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辣椒疫霉elicitins基因的克隆及瞬时表达分析

DOI: 10.7685/j.issn.1000-2030.2014.06.005, PP. 28-36

Keywords: 辣椒疫霉,elicitin,瞬时表达,植物抗病性

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Abstract:

通过挖掘辣椒疫霉中的外泌蛋白激发子elicitins的基因序列,分析其转录特征,克隆基因序列并进行瞬时表达,以阐明其在辣椒疫霉生长发育和侵染过程中的作用。本研究利用转录组测序结果和致病疫霉INF1序列作为比对序列,对辣椒疫霉基因组数据库进行elicitins序列挖掘。采用RT-PCR方法分析elicitins基因在辣椒疫霉生长发育(菌丝、游动孢子囊、游动孢子、萌发的休止孢)和侵染寄主阶段(本氏烟灌根1.5、3、6、12、24、36和72h后)的转录水平。克隆elicitins基因序列,并与其他卵菌的elicitins进行序列比对,分析进化关系。在本氏烟上进行elicitins的瞬时表达分析。结果表明经生物信息学分析获得14个含有信号肽编码序列的elicitins基因全长序列。新发现6个elicitins基因在辣椒疫霉生长发育和侵染寄主阶段差异表达。对这6个和之前报道的5个elicitins基因进行全长克隆,能够克隆出的10个elicitins都属于酸性elicitins,聚类分析可将其分在第一和第三聚类组。异源表达发现2个elicitins能够激发本氏烟产生过敏反应。本研究结果不仅为阐明elicitins在辣椒疫霉生长发育和侵染寄主过程中的作用提供了重要数据,也为植物抗疫病的基因工程提供了科学依据。

