胰岛素受体基因InR调控褐飞虱海藻糖代谢研究
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摘要
在昆虫中已发现成熟的典型胰岛素信号通路,但是其调控海藻糖代谢途径的机制还未清晰。为探讨胰岛素受体基因在褐飞虱海藻糖代谢平衡及其发育的调控作用,本文采用RNAi技术抑制胰岛素受体(InR)基因的表达,测定处理后海藻糖、糖原和葡萄糖含量及海藻糖酶活变化,并检测InR、类胰岛素多肽(Ilp)、海藻糖代谢途径中关键基因的表达。研究结果表明dsRNA注射后能够显著抑制Ilp和InR基因的表达;InR1低表达后72 h能够显著抑制3种糖类物质的含量;InR表达抑制后72 h可溶性海藻糖酶活性上升,而膜结合型海藻糖酶活性下降;当InR表达受抑制后3个海藻糖酶和2个海藻糖合成酶基因的表达都显著下降。这些结果说明InR能够影响海藻糖等糖类物质的平衡。从而为将来通过调控昆虫血糖平衡来控制害虫提供理论依据。
Mature insulin signaling pathways have been found in insects, but the mechanism of its regulation of trehalose metabolic pathways is not yet clear. These experiments were design for to investigate the effects of insulin signal pathways on the balance and development of trehalose metabolism in Nilaparvata lugens. After the expression of insulin receptor(InR) gene were inhibited by RNAi technology, the content of trehalose, glycogen and glucose, the trehalase activity changes were detected at 48 h and 72 h. In addition, the expression of InR, insulin-like polypeptide(Ilp), and some genes from trehalose metabolic pathway were also detected at 48 h and 72 h. These results showed that dsRNA could significantly inhibit the expression of Ilp and InR genes. The expression of InR1 was inhibited could significantly decreased the contents of three kinds of carbohydrates at 72 h, followed the membrane-bound trehalase activity decreased, but the activity of soluble trehalase increased at 72 h. The expression of three trehalases and two trehalose-6-phosphate synthase genes decreased significantly when the expressions of InR were inhibited. These results indicate that InR can affect the balance of carbohydrates, such as trehalose. These results will provide a theoretical basis for controlling insect pests by regulating insect blood glucose balance in the future.
Mature insulin signaling pathways have been found in insects, but the mechanism of its regulation of trehalose metabolic pathways is not yet clear. These experiments were design for to investigate the effects of insulin signal pathways on the balance and development of trehalose metabolism in Nilaparvata lugens. After the expression of insulin receptor(InR) gene were inhibited by RNAi technology, the content of trehalose, glycogen and glucose, the trehalase activity changes were detected at 48 h and 72 h. In addition, the expression of InR, insulin-like polypeptide(Ilp), and some genes from trehalose metabolic pathway were also detected at 48 h and 72 h. These results showed that dsRNA could significantly inhibit the expression of Ilp and InR genes. The expression of InR1 was inhibited could significantly decreased the contents of three kinds of carbohydrates at 72 h, followed the membrane-bound trehalase activity decreased, but the activity of soluble trehalase increased at 72 h. The expression of three trehalases and two trehalose-6-phosphate synthase genes decreased significantly when the expressions of InR were inhibited. These results indicate that InR can affect the balance of carbohydrates, such as trehalose. These results will provide a theoretical basis for controlling insect pests by regulating insect blood glucose balance in the future.
引文
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Chen XA, Yao HW, Ye GY. Research advances on insulin-like peptides and their functions in insects [J]. Chinese Journal of Biological Control, 2017, 33 (5): 699-712. [陈晓昂, 姚洪渭, 叶恭银. 昆虫类胰岛素及其功能研究进展 [J]. 中国生物防治学报, 2017, 33 (5): 699-712]
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He LM, Shentu XP, Li DT, et al. Effects of fungicide insecticide mixtures on population growth of brown planthopper, Nilaparvata lugens St?l (Hemiptera: Delphacidae) [J]. Chinese Journal of Biological Control, 2018, 2: 1-6. [何林懋, 申屠旭萍, 李丹婷, 等. 杀菌剂与吡虫啉混合配剂对褐飞虱种群增长的影响 [J]. 中国生物防治学报, 2018, 2: 1-6]
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Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method [J]. Methods, 2001, 25 (4): 402-408.
