| 363 | 2 | 169 |
| 下载次数 | 被引频次 | 阅读次数 |
谷胱甘肽S-转移酶（GST）可催化还原型谷胱甘肽（GSH）与底物连接，形成水溶性产物。植物受到胁迫时，GST会诱导表达，保护植物免受伤害，对植物抗逆起到重要作用。文章研究分析了拟南芥AtGST8的定位、表达及其过表达植株对甲基紫精（MV）胁迫的响应。结果表明：AtGST8定位在细胞膜。除种子和胚外，AtGST8在其余器官中均有表达，在莲座叶中表达较弱，而在花和果荚中的表达与生育进程有关。在果荚中表达量最高，叶中表达量最低。MV显著提高了所有供试材料幼苗中丙二醛（MDA）和抗坏血酸（AsA）含量，诱导超氧阴离子（O2·-）的产生，过表达AtGST8植株的MDA和O2·-的增幅小于野生型，AsA则相反。MV处理明显降低了野生型中的还原型谷胱甘肽/氧化型谷胱甘肽（GSH/GSSG）水平，在过表达AtGST8植株中的变化则不明显；MV处理显著增加了所有供试材料中氧化胁迫保护酶（超氧化物歧化酶（SOD）、过氧化物酶（POD）、过氧化氢酶（CAT）和抗坏血酸过氧化物酶（APX））的活性；过表达AtGST8植株中GSH过氧化物酶（GSH-Px）和GST活性明显增大，而野生型变化不明显；MV处理的保护酶相关基因（SAPX、PRX34、CAT1、GST8）的表达水平与酶活表现一致。过表达AtGST8植株遭受MV胁迫后能够维持较高的AsA含量、GSH/GSSG水平、抗氧化酶活性及转录水平，从而及时清除体内的活性氧以减轻MV胁迫对植物的伤害。
Abstract：Reduced glutathione(GSH) is connected to the substrate under the catalysis of glutathione S-transferase(GST) to form a water-soluble product. When plants are stressed, GST will induce expression to protect plants from damage and play an important role in plant stress resistance. In this study, the localization, expression of AtGST8 and the response of over-expressed plants to methyl viologen(MV) stress in Arabidopsis thaliana are analyzed. The results show that AtGST8 was localized in cell membrane and expressed in all organs except seeds and embryos. The expression was weak in rosette leaves, but was related to the reproductive process in flowers and pods. AtGST8showed the highest expression in pods and the lowest expression in leaves. MV significantly increased the content of malondialdehyde(MDA) and ascorbic acid(AsA) in seedlings, and induced the production of superoxide anion(O2·-). The increase of MDA and O2·-in the over-expressed lines of AtGST8 was less than that in the wild type, while the AsA showed the opposite trend. MV treatment significantly decreased GSH/GSSG level in wild type, but the change was not obvious in the over-expressed lines of AtGST8. The activities of oxidative stress protective enzymes,including superoxide dismutase(SOD), peroxidase(POD), catalase(CAT) and ascorbic acid peroxidase(APX), were significantly increased by MV in all materials. The activities of GSH peroxidase(GSH-Px) and GST in the over-expressed lines of AtGST8 were significantly increased, which was not found in the wild type. The expression level of protective enzyme related genes(SAPX, PRX34, CAT1, GST8) under MV treatment was consistent with the enzyme activity. This study suggests that over-expressed plants of AtGST8 can maintain high AsA content, GSH/GSSG level, antioxidant enzyme activities and transcription levels after MV stress, so as to remove reactive oxygen species in the body in time and reduce the damage of MV stress to plants.
[1] CUMMINS I, DIXON D P, FREITAG-POHL S, et al.Multiple roles for plant glutathione transferases in xenobiotic detoxification[J]. Drug Metabolism Reviews, 2011, 43(2):266-280.
[2] EDWARDS R, DIXON D P, WALBOT V. Plant glutathione S-transferases:enzymes with multiple functions in sickness and in health[J]. Trends in Plant Science, 2000,5(5):193-198.
[3] KUMAR S, ASIF M H, CHAKRABARTY D, et al. Expression of a rice Lambda class of glutathione S-transferase, OsGSTL2, in Arabidopsis provides tolerance to heavy metal and other abiotic stresses[J]. Journal of Hazardous Materials, 2013, 248/249:228-237.
[4] CUI F Q, BROSCH魪M, SHAPIGUZOV A, et al. Interaction of methyl viologen-induced chloroplast and mitochondrial signalling in Arabidopsis[J]. Free Radical Biology and Medicine, 2019, 134:555-566.
[5] WASZCZAK C, CARMODY M, KANGASJ?RVI J. Reactive oxygen species in plant signaling[J]. Annual Review of Plant Biology, 2018, 69:209-236.
[6] HAYES J D, FLANAGAN J U, JOWSEY I R. Glutathione transferases[J]. Annual Review of Pharmacology and Toxicology, 2005, 45:51-88.
