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本研究在我国主要冬瓜产区采集具有典型病毒病症状的病叶材料105份,根据葫芦科作物上常见的5种病毒病原的CP基因设计特异性引物,对105份待检冬瓜材料进行RT-PCR检测。检测结果表明:5对特异引物可分别在105份待检材料的95份中检测到小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)、西瓜花叶病毒(Watermelon mosaic virus,WMV)、黄瓜花叶病毒(Cucumber mosaic virus,CMV)、番木瓜环斑病毒(Papaya ringspot virus,PRSV)4种病毒,未检测到南瓜花叶病毒(Squash mosaic virus,SqMV);并且发现不同的冬瓜主产区致病的病毒种类有较大差异;同时还发现,在这些待检样品中4种病毒复合侵染现象较普遍,其中以PRSV与WMV组合最常见,占复合侵染现象的31.25%;未发现有4种病毒复合侵染。 相似文献
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Melon chlorotic mosaic virus and associated alphasatellite from Venezuela: genetic variation and sap transmission of a begomovirus–satellite complex 
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下载免费PDF全文 Begomoviruses represent one of the most damaging virus groups on many important crops worldwide. In Venezuela, the begomovirus Melon chlorotic mosaic virus (MeCMV) is the major constraint for melon and watermelon production. MeCMV has been associated with the satellite Melon chlorotic mosaic alphasatellite (MeCMA). Full‐length genome sequencing of 20 and 35 isolates of MeCMV and MeCMA, respectively, was carried out to estimate their genetic variability. Furthermore, mechanical transmission assays of MeCMV alone, or in conjunction with MeCMA, were performed. Genetic variation was low among MeCMV isolates, which exhibited 97–100% nucleotide identity for the DNA‐A component and 95–100% for the DNA‐B component. Alphasatellite isolates were highly variable ranging from 86·5 to 100% nucleotide identity. MeCMV isolates were phylogenetically related to begomoviruses belonging to the Squash leaf curl virus (SLCV) clade, while MeCMA isolates were clustered in two subgroups related to alphasatellites from the New World (Cuba and Brazil). MeCMV has a host range restricted to cucurbit species and two experimental hosts: Nicotiana benthamiana and Nicotiana clevelandii. MeCMV can be mechanically transmitted with up to 100% efficiency in melon. The physiological stage of the inoculated organ (cotyledon or leaf) represents a key factor for inoculation efficiency. This result provides a simple and reliable inoculation method to develop extensive screening for MeCMV resistance sources. In addition, the complex MeCMV + MeCMA was mechanically transmitted to melon, N. benthamiana and N. clevelandii plantlets and successfully back‐transmitted. To the authors’ knowledge, this finding is the first evidence of sap transmission for a begomovirus–satellite complex. 相似文献
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Identification et distribution des virus responsables de mosaiques chez les cucurbitacées en Tunisie
Les cucurbitacées sont économiquement importantes en Tunisie, elles sont sujettes à plusieurs maladies virales qui ont des effets néfastes sur les rendements et la valeur marchande de la production. Des symptômes de mosaïques sévères sur les feuilles ont été observés dans toutes les cultures de cucurbitacées de plein champ et sous abris. Des prospections (2000–2001, 2002–2003) ont été réalisées afin de tenter d’identifier les virus responsables de ces symptômes. Les échantillons symptomatiques ont été collectés dans les régions majeures de production des cucurbitacées et testés par DAS‐ELISA pour la présence des virus suivants: ZYMV, WMV, MWMV, PRSV‐W, ZYFV CMV, SqMV et MNSV. Cette étude a confirmé la présence de six virus (WMV, CMV, PRSV‐W, ZYMV, ZYFV et SqMV) déjà décrits avec une évolution très remarquable de la fréquence du ZYMV dans toutes les régions prospectées et sur toutes les espèces testées. Le pourcentage d’infection a atteint les 34% durant les prospections (2002–2003). Par contre le MWMV s’est révélé absent au niveau de tous les échantillons testés. Par