Lu B. Genetic and Functional Analysis of Insulin Receptor Genes of Brown Planthopper (Nilaparvata lugens) [D]. Zhejiang: Zhejiang University, 2015. [卢博. 褐飞虱胰岛素受体基因分析与功能研究 [D]. 浙江: 浙江大学硕士学位论文, 2015]
Luo J, Lushchak OV, Goergen P,et al. Drosophila insulin-producing cells are differentially modulated by serotonin and octopamine receptors and affect social behavior [J]. PLoS ONE, 2014, 9 (6): e99732.
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Qin JM, Luo SD, He SY, et al. Researching in characters and functions of trehalose and trehalase in insects [J]. Journal of Environmental Entomology, 2015, 37 (1): 163-169. [秦加敏, 罗术东, 和绍禹, 等. 昆虫海藻糖与海藻糖酶的特性及功能研究 [J]. 环境昆虫学报, 2015, 37 (1): 163-169]
Rao XJ. The Effect of Inhibiting FoxO1 on the Glucose Consumption of Insulin Resistant Cells and the Mechanism [D]. Zhengzhou: Zhengzhou University, 2009. [绕小娟. FoxO1对胰岛素抵抗细胞葡萄糖消耗的影响及机制研究 [D]. 郑州: 郑州大学硕士学位论文, 2009]
Ren XM, Xiang C, Lei DY, et al. Present status and prospect of resistance breeding of brown planthopper in rice [J]. Crop Research, 2017, 31 (4): 453-458, 462. [任西明, 向聪, 雷东阳, 等. 水稻抗褐飞虱育种研究进展与展望 [J]. 作物研究, 2017, 31 (4): 453-458, 462]
Rulifson EJ, Kim SK, Nusse R. Ablation of insulin-producing neurons in flies: Growth and diabetic phenotypes [J]. Science, 2002: 296.
Satake S, Masumura M, Ishizaki H,et al. Bombyxin, an insulin-related peptide of insects, reduces the major storage carbohydrates in the silk worm Bombyx mori [J]. Comparative Biochemistry and Physiology, Part B, Biochemistry and Molecular Biology, 1997, 118 (2): 349-357.
Satake S, Nagata K, Kataoka H,et al. Bombyx insecretion in the adult silk moth Bombyx mori: Sex-specificity and its correlation with metabolism [J]. Journal of Insect Physiology, 1999, 45 (10): 939-945.
Scott JG, Michel K, Bartholomay LC,et al. Towards the elements of successful insect RNAi [J]. Journal of Insect Physiology, 2013, 59 (12): 1212-1221.
Süren-Castillo S, Abrisqueta M, Maestro JL. FoxO is required for the activation of hypertrehalosemic hormone expression in cockroaches [J]. Biochimicaet Biophysica Acta, 2014, 1840: 86-94.
Tang B, Chen X, Liu Y, et al. Characterization and expression 90 patterns of a membrane-bound trehalase from Spodoptera exigua [J]. BMC Molecular Biology, 2008, 9: 51.
Tang B, Wei P, Zhao LN,et al. Knockdown of five trehalase genes using RNA interference regulates the gene expression of the chitin biosynthesis pathways in Tribolium castaneum [J]. BMC Biotechnology, 2016, 16: 67.
Tang B, Zhang L, Xiong XP, et al. Advances in trehalose metabolism and its regulation of insect chitin synthesis [J]. Scientia Agricultura Sinica, 2018, 51 (4): 697-707. [唐斌, 张露, 熊旭萍, 等. 海藻糖代谢及其调控昆虫几丁质合成研究进展 [J].中国农业科学, 2018, 51 (4): 697-707]
Tang B, Wei P, Chen J, et al. Progress in gene features and functions of insect trehalases [J]. Acta Entomologica Sinica, 2012, 55 (11): 1315-1321. [唐斌, 魏苹, 陈洁, 等. 昆虫海藻糖酶的基因特性及功能研究进展 [J]. 昆虫学报, 2012, 55 (11): 1315-1321]
Xi Y, Pan PL, Ye YX,et al. Chitinase-like gene family in the brown planthopper, Nilaparvata lugens [J]. Insect Molecular Biology, 2015, 24 (1): 29-40.
Xi Y, Pan PL, Ye YX,et al. Chitindeacetylase family genes in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae) [J]. Insect Molecular Biology, 2014, 23 (6): 695-705.
Xi Y, Pan PL, Zhang CX. The β-N-acetylhexosaminidase gene family in the brown planthopper, Nilaparvata lugens [J]. Insect Molecular Biology, 2015, 24 (6): 601-610.
