沙葱萤叶甲成虫越夏期间糖类、蛋白及脂肪含量的变化
|
摘要
【目的】明确沙葱萤叶甲Galeruca daurica成虫在越夏不同阶段主要生理生化指标变化的特点,为进一步揭示其越夏的机理提供基础。【方法】分别采用蒽酮硫酸法和氯仿-甲醇法测定越夏不同阶段(越夏前、越夏期和越夏后)的沙葱萤叶甲成虫体内总糖、海藻糖、糖原、脂肪及水分含量;采用考马斯亮蓝G-250染色法测定总蛋白含量。【结果】总糖、水分和总蛋白含量在沙葱萤叶甲成虫越夏前(分别为26.81μg/mg,77.47%和63.17μg/mg)和越夏后(分别为26.41~26.85μg/mg,75.29%~76.65%和59.53~64.93μg/mg)显著高于越夏期(分别为18.77~21.14μg/mg,61.50%~67.20%和39.82~52.54μg/mg),而糖原含量变化趋势正好相反(越夏前:8.43μg/mg;越夏后:5.91~6.14μg/mg;越夏期:10.18~11.58μg/mg),脂肪含量在越夏后(11.48%~11.65%)显著低于越夏前(42.48%)和越夏期(36.05%~64.79%);脂肪和总蛋白含量在越夏期间逐渐下降,而水分、总糖和糖原含量变化不明显;海藻糖含量在越夏不同阶段波动较大(2.66~25.31μg/mg)。【结论】脂肪和糖原是沙葱萤叶甲成虫越夏的主要能源物质。
【Aim】This study aims to clarify the characteristics of main physiological and biochemical change in Galeruca daurica adults at different oversummering stages so as to lay a foundation for revealing the oversummering mechanism. 【Methods】The contents of total sugar,trehalose,glycogen,lipid and water in G. daurica adults at different oversummering stages( pre-oversummering,during oversummering and post-oversummering) were determined by anthrone-sulfuric acid and methanol-chloroform methods,respectively,and the total protein content was detected by Coomassie brilliant blue G-250 method.【Results】The contents of total sugar,water and total protein in G. daurica adults were significantly higher at the pre-oversummering stage( 26. 81 μg/mg,77. 47% and 63. 17 μg/mg,respectively) and post-oversummering stage( 26. 41-26. 85 μg/mg,75. 29%-76. 65% and 59. 53-64. 93 μg/mg,respectively) than during oversummering( 18. 77-21. 14 μg/mg,61. 50%-67. 20% and 39. 82-52. 54 μg/mg,respectively),whereas the change trend of glycogen contents was the opposite,being8. 43 μg/mg at the pre-oversummering stage,5. 91-6. 14 μg/mg at the post-oversummering stage and10. 18-11. 58 μg/mg during oversummering,respectively,and the content of lipid was significantly less at the post-summering stage( 11. 48%-11. 65%) than at the pre-oversummering stage( 42. 48%) and during oversummering( 36. 05%-64. 79%). The contents of lipid and total protein decreased gradually in G. daurica adults during oversummering,while the water,total sugar and glycogen contents showed no obvious change. The trehalose content in G. daurica adults at different oversummering stages fluctuated greatly from 2. 66 μg/mg to 25. 31 μg/mg. 【Conclusion 】 Lipid and glycogen are the main energy materials of G. daurica adults during oversummering.