ailleurs, nous avons mis en évidence pour la première fois la présence du MNSV dans plus de 9% des échantillons testés et ce sur melon de plein champ et sous abris, sur courge et sur pastèque. En outre, nous rapportons pour la première fois l’infection d’une cucurbitacée sauvage, Ecballium elaterium, par le CMV, PRSV‐W et WMV. Les résultats ont montré aussi qu’un pourcentage d’échantillons montrant des symptômes de mosaïques n’a pas été expliqué par la présence de l’un ou l’autre des virus étudiés. Ceci pourrait s’expliquer par l’existence d’autres virus ou de facteurs abiotiques qui restent à identifier. 相似文献
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P. J. Mansilla A. G. Moreira A. P. O. A. Mello J. A. M. Rezende J. A. Ventura V. A. Yuki F. J. Levatti 《Plant pathology》2013,62(3):571-577
Papaya ringspot virus type P (PRSV‐P) systemically infects Carica papaya and species belonging to the family Cucurbitaceae. Attempts to recover PRSV‐P from naturally infected cucurbit plants grown near or among diseased papaya trees have shown conflicting results worldwide. This study aimed to evaluate the natural infection of cucurbit species grown among and near papaya trees infected with PRSV‐P in Brazil. Natural infection of cucurbits with PRSV‐P occurred in zucchini squash but not in watermelon and cucumber. However, several attempts to recover PRSV‐P from numerous Cucurbita pepo cv. Caserta (zucchini squash) plants grown 5–80 m from diseased papaya trees in the field failed. Mechanical inoculations of Cucurbita pepo cv. Caserta, Cucurbita maxima cv. Exposição (pumpkin), Cucumis sativus cv. Primepack Plus (cucumber) and Citrullus lanatus cv. Crimson Sweet (watermelon) with five Brazilian PRSV‐P isolates showed that zucchini squash was the most susceptible species followed by watermelon and cucumber, while pumpkin was not infected. The results confirmed the variable susceptibility of cucurbit species to experimental and natural PRSV‐P infection. Given these facts, the control of the disease through roguing should focus mainly on diseased papaya plants, as has been practised successfully in Brazil for many years, and on those cucurbits particularly known to be susceptible to natural infection with PRSV‐P. 相似文献
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基于外壳蛋白基因序列对3种葫芦科作物病毒的分子分析 总被引：2，自引：0，他引：2
从山西运城的南瓜(Cucurbita moschata)、河南开封和孟津的西葫芦(Cucurbita pepo)上通过ELISA分别检测到南瓜花叶病毒(Squash mosaic virus,SqMV)、西瓜花叶病毒(Watermelon mosaic virus,WMV)和番木瓜环斑病毒西瓜株系(Papaya ringspot virus-Watermelon type,PRSV.w),通过接种分离纯化获得其病毒毒原(CH99/211,CH99/113,CH99/69)。本研究克隆了这3个分离物的外壳蛋白基因,应用DNAStar、Clustal X、Phylip等分析软件将这些序列同已报道的相应序列进行分析并构建系统树。结果发现SqMV可以分为两个组,CH99/211分离物与美国的Arizona分离物接近,应属于z组;WMV分为两个组,CH99/69同国内黑龙江分离物(WMV-HLJ)和云南分离物(WMV-CHN)最接近,成一小簇;PRSV分成两个组。CH99/113属于Ⅱ组,与国内SP、MZ和BN分离物接近。序列分析表明PRSV和WMV在进化上具有地理相关性。SqMV和PRSV-W外壳蛋白基因序列分析属国内首次报道。 相似文献
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S. Yakoubi C. Desbiez H. Fakhfakh C. Wipf‐Scheibel F. Fabre M. Pitrat M. Marrakchi H. Lecoq 《Plant pathology》2008,57(6):1146-1154
A study was conducted to better understand the population structure of Zucchini yellow mosaic virus (ZYMV), a severe virus affecting cucurbit crops worldwide, in Tunisia and to estimate whether the use of resistant cultivars may provide durable control. Analysis of the polymerase and coat protein (NIb‐CP) partial sequences of 83 isolates collected in the three main cucurbit‐growing areas in Tunisia showed that ZYMV grouped into two distinct clusters within ZYMV molecular group A. An important variability was observed in the MREK motif of the P3 protein, a motif associated with tolerance breaking in ZYMV‐tolerant zucchini squash cultivars. Interestingly, significant differences were found in the distribution