莲草直胸跳甲OBP51的克隆及表达分析
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摘要
莲草直胸跳甲在防治入侵性杂草喜旱莲子草中发挥重要作用。莲草直胸跳甲主要通过气味寻找宿主植物;气味结合蛋白是昆虫气味反应的第一步。研究克隆得到莲草直胸跳甲OBP51的全长序列,进行生物信息学分析,并分析其在不同组织中的相对表达量。结果表明,OBP51的c DNA全长为489 bp,ORF为411 bp,编码136个氨基酸,具有18个氨基酸残基的信号肽;AhygOBP51编码的成熟蛋白为脂溶性蛋白,主要为α-螺旋结构,占68.6%,无规则卷曲占28.8%。定量PCR分析表明,其在触角、后足中高表达,且雄虫后足高于雌虫后足,提示AhygOBP51在触角和后足气味识别过程中发挥重要作用。
Agasicles hygrophila plays an important role in the prevention and control of invasive weeds, Alternanthera philoxeroides. Agasicles hygrophila is mainly used to find the host plant through the smell, and the odor-binding protein is the first step in the insect odor reaction. In this study, the full-length sequence of OBP51 was cloned, and bioinformatics analysis was performed, and its expression in different tissues was analyzed. The results showed that the OBP51 c DNA had a full length of 489 bp and an ORF of 411 bp,encoding a signal peptide of 136 amino acids with 18 amino acid residues. The mature protein encoded by AhygOBP51 was a fat-soluble protein, mainly composed of α-helical structure, accounting for 68.6%, and irregular curl accounting for 28.8%. Quantitative PCR analysis showed that it was highly expressed in the antennae and hind leg, and the male hind leg was higher than the female, which suggested that AhygOBP51 played an important role in the antennae and hind leg odor recognition process.
Agasicles hygrophila plays an important role in the prevention and control of invasive weeds, Alternanthera philoxeroides. Agasicles hygrophila is mainly used to find the host plant through the smell, and the odor-binding protein is the first step in the insect odor reaction. In this study, the full-length sequence of OBP51 was cloned, and bioinformatics analysis was performed, and its expression in different tissues was analyzed. The results showed that the OBP51 c DNA had a full length of 489 bp and an ORF of 411 bp,encoding a signal peptide of 136 amino acids with 18 amino acid residues. The mature protein encoded by AhygOBP51 was a fat-soluble protein, mainly composed of α-helical structure, accounting for 68.6%, and irregular curl accounting for 28.8%. Quantitative PCR analysis showed that it was highly expressed in the antennae and hind leg, and the male hind leg was higher than the female, which suggested that AhygOBP51 played an important role in the antennae and hind leg odor recognition process.
引文
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[28] GU S H,WANG S P,ZHANG X Y,et al. Identification and tissue distribution of odorant binding protein genes in the lucerne plant bug Adelphocoris lineolatus(Goeze)[J]. Insect Biochemistry and Molecular Biology,2011,41(4):254-263.
[2]马瑞燕,王韧.喜旱莲子草在中国的入侵机理及其生物防治[J].应用与环境生物学报,2005(2):246-250.
[3]BUCKINGHAM G R. Biological control of Alligator weed,Alternanthera philoxeroides,the world's first aquatic weed success story[J].Castanea,1996,61(3):232-243.
[4] VOSSHALL L B,STOCKER R F. Molecular architecture of smell and taste in Drosophila[J]. Annual Review of Neuroscience,2007,30:505-533.
[5] HALLEM E A,DAHANUKAR A,CARLSON J R. Insect odor and taste receptors[J]. Proceedings of the National Academy of Sciences of the United States of America,2006,51(10):113-135.
[6]詹文会,张苏芳,耿红卫,等.天牛嗅觉感受相关蛋白研究进展[J].河南农业科学,2018,47(3):1-6.
[7] YIN J,FENG H,SUN H,et al. Functional analysis of general odorant binding protein 2 from the meadow moth,Loxostege sticticalis L.(Lepidoptera:Pyralidae)[J]. PLo S One,2012,7(3):e33589.
[8] HE M,HE P. Molecular characterization,expression profiling,and binding properties of odorant binding protein genes in the whitebacked planthopper,Sogatella furcifera[J]. Comparative Biochemistry and Physiology Part B,Biochemistry&Molecular Biology,2014,174:1-8.