【Aim】This study aims to clarify the characteristics of main physiological and biochemical change in Galeruca daurica adults at different oversummering stages so as to lay a foundation for revealing the oversummering mechanism. 【Methods】The contents of total sugar,trehalose,glycogen,lipid and water in G. daurica adults at different oversummering stages( pre-oversummering,during oversummering and post-oversummering) were determined by anthrone-sulfuric acid and methanol-chloroform methods,respectively,and the total protein content was detected by Coomassie brilliant blue G-250 method.【Results】The contents of total sugar,water and total protein in G. daurica adults were significantly higher at the pre-oversummering stage( 26. 81 μg/mg,77. 47% and 63. 17 μg/mg,respectively) and post-oversummering stage( 26. 41-26. 85 μg/mg,75. 29%-76. 65% and 59. 53-64. 93 μg/mg,respectively) than during oversummering( 18. 77-21. 14 μg/mg,61. 50%-67. 20% and 39. 82-52. 54 μg/mg,respectively),whereas the change trend of glycogen contents was the opposite,being8. 43 μg/mg at the pre-oversummering stage,5. 91-6. 14 μg/mg at the post-oversummering stage and10. 18-11. 58 μg/mg during oversummering,respectively,and the content of lipid was significantly less at the post-summering stage( 11. 48%-11. 65%) than at the pre-oversummering stage( 42. 48%) and during oversummering( 36. 05%-64. 79%). The contents of lipid and total protein decreased gradually in G. daurica adults during oversummering,while the water,total sugar and glycogen contents showed no obvious change. The trehalose content in G. daurica adults at different oversummering stages fluctuated greatly from 2. 66 μg/mg to 25. 31 μg/mg. 【Conclusion 】 Lipid and glycogen are the main energy materials of G. daurica adults during oversummering.
引文
Adamek G,Fischer J,1985.The oxygen consumption of non-dormant and dormant larvae of Chironomus plumosus(Diptera).J.Insect Physiol.,31(10):767-772.
Bradford MM,1976.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal.Biochem.,72(1-2):248-254.
Cheng WN,Li YP,Li JJ,Li XL,Wu JX,Wang HL,2008.Protein contents and electrophoresis patterns of the pre-diapause,diapause and post-diapause larvae in the wheat blossom midge,Sitodiplosis mosellana Gehin.J.Plant Prot.,35(2):155-159.[成卫宁,李怡萍,李建军,李修炼,仵均祥,王洪亮,2008.小麦吸浆虫滞育前后和滞育期蛋白质含量及其电泳分析.植物保护学报,35(2):155-159]
Cheng Y,Han LL,Zhao KJ,Xu XZ,Li DP,Zhang XX,2016.Effect of host plants and temperature on the accumulation of carbohydrates and lipid in the soybean pod borer,Leguminivora glycinivorella(Mats.)Obraztsov.Chin.J.Appl.Entomol.,53(4):716-722.[程媛,韩岚岚,赵奎军,徐馨竹,李东坡,张旭霞,2016.寄主植物与温度对大豆食心虫滞育期间糖类和脂质含量的影响.应用昆虫学报,53(4):716-722]
Colineta H,Vernonb P,Hance T,2007.Does thermal-related plasticity in size and fat reserves influence super-cooling abilities and coldtolerance in Aphidius colemani(Hymenoptera:Aphidiinae)mummies?J.Therm.Biol.,32(7-8):374-382.
Danks HV,2000.Dehydration in dormant insects.J.Insect Physiol.,46(6):837-852.
de Kort CAD,1990.Thirty-five years of diapause research with the Colorado potato beetle.Entomol.Exp.Appl.,56(1):1-13.
Denekamp NY,Thorne MAS,Clark MS,Kube M,Reinhardt R,Lubzens E,2009.Discovering genes associated with dormancy in the monogonont rotifer Brachionus plicatilis.BMC Genomics,10(1):108.
Denlinger DL,2002.Regulation of diapause.Annu.Rev.Entomol.,47:93-122.
Gao YH,Zhao T,Guo XR,Luo MH,Jiang JW,2004.Changes of physiological and biochemical during insects diapause.In:Li DM et al.eds.Contemporary Entomological Research-Proceedings of the Entomological Society of China 60th Anniversary Conference and Symposium.China Agricultural Science and Technology Press,Beijing.[高玉红,赵特,郭线茹,罗梅浩,蒋金炜,2004.昆虫滞育期中生理生化的变化.见:李典漠等主编.当代昆虫学研究---中国昆虫学会成立60周年纪念大会暨学术讨论会论文集.北京:中国农业科学技术出版社]
Garcia R,Hagen KS,1987.Summer dormancy in adult Agabus disintegratus(Crotch)(Coleoptera:Dytiscidae)in dried ponds in California.Ann.Entomol.Soc.Am.,80(2):267-271.
Hahn DA,Denlinger DL,2007.Meeting the energetic demands of insect diapause nutrient storage and utilization.J.Insect Physiol.,53(8):760-773.
