Clone and expression analysis of three glutathione-S-transferase genes in different host-specific types of Aphis gossypii Glover

doi:10.7679/j.issn.2095-1353.2020.084

Abstract

Abstract:

[Objectives] To determine the profiles of three high expression glutathione-S-transferase (GST) genes in various developmental stages of Aphis. gossypii Glover, and compare the expression of these genes in different host-races of this pest. [Methods] Full-length cDNA sequences of GSTs genes were cloned based on transcriptome data of A. gossypii specimens collected from seedling and mid-summer growth stages of cotton. The relative expression levels in different developmental stages (nymph and adult) and host-races (cucumber and cotton host-races) were investigated with real-time quantitative PCR. [Results] Three high expression GSTs were identified and named AgoGST-s1 (GenBank accession no.: MN688789), AgoGST-d1 (GenBank accession no.: MN688790) and AgoGST-d2 (GenBank accession no.: MN688791). These 3 genes were expressed in all development stages and their temporal expression profiles all tended to increase over time. The relative expression of AgoGST-s1 gene was less in all developmental stages, whereas that of AgoGST-d1 and AgoGST-d2 was more variable. Expression of these 3 GSTs in the cucumber host-race was higher than in the cotton host-race for most age-classes. [Conclusion] Expression of the AgoGST-s1, AgoGST-d1 and AgoGST-d2 genes is significantly associated with the developmental stage of A. gossypii. The expression patterns of these 3 GST genes in two host-races differed, which may reflect adaptation to different host plants.

Key words: Aphis gossypii, glutathione-S-transferase (GST), host specialization, development stage, expression profile

Chuan-Peng WANG, Shuai ZHANG, Xue-Ke GAO, Jun-Yu LUO, Xiang-Zhen ZHU, Li WANG, Kai-Xin ZHANG, Yi-Hua YANG, Jin-Jie CUI. Clone and expression analysis of three glutathione-S-transferase genes in different host-specific types of Aphis gossypii Glover[J]. Chinese Journal of Applied Entomology, 2020, 57(4): 823-832.

Figures/Tables 5

Table 1 Primers used for real-time quantitative PCR

基因 Gene	引物序列 Primer sequence（5?-3?）	扩增效率（%） Amplification efficiency	相关系数（R²） Correlation coefficient
AgoGST-s1	F-TGATAATGGCACTTAACTACTA R-AACCACTATCCGAACAAC	90	0.994
AgoGST-d1	F-ACAGTCAGTCGTCATACAC R-TTCTGAATCGGTCGGTTC	91	0.997
AgoGST-d2	F-GTGATTGGCTTCGTTGTAAT R-ATCTGACCAGTTATCCTAATGT	94	0.995
AgoGAPDH	F-ATCATTCCAGCATCTACT R-TCCTTAACCTTATCCTTGA	90	0.999

Fig. 1 Amino acid sequence alignment of AgoGST-s1 from Aphis gossypii with Sigma-class homologues from other insects Black box region is the N-terminal domain of AgoGST-s1; Red box region is the C-terminal domain of AgoGST-s1; Predicted GSH binding sites (G-sites) and substrate binding sites (H-sites) are shaded red and green, respectively. The typical catalytic site residue “Y8” of Sigma-class is indicated in black arrow.

Fig. 2 Amino acid sequence alignment of AgoGST-d1, AgoGST-d2 from Aphis gossypii with Delta-class homologues from other insects Black box region is the N-terminal domain of AgoGST-d1, AgoGST-d2; Red box region is the C-terminal domain of AgoGST-d1, AgoGST-d2; Predicted GSH binding sites (G-sites) and substrate binding sites (H-sites) are shaded red and green, respectively. The typical sequence “T*AD” of Delta-class is marked by black arrow.

Fig. 3 Phylogenetic analysis of GSTs of Aphis gossypii and other insects The tree is constructed by using neighbor-joining method. Bootstrap support values are based on 1 000 replicates. The GSTs of cotton aphid is marked with the red. So: Sitophilus oryzae; Bg: Blattella germanica; Lm: Locusta migratoria; Cs: Chilo suppressalis; Px: Papilio xuthus; Aga: Anopheles gambiae; Dv: Daktulosphaira vitifoliae; Dm: Drosophila melanogaster; Dp: Danaus plexippus; Tm: Tenebrio molitor; Bm: Bombyx mori; Pa: Periplaneta americana; Zn: Zootermopsis nevadensis; Api: Acyrthosiphon pisum; Sl: Spodoptera littoralis; Ls: Lasioderma serricorne; Bt: Bemisia tabaci; Ape: Antheraea pernyi: Ago: Aphis gossypii.