of the MREK variants in the two clusters defined by the partial NIb‐CP sequences, MREK and MKEK sequences being more common in cluster 1 and cluster 2, respectively. When combining NIb‐CP and P3 sequence information, ZYMV molecular variability was shown to be significantly higher in the Cap Bon region than in the Bizerte area. An important biological variability was observed in a subset of 23 isolates regarding symptomatology in susceptible or resistant cucurbits. Some isolates overcame ZYMV tolerance or resistance in zucchini squash and melon, but not in cucumber. Three serotypes were differentiated using a set of 13 monoclonal antibodies (MAbs). Seven parameters characterizing the 23 isolates, including molecular, serological and biological properties, were used for a multiple component analysis (MCA). This analysis revealed that symptom intensity of a given isolate was similar in different susceptible cucurbit hosts, suggesting similar degrees of aggressiveness in different hosts. 相似文献
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侵染西瓜的5种病毒ZYMV、WMV、TMV、SqMV和CMV的多重RT-PCR检测体系的建立与检测应用 总被引：5，自引：1，他引：4
根据5种病毒小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)、西瓜花叶病毒(Watermelon mosaic virus,WMV)、烟草花叶病毒(Tobacco mosaic virus,TMV)、南瓜花叶病毒(Squash mosaic virus,SqMV)和黄瓜花叶病毒(Cucumber mosaic virus,CMV)的核苷酸保守区序列,设计特异性引物对,从影响多重RT-PCR (mRT-PCR)扩增的引物浓度、Mg2+浓度、Taq DNA聚合酶浓度、dNTPs浓度、退火温度等方面进行反应体系的优化,建立了一种能够同时检测ZYMV、WMV、TMV、SqMV和CMV的多重RT-PCR技术体系,并进行了实际应用。在一个体系中对上述5种病毒复合侵染的西瓜材料进行多重RT-PCR扩增,得到与试验设计相符的5条特异性条带,依次是542、485、410、354和293bp。该体系实现了对侵染西瓜的5种病毒的同时检测,极大地提高了检测效率,降低了检测成本,体现了多重RT-PCR的优越性。 相似文献
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Sang-Wook Kwon Mi-Soon Kim Hong-Soo Choi Kook-Hyung Kim 《Journal of General Plant Pathology》2005,71(1):80-85
Zucchini yellow mosaic virus (ZYMV) is one of the most economically important viruses of cucurbit crops, causing severe mosaic, necrosis, and malformation. Three ZYMV isolates were obtained from pumpkins at Andong (ZYMV-PA), Euiryung (ZYMV-PE), and Suwon (ZYMV-PS), and their biological variability was determined on different hosts, including cucurbit crops as well as other indicator plants. ZYMV-PA caused the most severe symptoms, including severe mosaic, size reduction, and deformation, in oriental melon (Cucumis melo) and cucumber (Cucumis sativus) leaves. In contrast, ZYMV-PE and ZYMV-PS caused mild mosaic symptoms on oriental melon and cucumber. The nucleotide sequences of the genomic RNAs were determined and compared to the sequences of other potyviruses, including ZYMV isolates Reunion Island and TW-TN3. Each ZYMV Isolate had a genome of 9593 nucleotides, excluding the poly(A) tail, and contained 139 and 214 nucleotides in the 5- and 3-untranslated regions, respectively. Each had one large open reading frame encoding a protein of about 351kDa. The nucleotide sequences of ZYMV-PA, ZYMV-PE, and ZYMV-PS were more than 96.0% and the deduced amino acid sequences were more than 98.1% identical. When compared with other ZYMV isolates in a phylogenetic analysis, these three viruses formed a distinct virus clade and were more distantly related to other potyviruses (43.5%–62.8% identity).The nucleotide sequence data reported are available in the GenBank database under the accession numbers AY278998 to AY279000 for ZYMV-PA, ZYMV-PE and ZYMV-PS 相似文献
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Survey of viruses infecting open‐field pepper crops in Côte d'Ivoire and diversity of Pepper veinal mottle virus and Cucumber mosaic virus 下载免费PDF全文
下载免费PDF全文 B. A. Bolou Bi B. Moury K. Abo F. Sorho M. Cherif G. Girardot N. P. Kouassi D. Kone 《Plant pathology》2018,67(6):1416-1425