Hahn DA,Denlinger DL,2011.Energetics of insect diapause.Annu.Rev.Entomol.,56:103-121.
Hao X,Zhou XR,Pang BP,Zhang ZR,Bao X,2015.Morphological and biological characteristics of Galeruca daurica Joannis(Coleoptera:Chrysomelidae).Acta Agrest.Sin.,23(5):1106-1108.[昊翔,周晓榕,庞保平,张卓然,包祥,2015.沙葱萤叶甲的形态特征和生物学特性研究.草地学报,23(5):1106-1108]
Hodek I,2012.Adult diapause in Coleoptera.Psyche,2012:ID249081.
Huang L,Sun LZ,Wang Y,Ru YT,Irfan M,Jiang YR,Shi SL,Yang RS,Li XS,Qin L,2016.Changes in the expression of trehalose-6-phosphate synthase gene in Antheraea pernyi(Lepidoptera:Saturniidae)during pupal diapause termination.Acta Entomol.Sin.,59(9):938-947.[黄伶,孙良振,王勇,汝玉涛,Irfan M,姜义仁,石生林,杨瑞生,李喜升,秦利,2016.柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化.昆虫学报,59(9):938-947]
Katsoyannos P,Kontodimas DC,Stathas GJ,1997.Summer diapause and winter quiescence of Coccinella septempunctata(Col.Coccinellidae)in central Greece.Entomophaga,42(4):483-491.
Kida Y,Numata H,Fujii H,1997.Summer diapause in females of Minois dryas(Lepidoptera:Satyridae).Environ.Entomol.,26(2):201-206.
Lee KY,Denlinger DL,1996.Diapause-regulated proteins in the gut of pharate first instar larvae of the gypsy moth,Lymantria dispar,and the effect of kk-42 and neck ligation on expression.J.Insect Physiol.,42(5):423-431.
Lees AD,1956.The physiology and biochemistry of diapause.Annu.Rev.Entomol.,1(1):1-16.
Liu ZD,Wu KJ,Gong PY,2002a.Summer diapause in insects.Entomol.Knowl.,39(3):234-237.[刘柱东,吴坤君,龚佩瑜,2002a.昆虫的夏滞育.昆虫知识,39(3):234-237]
Liu ZD,Wu KJ,Gong PY,2002b.Regulation and genetic basis of summer diapause in insects.Entomol.Knowl.,39(4):261-264.[刘柱东,吴坤君,龚佩瑜,2002b.昆虫夏滞育的调控及其遗传基础.昆虫知识,39(3):261-264]
Poelchau MF,Reynolds JA,Denlinger DL,Elsik CG,Armbruster PA,2011.A de novo transcriptome of the Asian tiger mosquito,Aedes albopictus,to identify candidate transcripts for diapause preparation.BMC Genomics,12(2):251-254.
Ragland GJ,Fuller J,Feder JL,Hahn DA,2009.Biphasic metabolic rate trajectory of pupal diapause termination and post-diapause development in a tephritid fly.J.Insect Physiol.,55(4):344-350.
Ren XY,Zhang LS,Qi XY,An T,Han YH,Chen HY,2015.Metabolic adaption and evaluation of cold hardiness on diapausing ladybird,Coccinella septempunctata L.J.Environ.Entomol.,37(6):1195-1202.[任小云,张礼生,齐晓阳,安涛,韩艳华,陈红印,2015.滞育七星瓢虫的代谢适应与抗寒性评价.环境昆虫学报,37(6):1195-1202]
Salama MS,Miller TA,1992.A diapause associated protein of the pink bollworm Pectinophora gossypiella Saunders.Arch.Insect Biochem.,21(1):1-11.
Simek P,Sula J,Kostal V,1998.Physiology of drought tolerance and cold hardiness of the Mediterranean tiger moth Cymbalophora pudica during summer diapause.J.Insect Physiol.,44(2):165-173.
Voss RH,Ferro DN,1992.Population dynamics of the Colorado potato beetle(Coleoptera:Chrysomelidae)in western Massachusetts.Am.Potato J.,69(8):473-482.