Xi Y. Analyses of Chitinolvtic Enzyme Gene Families in Nilaparvata lugens [D]. Zhejiang: Zhejiang University, 2014. [席羽. 褐飞虱几丁质降解酶系基因家族分析 [D]. 浙江: 浙江大学博士学位论文, 2014]
Xu HJ, Xue J, Lu B,et al. Two insulin receptors determine alternative wing morphs in planthoppers [J]. Nature, 2015, 519: 464-479.
Xu J. Molecular Mechanism of the Wing Dimorphism of Brown Plantopper, Nilaparvata lugens [D]. Hangzhou: Zhejiang University, 2015. [薛建. 褐飞虱翅型分化分子机理研究 [D]. 杭州: 浙江大学博士学位论文, 2015]
Xu KK. Studies on the Expression and Function of Six Insulin Signaling Pathway Genes from Bactrocera dorsalis [D]. Chongqing: Southwest University, 2014. [许抗抗. 桔小实蝇胰岛素信号通路六个重要基因的表达与功能研究 [D]. 重庆: 西南大学硕士学位论文, 2014]
Yang MM, Zhao LN, Shen QD,et al. Knockdown of two trehalose-6-phosphate synthases severely affects chitin metabolism gene expression in the rice brown planthopper Nilaparvata lugens [J]. Pest Management Science, 2016, 73: 206-216.
Yu CH, Lu D, Lin RH, et al. Trehalose-the blood sugar in insects [J]. Chinese Bulletin of Entomology, 2008, 45 (5): 832-837. [于彩虹, 卢丹, 林荣华, 等. 海藻糖-昆虫的血糖 [J]. 昆虫知识, 2008, 45 (5): 832-837]
Zhai YF, Sun ZX, Zhang JQ,et al. Activation of the TOR signaling pathway by glutamine regulates insect fecundity [J]. Scientific Reports, 2015: 1-9.
Zhang G, Li XJ, Yu JL, et al. The continuous outbreaks of Nilaparvata lugens(St?l) (Hemiptera: Delphacidae): A case study in Taihe county, Jiangxi province, China [J]. Acta Ecologica Sinica, 2018, 38 (9): 1-9. [张国, 李袭杰, 于居龙, 等. 褐飞虱持续暴发成因: 以江西泰和县为例 [J]. 生态学报, 2018, 38 (9): 1-9]
Zhang L, Zhu SC, Zheng H,et al. Regulatory function of trehalase genes on chitin metabolism in the cuticle of Nilaparvata lugens [J]. Scientia Agricultura Sinica, 2017, 50 (6): 1047-1056. [张露, 朱世城, 郑好, 等. 褐飞虱海藻糖酶基因在表皮几丁质代谢中的调控作用 [J]. 中国农业科学, 2017, 50 (6): 1047-1056]
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Zhao LN, Yang MM, Shen QD,et al. Knockdown of three trehalases regulating trehalose and chitin metabolism in the rice brown planthopper Nilaparvata lugens [J]. Scientific Reports, 2016, 6: 27841.
Zhu QS, Arakane Y, Beeman RW,et al. Functional specialization among insect chitinase family genes revealed by RNA interference [J]. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105 (18): 6650-6655.
Chen J, Zhang DW, Qian ZM. Characteristics and bacterial-induced expression analysis of two prophenoloxidase genes in the brown planthopper, Nilaparvata lugens (Hemiptera Delphacidae) [J]. Chinese Journal of Biological Control, 2018, 34 (1): 44-51. [陈静, 张道伟, 钱正敏. 褐飞虱两个酚氧化酶原基因的特性及细菌诱导后的表达分析 [J]. 中国生物防治学报, 2018, 34 (1): 44-51]
Chen XA, Yao HW, Ye GY. Research advances on insulin-like peptides and their functions in insects [J]. Chinese Journal of Biological Control, 2017, 33 (5): 699-712. [陈晓昂, 姚洪渭, 叶恭银. 昆虫类胰岛素及其功能研究进展 [J]. 中国生物防治学报, 2017, 33 (5): 699-712]
Deng CL, Xie HL, Zhang O. Measures of drug resistance monitoring and comprehensive control of rice brown planthopper, Nilaparvata lugens in Hubei province, 2010-2017 [J]. Hubei Plant Protection, 2018, 1: 26-29. [邓春林, 谢华伦, 张欧. 2010-2017年湖北省水稻褐飞虱抗药性监测及综合治理措施 [J]. 湖北植保, 2018, 1: 26-29]
Ghosh A, Chatterjee ML. Field evaluation of some new insecticides against brown plant hopper, Nilaparvata lugens (St?l) in rice [J]. Journal of Insect Science, 2012, 25 (3): 247-249.
