酸碱度对StRALF基因表达量的影响及初步分析
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摘要
快速碱化因子(RALF)是一种能够参与植物发育调控的植物肽激素,在真菌中也广泛存在并且影响真菌对寄主的侵染状况。通过外源添加酸碱来明确pH值对玉米大斑菌生长的影响,使用实时荧光定量PCR技术分析玉米大斑病菌中StRALF基因在外源添加酸、碱时表达量的变化。运用预测网站对克隆得到的StRALF基因编码区蛋白序列的基本性质进行分析。结果表明,玉米大斑瓋菌在碱性环境下生长状态较好。碱性条件下,StRALF基因比酸性条件下基因表达量高。在侵染寄主过程中,96 h及以后表达量达到最大。经预测分析,初步确定其为分泌蛋白或膜蛋白。在弱碱性条件下,StRALF基因表达量较高并参与侵染过程及侵染后的生长阶段。
Rapid alkalization factor(RALF) is a kind of plant peptide hormone that can participate in the regula-tion of plant development. It exists widely in fungi and affects the infectivity of fungi. In this paper, the effects of dif-ferent pH values on the growth of Setosphaeria turcica were determined by adding acid and alkali to cultural medi-um. The changes in the expression of StRALF gene in S. turcica were analyzed by real-time fluorescence quantita-tive PCR technology. The basic properties of StRALF gene coding region protein were analyzed at prediction web-site. The results showed that S. turcica grew better in alkaline environment; and in alkaline condition, the expression level of StRALF gene was higher than that in acid condition. During the process of S. turcica infection to host, the ex-pression of StRALF gene reached the maximum after 96 h, and it was preliminarily predicted as secretory protein or membrane protein. In total, under weak alkaline condition, StRALF gene was up-regulated and then involved in the infection process and growth period. This study can provide a reference for the further research on the action and ac-tion mechanism of RALF in S. turcica.
Rapid alkalization factor(RALF) is a kind of plant peptide hormone that can participate in the regula-tion of plant development. It exists widely in fungi and affects the infectivity of fungi. In this paper, the effects of dif-ferent pH values on the growth of Setosphaeria turcica were determined by adding acid and alkali to cultural medi-um. The changes in the expression of StRALF gene in S. turcica were analyzed by real-time fluorescence quantita-tive PCR technology. The basic properties of StRALF gene coding region protein were analyzed at prediction web-site. The results showed that S. turcica grew better in alkaline environment; and in alkaline condition, the expression level of StRALF gene was higher than that in acid condition. During the process of S. turcica infection to host, the ex-pression of StRALF gene reached the maximum after 96 h, and it was preliminarily predicted as secretory protein or membrane protein. In total, under weak alkaline condition, StRALF gene was up-regulated and then involved in the infection process and growth period. This study can provide a reference for the further research on the action and ac-tion mechanism of RALF in S. turcica.
引文
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[10]Hardtke C S.Phytohormone collaboration:zooming in on auxinbrassinosteroid interactions[J].Trends in Cell Biology.2007,17:485-492.
[11]Shiu S H,Bleecker A B.Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98:10763-10768.
[12]Lease K A,Walker J C.The Arabidopsis unannotated secreted pep-tide database,a resource for plant peptidomics[J].Plant Physiology,2006,142:831-838.
[13]Hirakawa Y.Non-cell-autonomous control of vascular stem cell fate by a CLE peptide/receptor system[J].Proceedings of the Na-tional Academy of Sciences of the United States of America.2008,105:15208-15213.
[14]DeYoung B J.The CLAVATA1-related BAM1,BAM2 and BAM3receptor kinase-like proteins are required for meristem function in Arabidopsis[J].Plant Journal 2006,45:1-16.
[15]Shpak E D.Stomatal patterning and differentiation by synergistic interactions of receptor kinases[J].Science 2005,309:290-293.
[16]Jeong S.The Arabidopsis CLAVATA2 gene encodes a receptorlike protein required for the stability of the CLAVATA1 receptorlike kinase[J].Plant Cell 1999,11:1925-1934.
