Cloning of glutathione S-transferases gene from Monochamus alternatus and its expression characteristics under heat stress

LI Zichun, HAO Dejun, LI Hui, LI Changyan, XU Danwenyi, YANG Hualei, ZHAO Peiyuan

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2025, Vol. 49 ›› Issue (1) : 28-36.

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南京林业大学学报（自然科学版）
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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2025, Vol. 49 ›› Issue (1) : 28-36. DOI: 10.12302/j.issn.1000-2006.202304018

Cloning of glutathione S-transferases gene from Monochamus alternatus and its expression characteristics under heat stress

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Abstract

【Objective】To clone the glutathione S-transferases (GST) genes of Monochamus alternatus (Coleoptera: Cerambycidae) and to determine the expression levels of these genes under heat stress in adult males and females, as well as at different developmental stages of M. alternatus, we used molecular biology and bioinformatics methods. 【Method】 Three GST cDNA sequences were cloned, and their structural characteristics were analyzed using DNAMAN 9.0 and I-TASSER software. qRT-PCR was employed to measure the relative expression levels of GST genes in adult and fourth instar larvae of M. alternatus under varying temperatures and treatment durations. The role of these GST genes in protecting the organism from oxidative stress was assessed through disc diffusion assays. 【Result】Three GST cDNA sequences were cloned and named MaltGSTe1, MaltGSTe2, and MaltGSTt1. MaltGSTe1 and MaltGSTe2 belong to the Epsilon family, while MaltGSTt1 belongs to the Theta family. Analysis of three-dimensional protein structures indicated that these GST are cytoplasmic. The relative expression levels of MaltGSTe1, MaltGSTe2 and MaltGSTt1 in fourth instar larvae significantly changed under heat stress, with MaltGSTe2 showing the greatest change. In males, the expression level of MaltGSTt1 was significantly down-regulated. Escherichia coli expressing the GST genes demonstrated enhanced antioxidant capacity, with MaltGSTe2 exhibiting the strongest activity. 【Conclusion】We successfully cloned three GST genes and investigated their expression characteristics under high temperature stress in M. alternatus. High temperature stress was found to induce up-regulation of GST gene expression. Disc diffusion assays confirmed that heterogeneously expressed GST proteins have antioxidant capabilities. These findings suggest that GST genes play a role in the response mechanism to high temperature stress by protecting against oxidative stress, providing a theoretical foundation for exploring the heat resistance mechanisms of M. alternatus in subtropical regions.

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Monochamus alternatus / heat stress / glutathione S-transferases / GST gene / oxidative stress / gene clone / express analysis / bioinformatics

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LI Zichun , HAO Dejun , LI Hui , et al . Cloning of glutathione S-transferases gene from Monochamus alternatus and its expression characteristics under heat stress[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2025, 49(1): 28-36 https://doi.org/10.12302/j.issn.1000-2006.202304018

