稻曲病菌UvHog1基因的克隆及表达分析
|
摘要
稻曲病由稻曲病菌引起,是水稻穗部的重要病害。利用同源克隆和RACE技术,根据丝状真菌相对保守的氨基酸序列设计简并引物,从稻曲病菌中分离了丝裂原活化蛋白激酶(MAPK)编码基因的全长cDNA序列,命名为UvHog1。UvHog1含有MAPK保守的蛋白激酶激活域(TGY),序列与稻瘟病菌MgOsm1(AAF09475.1)、球孢白僵菌BbHog1(AAS77871.1)、烟曲霉AfOsm1(XP752664.1)和隐球酵母CrHog1(AAM26267.1)等MAPK高度同源,相似性分别为95%、93%、88%和81%。系统聚类结果表明,UvHog1与酵母Hog1MAPK同源。在盐胁迫条件下,UvHog1的相对表达量明显降低,表明UvHog1参与了对盐胁迫的信号响应。
Rice false smut,caused by Ustilaginoidea virens,is an important disease in rice panicle.Degenerate PCR primers were designed according to the conserved amino acid sequence of several filamentous fungus mitogen activated protein kinase(MAPK)homologous to yeast Hog1MAPK,and partial DNA fragment encoding a MAPK was amplified fromU.virens through PCR.Then,a whole DNA sequence encoding the MAPK,designated as UvHog1,was obtained from U.virens by extending upstream and downstream sequence of the amplified fragment using RACE method.And its amino acid sequence shared 95%,93%,88% and 81%identity to MgOsm1(AAF09475.1)from Magnaporthe grisea,BbHog1(AAS77871.1)from Beauveria bassiana,AfOsm1(XP752664.1)from Aspergillus fumigatus and CrHog1(AAM26267.1)from Cryptococcus neoformans var.neoformans,respectively.Phylogenetic clustering suggested that UvHog1 is homologous to yeast Hog1MAPK.The results of the decreased transcript levels of UvHog1 by salinity suggest that UvHog1 may be involved in the specifically responses to salt stresses.
Rice false smut,caused by Ustilaginoidea virens,is an important disease in rice panicle.Degenerate PCR primers were designed according to the conserved amino acid sequence of several filamentous fungus mitogen activated protein kinase(MAPK)homologous to yeast Hog1MAPK,and partial DNA fragment encoding a MAPK was amplified fromU.virens through PCR.Then,a whole DNA sequence encoding the MAPK,designated as UvHog1,was obtained from U.virens by extending upstream and downstream sequence of the amplified fragment using RACE method.And its amino acid sequence shared 95%,93%,88% and 81%identity to MgOsm1(AAF09475.1)from Magnaporthe grisea,BbHog1(AAS77871.1)from Beauveria bassiana,AfOsm1(XP752664.1)from Aspergillus fumigatus and CrHog1(AAM26267.1)from Cryptococcus neoformans var.neoformans,respectively.Phylogenetic clustering suggested that UvHog1 is homologous to yeast Hog1MAPK.The results of the decreased transcript levels of UvHog1 by salinity suggest that UvHog1 may be involved in the specifically responses to salt stresses.
引文
[1]姜慎,唐春生,谭志琼.国内外稻曲病研究现状.热带农业科学,2010,30(3):62-66.
[2]Zhu H C,Xu X P,Xiao G Y,et al.Enhancing disease resistances of super hybrid rice with four antifungal genes.Sci China Ser C-Life Sci,2007,50(1):31-39.
[3]陈丽,胡东维,陈美军,等.稻曲球及稻曲病菌菌落微结构的SEM观察.菌物学报,2007,26(1):89-96.
[4]李西川.酿酒酵母菌和白念珠菌中RCK2和HOG1蛋白激酶在高渗胁迫、氧化胁迫和细胞壁完整性方面的功能研究[学位论文].天津:天津大学,2009:8-12
[5]吴雪昌,胡森杰,钱凯先.酵母HOG-MAPK途径.细胞生物学杂志,2005,27:247-252.
[6]杨洪强,接玉玲.植物MAPK及其在病原信号传递中的作用.植物病理学报,2003,33(1):8-13.
[7]Chen R E,Thorner J.Function and regulation in MAPK signaling pathways:Lessons learned from the yeast Saccharomyces cerevisiae.Biochim Biophys Acta,2007,1773(8):1311-1340.
[8]Gustin M C,Alberty N J,Alexander M.MAP kinase pathways in the yeast Saccharomyces cerevisiae.Microbiol Mol Biol Rev,1998,62:1264-1301.