[9]SUN L,GU S H,XIAO H J,et al. The preferential binding of a sensory organ specific odorant binding protein of the alfalfa plant bug Adelphocoris lineolatus AlinOBP10 to biologically active host plant volatiles[J]. Journal of Chemical Ecology,2013,39(9):1221-1231.
[10] LI H L,ZHANG L Y,NI C X,et al. Molecular recognition of floral volatile with two olfactory related proteins in the Eastern honeybee(Apis cerana)[J]. International Journal of Biological Macromolecules,2013,56:114-121.
[11] LI Z Q,ZHANG S,LUO J Y,et al. Two Minus-C odorant binding proteins from Helicoverpa armigera display higher ligand binding affinity at acidic pH than neutral pH[J]. Journal of Insect Physiology,2013,59(3):263-272.
[12] HUA J F,ZHANG S,CUI J J,et al. Identification and binding characterization of three odorant binding proteins and one chemosensory protein from Apolygus lucorum(Meyer-Dur)[J].Journal of Chemical Ecology,2012,38(9):1163-1170.
[13] LIU S J,LIU N Y,HE P,et al. Molecular characterization,expression patterns,and ligand-binding properties of two odorant-binding protein genes from Orthaga achatina(Butler)(Lepidoptera:Pyralidae)[J]. Archives of Insect Biochemistry and Physiology,2012,80(3):123-139.
[14]DENG S,YIN J,ZHONG T,et al. Function and immunocytochemical localization of two novel odorant-binding proteins in olfactory sensilla of the scarab beetle Holotrichia oblita Faldermann(Coleoptera:Scarabaeidae)[J]. Chemical Senses,2012,37(2):141-150.
[15]贾栋,赵龙龙,王森,等.莲草直胸跳甲Hsp70蛋白的生物信息学分析[J].山西农业科学,2015,43(4):397-400.
[16]KRIEGER J,VON NICKISCH-ROSENEGK E,MAMELI M,et al.Binding proteins from the antennae of Bombyx mori[J]. Insect Biochemistry and Molecular Biology,1996,26(3):297-307.
[17]GONG D P,ZHANG H J,ZHAO P,et al. The odorant binding protein gene family from the genome of silkworm,Bombyx mori[J].BMC Genomics,2009,10:332.
[18]张婷,刘乃勇,董双林.甜菜夜蛾触角结合蛋白Ⅱ的c DNA克隆、组织分布及配体结合特性分析[J].昆虫学报,2012,55(5):499-509.
[19]ZHU J Y,ZHANG L F,ZE S Z,et al. Identification and tissue distribution of odorant binding protein genes in the beet armyworm,Spodoptera exigua[J]. Journal of Insect Physiology,2013,59(7):722-728.
[20] SUN H Z,SONG Y Q,DU J,et al. Identification and tissue distribution of chemosensory protein and odorant binding protein genes in Athetis dissimilis(Lepidoptera:Noctuidae)[J]. Applied Entomology&Zoology,2016,51(3):409-420.
[21]马龙.茶尺蠖化学感受相关基因的克隆与功能研究[D].北京:中国农业科学院,2016.
[22]王娟.大草蛉嗅觉相关蛋白基因的鉴定及功能研究[D].北京:中国农业科学院,2016.
[23]王娟.松褐天牛气味结合蛋白基因的克隆与功能分析[D].武汉:华中农业大学,2015.
[24]赵雪.茄无网蚜气味结合蛋白OBP7的克隆、表达谱及其触角和腹管超微结构观察[D].哈尔滨:东北农业大学,2014.
[25] PELLETIER J,LEAL W S. Genome analysis and expression patterns of odorant-binding proteins from the Southern House mosquito Culex pipiens[J]. PLo S One,2009,4(7):e6237.
[26]XU W,CORNEL A J,LEAL W S. Odorant-binding proteins of the malaria mosquito Anopheles funestus sensu stricto[J]. PLo S One,2010,5(10):e15403.
[27] MITAKA H,MATSUO T,MIURA N,et al. Identification of odorant-binding protein genes from antennal expressed sequence tags of the onion fly,Delia antiqua[J]. Molecular Biology Reports,2011,38(3):1787-1792.
[28] GU S H,WANG S P,ZHANG X Y,et al. Identification and tissue distribution of odorant binding protein genes in the lucerne plant bug Adelphocoris lineolatus(Goeze)[J]. Insect Biochemistry and Molecular Biology,2011,41(4):254-263.