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山东地区樱桃绿环斑驳病毒(CGRMV)的RT-PCR检测及外壳蛋白基因的克隆
引用本文:王文文,宗晓娟,陈立伟,王甲威,魏海蓉,徐丽,孟艳玲,严雪瑞,刘庆忠.山东地区樱桃绿环斑驳病毒(CGRMV)的RT-PCR检测及外壳蛋白基因的克隆[J].山东农业大学学报(自然科学版),2012,43(2):206-210,219.
作者姓名:王文文  宗晓娟  陈立伟  王甲威  魏海蓉  徐丽  孟艳玲  严雪瑞  刘庆忠
作者单位:1. 沈阳农业大学植物保护学院,辽宁沈阳 110866;山东省果树研究所,山东泰安 271000;山东省果树生物技术育种重点实验室,山东泰安 271000
2. 山东省果树研究所,山东泰安 271000;山东省果树生物技术育种重点实验室,山东泰安 271000
3. 山东威海市农业科学院,山东威海,264200
4. 沈阳农业大学植物保护学院,辽宁沈阳,110866
基金项目:农业部948项目,公益性行业(农业)科研专项,%%
摘    要:为调查我省甜樱桃感染樱桃绿环斑驳病毒(Cherry Green Ring Mottle Virus,CGRMV)情况,本研究以甜樱桃(Prunus avium L.)品种"红灯"叶片总RNA为模板,根据CGRMV基因组序列设计特异引物,对山东地区37份甜樱桃"红灯"样品进行RT-PCR检测,共检测出19份阳性样品。利用CGRMV外壳蛋白基因序列引物,从阳性植物样本中分离到约800 bp的目的片段,克隆测序,序列分析显示该片段全长807 bp,编码268个氨基酸,与GenBank中已登录的CGRMV分离物的外壳蛋白基因序列一致性为87%~97%,氨基酸序列相似性为95%~99%。该结果表明山东地区甜樱桃生产园中感染CGRMV的病例较为普遍。

关 键 词:甜樱桃  樱桃绿环斑驳病毒  RT-PCR检测  外壳蛋白  基因克隆

DETECTION OF CHERRY GREEN RING MOTTLE VIRUS (CGRMV) AND ISOLATION OF ITS COAT PROTEIN GENES FROM SWEET CHERRY IN SHANDONG PROVINCE
WANG Wen-wen , ZONG Xiao-juan , CHEN Li-wei , WANG Jia-wei , WEI Hai-rong , XU Li , MENG Yan-ling , YAN Xue-rui , LIU Qing-zhong.DETECTION OF CHERRY GREEN RING MOTTLE VIRUS (CGRMV) AND ISOLATION OF ITS COAT PROTEIN GENES FROM SWEET CHERRY IN SHANDONG PROVINCE[J].Journal of Shandong Agricultural University,2012,43(2):206-210,219.
Authors:WANG Wen-wen  ZONG Xiao-juan  CHEN Li-wei  WANG Jia-wei  WEI Hai-rong  XU Li  MENG Yan-ling  YAN Xue-rui  LIU Qing-zhong
Institution:2.3(1.College of Plant Protection,Shenyang Agricultural University,Shenyang 110866,China; 2.Shandong Institute of Pomology,Taian 271000,China; 3.Shandong Key Laboratory of Fruit Biotechnology Breeding,Taian 271000,China; 4.Weihai Academy of Agricultural Science of Shandong Province,Weihai 264200,China)
Abstract:In order to investigate the infection of Cherry Green Ring Mottle Virus(CGRMV) from sweet cherry(Prunus avium L.) in Shandong province,total RNA was extracted from the leaves of the sweet cherry cultivar Red Lamp’.The specific primers were designed corresponding to the CGRMV genome sequence and used for the RT-PCR detection.Thirty-seven sweet cherry samples from six orchards were analyzed in the experiment and nineteen of them were detected as positive.The gene that encodes the virus coat protein was amplified and sequenced.Sequence analysis revealed that the fragment was 807 nucleotides in length,encoding 268 amino acids.It shared 87%~97% identity to the other CGRMV isolates reported in GenBank in the nucleotide level and 95%~99% similarity in the amino acids level.These results showed CGRMV had been wide spread on sweet cherry in Shandong district.
Keywords:Prunus avium L    Cherry green ring mottle virus  RT-PCR detection  coat protein  gene cloning
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