References

List( Publishing order | Descend order by publishing year | Descend order by cited within ) Chart analysis
[1]
GARRAD R, BOOTH D T, FURLONG M J. The effect of rearing temperature on development, body size, energetics and fecundity of the diamondback moth[J]. Bull Entomol Res, 2016, 106(2): 175-181. DOI: 10.1017/S000748531500098X.
Cited in this article [1]
[2]
钱雪, 王月莹, 谢欢欢, 等. 温度对西伯利亚蝗呼吸代谢关键酶活性的影响[J]. 昆虫学报, 2017, 60(5): 499-504.
QIAN X, WANG Y Y, XIE H H, et al. Effects of temperature on the activities of key enzymes related to respiratory metabolism in adults of Gomphocerus sibiricus (Orthoptera: Acrididae)[J]. Acta Entomol Sin, 2017, 60(5): 499-504. DOI: 10.16380/j.kcxb.2017.05.001.
Cited in this article [1]
[3]
TURRENS J F. Mitochondrial formation of reactive oxygen species[J]. J Physiol, 2003, 552(2): 335-344. DOI: 10.1113/jphysiol.2003.049478.
Cited in this article [1]
[4]
HALLIWELL B. Antioxidants: the basics, what they are and how to evaluate them[J]. Adv Pharmacol, 1996, 38: 3-20. DOI: 10.1016/s1054-3589(08)60976-x.
Cited in this article [1]
[5]
SLIMEN I B, NAJAR T, GHRAM A, et al. Reactive oxygen species, heat stress and oxidative-induced mitochondrial damage: a review[J]. Int J Hyperthermia, 2014, 30(7): 513-523. DOI: 10.3109/02656736.2014.971446.
Cited in this article [1]
[6]
HABASHY W S, MILFORT M C, REKAYA R, et al. Cellular antioxidant enzyme activity and biomarkers for oxidative stress are affected by heat stress[J]. Int J Biometeorol, 2019, 63(12): 1569-1584. DOI: 10.1007/s00484-019-01769-z.
Cited in this article [1]
[7]
李慧, 郝德君, 徐天, 等. 高温胁迫对植食性昆虫影响研究进展[J]. 南京林业大学学报(自然科学版), 2022, 46(6): 215-224.
LI H, HAO D J, XU T, et al. The effects of heat stress on herbivorous insects: an overview and future directions[J]. J Nanjing For Univ (Nat Sci Ed), 2022, 46(6): 215-224. DOI: 10.12302/j.issn.1000-2006.202209041.
Cited in this article [1]
[8]
RIX R R, CUTLER G C. Review of molecular and biochemical responses during stress induced stimulation and hormesis in insects[J]. Sci Total Environ, 2022, 827: 154085. DOI: 10.1016/j.scitotenv.2022.154085.
Cited in this article [1]
[9]
ZHANG S Z, FU W Y, LI N, et al. Antioxidant responses of Propylaea japonica (Coleoptera: Coccinellidae) exposed to high temperature stress[J]. J Insect Physiol, 2015, 73: 47-52. DOI: 10.1016/j.jinsphys.2015.01.004.
Cited in this article [1]
[10]
KANG Z W, LIU F H, LIU X, et al. The potential coordination of the heat-shock proteins and antioxidant enzyme genes of Aphidius gifuensis in response to thermal stress[J]. Front Physiol, 2017, 8: 976. DOI: 10.3389/fphys.2017.00976.
Cited in this article [2]
[11]
MOREIRA D C, PAULA D P, HERMES-LIMA M. Changes in metabolism and antioxidant systems during tropical diapause in the sunflower caterpillar Chlosyne lacinia (Lepidoptera: Nymphalidae)[J]. Insect Biochem Mol Biol, 2021, 134: 103581. DOI: 10.1016/j.ibmb.2021.103581.
Cited in this article [1]
[12]
WANG Y J, QIU L, RANSON H, et al. Structure of an insect epsilon class glutathione S-transferase from the malaria vector Anopheles gambiae provides an explanation for the high DDT-detoxifying activity[J]. J Struct Biol, 2008, 164(2): 228-235. DOI: 10.1016/j.jsb.2008.08.003.
Cited in this article [1]
[13]
OAKLEY A. Glutathione transferases: a structural perspective[J]. Drug Metab Rev, 2011, 43(2): 138-151. DOI: 10.3109/03602532.2011.558093.
Cited in this article [1]
[14]
MANNERVIK B. Five decades with glutathione and the GSTome[J]. J Biol Chem, 2012, 287(9): 6072-6083. DOI: 10.1074/jbc.X112.342675.
Cited in this article [1]
[15]
GALLÉ Á, BELA K, HAJNAL Á, et al. Crosstalk between the redox signalling and the detoxification:GSTs under redox control?[J]. Plant Physiol Biochem, 2021, 169: 149-159. DOI: 10.1016/j.plaphy.2021.11.009.
Cited in this article [1]
[16]
HAYES J D, FLANAGAN J U, JOWSEY I R. Glutathione transferases[J]. Annu Rev Pharmacol Toxicol, 2005, 45: 51-88. DOI: 10.1146/annurev.pharmtox.45.120403.095857.