References

[1]  Erwin D C,Ribeiro O K. Phytophthora Diseases Worldwide[M]. St Paul:APS Press,1996:262-268
[2]  Lamour K H,Stam R,Jupe J,et al. The oomycete broad-host-range pathogen Phytophthora capsici[J]. Mol Plant Pathol,2012,13:329-337
[3]  Yu L M. Elicitins from Phytophthora and basic resistance in tobacco[J]. Proc Natl Acad Sci USA,1995,92:4088-4094
[4]  Baillieul F,de Ruffray P,Kauffmann S. Molecular cloning and biological activity of α-,β- and γ-megaspermin,three elicitors secreted by Phytophthora megasperma H20[J]. Plant Physiol,2003,131:155-166
[5]  Kamoun S,Klucher K M,Coffey M D,et al. A gene encoding a host-specific elicitor protein of Phytophthora parasitica[J]. Mol Plant Microbe In,1993,6:573-581
[6]  Jiang R H Y,Tyler B M,Whisson S C,et al. Ancient origin of elicitin gene clusters in Phytophthora genomes[J]. Mol Biol Evol,2006,23:338-351
[7]  Kamoun S,van West P,Vleeshouwers V G,et al. Resistance of Nicotiana benthamiana to Phytophthora infestans is mediated by the recognition of the elicitor protein INF1[J]. Plant Cell,1998,10:1413-1426
[8]  Kamoun S. Nonhost resistance to Phytophthora:novel prospects for a classical problem[J]. Curr Opin Plant Biol,2001,4:295-300
[9]  Chen X R,Xing Y P,Li Y P,et al. RNA-Seq reveals infection-related gene expression changes in Phytophthora capsici[J]. PLoS ONE,2013,8:e74588
[10]  Kamoun S,Lindqvist H,Govers F. A novel class of elicitin-like genes from Phytophthora infestans[J]. Mol Plant Microbe In,1997,10:1028-1030
[11]  Panabières F,Marais A,Le Berre J Y,et al. Characterization of a gene cluster of Phytophthora cryptogea which codes for elicitins,proteins inducing a hypersensitive-like response in tobacco[J]. Mol Plant Microbe In,1995,8:996-1003
[12]  Mao Y,Tyler B M. Cloning and sequence analysis of elicitin genes of Phytophthora sojae[J]. Fungal Genet Biol,1996,20:169-172
[13]  申贵,王源超,郑小波. 棉疫病菌elicitin基因的克隆及其序列分析[J]. 植物病理学报,2003,33(6):559-562
[14]  李俊,张海峰,张正光,等. 棉疫病菌诱导非寄主过敏反应激发子基因的克隆[J]. 科学通报,2006,51(22):2638-2643 [Li J,Zhang H F,Zhang Z G,et al. Cloning of genes encoding nonhost hypersensitive response-inducing elicitors from Phytophthora boehmeriae[J]. Chinese Sci Bull,2006,51(22):2638-2643(in Chinese)]
[15]  Wang Q,Han C,Ferreira A O,et al. Transcriptional programming and functional interactions within the Phytophthora sojae RXLR effector repertoire[J]. Plant Cell,2011,23:2064-2086
[16]  Yousef L F,Yousef A F,Mymryk J S,et al. Stigmasterol and cholesterol regulate the expression of elicitin genes in Phytophthora sojae[J]. J Chem Ecol,2009,35:824-832
[17]  Lu R,Malcuit I,Moffett P,et al. High throughput virus-induced gene silencing implicates heat shock protein 90 in plant disease resistance[J]. EMBO J,2003,22:5690-5699
[18]  Kanneganti T D,Huitema E,Kamoun S. In planta expression of fungal and oomycete effectors[C]//Ronald P. Plant-Pathogen Interactions. Totowa:Humana Press,2007:35-43
[19]  陈孝仁,王源超. 卵菌基因功能分析方法的研究进展[J]. 农业生物技术学报,2012,20(5):568-575[Chen X R,Wang Y C. Advances on the methods used for the functional analysis of oomycete genes[J]. J Agric Biotech,2012,20(5):568-575(in Chinese with English abstract)]
[20]  Vleeshouwers V G A A,Rietman H,Krenek P,et al. Effector genomics accelerates discovery and functional profiling of potato disease resistance and Phytophthora infestans avirulence genes[J]. PLoS ONE,2008,3:e2875
[21]  Ricci P,Bonnet P,Huet J C,et al. Structure and activity of proteins from pathogenic fungi Phytophthora eliciting necrosis and acquired resistance in tobacco[J]. Eur J Biochem,1989,183:555-563
[22]  Rozen S,Skaletsky H. Primer 3 on the WWW for general users and for biologist programmers[J]. Methods Mol Biol,2000,132:365-386
[23]  Hall T A. BioEdit:a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT[J]. Nucleic Acids Symp Ser,1999,41:95-98
[24]  Bendtsen J D,Nielsen H,von Heijne G,et al. Improved prediction of signal peptides:SignalP 3.0[J]. J Mol Biol,2004,340:783-795
[25]  Tamura K,Peterson D,Peterson N,et al. MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods[J]. Mol Biol Evol,2011,28:2731-2739
[26]  Ye W,Wang X,Tao K,et al. Digital gene expression profiling of the Phytophthora sojae transcriptome[J]. Mol Plant Microbe In,2011,24:1530-1539
[27]  Kunjeti S G,Evans T A,Marsh A G,et al. RNA-Seq reveals infection-related global gene changes in Phytophthora phaseoli,the causal agent of lima bean downy mildew[J]. Mol Plant Pathol,2012,13:454-466
[28]  Keller H,Pamboukdjian N,Ponchet M,et al. Pathogen-induced elicitin production in transgenic tobacco generates a hypersensitive response and nonspecific disease resistance[J]. Plant Cell,1999,11:223-235
[29]  Dangl J L,Jones J D. Plant pathogens and integrated defence responses to infection[J]. Nature,2001,411:826-833
[30]  Osman H,Mikes V,Milat M L,et al. Fatty acids bind to the fungal elicitor cryptogein and compete with sterols[J]. FEBS Lett,2001,489:55-58

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