Wang FH,Gong H,1998.Measurement and eletrophoresis patterns of haemolymph protein from diapause-and non-diapause-destined Helicoverpa armigera.Acta Entomol.Sin.,41(4):426-430.[王方海,龚和,1998.滞育和非滞育棉铃虫血淋巴中蛋白质含量及图谱的比较.昆虫学报,41(4):426-430]
Wu JX,Yuan F,Su L,2004.Change of carbohydrate contents in larvae of the wheat midge,Sitodiplosis mosellana(Gehin)during mature and diapause stage.Acta Entomol.Sin.,47(2):178-183.[仵均祥,袁锋,苏丽,2004.麦红吸浆虫幼虫滞育期间糖类物质变化.昆虫学报,47(2):178-183]
Xu WH,1999.Advances in insect diapause studies.Acta Entomol.Sin.,42(1):100-106.[徐卫华,1999.昆虫滞育研究进展.昆虫学报,42(1):100-106]
Yang GP,Liu YD,Hou ML,2013.Changes of the protein and nucleic acid contents and the activities of protective enzymes in diapausing larvae of the rice stem borer,Chilo supprressalis(Lepidoptera:Crambidae).Acta Entomol.Sin.,56(3):251-256.[杨光平,刘玉娣,侯茂林,2013.二化螟滞育幼虫的蛋白和核酸含量以及保护酶活性的变化.昆虫学报,56(3):251-256]
Zhang JH,Luo LZ,Jiang XF,Zhang L,2012.Changes in protein and nucleic acid contents in diapause larvae of the beet webworm,Loxostege sticticalis(Lepidoptera:Pyralidae).Acta Entomol.Sin.,55(2):156-161.[张健华,罗礼智,江幸福,张蕾,2012.草地螟滞育幼虫的蛋白和核酸含量变化.昆虫学报,55(2):156-161]
Zhou G,Miesfeld RL,2009.Energy metabolism during diapause in Culex pipiens mosquitoes.J.Insect Physiol.,55(1):40-46.
Zhou XR,Gao JC,Pang BP,2016.Effects of temperature on the termination of egg diapause and post-diapause embryonic development of Galeruca daurica(Coleoptera:Chrysomelidae).Environ.Entomol.,45(4):1076-1080.
Zhu F,Li H,Wang Y,Wang XP,Zhou XM,Huang W,Lei CL,2008.Change of trehalose,glycogen and polyol contents of the diapausing larvae of Mylabris phalerata(Pallas)(Coleoptera:Meloidae)at different diapausing stages.Acta Entomol.Sin.,51(1):9-13.[朱芬,李红,王永,王小平,周兴苗,黄文,雷朝亮,2008.大斑芫菁滞育幼虫在滞育不同阶段体内糖类和醇类含量的变化.昆虫学报,51(1):9-13]
Bradford MM,1976.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Anal.Biochem.,72(1-2):248-254.
Cheng WN,Li YP,Li JJ,Li XL,Wu JX,Wang HL,2008.Protein contents and electrophoresis patterns of the pre-diapause,diapause and post-diapause larvae in the wheat blossom midge,Sitodiplosis mosellana Gehin.J.Plant Prot.,35(2):155-159.[成卫宁,李怡萍,李建军,李修炼,仵均祥,王洪亮,2008.小麦吸浆虫滞育前后和滞育期蛋白质含量及其电泳分析.植物保护学报,35(2):155-159]
Cheng Y,Han LL,Zhao KJ,Xu XZ,Li DP,Zhang XX,2016.Effect of host plants and temperature on the accumulation of carbohydrates and lipid in the soybean pod borer,Leguminivora glycinivorella(Mats.)Obraztsov.Chin.J.Appl.Entomol.,53(4):716-722.[程媛,韩岚岚,赵奎军,徐馨竹,李东坡,张旭霞,2016.寄主植物与温度对大豆食心虫滞育期间糖类和脂质含量的影响.应用昆虫学报,53(4):716-722]
Colineta H,Vernonb P,Hance T,2007.Does thermal-related plasticity in size and fat reserves influence super-cooling abilities and coldtolerance in Aphidius colemani(Hymenoptera:Aphidiinae)mummies?J.Therm.Biol.,32(7-8):374-382.
Danks HV,2000.Dehydration in dormant insects.J.Insect Physiol.,46(6):837-852.
de Kort CAD,1990.Thirty-five years of diapause research with the Colorado potato beetle.Entomol.Exp.Appl.,56(1):1-13.