[17]Torii K U.Leucine-rich repeat receptor kinases in plants:struc-ture,function,and signal transduction pathways[J].International Review of Cytology.2004,234:1-46.
[18]Butenko M A.Plant peptides in signalling:looking for new partners[J].Trends in Plant Science.2009,14:255-263.
[19]Atkinson N J.Identification of genes involved in the response of Arabidopsis to simultaneous biotic and abiotic stresses[J].Plant Physiology.2013,162:2028-2041.
[2]Evan M,Ive D S.Understanding the RALF family:a tale of many species[J].Trends in Plant Science,2014,19(10).
[3]Czyzewicz,N.Message in a bottle:small signalling peptide outputs during growth and development[J].Journal of Experimental Botany,2013,64:5281-5296.
[4]Masachis S,Segorbe D,TurràD,et al.Corrigendum:A fungal patho-gen secretes plant alkalinizing peptides to increase infection[J].Na-ture Microbiology,2016,1(6):16043.
[5]Pearce,G.RALF,a 5-kDa ubiquitous polypeptide in plants,arrests root growth and development[J].Proceedings of the National Acade-my of Sciences of the United States of America,2001,98:12843-12847.
[6]Rollins J A,Dickman M B.pH signaling in sclerotinia sclerotiorum:identification of pacC/R IM1 homolog[J].Applied and Environmen-tal Microbiology,2001,67:75-81.
[7]Eshel D,Lichter A,Dinoor A,et al.Characterization of Alternaria al-ternata glucanase genes expressed during infection of resistant and susceptible persimmon fruits[J].Molecular Plant Pathology.2002a,3:347-358.
[8]Manteau S,Abouna S,Lambert B,et al.Differential regulation by ambient p H of put ative virulence factors secretion by the phyto-pathogenic fungus Botrytis cinerea[J].Fems Microbiology Ecology.2003,43:359-366.
[9]Olsen A N.Peptomics,identification of novel cationic Arabidopsis peptides with conserved sequence motifs[J].In Silico Biology,2002,2:441-451.
[10]Hardtke C S.Phytohormone collaboration:zooming in on auxinbrassinosteroid interactions[J].Trends in Cell Biology.2007,17:485-492.
[11]Shiu S H,Bleecker A B.Receptor-like kinases from Arabidopsis form a monophyletic gene family related to animal receptor kinases[J].Proceedings of the National Academy of Sciences of the United States of America,2001,98:10763-10768.
[12]Lease K A,Walker J C.The Arabidopsis unannotated secreted pep-tide database,a resource for plant peptidomics[J].Plant Physiology,2006,142:831-838.
[13]Hirakawa Y.Non-cell-autonomous control of vascular stem cell fate by a CLE peptide/receptor system[J].Proceedings of the Na-tional Academy of Sciences of the United States of America.2008,105:15208-15213.
[14]DeYoung B J.The CLAVATA1-related BAM1,BAM2 and BAM3receptor kinase-like proteins are required for meristem function in Arabidopsis[J].Plant Journal 2006,45:1-16.
[15]Shpak E D.Stomatal patterning and differentiation by synergistic interactions of receptor kinases[J].Science 2005,309:290-293.
[16]Jeong S.The Arabidopsis CLAVATA2 gene encodes a receptorlike protein required for the stability of the CLAVATA1 receptorlike kinase[J].Plant Cell 1999,11:1925-1934.
[17]Torii K U.Leucine-rich repeat receptor kinases in plants:struc-ture,function,and signal transduction pathways[J].International Review of Cytology.2004,234:1-46.
[18]Butenko M A.Plant peptides in signalling:looking for new partners[J].Trends in Plant Science.2009,14:255-263.
[19]Atkinson N J.Identification of genes involved in the response of Arabidopsis to simultaneous biotic and abiotic stresses[J].Plant Physiology.2013,162:2028-2041.