小麦TaLEA3基因家族的全基因组鉴定及其响应赤霉病的表达模式分析
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摘要
胚胎晚期丰富(late embryogenesis abundant,LEA)蛋白是高等植物胚胎发育后期种子中大量积累的一类蛋白质,在植物逆境胁迫方面具有重要作用。其中第三组的LEA蛋白(LEA3)是该类蛋白家族的主要成员,因其在植物抗旱耐盐性方面的卓越表现而被广泛研究。在前期研究中,通过高通量蛋白质组学的方法,在禾谷镰刀菌接种的小麦穗轴中筛选到1个与抗病相关的差异积累蛋白Wrab17(LEA3基因家族)。为了揭示小麦中TaLEA3基因家族的序列特征及其对生物胁迫的响应机制,采用生物信息学方法对该基因家族进行了系统研究。结果表明,在小麦全基因组中共鉴定到26个TaLEA3基因,命名为TaG3LEA-1～TaG3LEA-26,分布于小麦的9条染色体上。串联重复和片段复制是该基因家族扩张的主要方式;MEME分析表明该家族成员含有motif 2或motif 5,是LEA3家族的典型特征。通过基因表达分析和qRT-PCR验证,发现TaG3LEA-25在干旱和高温胁迫下均高表达,并且在不同赤霉病抗感材料间存在显著差异,推测其可能参与了小麦赤霉病抗性调控。研究结果为阐明小麦TaLEA3家族基因的进化关系提供了有价值的信息,并且为进一步探究其功能机制奠定了理论基础。
Late embryogenesis abundant(LEA) proteins are a class of proteins accumulating in seeds during the late stage of embryonic development in higher plant, and they play important roles in plant resistance to adversity stress. LEA3, the third group of the LEA proteins, is the major member of this family and has been extensively studied for its excellent performance in drought and salt tolerance in plants. In previous study, a differential accumulation protein Wrab17(TaLEA3 gene family) associated with Fusarium head blight(FHB) resistance was screened from wheat cobs inoculated with Fusarium graminearum by high-throughput proteomics strategy. In order to reveal the sequence characteristics of TaLEA3 gene family in wheat and its response mechanism to biological stress, bioinformatics method was used to systematically study the gene family. The results showed that a total of 26 TaLEA3 genes were present in wheat genome named TaG3 LEA-1～TaG3 LEA-26, which were distributed on nine chromosomes of wheat. Tandem duplication and segmental duplication were probably the main expansion ways of this gene family. And MEME analysis showed that the members of this family contained either motif 2 or motif 5, which is the typical feature of LEA3 family. Through expression profiling analysis and qRT-PCR verification, TaG3 LEA-25 was highly expressed under both drought and high temperature stresses, and there was also a significant difference in FHB inoculation test, suggesting that it might be involved in FHB resistance. The results of this study provided valuable information for elucidation of the evolutionary relationships of TaLEA3 gene family in wheat and laid a theoretical foundation for further exploration of their functional mechanisms.
Late embryogenesis abundant(LEA) proteins are a class of proteins accumulating in seeds during the late stage of embryonic development in higher plant, and they play important roles in plant resistance to adversity stress. LEA3, the third group of the LEA proteins, is the major member of this family and has been extensively studied for its excellent performance in drought and salt tolerance in plants. In previous study, a differential accumulation protein Wrab17(TaLEA3 gene family) associated with Fusarium head blight(FHB) resistance was screened from wheat cobs inoculated with Fusarium graminearum by high-throughput proteomics strategy. In order to reveal the sequence characteristics of TaLEA3 gene family in wheat and its response mechanism to biological stress, bioinformatics method was used to systematically study the gene family. The results showed that a total of 26 TaLEA3 genes were present in wheat genome named TaG3 LEA-1～TaG3 LEA-26, which were distributed on nine chromosomes of wheat. Tandem duplication and segmental duplication were probably the main expansion ways of this gene family. And MEME analysis showed that the members of this family contained either motif 2 or motif 5, which is the typical feature of LEA3 family. Through expression profiling analysis and qRT-PCR verification, TaG3 LEA-25 was highly expressed under both drought and high temperature stresses, and there was also a significant difference in FHB inoculation test, suggesting that it might be involved in FHB resistance. The results of this study provided valuable information for elucidation of the evolutionary relationships of TaLEA3 gene family in wheat and laid a theoretical foundation for further exploration of their functional mechanisms.