Hall KD. A review of the carbohydrate-insulin model of obesity [J]. European Journal of Clinical Nutrition, 2017, 71 (3): 323-326.
He LM, Shentu XP, Li DT, et al. Effects of fungicide insecticide mixtures on population growth of brown planthopper, Nilaparvata lugens St?l (Hemiptera: Delphacidae) [J]. Chinese Journal of Biological Control, 2018, 2: 1-6. [何林懋, 申屠旭萍, 李丹婷, 等. 杀菌剂与吡虫啉混合配剂对褐飞虱种群增长的影响 [J]. 中国生物防治学报, 2018, 2: 1-6]
He SF. Genetic andFunctional Analysis of Insulin-like Genes of Brown Planthopper (Nilaparvata lugens) [D]. Zhejiang: Zhejiang University, 2015. [何叔舫. 褐飞虱类胰岛素基因分析与功能研究 [D]. 浙江: 浙江大学硕士学位论文, 2015]
Kim Y, Hong Y. Regulation of hemolymph trehalose level by an insulin-like peptide through diel feeding rhythm of the beet armyworm, Spodoptera exigua [J]. Peptides, 2015, 68: 91-98.
Li ZH. Study on the Effects of Sublethal Concentration of Insecticides on Juvenile Hormone Metabolic Enzymes and Insulin-like Genes of Nilaparvata lugens St?l (Hemiptera: Delphacidae) [D]. Jiangsu: Yangzhou University, 2017. [李正辉. 杀虫剂亚致死浓度对褐飞虱保幼激素水解酶及类胰岛素基因的影响研究 [D]. 江苏: 扬州大学硕士学位论文, 2017]
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method [J]. Methods, 2001, 25 (4): 402-408.
Lu B. Genetic and Functional Analysis of Insulin Receptor Genes of Brown Planthopper (Nilaparvata lugens) [D]. Zhejiang: Zhejiang University, 2015. [卢博. 褐飞虱胰岛素受体基因分析与功能研究 [D]. 浙江: 浙江大学硕士学位论文, 2015]
Luo J, Lushchak OV, Goergen P,et al. Drosophila insulin-producing cells are differentially modulated by serotonin and octopamine receptors and affect social behavior [J]. PLoS ONE, 2014, 9 (6): e99732.
N?ssel DR, Broeck JV. Insulin/IGF signaling in Drosophila and other insects: Factors that regulate production, release and post-release action of the insulinlike peptides [J]. Cellular and Molecular Life Sciences, 2016, 73 (2): 271-290.
Qin JM, Luo SD, He SY, et al. Researching in characters and functions of trehalose and trehalase in insects [J]. Journal of Environmental Entomology, 2015, 37 (1): 163-169. [秦加敏, 罗术东, 和绍禹, 等. 昆虫海藻糖与海藻糖酶的特性及功能研究 [J]. 环境昆虫学报, 2015, 37 (1): 163-169]
Rao XJ. The Effect of Inhibiting FoxO1 on the Glucose Consumption of Insulin Resistant Cells and the Mechanism [D]. Zhengzhou: Zhengzhou University, 2009. [绕小娟. FoxO1对胰岛素抵抗细胞葡萄糖消耗的影响及机制研究 [D]. 郑州: 郑州大学硕士学位论文, 2009]
Ren XM, Xiang C, Lei DY, et al. Present status and prospect of resistance breeding of brown planthopper in rice [J]. Crop Research, 2017, 31 (4): 453-458, 462. [任西明, 向聪, 雷东阳, 等. 水稻抗褐飞虱育种研究进展与展望 [J]. 作物研究, 2017, 31 (4): 453-458, 462]
Rulifson EJ, Kim SK, Nusse R. Ablation of insulin-producing neurons in flies: Growth and diabetic phenotypes [J]. Science, 2002: 296.
Satake S, Masumura M, Ishizaki H,et al. Bombyxin, an insulin-related peptide of insects, reduces the major storage carbohydrates in the silk worm Bombyx mori [J]. Comparative Biochemistry and Physiology, Part B, Biochemistry and Molecular Biology, 1997, 118 (2): 349-357.