Cited in this article [1]
[17]
RAZA H. Dual localization of glutathione S-transferase in the cytosol and mitochondria: implications in oxidative stress, toxicity and disease[J]. FEBS J, 2011, 278(22): 4243-4251. DOI: 10.1111/j.1742-4658.2011.08358.x.
Cited in this article [1]
[18]
LABORDE E. Glutathione transferases as mediators of signaling pathways involved in cell proliferation and cell death[J]. Cell Death Differ, 2010, 17(9): 1373-1380. DOI: 10.1038/cdd.2010.80.
Cited in this article [1]
[19]
ZHU G D, XUE M, LUO Y, et al. Effects of short-term heat shock and physiological responses to heat stress in two Bradysia adults, Bradysia odoriphaga and Bradysia difformis[J]. Sci Rep, 2017, 7(1): 13381. DOI: 10.1038/s41598-017-13560-4.
Cited in this article [1]
[20]
ZHAO Y, LI Y Y, HE M, et al. Antioxidant responses of the pest natural enemy Hylyphantes graminicola (Araneae: Linyphiidae) exposed to short-term heat stress[J]. J Therm Biol, 2020, 87: 102477. DOI: 10.1016/j.jtherbio.2019.102477.
Cited in this article [1]
[21]
ALVES M, PEREIRA A, MATOS P, et al. Bacterial community associated to the pine wilt disease insect vectors Monochamus galloprovincialis and Monochamus alternatus[J]. Sci Rep, 2016, 6: 23908. DOI: 10.1038/srep23908.
Cited in this article [1]
[22]
TOGASHI K. Population density of Monochamus alternatus adults (Coleoptera: Cerambycidae) and incidence of pine wilt disease caused by Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)[J]. Res Popul Ecol, 1988, 30(2): 177-192. DOI: 10.1007/BF02513243.
Cited in this article [1]
[23]
王立超, 陈凤毛, 董晓燕, 等. 松墨天牛取食和产卵特性研究[J]. 南京林业大学学报(自然科学版), 2023, 47(2): 219-224.
WANG L C, CHEN F M, DONG X Y, et al. A study on feeding and oviposition characteristics of Monochamus alternatus[J]. J Nanjing For Univ (Nat Sci Ed), 2023, 47(2): 219-224. DOI: 10.12302/j.issn.1000-2006.202103022.
Cited in this article [1]
[24]
陈宏健, 郝德君, 田敏, 等. 室内饲养松墨天牛幼虫不同肠段细菌的群落结构及功能分析[J]. 南京林业大学学报(自然科学版), 2021, 45(3): 143-151.
CHEN H J, HAO D J, TIAN M, et al. The community structure and functional analysis of intestinal bacteria in Monochamus alternatus larvae reared indoors[J]. J Nanjing For Univ (Nat Sci Ed), 2021, 45(3): 143-151. DOI: 10.12302/j.issn.1000-2006.202004009.
Cited in this article [1]
[25]
田开慧, 陈怡帆, 周宏威. 湖南湘西州马尾松毛虫和松材线虫病发生非线性建模与预测[J]. 森林工程, 2022, 38 (6): 38-44.
TIAN K H, CHEN Y F, ZHOU H W. Prediction of occurrence trend of Dendrolimus punctatus and pine wilt disease in Xiangxi Prefecture[J]. For Eng, 2022, 38 (6): 38-44.
Cited in this article [1]
[26]
王洋, 陈军, 陈凤毛, 等. 松墨天牛取食期间传播松材线虫的特性[J]. 南京林业大学学报(自然科学版), 2019, 43(6): 1-10.
WANG Y, CHEN J, CHEN F M, et al. Transmission of Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae) through feeding activity of Monochamus alternatus (Coleoptera: Cerambycidae)[J]. J Nanjing For Univ (Nat Sci Ed), 2019, 43(6): 1-10. DOI: 10.3969/j.issn.1000-2006.201903001.
Cited in this article [1]
[27]
ZHAO L L, ZHANG S, WEI W, et al. Chemical signals synchronize the life cycles of a plant-parasitic nematode and its vector beetle[J]. Curr Biol, 2013, 23(20): 2038-2043. DOI: 10.1016/j.cub.2013.08.041.
Cited in this article [1]
[28]
OHSAWA M, AKIBA M. Possible altitude and temperature limits on pine wilt disease: the reproduction of vector sawyer beetles(Monochamus alternatus), survival of causal nematode (Bursaphelenchus xylophilus), and occurrence of damage caused by the disease[J]. Eur J For Res, 2014, 133(2): 225-233. DOI: 10.1007/s10342-013-0742-x.
Cited in this article [1]
[29]
吴佳雯, 尹艳楠, 谈家金, 等. 蜡样芽孢杆菌NJSZ-13菌株诱导马尾松抗松材线虫病研究[J]. 南京林业大学学报(自然科学版), 2022, 46(4): 53-58.
WU J W, YIN Y N, TAN J J, et al. A preliminary study on resistance of Pinus massoniana induced by Bacillus cereus NJSZ-13 strain to Bursaphelenchus xylophilus[J]. J Nanjing For Univ (Nat Sci Ed), 2022, 46(4): 53-58. DOI: 10.12302/j.issn.1000-2006.202006051.
Cited in this article [1]
[30]
