茶树CsGPX基因全基因组鉴定和表达分析
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摘要
为明确茶树谷胱甘肽过氧化物酶(CsGPX)基因与非生物胁迫的关系,本研究利用生物信息学手段在茶树基因组中筛选得到3条CsGPX基因,对其编码蛋白理化性质、基因结构、系统进化树、顺式作用元件进行了分析。结果表明,CsGPX包含完整GPX结构域和3段保守序列,都具6个外显子。预测启动子上有参与植物激素和非生物胁迫响应的顺式作用元件。使用实时荧光定量PCR测定该基因的表达谱,发现它们在不同组织中的表达有显著差异。进一步分析表明,CsGPX被低温、ABA、盐以及干旱处理上调表达,但CsGPX2和CsGPX3的表达受低温抑制。本研究为茶树CsGPX家族基因克隆和功能验证提供基础。
To reveal the possible involvement of glutathione peroxidase genes(CsGPX) in the responses of tea plants to abiotic stresses, three CsGPX genes were identified by bioinformatics tools from the tea genome data, and the gene structures, the cis-elements on their promoters, the physiochemical characteristics of the encoded proteins and the phylogenetic trees were also studied. The results showed that all the CsGPX genes contained 6 exons and shared the gene family-specific GPX domain in addition to three other conserved domains. Cis-elements for the responses of plants to hormones and abiotic stresses were identified on the promoters. Quantitative real-time PCR analysis of the expression profiles revealed that they were significantly different in expression in different tissues. Further analysis showed that the three CsGPX genes were, in general, up-regulated by treatments of low temperature, ABA, salt and drought except that CsGPX2 and CsGPX3 were down-regulated by treatment of low-temperature. The results should have laid down a basis for further characterization of CsGPX family genes in tea plant.
To reveal the possible involvement of glutathione peroxidase genes(CsGPX) in the responses of tea plants to abiotic stresses, three CsGPX genes were identified by bioinformatics tools from the tea genome data, and the gene structures, the cis-elements on their promoters, the physiochemical characteristics of the encoded proteins and the phylogenetic trees were also studied. The results showed that all the CsGPX genes contained 6 exons and shared the gene family-specific GPX domain in addition to three other conserved domains. Cis-elements for the responses of plants to hormones and abiotic stresses were identified on the promoters. Quantitative real-time PCR analysis of the expression profiles revealed that they were significantly different in expression in different tissues. Further analysis showed that the three CsGPX genes were, in general, up-regulated by treatments of low temperature, ABA, salt and drought except that CsGPX2 and CsGPX3 were down-regulated by treatment of low-temperature. The results should have laid down a basis for further characterization of CsGPX family genes in tea plant.
引文
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[24]Yue C,Cao H,Wang L,et al.Molecular cloning and expression analysis of tea plant aquaporin(AQP)gene family[J].Plant Physiology and Biochemistry,2014,83:65-76.
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[26]Cao H,Wang L,Yue C,et al.Isolation and expression analysis of 18 CsbZIP genes implicated in abiotic stress responses in the tea plant(Camellia sinensis)[J].Plant Physiology and Biochemistry,2015,97:432-442.
[27]Ramos J,Matamoros M A,Naya L,et al.The glutathione peroxidase gene family of Lotus japonicus:characterization of genomic clones,expression analyses and immunolocalization in legumes[J].The New Phytologist 2009,181(1):103-114.
[28]Zhai C,Zhao L,Yin L,et al.Two wheat glutathione peroxidase genes whose products are located in chloroplasts improve salt and H2O2 tolerances in Arabidopsis[J].PLoS One,2013,8(10):e73989.
[29]Liu W,Zhao C,Wang P,et al.The response of glutathione peroxidase 1 and glutathione peroxidase 7 under different oxidative stresses in black tiger shrimp,Penaeus monodon[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2018,217:1-13.
[30]Attacha S,Solbach D,Bela K,et al.Glutathione peroxidase-like enzymes cover five distinct cell compartments and membrane surfaces in Arabidopsis thaliana[J].Plant,Cell&Environment,2017,40(8):1281-1295.
[31]Navrot N,Collin V,Gualberto J,et al.Plant glutathione peroxidases are functional peroxiredoxins distributed in several subcellular compartments and regulated during biotic and abiotic stresses[J].Plant Physiology,2006,142(4):1364-1379.