[9]Stock A M,Robinson V L,Goudreau P N.Two-component signal transduction.Annu Rev Biochem,2000,69:183-215.
[10]O′Rourke S M,Herskowitz I,O′Shea E K.Yeast go the whole HOG for the hyperosmotic response.Trends Genet,2002,18:405-412.
[11]Posas F,Saito H.Activation of the yeast SSK2 MAP kinase kinase kinase by the SSK1two-component response regulator.EMBO J,1998,17:1385-1394.
[12]Posas F,Witten E A,Saito H.Requirement of STE50for osmostress-induced activation of the STE11 mitogen-activated protein kinase kinase kinase in the high-osmolarity glycerol response pathway.Mol Cell Biol,1998,18:5788-5796.
[13]van Wuytswinkel O,Reiser V,Siderius M,et al.Response of Saccharomyces cerevisiae to severe osmotic stress:Evidence for a novel activation mechanism of the HOG MAP kinase pathway.Mol Microbiol,2000,37:382-397.
[14]Ramezani Rad M,Jansen G,Buhring F,et al.Ste50p is involved in regulating filamentous growth in the yeast Saccharomyces cerevisiae and associates with Stel lp.Mol Gen Genet,1998,259:29-38.
[15]Wu X C,Chi X Q,Wang P M,et al.The evolutionary rate variation among genes of HOG-signaling pathway in yeast genomes.Biol Direct,2010,5:46.
[16]Arana D M,Alonso-Monge R,Du C,et al.Differential susceptibility of mitogen-activated protein kinase pathway mutants to oxidative-mediated killing by phagocytes in the fungal pathogen Candida albicans.Cell Microbiol,2007,9:1647-1659.
[17]Dixon K P,Xu J R,Smirnoff N,et al.Independent signaling pathways regulate cellular turgor during hyperosmotic stress and appressorium-mediated plant infection by Magnaporthe grisea.Plant Cell,1999,11:2045-2058.
[18]Schuller C,Brewster J L,Alexander M R,et al.The HOG pathway controls osmotic regulation of transcription via the stress response element(STRE)of the Saccharomyces cerevisiae CTT1gene.J EMBO,1994,13:4382-4389.
[19]Horwitz B A,Sharon A,Lu S,et al.A G protein alpha subunit from Cochliobolus heterostrophus involved in mating and appressorium formation.Fungal Genet Biol,1999,26:19-32.
[20]Zhang Y,Neo S Y,Wang X H,et al.Axin forms a complex with MEKK1and activates c-Jun NH2-terminal kinase/stressactivated protein kinase through domains distinct from Wnt signaling.J Biol Chem,1999,274:35247-35254.
[21]Kojima K,Takano Y,Yoshimi A,et al.Fungicide activity through activation of a fungal signaling pathway.Mol Microbiol,2004,53(6):1785-1796.
[22]Zhang Y J,Zhao J H,Fang W G,et al.Mitogen-activated protein kinase hog1in the entomopathogenic fungus Beauveria bassiana regulates environmental stress responses and virulence to insects.Appl Environ Microbiol,2009,75(11):3787-3795.
[23]张震,王教瑜,杜新法,等.稻曲病菌cDNA文库的构建.植物病理学报,2008,38(5):462-467.
[24]张震,杜新法,柴荣耀,等.稻曲病菌PMK1类同源基因克隆及在稻瘟病菌遗传互补中的功能验证.微生物学报,2008,48(11):1473-1478.
[25]冯飞.极细链格袍菌HOG1、PBS2基因克隆及功能初步分析[学位论文].北京:中国农业科学院,2007:50-64.
[26]周洁,郑祥梓,兰斓,等.稻瘟病菌假定的糖基水解酶62家族初步研究.中国农业科学,2009,42(8):2754-2762.
[27]冯飞,纪春艳,杨秀芬,等.细极链格孢菌Hog1 MAPK(酵母)同源基因AtHOG1的克隆与功能分析.华北农学报,2009,24(4):74-79.
[2]Zhu H C,Xu X P,Xiao G Y,et al.Enhancing disease resistances of super hybrid rice with four antifungal genes.Sci China Ser C-Life Sci,2007,50(1):31-39.
[3]陈丽,胡东维,陈美军,等.稻曲球及稻曲病菌菌落微结构的SEM观察.菌物学报,2007,26(1):89-96.
[4]李西川.酿酒酵母菌和白念珠菌中RCK2和HOG1蛋白激酶在高渗胁迫、氧化胁迫和细胞壁完整性方面的功能研究[学位论文].天津:天津大学,2009:8-12
[5]吴雪昌,胡森杰,钱凯先.酵母HOG-MAPK途径.细胞生物学杂志,2005,27:247-252.