Denekamp NY,Thorne MAS,Clark MS,Kube M,Reinhardt R,Lubzens E,2009.Discovering genes associated with dormancy in the monogonont rotifer Brachionus plicatilis.BMC Genomics,10(1):108.
Denlinger DL,2002.Regulation of diapause.Annu.Rev.Entomol.,47:93-122.
Gao YH,Zhao T,Guo XR,Luo MH,Jiang JW,2004.Changes of physiological and biochemical during insects diapause.In:Li DM et al.eds.Contemporary Entomological Research-Proceedings of the Entomological Society of China 60th Anniversary Conference and Symposium.China Agricultural Science and Technology Press,Beijing.[高玉红,赵特,郭线茹,罗梅浩,蒋金炜,2004.昆虫滞育期中生理生化的变化.见:李典漠等主编.当代昆虫学研究---中国昆虫学会成立60周年纪念大会暨学术讨论会论文集.北京:中国农业科学技术出版社]
Garcia R,Hagen KS,1987.Summer dormancy in adult Agabus disintegratus(Crotch)(Coleoptera:Dytiscidae)in dried ponds in California.Ann.Entomol.Soc.Am.,80(2):267-271.
Hahn DA,Denlinger DL,2007.Meeting the energetic demands of insect diapause nutrient storage and utilization.J.Insect Physiol.,53(8):760-773.
Hahn DA,Denlinger DL,2011.Energetics of insect diapause.Annu.Rev.Entomol.,56:103-121.
Hao X,Zhou XR,Pang BP,Zhang ZR,Bao X,2015.Morphological and biological characteristics of Galeruca daurica Joannis(Coleoptera:Chrysomelidae).Acta Agrest.Sin.,23(5):1106-1108.[昊翔,周晓榕,庞保平,张卓然,包祥,2015.沙葱萤叶甲的形态特征和生物学特性研究.草地学报,23(5):1106-1108]
Hodek I,2012.Adult diapause in Coleoptera.Psyche,2012:ID249081.
Huang L,Sun LZ,Wang Y,Ru YT,Irfan M,Jiang YR,Shi SL,Yang RS,Li XS,Qin L,2016.Changes in the expression of trehalose-6-phosphate synthase gene in Antheraea pernyi(Lepidoptera:Saturniidae)during pupal diapause termination.Acta Entomol.Sin.,59(9):938-947.[黄伶,孙良振,王勇,汝玉涛,Irfan M,姜义仁,石生林,杨瑞生,李喜升,秦利,2016.柞蚕蛹解除滞育过程中海藻糖合成酶基因的表达变化.昆虫学报,59(9):938-947]
Katsoyannos P,Kontodimas DC,Stathas GJ,1997.Summer diapause and winter quiescence of Coccinella septempunctata(Col.Coccinellidae)in central Greece.Entomophaga,42(4):483-491.
Kida Y,Numata H,Fujii H,1997.Summer diapause in females of Minois dryas(Lepidoptera:Satyridae).Environ.Entomol.,26(2):201-206.
Lee KY,Denlinger DL,1996.Diapause-regulated proteins in the gut of pharate first instar larvae of the gypsy moth,Lymantria dispar,and the effect of kk-42 and neck ligation on expression.J.Insect Physiol.,42(5):423-431.
Lees AD,1956.The physiology and biochemistry of diapause.Annu.Rev.Entomol.,1(1):1-16.
Liu ZD,Wu KJ,Gong PY,2002a.Summer diapause in insects.Entomol.Knowl.,39(3):234-237.[刘柱东,吴坤君,龚佩瑜,2002a.昆虫的夏滞育.昆虫知识,39(3):234-237]
Liu ZD,Wu KJ,Gong PY,2002b.Regulation and genetic basis of summer diapause in insects.Entomol.Knowl.,39(4):261-264.[刘柱东,吴坤君,龚佩瑜,2002b.昆虫夏滞育的调控及其遗传基础.昆虫知识,39(3):261-264]
Poelchau MF,Reynolds JA,Denlinger DL,Elsik CG,Armbruster PA,2011.A de novo transcriptome of the Asian tiger mosquito,Aedes albopictus,to identify candidate transcripts for diapause preparation.BMC Genomics,12(2):251-254.