杨帆, 零雅茗, 舒红, 等. 松材线虫Bx-TIMP克隆及功能研究[J]. 森林工程, 2022, 38 (2): 14-19.
YANG F, LING Y M, SHU H, et al. Cloning and functional analysis of Bursaphelenchus xylophilus Bx-TIMP[J]. For Eng, 2022, 38 (2): 14-19.
Cited in this article [1]
[31]
李慧, 何玄玉, 陶蓉, 等. 松墨天牛小热激蛋白基因的克隆、表达谱及对温度胁迫的响应[J]. 昆虫学报, 2018, 61(7): 749-760.
LI H, HE X Y, TAO R, et al. cDNA cloning and expression profiling of small heat shock protein genes and their response to temperature stress in Monochamus alternatus(Coleoptera: Cerambycidae)[J]. Acta Entomol Sin, 2018, 61(7): 749-760. DOI: 10.16380/j.kcxb.2018.07.001.
Cited in this article [1]
[32]
LI H, QIAO H, LIU Y J, et al. Characterization, expression profiling, and thermal tolerance analysis of heat shock protein 70 in pine sawyer beetle,Monochamus alternatus Hope (Coleoptera: Cerambycidae)[J]. Bull Entomol Res, 2021, 111(2): 217-228. DOI: 10.1017/S0007485320000541.
Cited in this article [1]
[33]
CAI Z L, CHEN J X, CHENG J, et al. Overexpression of three heat shock proteins protects Monochamus alternatus (Coleoptera: Cerambycidae) from thermal stress[J]. J Insect Sci, 2017, 17(6): 113. DOI: 10.1093/jisesa/iex082.
Cited in this article [1]
[34]
李慧. 热激蛋白在松墨天牛响应高温胁迫中的功能研究[D]. 南京: 南京林业大学, 2021.
LI H. Function analysis of heat shock protein in Monochamus alternatus response to high temperature[D]. Nanjing: Nanjing For Univ, 2021. DOI: 10.27242/d.cnki.gnjlu.2021.000015.
Cited in this article [3]
[35]
LI H, ZHAO X Y, QIAO H, et al. Comparative transcriptome analysis of the heat stress response in Monochamus alternatus Hope (Coleoptera: Cerambycidae)[J]. Front Physiol, 2020, 10: 1568. DOI: 10.3389/fphys.2019.01568.
Cited in this article [2]
[36]
LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR[J]. Methods, 2002, 25(4): 402-408. DOI: 10.1006/meth.2001.1262.
Cited in this article [2]
[37]
RAJARAPU S P, MAMIDALA P, HERMS D A, et al. Antioxidant genes of the emerald ash borer(Agrilus planipennis): gene characterization and expression profiles[J]. J Insect Physiol, 2011, 57(6): 819-824. DOI: 10.1016/j.jinsphys.2011.03.017.
Cited in this article [2]
[38]
SANDAMALIKA W M G, PRIYATHILAKA T T, LEE S, et al. Immune and xenobiotic responses of glutathione S-Transferase theta (GST-θ) from marine invertebrate disk abalone (Haliotis discus discus): with molecular characterization and functional analysis[J]. Fish Shellfish Immunol, 2019, 91: 159-171. DOI: 10.1016/j.fsi.2019.04.004.
Cited in this article [2]
[39]
SUN L L, YIN J J, DU H, et al. Characterisation of GST genes from the Hyphantria cunea and their response to the oxidative stress caused by the infection of Hyphantria cunea nucleopolyhedrovirus (HcNPV)[J]. Pestic Biochem Physiol, 2020, 163: 254-262. DOI: 10.1016/j.pestbp.2019.11.019.
Cited in this article [1]
[40]
李长春, 宁青, 戴余军, 等. 拟环纹豹蛛谷胱甘肽S-转移酶基因的克隆及表达分析[J]. 江苏农业学报, 2019, 35(5): 1068-1074.
LI C C, NING Q, DAI Y J, et al. Cloning and expression analysis of glutathione S-transferase gene in Pardosa pseudoannulata[J]. Jiangsu J Agri Sci, 2019, 35(5): 1068-1074. DOI: 10.3969/j.issn.1000-4440.2019.05.010.
Cited in this article [1]
[41]
YANG Q, LIU J P, WYCKHUYS K A G, et al. Impact of heat stress on the predatory ladybugs Hippodamia variegata and Propylaea quatuordecimpunctata[J]. Insects, 2022, 13(3): 306. DOI: 10.3390/insects13030306.
Cited in this article [1]
[42]
YANG X J, ZHENG H L, LIU Y Y, et al. Selection of reference genes for quantitative real-time PCR in Aquatica leii (Coleoptera: Lampyridae) under five different experimental conditions[J]. Front Physiol, 2020, 11: 555233. DOI: 10.3389/fphys.2020.555233.
Cited in this article [1]
[43]
张媛英. 中华蜜蜂谷胱甘肽S-转移酶和小分子热激蛋白基因的生物学功能分析[D]. 泰安: 山东农业大学, 2014.
ZHANG Y Y. Biological analysis of glutathione S-transferase and small heat shock protein genes in Apis cerana cerana[D]. Tai’an: Shandong Agricul Univ, 2014. DOI: 10.7666/d.Y2587648.
Cited in this article [1]
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