[6]杨洪强,接玉玲.植物MAPK及其在病原信号传递中的作用.植物病理学报,2003,33(1):8-13.
[7]Chen R E,Thorner J.Function and regulation in MAPK signaling pathways:Lessons learned from the yeast Saccharomyces cerevisiae.Biochim Biophys Acta,2007,1773(8):1311-1340.
[8]Gustin M C,Alberty N J,Alexander M.MAP kinase pathways in the yeast Saccharomyces cerevisiae.Microbiol Mol Biol Rev,1998,62:1264-1301.
[9]Stock A M,Robinson V L,Goudreau P N.Two-component signal transduction.Annu Rev Biochem,2000,69:183-215.
[10]O′Rourke S M,Herskowitz I,O′Shea E K.Yeast go the whole HOG for the hyperosmotic response.Trends Genet,2002,18:405-412.
[11]Posas F,Saito H.Activation of the yeast SSK2 MAP kinase kinase kinase by the SSK1two-component response regulator.EMBO J,1998,17:1385-1394.
[12]Posas F,Witten E A,Saito H.Requirement of STE50for osmostress-induced activation of the STE11 mitogen-activated protein kinase kinase kinase in the high-osmolarity glycerol response pathway.Mol Cell Biol,1998,18:5788-5796.
[13]van Wuytswinkel O,Reiser V,Siderius M,et al.Response of Saccharomyces cerevisiae to severe osmotic stress:Evidence for a novel activation mechanism of the HOG MAP kinase pathway.Mol Microbiol,2000,37:382-397.
[14]Ramezani Rad M,Jansen G,Buhring F,et al.Ste50p is involved in regulating filamentous growth in the yeast Saccharomyces cerevisiae and associates with Stel lp.Mol Gen Genet,1998,259:29-38.
[15]Wu X C,Chi X Q,Wang P M,et al.The evolutionary rate variation among genes of HOG-signaling pathway in yeast genomes.Biol Direct,2010,5:46.
[16]Arana D M,Alonso-Monge R,Du C,et al.Differential susceptibility of mitogen-activated protein kinase pathway mutants to oxidative-mediated killing by phagocytes in the fungal pathogen Candida albicans.Cell Microbiol,2007,9:1647-1659.
[17]Dixon K P,Xu J R,Smirnoff N,et al.Independent signaling pathways regulate cellular turgor during hyperosmotic stress and appressorium-mediated plant infection by Magnaporthe grisea.Plant Cell,1999,11:2045-2058.
[18]Schuller C,Brewster J L,Alexander M R,et al.The HOG pathway controls osmotic regulation of transcription via the stress response element(STRE)of the Saccharomyces cerevisiae CTT1gene.J EMBO,1994,13:4382-4389.
[19]Horwitz B A,Sharon A,Lu S,et al.A G protein alpha subunit from Cochliobolus heterostrophus involved in mating and appressorium formation.Fungal Genet Biol,1999,26:19-32.
[20]Zhang Y,Neo S Y,Wang X H,et al.Axin forms a complex with MEKK1and activates c-Jun NH2-terminal kinase/stressactivated protein kinase through domains distinct from Wnt signaling.J Biol Chem,1999,274:35247-35254.
[21]Kojima K,Takano Y,Yoshimi A,et al.Fungicide activity through activation of a fungal signaling pathway.Mol Microbiol,2004,53(6):1785-1796.
[22]Zhang Y J,Zhao J H,Fang W G,et al.Mitogen-activated protein kinase hog1in the entomopathogenic fungus Beauveria bassiana regulates environmental stress responses and virulence to insects.Appl Environ Microbiol,2009,75(11):3787-3795.
[23]张震,王教瑜,杜新法,等.稻曲病菌cDNA文库的构建.植物病理学报,2008,38(5):462-467.
[24]张震,杜新法,柴荣耀,等.稻曲病菌PMK1类同源基因克隆及在稻瘟病菌遗传互补中的功能验证.微生物学报,2008,48(11):1473-1478.
[25]冯飞.极细链格袍菌HOG1、PBS2基因克隆及功能初步分析[学位论文].北京:中国农业科学院,2007:50-64.
[26]周洁,郑祥梓,兰斓,等.稻瘟病菌假定的糖基水解酶62家族初步研究.中国农业科学,2009,42(8):2754-2762.
[27]冯飞,纪春艳,杨秀芬,等.细极链格孢菌Hog1 MAPK(酵母)同源基因AtHOG1的克隆与功能分析.华北农学报,2009,24(4):74-79.