[7] SAPPL P G, O?ATE-SáNCHEZ L, SINGH K B, et al.Proteomic analysis of glutathione S-transferases of Arabidopsis thaliana reveals differential salicylic acid-induced expression of the plant-specific phi and tau classes[J].Plant Molecular Biology, 2004, 54(2):205-219.
[8] ZHAO Y, LIU M, HE L, et al. A cytosolic NAD+-dependent GPDH from maize(ZmGPDH1)is involved in conferring salt and osmotic stress tolerance[J]. BMC Plant Biology, 2019, 19(1):16.
[9] POLIDOROS A N, SCANDALIOS J G. Role of hydrogen peroxide and different classes of antioxidants in the regulation of catalase and glutathione S-transferase gene expression in maize(Zea mays L.)[J]. Physiologia Plantarum,1999, 106(1):112-120.
[10]胡廷章，周大祥，罗凯.植物谷胱甘肽转移酶的结构与功能及其基因表达[J].植物生理学通讯，2007, 43(1):195-200.HU T Z, ZHOU D X, LUO K. Structure and biological function of glutathione transferases and their genes in plants[J]. Plant Physiology Communications, 2007, 43(1):195-200.(in Chinese)
[11] XU J, TIAN Y S, XING X J, et al. Over-expression of AtGSTU19 provides tolerance to salt, drought and methyl viologen stresses in Arabidopsis[J]. Physiologia Plantarum,2016, 156(2):164-175.
[12] THOM R, DIXON D P, EDWARDS R, et al. The structure of a zeta class glutathione S-transferase from Arabidopsis thaliana:characterisation of a GST with novel active-site architecture and a putative role in tyrosine catabolism[J]. Journal of Molecular Biology, 2001, 308(5):949-962.
[13] MCGONIGLE B, KEELER S J, LAU S M C, et al. A genomics approach to the comprehensive analysis of the glutathione S-transferase gene family in soybean and maize[J].Plant Physiology, 2000, 124(3):1105-1120.
[14] SORANZO N, GORLA M S, MIZZI L, et al. Organisation and structural evolution of the rice glutathione S-transferase gene family[J]. Molecular Genetics and Genomics,2004, 271(5):511-521.
[15] DENG F, HATZIOS K K. Purification and characterization of two glutathione S-transferase isozymes from Indica-type rice involved in herbicide detoxification[J]. Pesticide Biochemistry and Physiology, 2002, 72(1):10-23.
[16]史倩倩，周琳，王雁.云南野生黄牡丹谷胱甘肽转移酶（GST）基因的分离及表达分析[J].林业科学，2014, 50(12):63-72.SHI Q Q, ZHOU L, WANG Y. Isolation and expression analysis of GST gene encoding glutathione S-transferase of Paeonia delavayi var. lutea wild population in Yunnan[J].Scientia Silvae Sinicae, 2014, 50(12):63-72.(in Chinese)
[17] AGRAWAL G K, JWA N S, RAKWAL R. A pathogeninduced novel rice(Oryza sativa L.)gene encodes a putative protein homologous to type II glutathione S-transferases[J]. Plant Science, 2002, 163(6):1153-1160.
[18] BIANCHI M W, ROUX C, VARTANIAN N. Drought regulation of GST8, encoding the Arabidopsis homologue of ParC/Nt107 glutathione transferase/peroxidase[J]. Physiologia Plantarum, 2002, 116(1):96-105.
[19] DALAL A, KUMAR A, YADAV D, et al. Alleviation of methyl viologen-mediated oxidative stress by Brassica juncea annexin-3 in transgenic Arabidopsis[J]. Plant Science, 2014, 219/220:9-18.
[20] CLOUGH S J, BENT A F. Floral dip:a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J]. The Plant Journal:for Cell and Molecular Biology, 1998, 16(6):735-743.
[21] JEFFERSON R A, KAVANAGH T A, BEVAN M W. GUS fusions:β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants[J]. The EMBO Journal,1987, 6(13):3901-3907.
[22] CAO Y Y, ZHANG Q, CHEN Y H, et al. Identification of differential expression genes in leaves of rice(Oryza sativa L.)in response to heat stress by cDNA-AFLP analysis[J].BioMed Research International, 2013, 2013:576189.
[23]许长成，邹琦，程炳嵩.硫代巴比妥酸（TBA）法检测脂质过氧化水平的探讨[J].植物生理学通讯，1989, 25(6):58-60.XU C C, ZOU Q, CHENG B S. Stady on thiobarbiburic acid(TBA)method for determination of lipid peroxidation[J]. Plant Physiology Communications, 1989, 25(6):58-60.(in Chinese)
[24] ZHOU R, HU Q J, PU Q, et al. Spermidine enhanced free polyamine levels and expression of polyamine biosynthesis enzyme gene in rice spikelets under heat tolerance before heading[J]. Scientific Reports, 2020, 10:8976.
