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园艺学报 ›› 2015, Vol. 42 ›› Issue (1): 167-173.doi: 10.16420/j.issn.0513-353x.2014-0557

• 研究报告 • 上一篇    下一篇

番茄黄化曲叶病毒石家庄分离物的分子鉴定及序列分析

白晓娟1,2,李亚栋1,*,王国华1,尹庆珍1,耿保进1,董文琦1,**,周春江2,吴志明1,**   

  1. 1河北省农林科学院经济作物研究所,石家庄 050051;2河北师范大学生命科学学院,石家庄 050024
  • 出版日期:2015-01-25 发布日期:2015-01-25
  • 基金资助:

    河北省财政专项计划项目(2013055003);河北省农林科学院院士合作项目(A2012050601);河北省科技支撑计划项目(14226310D)

Molecular Identification and Sequence Analysis of Tomato yellow leaf curl virus(TYLCV)Isolate from Shijiazhuang

BAI Xiao-juan1,2,LI Ya-dong1,*,WANG Guo-hua1,YIN Qing-zhen1,GENG Bao-jin1,DONG Wen-qi1,**,ZHOU Chun-jiang2,and WU Zhi-ming1,**   

  1. 1Institute of Cash Crops,Hebei Academy of Agriculture and Forestry Sciences,Shijiazhuang 050051,China;2College of Life Sciences,Hebei Normal University,Shijiazhuang 050024,China
  • Online:2015-01-25 Published:2015-01-25

摘要: 根据番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)基因组序列的复制酶保守区设计1对引物JC1,对石家庄具有典型番茄黄化曲叶病毒症状的番茄样品进行检测,经PCR扩增得到626 bp的片段,序列分析比对表明其为TYLCV的一部分。根据上述序列设计1对特异引物QC,通过PCR分离石家庄的TYLCV全长序列并进行测序。同时利用已报道的DNA-B的通用引物CR01/CR02进行PCR扩增。结果表明石家庄分离物只含有DNA-A组分,全长为2 781 bp(GenBank登录号:KF612971),命名为TYLCV-Shijiazhuang。基因组序列比较发现,该分离物与TYLCV-Israel株系相似性为99.0%。基因组序列系统进化分析表明,石家庄病毒分离物与北京分离物TYLCV-Beijing3(GenBank登录号:GU983859)、山东分离物TYLCV-SDSG-XC(GenBank登录号:KC999851)序列相似性很高,均属于TYLCV-Israel分支。

关键词: 番茄, 番茄黄化曲叶病毒(TYLCV), DNA-A, 分子鉴定, 序列分析

Abstract: A 626 bp fragment of replication protein region of TYLCV(Tomato yellow leaf curl virus) was cloned using PCR with primers JC1 from Shijiazhuang samples with typical symptoms of yellow leaf curl,and sequence analysis confirmed that it was a part of TYLCV. Then the full-length of TYLCV was obtained with a pair of primers QC and sequenced. The already reported universary primers CR01/CR02 for DNA-B were used for amplification and the results showed that only DNA-A containing 2 781 bp nucleotides and it was named as TYLCV-shijiazhuang(GenBank accession No. KF612971). Sequenceanalysis showed that TYLCV-shijiazhuang was similar to TYLCV-Israel for 99.0%. However,it was sometimes expceptional that TYLCV-Shijiazhuang isolate was most closely related to TYLCV-Beijing3(GenBank accession No. GU983859),TYLCV-SDSG-XC(GenBank accession No. KC999851),and they all belonged to the same monophyletic clusters of TYLCV-Israel isolate.

Key words: tomato, TYLCV, DNA-A, molecular identifcation, sequence analysis

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