引文
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[3] Dure L.A repeating 11-mer amino acid motif and plant desiccation[J].Plant J.,1993,3(3):363-369.
[4] Tolleter D,Jaquinod M,Mangavel C,et al..Structure and function of a mitochondrial late embryogenesis abundant protein are revealed by desiccation[J].Plant Cell,2007,19(5):1580-1589.
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[6] Gal T Z,Glazer I,Koltai H.An LEA group 3 family member is involved in survival of C.elegans during exposure to stress[J].FEBS Lett.,2004,577(1):21-26.
[7] 张林生,赵文明.LEA蛋白与植物的抗旱性[J].植物生理学报,2003,39(1):61-66.
[8] Xu D,Duan X,Wang B,et al..Expression of a late embryogenesis abundant protein gene,HVA1,from barley confers tolerance to water deficit and salt stress in transgenic rice[J].Plant Physiol.,1996,110(1):249-257.
[9] 王瑛,朱宝成,孙毅,等.外源lea3基因转化紫花苜蓿的研究[J].核农学报,2007,21(3):249-252.
[10] 刘昀,李冉辉,汪为茂,等.植物抗逆蛋白(LEA3)22-氨基酸耐盐结构域在酵母细胞中的鉴定[J].植物研究,2009,29(1):74-79.
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[12] Liu J,Li L,Foroud N A,et al..Proteomics of bulked rachides combined with documented QTLs uncovers genotype non-specific players of the Fusarium head blight responses in wheat[J].Phytopathology,2018,109(1):111-119.
[13] Wang X S,Zhu H B,Jin G L,et al..Genome-scale identification and analysis of LEA genes in rice (Oryza sativa L.)[J].Plant Sci.,2007,172(2):414-420.
[14] Filiz E,Ozyigit I I,Tombuloglu H,et al..In silico comparative analysis of LEA (late embryogenesis abundant) proteins in Brachypodium distachyon L.[J].Plant Omics,2013,6(6):433-440.
[15] Finn R D,Bateman A,Clements J,et al..Pfam:The protein families database[J].Nucl.Acids Res.,2014,42(D1):D222-D230.
[16] Sisson S.Hidden Markov models for bioinformatics[J].J.Roy.Stati.Soc.Sta.,2004,167(1):194-195.
[17] Hundertmark M,Hincha D K.LEA (late embryogenesis abundant) proteins and their encoding genes in Arabidopsis thaliana[J].BMC Genomics,2008,9(1):118.
[18] Finn R D,Coggill P,Eberhardt R Y,et al..The Pfam protein families database:Towards a more sustainable future[J].Nucl.Acids Res.,2016,44(D1):D279-D285.
[19] Rey S,Acab M,Gardy J L,et al..PSORTdb:A protein subcellular localization database for bacteria[J].Nucl.Acids Res.,2005,33(D1):D164-D168.
[20] Chen C,Xia R,Chen H,et al..TBtools,a toolkit for Biologists integrating various HTS-data handling tools with a user-friendly interface[J].BioRxiv,2018,289660.
[21] Wang Y P,Tang H B,DeBarry J D,et al..MCScanX:A toolkit for detection and evolutionary analysis of gene synteny and collinearity[J].Nucl.Acids Res.,2012,40(7):e49.
[22] Wang D,Zhang Y,Zhang Z,et al..KaKs_Calculator 2.0:A toolkit incorporating gamma-series methods and sliding window strategies[J].Genom.Proteome.Bioinf.,2010,8(1):77-80.
[23] 李乐,许红亮,杨兴露,等.大豆LEA基因家族全基因组鉴定、分类和表达[J].中国农业科学,2011,44(19):3945-3954.
[24] Kumar S,Stecher G,Tamura K.MEGA7:Molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Mol.Boil.Evol.,2016,33(7):1870-1874.