Ragland GJ,Fuller J,Feder JL,Hahn DA,2009.Biphasic metabolic rate trajectory of pupal diapause termination and post-diapause development in a tephritid fly.J.Insect Physiol.,55(4):344-350.
Ren XY,Zhang LS,Qi XY,An T,Han YH,Chen HY,2015.Metabolic adaption and evaluation of cold hardiness on diapausing ladybird,Coccinella septempunctata L.J.Environ.Entomol.,37(6):1195-1202.[任小云,张礼生,齐晓阳,安涛,韩艳华,陈红印,2015.滞育七星瓢虫的代谢适应与抗寒性评价.环境昆虫学报,37(6):1195-1202]
Salama MS,Miller TA,1992.A diapause associated protein of the pink bollworm Pectinophora gossypiella Saunders.Arch.Insect Biochem.,21(1):1-11.
Simek P,Sula J,Kostal V,1998.Physiology of drought tolerance and cold hardiness of the Mediterranean tiger moth Cymbalophora pudica during summer diapause.J.Insect Physiol.,44(2):165-173.
Voss RH,Ferro DN,1992.Population dynamics of the Colorado potato beetle(Coleoptera:Chrysomelidae)in western Massachusetts.Am.Potato J.,69(8):473-482.
Wang FH,Gong H,1998.Measurement and eletrophoresis patterns of haemolymph protein from diapause-and non-diapause-destined Helicoverpa armigera.Acta Entomol.Sin.,41(4):426-430.[王方海,龚和,1998.滞育和非滞育棉铃虫血淋巴中蛋白质含量及图谱的比较.昆虫学报,41(4):426-430]
Wu JX,Yuan F,Su L,2004.Change of carbohydrate contents in larvae of the wheat midge,Sitodiplosis mosellana(Gehin)during mature and diapause stage.Acta Entomol.Sin.,47(2):178-183.[仵均祥,袁锋,苏丽,2004.麦红吸浆虫幼虫滞育期间糖类物质变化.昆虫学报,47(2):178-183]
Xu WH,1999.Advances in insect diapause studies.Acta Entomol.Sin.,42(1):100-106.[徐卫华,1999.昆虫滞育研究进展.昆虫学报,42(1):100-106]
Yang GP,Liu YD,Hou ML,2013.Changes of the protein and nucleic acid contents and the activities of protective enzymes in diapausing larvae of the rice stem borer,Chilo supprressalis(Lepidoptera:Crambidae).Acta Entomol.Sin.,56(3):251-256.[杨光平,刘玉娣,侯茂林,2013.二化螟滞育幼虫的蛋白和核酸含量以及保护酶活性的变化.昆虫学报,56(3):251-256]
Zhang JH,Luo LZ,Jiang XF,Zhang L,2012.Changes in protein and nucleic acid contents in diapause larvae of the beet webworm,Loxostege sticticalis(Lepidoptera:Pyralidae).Acta Entomol.Sin.,55(2):156-161.[张健华,罗礼智,江幸福,张蕾,2012.草地螟滞育幼虫的蛋白和核酸含量变化.昆虫学报,55(2):156-161]
Zhou G,Miesfeld RL,2009.Energy metabolism during diapause in Culex pipiens mosquitoes.J.Insect Physiol.,55(1):40-46.
Zhou XR,Gao JC,Pang BP,2016.Effects of temperature on the termination of egg diapause and post-diapause embryonic development of Galeruca daurica(Coleoptera:Chrysomelidae).Environ.Entomol.,45(4):1076-1080.
Zhu F,Li H,Wang Y,Wang XP,Zhou XM,Huang W,Lei CL,2008.Change of trehalose,glycogen and polyol contents of the diapausing larvae of Mylabris phalerata(Pallas)(Coleoptera:Meloidae)at different diapausing stages.Acta Entomol.Sin.,51(1):9-13.[朱芬,李红,王永,王小平,周兴苗,黄文,雷朝亮,2008.大斑芫菁滞育幼虫在滞育不同阶段体内糖类和醇类含量的变化.昆虫学报,51(1):9-13]