Mgbechi-Ezeri J U, Alabi O J, Naidu R A, Kumar P Lava
International Institute of Tropical Agriculture (IITA), Ibadan - PMB 5320, Nigeria.
Department of Plant Pathology, Washington State University, Irrigated Agriculture Research and Extension Center, Prosser, 99350.
Plant Dis. 2008 Dec;92(12):1709. doi: 10.1094/PDIS-92-12-1709B.
African cassava mosaic virus (ACMV; genus Begomovirus, family Geminiviridae) is one of six viruses documented in cassava (Manihot esculenta Crantz.) plants showing cassava mosaic disease in sub-Saharan Africa (SSA). In addition to cassava, the natural host range of ACMV includes a few wild Manihot species, Jatropha multifida, and Ricinus communis L. in Euphorbiaceae, and Hewittia sublobata in Convolvulaceae. The experimental host range of ACMV includes Nicotiana sp. and Datura sp. in the Solanaceae (2). Recently, natural occurrence of ACMV was reported in Combretum confertum (Benth.), Leucana leucocephala (Lam.) De Witt, and Senna occidentalis (L.) Link belonging to Leguminasae from Nigeria (1,3). During reconnaissance studies conducted on soybean (Glycine max L. Merr.) in September and October of 2007 in the Ibadan (N = 19) and Benue (N = 23) regions and in February of 2008 in Ibadan (N = 16), we observed soybean showing yellow mosaic and mottling symptoms. Samples from these plants (N = 58) were tested by indirect ELISA and symptomatic leaves tested negative to Cucumber mosaic virus, Cowpea mottle virus, Southern bean mosaic virus, Tobacco ringspot virus, Soybean dwarf virus, Cowpea aphid-borne mosaic virus, Blackeye cowpea mosaic virus, Peanut mottle virus, and Broad bean mosaic virus, which have been documented in soybean in SSA. However, 8.6% of these samples (5 of 58) (one each from Ibadan and Benue in the 2007 survey and three from Ibadan in the 2008 survey) tested positive in triple-antibody sandwich-ELISA with a monoclonal antibody (SCR33) to ACMV. ELISA results were further confirmed by PCR with ACMV specific primers AL1/F and AR0/R that amplified a 987-bp DNA fragment corresponding to the intergenic region, AC-4 and AC-1 genes of DNA-A segment (4). The PCR product was cloned into pCR2.1 (Invitrogen, Carlsbad, CA) and three independent clones were sequenced in both orientations. Pairwise comparison of the derived consensus sequence (GenBank Accession No. EU367500) with corresponding ACMV sequence of ACMV isolate from Nigeria (GenBank Accession No. X17095) showed 98% identity at the nucleotide level. To further confirm the virus identity, complete nucleotide sequence of the DNA-A segment was determined by PCR amplification of viral DNA with four primers, cloning of overlapping products into pCR2.1 vector and sequencing. The derived sequence (2,781 nucleotides; GenBank Accession No. EU685385) was compared with the DNA sequences available at NCBI database using BLAST. This revealed 97% nucleotide sequence identity with ACMV-[NG:Ogo:90] (Accession No. AJ427910) and ACMV-[NG] (Accession No. X17095) from Nigeria. These results confirm the presence of ACMV in symptomatic soybean leaves. To our knowledge, this is the first report of soybean as a natural host of ACMV in SSA. On the basis of previous reports (1) and the results currently presented it seems that ACMV has a wide host range. References: (1) O. J. Alabi et al. Phytopathology (Abstr.) 97(suppl.):S3, 2007. (2) A. A. Brunt et al., eds. Plant viruses online: Descriptions and lists from the VIDE database. Version 20. Online publication, 1996. (3) F. O. Ogbe et al. Plant Dis. 90:548, 2006; (4) X. Zhou et al. J. Gen. Virol. 78:2101, 1997.
非洲木薯花叶病毒(ACMV;菜豆金色花叶病毒属,双生病毒科)是撒哈拉以南非洲(SSA)木薯(Manihot esculenta Crantz.)植株上出现木薯花叶病症状的六种病毒之一。除木薯外,ACMV的自然寄主范围包括一些野生木薯属物种、大戟科的珊瑚花、蓖麻以及旋花科的浅裂月光花。ACMV的实验寄主范围包括茄科的烟草属和曼陀罗属植物(2)。最近,在尼日利亚豆科的团花、银合欢和望江南中发现了ACMV的自然发病情况((1,3)。在2007年9月和10月对伊巴丹地区(N = 19)和贝努埃地区(N = 23)的大豆(Glycine max L. Merr.)进行的勘查研究以及2008年2月对伊巴丹地区(N = 16)的大豆进行的勘查研究中,我们观察到大豆出现黄色花叶和斑驳症状。对这些植株的样本(N = 58)进行间接ELISA检测,有症状叶片对黄瓜花叶病毒、豇豆花叶病毒、南方菜豆花叶病毒、烟草环斑病毒、大豆矮缩病毒、豇豆蚜传花叶病毒、黑眼豇豆花叶病毒、花生斑驳病毒和蚕豆花叶病毒检测呈阴性,这些病毒在SSA的大豆中均有记录。然而,这些样本中有8.6%(58个样本中的5个)(2007年调查中伊巴丹和贝努埃各1个,2008年调查中伊巴丹3个)在用针对ACMV的单克隆抗体(SCR33)进行的三抗体夹心ELISA检测中呈阳性。通过使用ACMV特异性引物AL1/F和AR0/R进行PCR进一步证实了ELISA结果,该引物扩增出一个987 bp的DNA片段,对应于DNA - A片段的基因间隔区、AC - 4和AC - 1基因(4)。将PCR产物克隆到pCR2.1(Invitrogen,卡尔斯巴德,加利福尼亚州)中,并对三个独立克隆进行双向测序。将推导的共有序列(GenBank登录号EU367500)与来自尼日利亚的ACMV分离株的相应ACMV序列(GenBank登录号X17095)进行成对比较,在核苷酸水平上显示出98%的同一性。为了进一步确认病毒的身份,通过用四种引物对病毒DNA进行PCR扩增、将重叠产物克隆到pCR2.1载体中并测序,确定了DNA - A片段的完整核苷酸序列。使用BLAST将推导的序列(2781个核苷酸;GenBank登录号EU685385)与NCBI数据库中可用的DNA序列进行比较。这显示与来自尼日利亚的ACMV - [NG:Ogo:90](登录号AJ427910)和ACMV - [NG](登录号X17095)的核苷酸序列同一性为97%。这些结果证实了有症状的大豆叶片中存在ACMV。据我们所知,这是SSA中大豆作为ACMV自然寄主的首次报道。根据先前的报道(1)和目前呈现的结果,似乎ACMV具有广泛的寄主范围。参考文献:(1)O. J.Аlabi等人,《植物病理学(摘要)》97(增刊):S3,2007。(2)A. A. Brunt等人编,《植物病毒在线:来自VIDE数据库的描述和列表》。第20版。在线出版物,1996。(3)F. O. Ogbe等人,《植物病害》90:548,2006;(4)X. Zhou等人,《普通病毒学杂志》78:2101,1997。