[25] Zhang H,Gao S,Lercher M J,et al..EvolView,an online tool for visualizing,annotating and managing phylogenetic trees[J].Nucl.Acids Res.,2012,40(W1):W569-W572.
[26] Bailey T L,Boden M,Buske F A,et al..MEME SUITE:Tools for motif discovery and searching[J].Nucl.Acids Res.,2009,37(W1):W202-W208.
[27] Lescot M,Déhais P,Thijs G,et al..PlantCARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J].Nucl.Acids Res.,2002,30(1):325-327.
[28] Liu Z,Xin M,Qin J,et al..Temporal transcriptome profiling reveals expression partitioning of homeologous genes contributing to heat and drought acclimation in wheat (Triticum aestivum L.)[J].BMC Plant Biol.,2015,15(1):152-162.
[29] Gou L,Hattori J,Fedak G,et al..Development and validation of Thinopyrum elongatum:Expressed molecular markers specific for the long arm of chromosome 7E[J].Crop Sci.,2016,56(1):354-364.
[30] Holub E B.The arms race is ancient history in Arabidopsis,the wildflower[J].Nat.Rev.Genet.,2001,2(7):516-527.
[31] Gordon C S,Rajagopalan N,Risseeuw E P,et al..Characterization of Triticum aestivum abscisic acid receptors and a possible role for these in mediating fusairum head blight susceptibility in wheat[J].PLoS ONE,2016,11(10):e0164996.
[32] Sun Y,Jin X,Jia X,et al..The role of wheat jasmonic acid and ethylene pathways in response to Fusarium graminearum infection[J].Plant Growth Regul.,2016,80(1):69-77.
[33] Makandar R,Nalam V J,Lee H,et al..Salicylic acid regulates basal resistance to Fusarium head blight in wheat[J].Mol.Plant Microbe Interact.,2012,25(3):431-439.
[34] Ried J L,Walker-Simmons M K.Group 3 late embryogenesis abundant proteins in desiccation-tolerant seedlings of wheat (Triticum aestivum L.) [J].Plant Physiol.,1993,102(1):125-131.
[35] 汤晓倩,于丽霞,武晓璐,等.第三组胚胎晚期丰富蛋白lea3基因研究概述[J].生命科学,2010,22(6):551-555.
[36] Talanova V V,Titov A F,Topchieva L V,et al..Expression patterns of ABA-dependent and ABA-independent genes during wheat cold adaptation[J].Russ.J.Plant Physiol.,2011,58(6):1005.
[37] Gaoshan Y,Na L,Dong D,et al..Functional characterization of the Wrab17 gene in the interaction process between wheat and Puccinia triticina[J].Plant Physiol.Biochem.,2018,133(1):100-106.
[38] 王莲哲,王渊,柳超奇,等.小麦TaLEA3-1基因的克隆、表达与渗透胁迫功能分析[J/OL].分子植物育种,http://kns.cnki.net/kcms/detail/46.1068.s.20181220.1039.002.html,2018.
[39] Oraby H F,Ransom C B,Kravchenko A N,et al..Barley HVA1 gene confers salt tolerance in R3 transgenic oat[J].Crop Sci.,2005,45(6):2218-2227.
[40] Hu T,Yang J,Yang Y,et al..Molecular characterization,heterologous expression and resistance analysis of OsLEA3-1 from Oryza sativa[J].Biologia,2014,69(5):625-634.
[41] Yang L,Li W,Xin X,et al..ZmLEA3,a multifunctional group 3 LEA protein from maize (Zea mays L.),is involved in biotic and abiotic stresses[J].Plant Cell Physiol.,2013,54(6):944-959.
[42] Liu Y,Zheng Y.PM2,a group 3 LEA protein from soybean,and its 22-mer repeating region confer salt tolerance in Escherichia coli[J].Biochem.Bioph.Res.Co.,2005,331(1):325-332.
[43] 赵咏梅,杨建雄,俞嘉宁.第3组LEA蛋白及其基因研究进展[J].西安文理学院学报(自然科学版),2006,9(4):27-30.
[2] Galau G A,Hughes D W,Dure L.Abscisic acid induction of cloned cotton late embryogenesis-abundant (Lea) mRNAs[J].Plant Mol.Biol.,1986,7(3):155-170.