Fig. 4 Relative expression levels of AgoGST-s1, AgoGST-d1, AgoGST-d2 in cotton specialized and cucumber specialized aphids 2 L: 2nd instar nymph; 3 L: 3rd instar nymph; 4 L: 4th instar nymph; 1 D: 1st adult; 3 D: 3rd adult; 5 D: 5th adult; 7 D: 7th adult. Different lowercase letters above histograms indicate significant difference among various genes (ANOVA, P<0.05).

References 49

[1]	Broeders S, Huber I, Grohmann L, Berben G, Taverniers I, Mazzara M, Roosens N, Morisset D , 2014. Guidelines for validation of qualitative real-time PCR methods. Trends in Food Science & Technology, 37(2):115-126. doi: 10.1016/j.tifs.2014.03.008 URL
[2]	Cao DP, Liu Y, Walker WB, Li JH, Wang GR , 2014. Molecular Characterization of the Aphis gossypii olfactory receptor gene families. PLoS ONE, 9(6):e101187. doi: 10.1371/journal.pone.0101187 URL pmid: 24971460
[3]	Chen WS, Gu DJ, Li W, Chen ZP, Zhang WQ , 1997. Study on the host specialisation of Myzus persicae in south china. Journal of South China Agricultural University, 18(4):54-56.
	[ 陈文胜, 古德就, 李卫, 陈泽鹏, 张维球 , 1997. 烟蚜寄主专化性的研究. 华南农业大学学报, 18(4):54-56.]
[4]	Dixon DP, Edwards R , 2005. Glutathione transferases. The Arabidopsis Book, 8(45):125-131.
[5]	Enayati AA, Ranson H, Hemingway J , 2005. Insect glutathione transferases and insecticide resistance. Insect Molecular Biology, 14(1):3-8. URL pmid: 15663770
[6]	Fang Y, Qiao GX, Zhang GX , 2011. Morphological adaptation of aphid species on different host plant leaves. Acta Entomologica Sinica, 54(2):157-178.
	[ 方燕, 乔格侠, 张广学 , 2011. 不同寄主植物叶片上蚜虫的形态适应. 昆虫学报, 54(2):157-178.]
[7]	Gao F, Zhu SR, Sun YC, Du L, Parajulee M, Kang L, Ge F , 2008. Interactive effects of elevated CO₂ and cotton cultivar on tri-trophic interaction of Gossypium hirsutum, Aphis gossypii and Propylaea japonica. Environmental Entomology, 37(1):29-37. doi: 10.1603/0046-225X(2008)37[29:IEOECA]2.0.CO;2 URL pmid: 18348793
[8]	Hayaoka T, Dauterman WC , 1983. The effect of phenobarbital induction on glutathione conjugation of diazinon in susceptible and resistant house flies. Pesticide Biochemistry and Physiology, 19(3):344-349. doi: 10.1016/0048-3575(83)90063-9 URL
[9]	He C, Xie W, Zhang YJ , 2017. Comparson of the responses of glutathione- S-transferase genes in Bemisia tabaci B to different host plant transfers. Plant Protection, 43(6):72-77.
	[ 何超, 谢文, 张友军 , 2017. 谷胱甘肽-S-转移酶基因在B型烟粉虱寄主转换中的差异比较. 植物保护, 43(6):72-77.]
[10]	Hedin PA, Parrott WL, Jenkins JN , 1992. Relationships of glands, cotton square terpenoid aldehydes, and other allelochemicals to larval growth of Heliothis virescens(Lepidoptera: Noctuidae). Journal of Economic Entomology, 85(2):359-364. doi: 10.1093/jee/85.2.359 URL
[11]	Jin YL, Zhang BX, Lin HF , 2018. Identifying and measuring the expression profiles, of Chilo suppressalis glutathione-S-transferase genes. Chinese Jounrnal of Applied Entomology, 55(3):349-359.
	[ 金燕璐, 张邦贤, 林华峰 , 2018. 二化螟谷胱甘肽-S-转移酶基因的鉴定与表达模式分析. 应用昆虫学报, 55(3):349-359.]
[12]	Ketterman AJ, Saisawang C, Wongsantichon J , 2011. Insect glutathione transferases. Drug Metabolism Reviews, 43(2):253-265. doi: 10.3109/03602532.2011.552911 URL