[25]支添添，周舟，韩成云，等.台盼蓝染色鉴定拟南芥sdl1突变体的细胞死亡[J].作物研究，2013, 27(3):217-218.ZHI T T, ZHOU Z, HAN C Y, et al. Identification of cell death of Arabidopsis thaliana mutant sdl1 by trypan blue staining[J]. Crop Research, 2013, 27(3):217-218.(in Chinese)
[26]汪承润，何梅，李月云，等.植物体超氧阴离子自由基不同检测方法的比较[J].环境化学，2012, 31(5):726-730.WANG C R, HE M, LI Y Y, et al. Comparative study on different detection methods of superoxide radicals in plant tissues[J]. Environmental Chemistry, 2012, 31(5):726-730.(in Chinese)
[27] KN魻RZER O C, DURNER J, B魻GER P. Alterations in the antioxidative system of suspension-cultured soybean cells(Glycine max)induced by oxidative stress[J]. Physiologia Plantarum, 1996, 97(2):388-396.
[28]王秋林，王浩毅，丁怡，等.血浆中游离还原型及氧化型谷胱甘肽的荧光测定法[J].光谱学与光谱分析，2005,25(11):1834-1838.WANG Q L, WANG H Y, DING Y, et al. A novel fluorophotometric method for assaying free glutathione of both reduced and oxidized in plasma[J]. Spectroscopy and Spectral Analysis, 2005, 25(11):1834-1838.(in Chinese)
[29]赵雁，马向丽，毕玉芬，等.不同提取系统中分析紫花苜蓿抗氧化酶活性和同工酶的比较[J].云南农业大学学报（自然科学），2014, 29(5):636-641.ZHAO Y, MA X L, BI Y F, et al. Comparison of alfalfa antioxidant enzyme activity and isozyme in different extraction systems[J]. Journal of Yunnan Agricultural University(Natural Science), 2014, 29(5):636-641.(in Chinese)
[30] ALFENITO M R, SOUER E, GOODMAN C D, et al.Functional complementation of anthocyanin sequestration in the vacuole by widely divergent glutathione S-transferases[J]. The Plant Cell, 1998, 10(7):1135-1149.
[31] ALSCHER R G, ERTURK N, HEATH L S. Role of superoxide dismutases(SODs)in controlling oxidative stress in plants[J]. Journal of Experimental Botany, 2002, 53(372):1331-1341.
[32] GONG H B, JIAO Y X, HU W W, et al. Expression of glutathione-S-transferase and its role in plant growth and development in vivo and shoot morphogenesis in vitro[J].Plant Molecular Biology, 2005, 57(1):53-66.
[33] YU T, LI Y S, CHEN X F, et al. Transgenic tobacco plants overexpressing cotton glutathione S-transferase(GST)show enhanced resistance to methyl viologen[J]. Journal of Plant Physiology, 2003, 160(11):1305-1311.
[34] YANG G Y, WANG Y C, XIA D A, et al. Overexpression of a GST gene(ThGSTZ1)from Tamarix hispida improves drought and salinity tolerance by enhancing the ability to scavenge reactive oxygen species[J]. Plant Cell, Tissue and Organ Culture, 2014, 117(1):99-112.
[35]刘新仿，李家洋.紫外线强烈诱导的谷胱甘肽转移酶基因的功能鉴定[J].遗传学报，2002, 29(5):458-460.LIU X F, LI J Y. Characterization of an ultra-violet inducible gene that encodes glutathione S-transferase in Arabidopsis thaliana[J]. Acta Genetica Sinica, 2002, 29(5):458-460.(in Chinese)
[36] LIM J D, HAHN S J, YU C Y, et al. Expression of the glutathione S-transferase gene(NT107)in transgenic Dianthus superbus[J]. Plant Cell, Tissue and Organ Culture,2005, 80(3):277-286.
[37] HORVáTH E, BELA K, HOLINKA B, et al. The Arabidopsis glutathione transferases, AtGSTF8 and AtGSTU19are involved in the maintenance of root redox homeostasis affecting meristem size and salt stress sensitivity[J]. Plant Science, 2019, 283:366-374.
[38] DE GARA L, DE PINTO M C, ARRIGONI O. Ascorbate synthesis and ascorbate peroxidase activity during the early stage of wheat germination[J]. Physiologia Plantarum, 1997,100:894-900.
[39] GULLNER G, KOMIVES T, KIRáLY L, et al. Glutathione S-transferase enzymes in plant-pathogen interactions[J]. Frontiers in Plant Science, 2018, 9:1836.
[40] JIANG Y Q, YANG B, HARRIS N S, et al. Comparative proteomic analysis of NaCl stress-responsive proteins in Arabidopsis roots[J]. Journal of Experimental Botany, 2007,58(13):3591-3607.
基本信息:
DOI：10.12194/j.ntu.20220331001
中图分类号:Q945.78
引用信息:
[1]王玲娟,鲁帅,高聪,等.拟南芥AtGST8的功能及其过表达植株对甲基紫精胁迫的响应[J],2023,22(01):44-53.DOI:10.12194/j.ntu.20220331001.
基金信息:
国家自然科学基金面上项目（32172104）