三七Y病毒部分ci基因和cp基因的克隆及序列分析
|
摘要
对云南文山地区三七上获得的三七Y病毒(Panax virus Y,Pn VY)不同分离物进行了部分ci基因和cp基因的克隆及序列分析,获得的部分ci基因长657 nt,编码219个氨基酸;获得的部分cp基因长834 nt,编码278个氨基酸。所获得的11个Pn VY分离物的ci基因之间核苷酸和氨基酸的同源性分别为80.8%~99.8%和91.3%~100%,与其他Pn VY分离物的同源性分别为80.8%~98.8%和92.7%~99.5%。所获得的18个Pn VY分离物的cp基因核苷酸和氨基酸序列同源性分别为91.8%~99.9%和93.9%~100%,与其他Pn VY分离物的同源性分别为92.0%~100%和95.7%~100%。系统进化树分析表明,基于ci基因可以将Pn VY分为2个大组,第Ⅱ组分离物之间的差异相比第Ⅰ组的复杂;基于cp基因可以将Pn VY分为3个大组,第Ⅲ组分离物之间差异最大。基于ci基因和cp基因的分析结果均表明,Pn VY不同种群间存在较为丰富的遗传多样性,但尚不清楚Pn VY的分子变异是否与Pn VY引起的多种症状类型有关。
Partial ci and cp genes of different Panax virus Y( Pn VY) isolates from Wenshan in Yunnan Province were amplified,cloned and sequenced. The partial ci gene obtained in this study consists of 657 nt,encoding 219 amino acid,while the partial cp gene consists of 834 nt,encoding 278 amino acid. The results showed that the nucleotide( nt) and amino acid( aa) sequences of ci gene of 11 Pn VY Yunnan isolates shared 80. 8% ~ 99. 8% and 91. 3% ~ 100% identity with each other,and shared 80. 8% ~ 98. 8% and 92. 7% ~ 99. 5% identity with the recorded Pn VY isolates. The nt andaa sequences of cp gene of 18 Pn VY Yunnan isolates shared 91. 8% ~ 99. 9% and 93. 9% ~ 100% identity with each other,and shared 92. 0% ~ 100% and 95. 7% ~ 100% identity with the Pn VY isolates deposited in Gen Bank. The available Pn VY isolates could be divided into two groups according to the phylogenetic tree based on the nucleotide sequences of partial ci gene,and the isolates in groupⅡshowed more diversity compared with those in groupⅠ. While the Pn VY isolates could be divided into three groups base on the cp nt sequence,and the isolates of group Ⅲ showed the highest diversity. The results indicated that high genetic diversity exists among different Pn VY populations,but whether the molecular variations on Pn VY genome related to the different symptoms of the disease is still unknown.
Partial ci and cp genes of different Panax virus Y( Pn VY) isolates from Wenshan in Yunnan Province were amplified,cloned and sequenced. The partial ci gene obtained in this study consists of 657 nt,encoding 219 amino acid,while the partial cp gene consists of 834 nt,encoding 278 amino acid. The results showed that the nucleotide( nt) and amino acid( aa) sequences of ci gene of 11 Pn VY Yunnan isolates shared 80. 8% ~ 99. 8% and 91. 3% ~ 100% identity with each other,and shared 80. 8% ~ 98. 8% and 92. 7% ~ 99. 5% identity with the recorded Pn VY isolates. The nt andaa sequences of cp gene of 18 Pn VY Yunnan isolates shared 91. 8% ~ 99. 9% and 93. 9% ~ 100% identity with each other,and shared 92. 0% ~ 100% and 95. 7% ~ 100% identity with the Pn VY isolates deposited in Gen Bank. The available Pn VY isolates could be divided into two groups according to the phylogenetic tree based on the nucleotide sequences of partial ci gene,and the isolates in groupⅡshowed more diversity compared with those in groupⅠ. While the Pn VY isolates could be divided into three groups base on the cp nt sequence,and the isolates of group Ⅲ showed the highest diversity. The results indicated that high genetic diversity exists among different Pn VY populations,but whether the molecular variations on Pn VY genome related to the different symptoms of the disease is still unknown.
引文
[1]李晓静,刘芳,李梅蓉,等.4种提取试剂对三七Y病毒RNA提取效果的比较[J].云南农业大学学报(自然科学),2014,29(6):822.
[2]宋丽敏,梁文星,姜辛,等.三七上番茄花叶病毒的初步鉴定[J].云南农业大学学报,2003,18(增):111.
[3]金羽.三七病毒病病原的初步研究[D].哈尔滨:东北农业大学,2005.
[4]杨建忠,王勇,张葵,等.蓟马危害三七调查初报[J].中药材,2008,31(5):636.
[5]YAN Z L,SONG L M,ZHOU T,et al.Identification and molecular characterization of a new potyvirus from Panax notoginseng[J].Archives of Virology,2010,155(6):949.