[3] Dure L.A repeating 11-mer amino acid motif and plant desiccation[J].Plant J.,1993,3(3):363-369.
[4] Tolleter D,Jaquinod M,Mangavel C,et al..Structure and function of a mitochondrial late embryogenesis abundant protein are revealed by desiccation[J].Plant Cell,2007,19(5):1580-1589.
[5] Tunnacliffe A,Wise M J.The continuing conundrum of the LEA proteins[J].Naturwissenschaften,2007,94(10):791-812.
[6] Gal T Z,Glazer I,Koltai H.An LEA group 3 family member is involved in survival of C.elegans during exposure to stress[J].FEBS Lett.,2004,577(1):21-26.
[7] 张林生,赵文明.LEA蛋白与植物的抗旱性[J].植物生理学报,2003,39(1):61-66.
[8] Xu D,Duan X,Wang B,et al..Expression of a late embryogenesis abundant protein gene,HVA1,from barley confers tolerance to water deficit and salt stress in transgenic rice[J].Plant Physiol.,1996,110(1):249-257.
[9] 王瑛,朱宝成,孙毅,等.外源lea3基因转化紫花苜蓿的研究[J].核农学报,2007,21(3):249-252.
[10] 刘昀,李冉辉,汪为茂,等.植物抗逆蛋白(LEA3)22-氨基酸耐盐结构域在酵母细胞中的鉴定[J].植物研究,2009,29(1):74-79.
[11] 杨高山.小麦与叶锈菌互作中Wrab17基因的克隆、表达及功能研究[D].河北保定:河北农业大学,硕士学位论文,2015.
[12] Liu J,Li L,Foroud N A,et al..Proteomics of bulked rachides combined with documented QTLs uncovers genotype non-specific players of the Fusarium head blight responses in wheat[J].Phytopathology,2018,109(1):111-119.
[13] Wang X S,Zhu H B,Jin G L,et al..Genome-scale identification and analysis of LEA genes in rice (Oryza sativa L.)[J].Plant Sci.,2007,172(2):414-420.
[14] Filiz E,Ozyigit I I,Tombuloglu H,et al..In silico comparative analysis of LEA (late embryogenesis abundant) proteins in Brachypodium distachyon L.[J].Plant Omics,2013,6(6):433-440.
[15] Finn R D,Bateman A,Clements J,et al..Pfam:The protein families database[J].Nucl.Acids Res.,2014,42(D1):D222-D230.
[16] Sisson S.Hidden Markov models for bioinformatics[J].J.Roy.Stati.Soc.Sta.,2004,167(1):194-195.
[17] Hundertmark M,Hincha D K.LEA (late embryogenesis abundant) proteins and their encoding genes in Arabidopsis thaliana[J].BMC Genomics,2008,9(1):118.
[18] Finn R D,Coggill P,Eberhardt R Y,et al..The Pfam protein families database:Towards a more sustainable future[J].Nucl.Acids Res.,2016,44(D1):D279-D285.
[19] Rey S,Acab M,Gardy J L,et al..PSORTdb:A protein subcellular localization database for bacteria[J].Nucl.Acids Res.,2005,33(D1):D164-D168.
[20] Chen C,Xia R,Chen H,et al..TBtools,a toolkit for Biologists integrating various HTS-data handling tools with a user-friendly interface[J].BioRxiv,2018,289660.
[21] Wang Y P,Tang H B,DeBarry J D,et al..MCScanX:A toolkit for detection and evolutionary analysis of gene synteny and collinearity[J].Nucl.Acids Res.,2012,40(7):e49.
[22] Wang D,Zhang Y,Zhang Z,et al..KaKs_Calculator 2.0:A toolkit incorporating gamma-series methods and sliding window strategies[J].Genom.Proteome.Bioinf.,2010,8(1):77-80.
[23] 李乐,许红亮,杨兴露,等.大豆LEA基因家族全基因组鉴定、分类和表达[J].中国农业科学,2011,44(19):3945-3954.
[24] Kumar S,Stecher G,Tamura K.MEGA7:Molecular evolutionary genetics analysis version 7.0 for bigger datasets[J].Mol.Boil.Evol.,2016,33(7):1870-1874.