[32]Kim Y,Jang M,Noh H,et al.Molecular characterization of two glutathione peroxidase genes of Panax ginseng and their expression analysis against environmental stresses[J].Gene,2014,535(1):33-41.
[33]Gao F,Chen J,Ma T,et al.The glutathione peroxidase gene family in Thellungiella salsuginea:genome-wide identification,classification,and gene and protein expression analysis under stress conditions[J].International Journal of Molecular Sciences,2014,15(2):3319-3335.
[34]Passaia G,Queval G,Bai J,et al.The effects of redox controls mediated by glutathione peroxidases on root architecture in Arabidopsis thaliana[J].Journal of Experimental Botany,2014,65(5):1403-1413.
[35]Miao Y,Lv D,Wang P,et al.An Arabidopsis glutathione peroxidase functions as both a redox transducer and a scavenger in abscisic acid and drought stress responses[J].The Plant Cell,2006,18(10):2749-2766.
[36]Yu L,Ma J,Niu Z,et al.Tissue-specific transcriptome analysis reveals multiple responses to salt stress in populus euphratica seedlings[J].Genes,2017,8(12):372.
[37]Islam T,Manna M,Reddy M K.Glutathione peroxidase of pennisetum glaucum(PgGPx)Is a functional Cd2+dependent peroxiredoxin that enhances tolerance against salinity and drought stress[J].PLoS One,2015,10(11):e143344.
[38]Koua D,Cerutti L,Falquet L,et al.PeroxiBase:a database with new tools for peroxidase family classification[J/OL].Nucleic Acids Research,2009,37(suppl_1):D261-D266.[2018-10-11].https://doi.org/10.1093/nar/gkn680
[39]Yoo Y,Anil Kumar N C,Park J,et al.Global analysis of differentially expressed genes between japonica and indica rice roots reveals the molecular basis for enhanced cold tolerance in japonica rice[J].Plant Biotechnology Reports,2017,11(6):461-473.
[40]Zhao J,Zhang S,Yang T,et al.Global transcriptional profiling of a cold-tolerant rice variety under moderate cold stress reveals different cold stress response mechanisms[J].Physiologia Plantarum,2015,154(3):381-394.
[41]Lim C W,Baek W,Jung J,et al.Function of ABA in stomatal defense against biotic and drought stresses[J].International Journal of Molecular Sciences,2015,16(12):15251-15270.
[42]Dong W,Wang M,Xu F,et al.Wheat oxophytodienoate reductase gene TaOPR1 confers salinity tolerance via enhancement of abscisic acid signaling and reactive oxygen species scavenging[J].Plant Physiology 2013,161(3):1217-1228.
[43]Gaber A.The importance of Arabidopsis glutathione peroxidase 8 for protecting Arabidopsis plant and E.coli cells against oxidative stress[J].GM Crops&Food,2014,5(1):20-26.
[2]Shakir S K,Irfan S,Akhtar B,et al.Pesticide-induced oxidative stress and antioxidant responses in tomato(Solanum lycopersicum)seedlings[J].Ecotoxicology,2018(9):1-17.
[3]Kumar D,Yusuf M A,Singh P,et al.Modulation of antioxidant machinery in a-tocopherol-enriched transgenic Brassica juncea plants tolerant to abiotic stress conditions[J].Protoplasma,2013,250(5):1079-1089.
[4]Manquián-Cerda K,Cruces E,Escudey M,et al.Interactive effects of aluminum and cadmium on phenolic compounds,antioxidant enzyme activity and oxidative stress in blueberry(Vaccinium corymbosum L.)plantlets cultivated in vitro[J].Ecotoxicology and Environmental Safety,2018,150:320-326.
[5]Mates J M.Effects of antioxidant enzymes in the molecular control of reactive oxygen species toxicology[J].Toxicology,2000,153(1-3):83-104.
[6]Gill S S,Tuteja N.Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants[J].Plant Physiology and Biochemistry,2010,48(12):909-930.
[7]You J,Chan Z.ROS regulation during abiotic stress responses in crop plants[J].Front Plant Science,2015,6:1092.