[13]	Lee K , 1991. Glutathione- S-transferase activities in phytophagous insects: Induction and inhibition by plant phototoxins and phenols. Insect Biochemistry, 21(4):353-361. doi: 10.1016/0020-1790(91)90001-U URL
[14]	Li F, Han ZJ, Wu ZF , 2002. Comparison of esterase and acetylcholinesterase of cotton aphid, Aphis gossypii Glover reared on different host plants. Journal of Nanjing Agricultural University, 25(2):57-60.
	[ 李飞, 韩召军, 吴智锋 , 2002. 取食不同寄主棉蚜的羧酸酯酶和乙酰胆碱酯酶特性比较. 南京农业大学学报, 25(2):57-60.]
[15]	Li YH , 2014. The progress in the research of insect GST on insecticide detoxification and endogenous metabolism, Journal of Shanxi Datong University (Natural Science), 30(4):49-51.
	[ 李亚红 , 2014. 昆虫谷胱甘肽硫转移酶农药解毒与内源代谢研究进展. 山西大同大学学报(自然科学版), 30(4):49-51.]
[16]	Li ZQ, Zhang S, Luo JY, Wang CY, Lv LM, Dong SL, Cui JJ , 2013. Ecological adaption analysis of the cotton aphid ( Aphis gossypii) in different phenotypes by transcriptome comparison. PLoS ONE, 8(12):e83180. doi: 10.1371/journal.pone.0083180 URL pmid: 24376660
[17]	Liu J, Li RM, Zhou XM , 2013. Characteristics of mRNA expression of GSTs1 gene in Spodoptera exigua and the effect of chlorantraniliprole on its expression. Plant Protection, 39(3):18-21.
	[ 刘佳, 李如美, 周小毛 , 2013. 甜菜夜蛾GSTs1基因mRNA表达特征及氯虫苯甲酰胺对其表达量的影响. 植物保护, 39(3):18-21.]
[18]	Liu S, Zhang YX, Wang WL, Zhang BX, Li SG , 2017. Identification and characterisation of seventeen glutathione- S-transferase genes from the cabbage white butterfly Pieris rapae. Pesticide Biochemistry and Physiology, 143:102-110. doi: 10.1016/j.pestbp.2017.09.001 URL pmid: 29183577
[19]	Liu XD, Zhai BP, Zhang XX , 2003. Studies on the host biotypes and its cause of cotton aphid in Nanjing, China. Scientia Agricultura Sinica, 36(1):54-58.
	[ 刘向东, 翟保平, 张孝羲 , 2003. 南京地区棉蚜寄主专化型及其成因研究. 中国农业科学, 36(1):54-58.]
[20]	Liu XD, Zhang XX, Zhai BP , 2004. Host biotypes and their formation causes in aphids. Acta Entomologica Sinica, 47(4):499-506.
	[ 刘向东, 张孝羲, 翟保平 , 2004. 蚜虫寄主专化型及其成因. 昆虫学报, 47(4):499-506.]
[21]	Liu ZH, Zhao GH, Lu JS, Shui Y, Wu G , 2008. Studies on content of cotton gossypol and characteristics of pest resistance. Xinjiang Agricultural Sciences, 45(3):409-413.
	[ 刘泽辉, 赵国虎, 陆敬善, 水涌, 武刚 , 2008. 棉花棉酚含量与抗虫特性的研究. 新疆农业科学, 45(3):409-413.]
[22]	Livak KJ, Schmittgen TD , 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2 ^-ΔΔCT method . Methods, 25(4):402-408. doi: 10.1006/meth.2001.1262 URL pmid: 11846609
[23]	Mao YB, Cai WJ, Wang JW, Hong GJ, Tao XY, Wang LJ, Huang YP, Chen XY , 2007. Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol. Nature Biotechnology, 25(11):1307-1313. doi: 10.1038/nbt1352 URL pmid: 17982444
[24]	Masutti FV, Chavigny P , 1998. Host-based genetic differentiation in the aphid Aphis gossypii Glover, evidenced from RAPD fingerprints. Molecular Ecology, 7(7):905-914. doi: 10.1046/j.1365-294x.1998.00421.x URL
[25]	Masutti FV, Chavigny P, Fuller SJ , 1999. Characterization of microsatellite loci in the aphid species Aphis gossypii Glover. Molecular Ecology, 8(4):693-695. doi: 10.1046/j.1365-294x.1999.00876.x URL pmid: 10327667
