞虱体内保幼激素(juvenile hormone, JH)和蜕皮激素(20-hydroxyecdysone, 20E)滴度。【结果】与饲喂100%浓度人工饲料的对照相比,用低浓度人工饲料饲喂褐飞虱至成虫导致5龄第2天至成虫体重显著降低,从3龄第2天若虫至成虫发育历期缩短,死亡率提高,每卵巢成熟卵量显著降低。取食低浓度人工饲料后,羽化后第2天成虫IIS, TOR以及AMPK信号通路相关基因InR2, TOR, 4EBP, AMPKα在卵巢、脂肪体和其他组织中, AKT, S6KAMPKγ在脂肪体和其他组织中,以及InR1, FoxOAMPKβ在其他组织中表达量较对照均显著下降,但卵巢中InR1, AKT, FoxOAMPKγ的表达量相对稳定;羽化后第6天成虫卵巢和其他组织中AKT, FoxO以及AMPK的磷酸化水平显著增加。随着人工饲料饲喂浓度的降低,羽化后第6天褐飞虱成虫不同组织中的ROS水平显著增加。不同营养条件下褐飞虱4龄第2天若虫体内JH滴度无显著差异,低营养条件下5龄第2天若虫体内JH滴度较对照显著降低,而在羽化后第2天成虫体内JH滴度则较对照显著增加;低营养条件导致褐飞虱若虫以及成虫体内20E滴度均显著增加。【结论】褐飞虱在面临营养缺失的情况下,营养信号传导通路(IIS, TOR以及APMK通路)的关键基因表达均下降,而ROS水平显著上升,通过调节AKT, FoxO以及AMPK的磷酸化水平,进而影响JH以及20E的生物合成。

关键词: 褐飞虱, IIS/TOR通路, AMPK, ROS, 蜕皮激素, 保幼激素

Abstract: 【Aim】 Insects perceive external nutritional status, and nutritional signals in vivo are transmitted through many signal pathways to regulate insect growth and reproduction. This study aims to tentatively explore the molecular mechanism of nutritional regulation on the growth and reproduction of the rice brown planthopper, Nilaparvata lugens. 【Methods】 The day-2 3rd instar nymphs of N. lugens were fed to adults under different nutrient conditions [fed with 100% artificial diet (pure artificial diet D-97, the control), 50% artificial diet and 25% artificial diet, respectively], and the body weight, developmental duration, survival rate and number of mature eggs per ovary were counted. Then, the relative expression levels of IIS, TOR and AMPK signaling pathway related genes (InR1, InR2, AKT, FoxO, TOR, S6K, 4EBP, AMPKα, AMPKβ and AMPKγ) in different tissues (ovary, body fat and other tissues) of the 2-day-old adults were detected by quantitative PCR, and the phosphorylation levels of Akt, FoxO and AMPK in the ovary and other tissues of the 6-day-old adults were detected by Western blot. Meanwhile, the ROS levels in different tissues of the 6-day-old adults and the titers of juvenile hormone (JH) and 20-hydroxyecdysone (20E) in N. lugens at different developmental stages were also detected. 【Results】 Compared with the control fed with 100% artificial diet, the artificial diet of lower concentrations resulted in a significant decrease in body weight from the day-2 5th instar nymph to adult, shortened developmental duration from day-2 3rd instar nymphs to adult, an increased mortality, and a significant decrease in the number of mature eggs per ovary of N. lugens. The expression levels of IIS, TOR and AMPK signaling pathway related genes InR2, TOR, 4EBP and AMPKα in the ovary, body fat and other tissues, AKT, S6K and AMPKγ in the fat body and other tissues, and InR1, FoxO and AMPKβ in the other tissues of the 2-day-old adults fed with the artificial diet of lower concentrations decreased significantly as compared to those in the control, while those of InR1, AKT, FoxO, and AMPKγ  in the ovary were relatively stable. The phosphorylation levels of AKT, FoxO, and AMPK in the ovary and other tissues of the 6-day-old adults fed with the artificial diet of lower concentrations were significantly increased as compared with those of the control. With the decrease of nutrient concentration in the artificial diet, the ROS levels in different tissues of the 6-day-old adults increased significantly as compared to that in the control. The JH titers in the day-2 4th instar nymphs under different nutrient conditions showed no significant difference, while the JH titers in the day-2 5th instar nymphs and 2-day-old adults under low nutrient conditions were significantly decreased and increased, respectively, as compared to those in the control. Low nutrient conditions resulted in a significant increase in 20E titer both in nymphs and adults. 【Conclusion】 Under conditions of nutrient deficiency, the expression levels of key genes in the nutritional signal transduction pathways including IIS, TOR and APMK pathways in N. lugens decrease, while the ROS level significantly increases, and the biosynthesis of JH and 20E is affected by regulating the phosphorylation levels of AKT, FoxO and AMPK.

Key words: Nilaparvata lugens, IIS/TOR pathway, AMPK, ROS, 20-hydroxyecdysone, juvenile hormone