[8]Zhou Y,Hu L,Ye S,et al.Genome-wide identification of glutathione peroxidase(GPX)gene family and their response to abiotic stress in cucumber[J].3 Biotech,2018,8(3):159.
[9]Herbette S,Menn A L,Rousselle P,et al.Modification of photosynthetic regulation in tomato overexpressing glutathione peroxidase[J].Biochimica et Biophysica Acta(BBA)-General Subjects,2005,1724(1-2):108-118.
[10]Iqbal A,Yabuta Y,Takeda T,et al.Hydroperoxide reduction by thioredoxin-specific glutathione peroxidase isoenzymes of Arabidopsis thaliana[J].The Federation of European Biochemical Societies Journal,2006,273(24):5589-5597.
[11]Milla M A R,Maurer A,Huete A R,et al.Glutathione peroxidase genes in Arabidopsis are ubiquitous and regulated by abiotic stresses through diverse signaling pathways[J].The Plant Journal,2003,36(5):602-615.
[12]Islam T,Manna M,Kaul T,et al.Genome-wide dissection of Arabidopsis and rice for the identification and expression analysis of glutathione peroxidases reveals their stress-specific and overlapping response patterns[J].Plant Molecular Biology Reporter,2015,33(5):1413-1427.
[13]Chen M,Li K,Li H,et al.The glutathione peroxidase gene family in Gossypium hirsutum:genome-wide identification,classification,gene expression and functional analysis[J].Scientific Reports,2017,7:44743.
[14]Martinez V,Nieves-Cordones M,Lopez-Delacalle M,et al.Tolerance to stress combination in tomato plants:New insights in the protective role of melatonin[J].Molecules,2018,23(3):535.
[15]Xing X,Fang C,Li L,et al.Improved drought tolerance byα-naphthaleneacetic acid-induced ROS accumulation in two soybean cultivars[J].Journal of Integrative Agriculture,2016,15(8):1770-1784.
[16]Ren L,Zhang D,Chen G,et al.Transcriptomic profiling revealed the regulatory mechanism of Arabidopsis seedlings response to oxidative stress from cryopreservation[J].Plant Cell Reports,2015,34(12):2161-2178.
[17]Wang X,Fang G,Yang J,et al.A thioredoxin-dependent glutathione peroxidase(OsGPX5)is required for rice normal development and salt stress tolerance[J].Plant Molecular Biology Reporter,2017,35(3):333-342.
[18]Zhou X,Chen S,Wu H,et al.Biochemical and proteomics analyses of antioxidant enzymes reveal the potential stress tolerance in Rhododendron chrysanthum Pall[J].Biology Direct,2017,12:10.
[19]Liu S C,Jin J Q,Ma J Q,et al.Transcriptomic analysis of tea plant responding to drought stress and recovery[J].PLoSONE,2016,11(1):e0147306.
[20]Hao X,Yang Y,Yue C,et al.Comprehensive transcriptome analyses reveal differential gene expression profiles of Camellia sinensis axillary buds at para-,endo-,ecodormancy,and bud flush stages[J].Frontiers in Plant Science,2017,8:553.
[21]Cheruiyot E K,Mumera L M,Ng'etich W K,et al.High fertilizer rates increase susceptibility of tea to water stress[J].Journal of Plant Nutrition,2009,33(1):115-129.
[22]Xia E,Zhang H,Sheng J,et al.The tea tree genome provides insights into tea flavor and independent evolution of caffeine biosynthesis[J].Molecular Plant,2017,10(6):866-877.
[23]Wei C L,Yang H,Wang S,et al.Draft genome sequence of Camellia sinensis var.sinensis provides insights into the evolution of the tea genome and tea quality[J].Proceedings of the National Academy of Sciences,2018,115(18):E4151-E4158.
[24]Yue C,Cao H,Wang L,et al.Molecular cloning and expression analysis of tea plant aquaporin(AQP)gene family[J].Plant Physiology and Biochemistry,2014,83:65-76.
[25]Hao X,Horvath D,Chao W,et al.Identification and evaluation of reliable reference genes for quantitative real-time PCR analysis in tea plant(Camellia sinensis(L.)O.Kuntze)[J].International Journal of Molecular Sciences,2014,15(12):22155-22172.
[26]Cao H,Wang L,Yue C,et al.Isolation and expression analysis of 18 CsbZIP genes implicated in abiotic stress responses in the tea plant(Camellia sinensis)[J].Plant Physiology and Biochemistry,2015,97:432-442.