[6]LI X J,LIU F,LI Y Y,et al.First report of tomato yellow leaf curl China virus with betasatellite infecting Panax notoginseng[J].Plant Disease,2014,98(9):1284.
[7]李晓静.三七病毒病病原种类鉴定及其相关病毒分子变异分析[D].昆明:云南农业大学,2014.
[8]CARRINGTON J C,JENSEN P E,SCHAAD M C.Genetic evidence for an essential role for potyvirus CI protein in cell-to-cell movement[J].The Plant Journal:for Cell and Molecular Biology,1998,14(4):393.
[9]杨炳辉.衣壳蛋白对烟草脉带花叶病毒侵染和致病力的影响[D].泰安:山东农业大学,2012.
[10]WARD C W,SHUKLA D D.Taxonomy of potyviruses:current problems and some solutions[J].Intervirology,1991,32(5):269.
[11]CHOI I R,HORKEN K M,STENGER D C,et al.Mapping of the P1 proteinase cleavage site in the polyprotein of Wheat streak Mosaic virus(genus Tritimovirus)[J].The Journal of General Virology,2002,83(Pt 2):443.
[12]DUNOYER P,THOMAS C,HARRISON S,et al.A cysteine-rich plant protein potentiates Potyvirus movement through an interaction with the virus genome-linked protein VPg[J].Journal of Virology,2004,78(5):2301.
[13]HA C,COOMBS S,REVILL P A,et al.Design and application of two novel degenerate primer pairs for the detection and complete genomic characterization of potyviruses[J].Archives of Virology,2008,153(1):25.
[14]LI R,MOCK R,HUANG Q,et al.A reliable and inexpensive method of nucleic acid extraction for the PCRbased detection of diverse plant pathogens[J].Journal of Virological Methods,2008,154(1/2):48.
[15]叶荣,陈剑平,于善谦.植物病毒介体传播的分子机制[J].中国病毒学,1999,4(4):285.
[16]李向东,于晓庆,古勤生,等.马铃薯Y病毒属病毒基因功能研究进展[J].山东科学,2006,19(3):1.
[2]宋丽敏,梁文星,姜辛,等.三七上番茄花叶病毒的初步鉴定[J].云南农业大学学报,2003,18(增):111.
[3]金羽.三七病毒病病原的初步研究[D].哈尔滨:东北农业大学,2005.
[4]杨建忠,王勇,张葵,等.蓟马危害三七调查初报[J].中药材,2008,31(5):636.
[5]YAN Z L,SONG L M,ZHOU T,et al.Identification and molecular characterization of a new potyvirus from Panax notoginseng[J].Archives of Virology,2010,155(6):949.
[6]LI X J,LIU F,LI Y Y,et al.First report of tomato yellow leaf curl China virus with betasatellite infecting Panax notoginseng[J].Plant Disease,2014,98(9):1284.
[7]李晓静.三七病毒病病原种类鉴定及其相关病毒分子变异分析[D].昆明:云南农业大学,2014.
[8]CARRINGTON J C,JENSEN P E,SCHAAD M C.Genetic evidence for an essential role for potyvirus CI protein in cell-to-cell movement[J].The Plant Journal:for Cell and Molecular Biology,1998,14(4):393.
[9]杨炳辉.衣壳蛋白对烟草脉带花叶病毒侵染和致病力的影响[D].泰安:山东农业大学,2012.
[10]WARD C W,SHUKLA D D.Taxonomy of potyviruses:current problems and some solutions[J].Intervirology,1991,32(5):269.
[11]CHOI I R,HORKEN K M,STENGER D C,et al.Mapping of the P1 proteinase cleavage site in the polyprotein of Wheat streak Mosaic virus(genus Tritimovirus)[J].The Journal of General Virology,2002,83(Pt 2):443.
[12]DUNOYER P,THOMAS C,HARRISON S,et al.A cysteine-rich plant protein potentiates Potyvirus movement through an interaction with the virus genome-linked protein VPg[J].Journal of Virology,2004,78(5):2301.
[13]HA C,COOMBS S,REVILL P A,et al.Design and application of two novel degenerate primer pairs for the detection and complete genomic characterization of potyviruses[J].Archives of Virology,2008,153(1):25.
[14]LI R,MOCK R,HUANG Q,et al.A reliable and inexpensive method of nucleic acid extraction for the PCRbased detection of diverse plant pathogens[J].Journal of Virological Methods,2008,154(1/2):48.
[15]叶荣,陈剑平,于善谦.植物病毒介体传播的分子机制[J].中国病毒学,1999,4(4):285.
[16]李向东,于晓庆,古勤生,等.马铃薯Y病毒属病毒基因功能研究进展[J].山东科学,2006,19(3):1.