[26]	Ranson H, Claudianos C, Ortelli F, Abgrall C, Hemingway J, Sharakhova MV, Unger MF, Collins FH, Feyereisen R , 2002. Evolution of supergene families associated with insecticide resistance. Science, 298(5591):179-181. doi: 10.1126/science.1076781 URL pmid: 12364796
[27]	Ranson H, Rossiter L, Ortelli F, Jensen B, Wang Xl, Roth CW, Collins FH, Hemingway J , 2001. Identification of a novel class of insect glutathione S-transferases involved in resistance to DDT in the malaria vector Anopheles gambiae. Biochemical Journal, 359(2):295-304. doi: 10.1042/bj3590295 URL
[28]	Reidy GF, Rose HA, Visetson S, Murray M , 1990. Increased glutathione- S-transferase activity and glutathione content in an insecticide-resistant strain of Tribolium castaneum(Herbst). Pesticide Biochemistry and Physiology, 36(3):269-276. doi: 10.1016/0048-3575(90)90035-Z URL
[29]	Ren KY, Zhang S, Luo JY, Wang CY, Lv LM, Zhang LJ, Zhu XZ, Wang L, Cui JJ, Zhu JB , 2018. Clone and expression analysis of salivary gland protein C002 in different host-specific types of Aphis gossypii Glover. Journal of Environmental Entomology, 40(3):617-623.
	[ 任柯昱, 张帅, 雒珺瑜, 王春义, 吕丽敏, 张利娟, 朱香镇, 王丽, 崔金杰, 祝建波 , 2018. 棉蚜唾液蛋白C002基因克隆及在棉蚜不同寄主专化型中的表达分析. 环境昆虫学报, 40(3):617-623.]
[30]	Saito T , 1990. Insecticide resistance of the cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae): Variation of esterase activity by host plants. Japanese Journal of Applied Entomology and Zoology, 34(1):37-41. doi: 10.1303/jjaez.34.37 URL
[31]	Tang DL , 1999. The secondary metabolism of plant insect resistant. World Agriculture, 239(3):32-33.
	[ 汤德良 , 1999. 植物抗虫的次生代谢物质. 世界农业, 239(3):32-33.]
[32]	Tang F, Liang P, Gao XW , 2005. 2-tridecylketone and quercetin induce tissue-specific expression of glutathione- S-transferase in Helicoverpa armigera. Natural Science Exhibition, 15(7):805-810.
	[ 汤方, 梁沛, 高希武 , 2005. 2-十三烷酮和槲皮素诱导棉铃虫谷胱甘肽-S-转移酶组织特异性表达. 自然科学进展, 15(7):805-810.]
[33]	Vontas JG, Small GJ, Hemingway J , 2001. Glutathione- S-transferases as antioxidant defence agents confer pyrethroid resistance in Nilaparvata lugens. Biochemical Journal, 357(1):65-72. doi: 10.1042/bj3570065 URL
[34]	Wadleigh RW, Yu SJ , 1987. Glutathione transferase activity of fall armyworm larvae toward α, β-unsaturated carbonyl allelochemicals and its induction by allelochemicals. Insect Biochemistry, 17(5):759-764. doi: 10.1016/0020-1790(87)90046-1 URL
[35]	Wadleigh RW, Yu SJ , 1988. Detoxification of isothiocyanate allelochemicals by gluthione- S-transferase in three lepidopterous species. Journal of Chemistry Ecology, 14(5):1279-1288. doi: 10.1007/BF01019352 URL
[36]	Wang L , 2015. Preliminary studies on the host biotypes and formation mechanism of cotton aphid in cotton areas of north China. Master dissertation. Anyang: Chinese Academy of Agricultural Sciences.
	[ 王丽 , 2015. 华北棉区棉蚜寄主专化型及其形成机制初步研究. 硕士学位论文. 安阳: 中国农业科学院.]