[25] Zhang H,Gao S,Lercher M J,et al..EvolView,an online tool for visualizing,annotating and managing phylogenetic trees[J].Nucl.Acids Res.,2012,40(W1):W569-W572.
[26] Bailey T L,Boden M,Buske F A,et al..MEME SUITE:Tools for motif discovery and searching[J].Nucl.Acids Res.,2009,37(W1):W202-W208.
[27] Lescot M,Déhais P,Thijs G,et al..PlantCARE,a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences[J].Nucl.Acids Res.,2002,30(1):325-327.
[28] Liu Z,Xin M,Qin J,et al..Temporal transcriptome profiling reveals expression partitioning of homeologous genes contributing to heat and drought acclimation in wheat (Triticum aestivum L.)[J].BMC Plant Biol.,2015,15(1):152-162.
[29] Gou L,Hattori J,Fedak G,et al..Development and validation of Thinopyrum elongatum:Expressed molecular markers specific for the long arm of chromosome 7E[J].Crop Sci.,2016,56(1):354-364.
[30] Holub E B.The arms race is ancient history in Arabidopsis,the wildflower[J].Nat.Rev.Genet.,2001,2(7):516-527.
[31] Gordon C S,Rajagopalan N,Risseeuw E P,et al..Characterization of Triticum aestivum abscisic acid receptors and a possible role for these in mediating fusairum head blight susceptibility in wheat[J].PLoS ONE,2016,11(10):e0164996.
[32] Sun Y,Jin X,Jia X,et al..The role of wheat jasmonic acid and ethylene pathways in response to Fusarium graminearum infection[J].Plant Growth Regul.,2016,80(1):69-77.
[33] Makandar R,Nalam V J,Lee H,et al..Salicylic acid regulates basal resistance to Fusarium head blight in wheat[J].Mol.Plant Microbe Interact.,2012,25(3):431-439.
[34] Ried J L,Walker-Simmons M K.Group 3 late embryogenesis abundant proteins in desiccation-tolerant seedlings of wheat (Triticum aestivum L.) [J].Plant Physiol.,1993,102(1):125-131.
[35] 汤晓倩,于丽霞,武晓璐,等.第三组胚胎晚期丰富蛋白lea3基因研究概述[J].生命科学,2010,22(6):551-555.
[36] Talanova V V,Titov A F,Topchieva L V,et al..Expression patterns of ABA-dependent and ABA-independent genes during wheat cold adaptation[J].Russ.J.Plant Physiol.,2011,58(6):1005.
[37] Gaoshan Y,Na L,Dong D,et al..Functional characterization of the Wrab17 gene in the interaction process between wheat and Puccinia triticina[J].Plant Physiol.Biochem.,2018,133(1):100-106.
[38] 王莲哲,王渊,柳超奇,等.小麦TaLEA3-1基因的克隆、表达与渗透胁迫功能分析[J/OL].分子植物育种,http://kns.cnki.net/kcms/detail/46.1068.s.20181220.1039.002.html,2018.
[39] Oraby H F,Ransom C B,Kravchenko A N,et al..Barley HVA1 gene confers salt tolerance in R3 transgenic oat[J].Crop Sci.,2005,45(6):2218-2227.
[40] Hu T,Yang J,Yang Y,et al..Molecular characterization,heterologous expression and resistance analysis of OsLEA3-1 from Oryza sativa[J].Biologia,2014,69(5):625-634.
[41] Yang L,Li W,Xin X,et al..ZmLEA3,a multifunctional group 3 LEA protein from maize (Zea mays L.),is involved in biotic and abiotic stresses[J].Plant Cell Physiol.,2013,54(6):944-959.
[42] Liu Y,Zheng Y.PM2,a group 3 LEA protein from soybean,and its 22-mer repeating region confer salt tolerance in Escherichia coli[J].Biochem.Bioph.Res.Co.,2005,331(1):325-332.
[43] 赵咏梅,杨建雄,俞嘉宁.第3组LEA蛋白及其基因研究进展[J].西安文理学院学报(自然科学版),2006,9(4):27-30.