Satake S, Nagata K, Kataoka H,et al. Bombyx insecretion in the adult silk moth Bombyx mori: Sex-specificity and its correlation with metabolism [J]. Journal of Insect Physiology, 1999, 45 (10): 939-945.
Scott JG, Michel K, Bartholomay LC,et al. Towards the elements of successful insect RNAi [J]. Journal of Insect Physiology, 2013, 59 (12): 1212-1221.
Süren-Castillo S, Abrisqueta M, Maestro JL. FoxO is required for the activation of hypertrehalosemic hormone expression in cockroaches [J]. Biochimicaet Biophysica Acta, 2014, 1840: 86-94.
Tang B, Chen X, Liu Y, et al. Characterization and expression 90 patterns of a membrane-bound trehalase from Spodoptera exigua [J]. BMC Molecular Biology, 2008, 9: 51.
Tang B, Wei P, Zhao LN,et al. Knockdown of five trehalase genes using RNA interference regulates the gene expression of the chitin biosynthesis pathways in Tribolium castaneum [J]. BMC Biotechnology, 2016, 16: 67.
Tang B, Zhang L, Xiong XP, et al. Advances in trehalose metabolism and its regulation of insect chitin synthesis [J]. Scientia Agricultura Sinica, 2018, 51 (4): 697-707. [唐斌, 张露, 熊旭萍, 等. 海藻糖代谢及其调控昆虫几丁质合成研究进展 [J].中国农业科学, 2018, 51 (4): 697-707]
Tang B, Wei P, Chen J, et al. Progress in gene features and functions of insect trehalases [J]. Acta Entomologica Sinica, 2012, 55 (11): 1315-1321. [唐斌, 魏苹, 陈洁, 等. 昆虫海藻糖酶的基因特性及功能研究进展 [J]. 昆虫学报, 2012, 55 (11): 1315-1321]
Xi Y, Pan PL, Ye YX,et al. Chitinase-like gene family in the brown planthopper, Nilaparvata lugens [J]. Insect Molecular Biology, 2015, 24 (1): 29-40.
Xi Y, Pan PL, Ye YX,et al. Chitindeacetylase family genes in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae) [J]. Insect Molecular Biology, 2014, 23 (6): 695-705.
Xi Y, Pan PL, Zhang CX. The β-N-acetylhexosaminidase gene family in the brown planthopper, Nilaparvata lugens [J]. Insect Molecular Biology, 2015, 24 (6): 601-610.
Xi Y. Analyses of Chitinolvtic Enzyme Gene Families in Nilaparvata lugens [D]. Zhejiang: Zhejiang University, 2014. [席羽. 褐飞虱几丁质降解酶系基因家族分析 [D]. 浙江: 浙江大学博士学位论文, 2014]
Xu HJ, Xue J, Lu B,et al. Two insulin receptors determine alternative wing morphs in planthoppers [J]. Nature, 2015, 519: 464-479.
Xu J. Molecular Mechanism of the Wing Dimorphism of Brown Plantopper, Nilaparvata lugens [D]. Hangzhou: Zhejiang University, 2015. [薛建. 褐飞虱翅型分化分子机理研究 [D]. 杭州: 浙江大学博士学位论文, 2015]
Xu KK. Studies on the Expression and Function of Six Insulin Signaling Pathway Genes from Bactrocera dorsalis [D]. Chongqing: Southwest University, 2014. [许抗抗. 桔小实蝇胰岛素信号通路六个重要基因的表达与功能研究 [D]. 重庆: 西南大学硕士学位论文, 2014]
Yang MM, Zhao LN, Shen QD,et al. Knockdown of two trehalose-6-phosphate synthases severely affects chitin metabolism gene expression in the rice brown planthopper Nilaparvata lugens [J]. Pest Management Science, 2016, 73: 206-216.
Yu CH, Lu D, Lin RH, et al. Trehalose-the blood sugar in insects [J]. Chinese Bulletin of Entomology, 2008, 45 (5): 832-837. [于彩虹, 卢丹, 林荣华, 等. 海藻糖-昆虫的血糖 [J]. 昆虫知识, 2008, 45 (5): 832-837]
Zhai YF, Sun ZX, Zhang JQ,et al. Activation of the TOR signaling pathway by glutamine regulates insect fecundity [J]. Scientific Reports, 2015: 1-9.
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