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一种新型双生病毒在墨西哥南下加利福尼亚州引发番茄卷叶病。

A New Begomovirus Causes Tomato Leaf Curl Disease in Baja California Sur, Mexico.

作者信息

Holguín-Peña R J, Vázquez-Juárez R, Rivera-Bustamante R F

机构信息

Centro de Investigaciones Biológicas del Noroeste, La Paz, B.C.S. 23000, Mexico.

Centro de Investigación y de Estudios Avanzados del IPN, Irapuato, Guanajuato 36500, Mexico.

出版信息

Plant Dis. 2005 Mar;89(3):341. doi: 10.1094/PD-89-0341A.

Abstract

More than 10,000 ha of tomatoes are grown in the field and greenhouses on the Baja California Peninsula of Mexico. Information about the etiology of geminivirus-like diseases affecting tomato crops in all horticultural regions in the area has been difficult to obtain and assess. From 2001 through 2003, stunting, foliar discoloration, reduced leaf size, and leaf crumpling symptoms were observed and analyzed in one large area of tomato plantings in El Carrizal (near the city of La Paz in Baja California Sur). This leaf curl disease resembled that caused by Chino del tomate virus and has been observed at levels of incidence ranging from 60 to 90%. DNA isolated from symptomatic plants was analyzed using DNA hybridizaton and polymerase chain reaction (PCR) amplification of the 5' regions of the replication and coat protein genes, including the intergenic region (3). Comparisons of the nucleotide sequence (GenBank Accession No. AY339619) with corresponding sequences in GenBank resulted in 84.2% identity with Tomato mild mottle virus and 61.7% with Tomato severe leaf curl virus; both isolates originate from Central America. The relatively low nucleotide sequence identities from its closest relatives suggested that the virus may be a new begomovirus species of unambiguous American ancestry. In a phylogenetic analysis using PAUP 4.0 software, the Baja California isolate clustered in a separate group from other Mexican sequences. Moreover, the iteron (iterative sequences motifs associated in virus replication) arrangements (1) are unique among known New World begomoviruses, but identical to analogous elements from a tobacco-infecting begomovirus from China. On the other hand, it is well known that there are interactions between geminiviruses in mixed infections in some horticultural areas of Mexico (2). To determine the identity of the putative geminivirus involved in the disease, we used selected restriction enzyme (EcoRI, HindIII and XbaI) analysis and PCR with specific primers. No evidence of mixed infections with other geminiviruses was obtained. DNA fragments of the expected size (1.1 kb) showed different digestion patterns compared with other well-characterized geminiviruses isolated from Mexico such as Chino del tomate virus, Pepper huasteco yellow vein virus, Tomato leaf curl Sinaloa virus, and Pepper golden mosaic virus. Epidemiological, experimental, and natural host range studies indicated that the Baja California isolate has a relatively narrow host range infecting tomatoes, peppers (Capsicum annuum L.), and Peruvian apple (Nicandra physalodes L.). Reproduction of characteristic leaf curling symptoms in tomato seedlings infected with viruliferous whiteflies (Bemisia tabaci Genn.) and inoculated biolistically using infectious DNA (0.5 μg/ml) as inoculum were obtained. Koch's postulates were completed using PCR and DNA hybridization to confirm virus identity. These results confirm that the Baja California isolate is different from other begomoviruses isolated from Mexico. The virus is tentatively named Tomato chino Baja California virus (ToChBCV), genus Begomovirus, family Geminiviridae. References: (1) G. R. Arguello-Astorga et al. Arch. Virol. 146:1465, 2001. (2) J. Mendez-Lozano et al. Phytopathology 93:270, 2003. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993.

摘要

墨西哥下加利福尼亚半岛的农田和温室里种植着超过10000公顷的番茄。一直以来,很难获取和评估该地区所有园艺种植区影响番茄作物的类双生病毒病的病因信息。2001年至2003年期间,在埃尔卡里萨尔(南下加利福尼亚州拉巴斯市附近)的一大片番茄种植区观察并分析了植株发育迟缓、叶片变色、叶片尺寸减小和叶片皱缩等症状。这种卷叶病与番茄奇诺病毒引起的症状相似,发病率在60%至90%之间。使用DNA杂交和聚合酶链反应(PCR)扩增复制和外壳蛋白基因的5'区域(包括基因间隔区),对从有症状植株中分离的DNA进行了分析(3)。将核苷酸序列(GenBank登录号AY339619)与GenBank中的相应序列进行比较,结果显示与番茄轻度斑驳病毒的同一性为84.2%,与番茄严重卷叶病毒的同一性为61.7%;这两种分离株均源自中美洲。与其亲缘关系最近的病毒的核苷酸序列同一性相对较低,这表明该病毒可能是一种起源明确来自美洲的新型双生病毒。在使用PAUP 4.0软件进行的系统发育分析中,下加利福尼亚分离株与其他墨西哥序列聚在一个单独的组中。此外,重复序列(与病毒复制相关的重复序列基序)排列(1)在已知的新大陆双生病毒中是独特的,但与来自中国的一种感染烟草的双生病毒的类似元件相同。另一方面,众所周知,在墨西哥的一些园艺种植区,双生病毒在混合感染中存在相互作用(2)。为了确定与该病相关的假定双生病毒的身份,我们使用了选定的限制性内切酶(EcoRI、HindIII和XbaI)分析以及用特异性引物进行的PCR。未获得与其他双生病毒混合感染的证据。与从墨西哥分离的其他特征明确的双生病毒(如番茄奇诺病毒、辣椒瓦斯特科黄脉病毒、番茄卷叶锡那罗亚病毒和辣椒金色花叶病毒)相比,预期大小(1.1 kb)的DNA片段显示出不同消化模式。流行病学、实验和自然寄主范围研究表明,下加利福尼亚分离株的寄主范围相对较窄,可感染番茄、辣椒(辣椒属)和秘鲁苹果(酸浆属)。在用带毒粉虱(烟粉虱)感染并使用感染性DNA(0.5 μg/ml)作为接种物进行基因枪接种的番茄幼苗中再现了典型的卷叶症状。使用PCR和DNA杂交完成了科赫法则以确认病毒身份。这些结果证实,下加利福尼亚分离株与从墨西哥分离的其他双生病毒不同。该病毒暂定名为番茄奇诺下加利福尼亚病毒(ToChBCV),属于双生病毒属,双生病毒科。参考文献:(1)G. R. Arguello-Astorga等人,《病毒学档案》146:1465,2001年。(2)J. Mendez-Lozano等人,《植物病理学》93:270,2003年。(3)M. R. Rojas等人,《植物病害》77:340,1993年。

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