Chen L, Shang Q X, Chen X Y, Xing D M, Yang R, Han C G, Ran C, Wei Y M, Zhao X Y, Liu Z P
Beijing Key Laboratory of New Technology in Agricultural Application, College of Plant Science and Technology, Beijing University of Agriculture, Beijing 102206, P. R. China.
Beijing Plant Protection Station, Beijing 100029, P. R. China.
Plant Dis. 2014 Jul;98(7):1015. doi: 10.1094/PDIS-11-13-1173-PDN.
More than 20 viruses are known to infect strawberry (Fragaria ananassa), and a substantial number of these include new viruses identified since 2000 that can contribute to disease complexes (2). The most serious virus related losses in commercial strawberries are caused by aphid transmitted viruses (3,4,5). A survey was undertaken from 2012 to 2013 to investigate virus prevalence in commercial strawberries in rural areas of Hebei Province around Beijing, China, that were exhibiting virus symptoms. Visual observations revealed that the incidence of virus-like symptoms ranged from 30 to 50% of the plants and these symptoms included yellowing, leaf malformation, sometimes combined with severe stunting and deformed flowers or fruits. Leaf samples were tested for Strawberry vein banding virus (SVBV), Strawberry mottle virus (SMoV), Strawberry mild yellow edge virus (SMYEV), and Strawberry crinkle virus (SCV), which are the four most prevalent aphid-transmitted viruses in single or mixed infections (2). Testing was conducted by RT-PCR using total RNA extracted from fresh symptomatic strawberry leaves (3). SVBV was detected in 58 of 190 samples, but all of the samples tested negative for SMoV, SMYEV, and SCV. Aphids were present on many of the plants, so the samples were tested for Cucumber mosaic virus (CMV) because CMV is prevalent in Beijing gardens and farms, and recently had been shown to infect maize in China (5). This RT-PCR was carried out with the CMV primer pair CM420-F (5'-TGATTCTACCGTGTGGGTGA-3') and CM420-R (5'-CCGTAAGCTGGATGGACAAC-3') to amplify a portion of the capsid protein coding region and the conserved 3'non-translated regions of the genomic RNAs. This test revealed the presence of 43 CMV-positives out of 190 samples, and only 16 of these samples were co-infected with both SVBV and CMV. Samples infected with CMV only had leaf malformations and yellowing, while no CMV was found in symptomless samples. One of the amplified, CMV-specific DNA fragments was sequenced directly from the PCR product and showed 93.8% nucleotide sequence identity and 100% amino acid sequence identity to the CMV subgroup I (GenBank Accession No. D10538) (1). Subsequent ELISA tests for the CMV presence verified the RT-PCR results (Agdia, Elkhart, IN), and transmission electron microscopy observations revealed 28 nm spherical particles characteristic of CMV in strawberry samples tested positive for CMV. However, we were unable to detect either CMV or SVBV in 89 of the 169 samples from symptomatic plants, which suggested possible presence of the other pathogen(s). To the best of our knowledge, this is the first report of natural infections of CMV in strawberry plants. These data suggests that CMV is a potential threat to strawberry production. References: (1) M. Q. K. Andrew et al. Virus taxonomy: IXth Report of the ICTV, 970, Elsevier, 2012. (2) R. R. Martin and I. E. Tzanetakis. Plant Dis. 97:1358, 2013. (3) J. R. Thompson et al. J. Virol. Methods 111:85, 2003. (4) I. E. Tzanetakis et al. Plant Dis. 90:1343, 2006. (5) R. Wang et al. J. Phytopathol. 161: 880, 2013.
已知有20多种病毒可感染草莓(凤梨草莓),其中相当一部分是2000年以来鉴定出的新病毒,它们可引发复合病害(2)。商业种植的草莓中,与病毒相关的最严重损失是由蚜虫传播的病毒造成的(3,4,5)。2012年至2013年,我们对中国北京周边河北省农村地区表现出病毒症状的商业种植草莓进行了一项调查,以研究病毒的流行情况。肉眼观察发现,有病毒样症状的植株发生率在30%至50%之间,这些症状包括黄化、叶片畸形,有时还伴有严重矮化以及花或果实畸形。对叶片样本进行了草莓脉带病毒(SVBV)、草莓斑驳病毒(SMoV)、草莓轻型黄边病毒(SMYEV)和草莓皱缩病毒(SCV)的检测,这四种是最常见的蚜虫传播病毒,可单独或混合感染(2)。检测采用RT-PCR,使用从新鲜的有症状草莓叶片中提取的总RNA(3)。在190个样本中,有58个检测出SVBV,但所有样本的SMoV、SMYEV和SCV检测均为阴性。许多植株上都有蚜虫,因此对样本进行了黄瓜花叶病毒(CMV)检测,因为CMV在北京的花园和农场中很普遍,并且最近已被证明可感染中国的玉米(5)。使用CMV引物对CM420-F(5'-TGATTCTACCGTGTGGGTGA-3')和CM420-R(5'-CCGTAAGCTGGATGGACAAC-3')进行该RT-PCR,以扩增衣壳蛋白编码区的一部分以及基因组RNA的保守3'非翻译区。该检测显示,190个样本中有43个CMV呈阳性,其中只有16个样本同时感染了SVBV和CMV。仅感染CMV的样本有叶片畸形和黄化症状,而无症状样本中未检测到CMV。从PCR产物中直接对扩增出的一个CMV特异性DNA片段进行测序,结果显示其与CMV亚组I(GenBank登录号D10538)的核苷酸序列同一性为93.8%,氨基酸序列同一性为100%(1)。随后针对CMV存在情况的ELISA检测验证了RT-PCR结果(Agdia公司,美国印第安纳州埃尔克哈特),透射电子显微镜观察在CMV检测呈阳性的草莓样本中发现了具有CMV特征的28纳米球形颗粒。然而,在169个有症状植株的样本中,我们有89个未检测到CMV或SVBV,这表明可能存在其他病原体。据我们所知,这是草莓植株自然感染CMV的首次报道。这些数据表明CMV对草莓生产构成潜在威胁。参考文献:(1)M. Q. K. Andrew等人,《病毒分类学:国际病毒分类委员会第九次报告》,970页,爱思唯尔出版社,2012年。(2)R. R. Martin和I. E. Tzanetakis,《植物病害》97:1358,2013年。(3)J. R. Thompson等人,《病毒学方法杂志》111:85,2003年。(4)I. E. Tzanetakis等人,《植物病害》90:1343,2006年。(5)R. Wang等人,《植物病理学杂志》161: 880,2013年。