[27]Ramos J,Matamoros M A,Naya L,et al.The glutathione peroxidase gene family of Lotus japonicus:characterization of genomic clones,expression analyses and immunolocalization in legumes[J].The New Phytologist 2009,181(1):103-114.
[28]Zhai C,Zhao L,Yin L,et al.Two wheat glutathione peroxidase genes whose products are located in chloroplasts improve salt and H2O2 tolerances in Arabidopsis[J].PLoS One,2013,8(10):e73989.
[29]Liu W,Zhao C,Wang P,et al.The response of glutathione peroxidase 1 and glutathione peroxidase 7 under different oxidative stresses in black tiger shrimp,Penaeus monodon[J].Comparative Biochemistry and Physiology Part B:Biochemistry and Molecular Biology,2018,217:1-13.
[30]Attacha S,Solbach D,Bela K,et al.Glutathione peroxidase-like enzymes cover five distinct cell compartments and membrane surfaces in Arabidopsis thaliana[J].Plant,Cell&Environment,2017,40(8):1281-1295.
[31]Navrot N,Collin V,Gualberto J,et al.Plant glutathione peroxidases are functional peroxiredoxins distributed in several subcellular compartments and regulated during biotic and abiotic stresses[J].Plant Physiology,2006,142(4):1364-1379.
[32]Kim Y,Jang M,Noh H,et al.Molecular characterization of two glutathione peroxidase genes of Panax ginseng and their expression analysis against environmental stresses[J].Gene,2014,535(1):33-41.
[33]Gao F,Chen J,Ma T,et al.The glutathione peroxidase gene family in Thellungiella salsuginea:genome-wide identification,classification,and gene and protein expression analysis under stress conditions[J].International Journal of Molecular Sciences,2014,15(2):3319-3335.
[34]Passaia G,Queval G,Bai J,et al.The effects of redox controls mediated by glutathione peroxidases on root architecture in Arabidopsis thaliana[J].Journal of Experimental Botany,2014,65(5):1403-1413.
[35]Miao Y,Lv D,Wang P,et al.An Arabidopsis glutathione peroxidase functions as both a redox transducer and a scavenger in abscisic acid and drought stress responses[J].The Plant Cell,2006,18(10):2749-2766.
[36]Yu L,Ma J,Niu Z,et al.Tissue-specific transcriptome analysis reveals multiple responses to salt stress in populus euphratica seedlings[J].Genes,2017,8(12):372.
[37]Islam T,Manna M,Reddy M K.Glutathione peroxidase of pennisetum glaucum(PgGPx)Is a functional Cd2+dependent peroxiredoxin that enhances tolerance against salinity and drought stress[J].PLoS One,2015,10(11):e143344.
[38]Koua D,Cerutti L,Falquet L,et al.PeroxiBase:a database with new tools for peroxidase family classification[J/OL].Nucleic Acids Research,2009,37(suppl_1):D261-D266.[2018-10-11].https://doi.org/10.1093/nar/gkn680
[39]Yoo Y,Anil Kumar N C,Park J,et al.Global analysis of differentially expressed genes between japonica and indica rice roots reveals the molecular basis for enhanced cold tolerance in japonica rice[J].Plant Biotechnology Reports,2017,11(6):461-473.
[40]Zhao J,Zhang S,Yang T,et al.Global transcriptional profiling of a cold-tolerant rice variety under moderate cold stress reveals different cold stress response mechanisms[J].Physiologia Plantarum,2015,154(3):381-394.
[41]Lim C W,Baek W,Jung J,et al.Function of ABA in stomatal defense against biotic and drought stresses[J].International Journal of Molecular Sciences,2015,16(12):15251-15270.
[42]Dong W,Wang M,Xu F,et al.Wheat oxophytodienoate reductase gene TaOPR1 confers salinity tolerance via enhancement of abscisic acid signaling and reactive oxygen species scavenging[J].Plant Physiology 2013,161(3):1217-1228.
[43]Gaber A.The importance of Arabidopsis glutathione peroxidase 8 for protecting Arabidopsis plant and E.coli cells against oxidative stress[J].GM Crops&Food,2014,5(1):20-26.