[37]	Wang L, Zhang S, Luo JY, Wang CY, Lü LM, Zhu XZ, Li CH, Cui JJ , 2016. Identification of Aphis gossypii Glover (Hemiptera: Aphididae) biotypes from different host plants in north China. PLoS ONE, 11(1):34-39.
[38]	Wang SY, Zhou XH, Zhang AS, Li LL, Men XY, Liu YJ, Li XK, Yu Y , 2013. Mechanisms and cross-resistance of imidacloprid resistance in Frankliniella occidentalis. Chinese Jounrnal of Applied Entomology, 50(1):167-172.
	[ 王圣印, 周仙红, 张安盛, 李丽莉, 门兴元, 刘永杰, 李许可, 于毅 , 2013. 西花蓟马对吡虫啉的抗性机制及交互抗性研究. 应用昆虫学报, 50(1):167-172.]
[39]	Wu YQ, Guo YY, Yang J , 2000. Analysis of flavonoid substance in cotton plants for resistance to pests by HPLC. Plant Protection, 26(5):1-3.
	[ 武予清, 郭予元, 杨舰 , 2000. 棉株中抗虫物质黄酮类化合物的高效液相色谱分析. 植物保护, 26(5):1-3.]
[40]	Yang J, Sun XQ, Yan SY, Pan WJ, Zhang MX, Ca QN , 2017. Interaction of ferulic acid with glutathione- S-transferase and carboxylesterase genes in the brown planthopper, Nilaparvata lugens. Journal of Chemical Ecology, 43(7):1-10. doi: 10.1007/s10886-016-0812-x URL
[41]	You YC, Xie M, You MS , 2013. The diversity and role of glutathione- S-transferase in insecticide resistance in insects. Chinese Journal of Applied Entomology, 50(3):831-840.
	[ 尤燕春, 谢苗, 尤民生 , 2013. 昆虫谷胱甘肽-S-转移酶的多样性及其介导的抗药性. 应用昆虫学报, 50(3):831-840.]
[42]	Yu SJ , 1982. Host plant induction of glutathione- S-transferase in the fall armyworm. Pesticide Biochemistry and Physiology, 18(1):101-106. doi: 10.1016/0048-3575(82)90092-X URL
[43]	Yu SJ , 1984. Interactions of allelochemicals with detoxication enzymes of insecticide-susceptible and resistant fall armyworms [ Spodoptera frugiperda, host plants]. Pesticide Biochemistry and Physiology, 22(1):60-68. doi: 10.1016/0048-3575(84)90010-5 URL
[44]	Yu SJ , 1999. Induction of new glutathione- S-transferase isozymes by allelochemicals in the fall armyworm. Pesticide Biochemistry and Physiology, 63(3):163-171. doi: 10.1006/pest.1999.2402 URL
[45]	Yuan LB, Gao ZL, Dang ZH, Li YF, Chi GT, Pan WL , 2008. Fitness differentiation in Aphids gossypii populations to cotton and cucumber. Chinese Bulletin of Entomolog, 45(6):896-899.
	[ 袁立兵, 高占林, 党志红, 李耀发, 赤国彤, 潘文亮 , 2008. 不同棉蚜种群对棉花和黄瓜的适合度分化. 昆虫知识, 45(6):896-899.]
[46]	Zhai YT , 2015. Defense function analysis of three host secondary substances for different biotypes of Sitobion avenae. Master dissertation. Xianyang: Northwest A&F University.
	[ 翟颖婷 , 2019. 三种寄主次生物质对不同生物型麦长管蚜的防御功能分析. 硕士学位论文. 咸阳:西北农林科技大学.]
[47]	Zhang S, Luo JY, Wang L, Wang CY, Lv LM, Zhang LJ, Zhu XZ, Cui JJ , 2018. The biotypes and host shifts of cotton-melon aphis Aphis gossypii in northern China. Journal of Integrative Agriculture, 17(9):2066-2073. doi: 10.1016/S2095-3119(17)61817-3 URL
[48]	Zhang YC , 2016. The regulation mechanisms of population density of Buchnera aphidicola in Aphis gossypii Glover. Doctoral dissertation. Nanjing: Nanjing Agricultural University.
	[ 张元臣 , 2016. 棉蚜体内原生共生菌Buchnera aphidicola种群密度的调控机制. 博士学位论文. 南京: 南京农业大学.]
[49]	Zhu JY , 2017. Host switching affects the fitness and metabolism of Myzus persicae. Master dissertation. Xianyang:Northwest A&F University.
	[ 朱经云 , 2017. 寄主转换对桃蚜适合度及生理代谢的影响. 硕士学位论文. 咸阳: 西北农林科技大学.]