The prevalence of viruses in pepper crops grown in open fields in the different agro‐ecological zones (AEZs) of Côte d'Ivoire was surveyed. Pepper veinal mottle virus (PVMV; genus Potyvirus) and Cucumber mosaic virus (CMV; genus Cucumovirus) were the most frequent viruses among those surveyed, while tobamoviruses (genus Tobamovirus) were detected at low frequency. PVMV showed a high heterogeneity across AEZs, which may be related to climatic, ecological or agronomical conditions, whereas CMV was more homogeneously distributed. The molecular diversity of CMV and PVMV were analysed from partial genome sequences. Despite the low number of CMV isolates characterized, two molecular groups were revealed, one corresponding to subgroup IA and the other to reassortants between subgroups IA and IB. RNAs 1 and 3 of the reassortants clustered with the IB subgroup of CMV isolates, whereas their RNA 2 clustered with the IA subgroup. Importantly, RNA 1 of CMV isolates of the IB subgroup has been shown to be responsible for adaptation to pepper resistance. The diversity of PVMV in the VPg‐ and coat protein‐coding regions revealed multiple clades. The central part of the VPg showed a high level of amino acid diversity and evidence of positive selection, which may be a signature of adaptation to plant recessive resistance. As a consequence, for efficient deployment of resistant pepper cultivars, it would be desirable to examine the occurrence of virulent isolates in the CMV or PVMV populations in Côte d'Ivoire and to follow their evolution as the resistance becomes more widely deployed. 相似文献
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西瓜花叶病毒(Watermelon mosaic virus, WMV)是危害葫芦科作物的重要病毒。制备特异性强、效价高的抗血清对快速准确检测和鉴定WMV具有重要意义。本研究将WMV外壳蛋白(Coat protein,CP)基因克隆到原核表达载体pEHISTEV获得pEHISTEV-WMV-CP。将pEHISTEV-WMV-CP转化大肠杆菌Rosetta,经IPTG诱导,成功表达出分子量约为36 kDa的蛋白,与预期WMV CP大小一致。切胶回收WMV CP,与等体积弗氏不完全佐剂充分乳化后免疫健康新西兰大白兔。Western blotting结果表明,制备的抗血清与WMV CP有反应,与同属的番木瓜环斑病毒(Papaya ringspot virus,PRSV)、马铃薯Y病毒(Potato virus Y,PVY)、小西葫芦黄花叶病毒(Zucchini yellow mosaic virus, ZYMV)和烟草花叶病毒属的黄瓜绿斑驳花叶病毒(Cucumber green mottle mosaic virus,CGMMV)均无反应。酶联免疫吸附测定(PTA-ELISA)结果表明,本研究制备的WMV抗血清效价为1∶8 192,并且能够检测稀释512倍的病毒汁液。利用该抗血清对田间采集的10个RT-PCR检测为阳性的样品进行检测,全部呈现阳性。本研究利用大肠杆菌表达的CP制备的WMV抗血清具有较高的特异性和灵敏度。研究结果为WMV的快速检测和鉴定奠定了基础。 相似文献
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Bo-Song Ryang Tadashi Matsumoto Takashi Kobori Yoshitaka Kosaka Satoshi T. Ohki 《Journal of General Plant Pathology》2005,71(4):308-313
Cucumber cotyledons inoculated with Cucumber mosaic virus (CMV, Pepo strain) or Zucchini yellow mosaic virus (ZYMV, Z5-1 isolate) developed either mild chlorotic spots or no symptoms. Cotyledons treated with CMV plus ZYMV also developed mild chlorotic spots. However, plants ZYMV-inoculated cotyledons had veinal yellowing and gradual cell death by 20 days postinoculation (dpi) when co-inoculated with CMV on the other cotyledon. When analyzing this synergism, an enzyme-linked immunosorbent assay showed that CMV gradually increased in CMV-inoculated cotyledons of plants, with the other cotyledon mock- or ZYMV-inoculated. However, CMV significantly increased at 9 to 14 dpi in the ZYMV-inoculated cotyledons of plants co-infected with CMV. ZYMV similarly increased in cotyledon pairs of both co-infected and singly infected plants. Inoculation with PepoΔ2b, a modified Pepo-CMV that lacks translation of the 2b protein, revealed that PepoΔ2b without the 2b protein systemically infected cucumber but induced no symptoms on cotyledons or true leaves. Plants with a ZYMV-inoculated cotyledon and co-infected with PepoΔ2b did not undergo cell death; nevertheless, PepoΔ2b was at high levels comparable to levels of CMV in the ZYMV-inoculated cotyledon. The 2b protein thus seems essential for induction of the novel gradual cell death in ZYMV-inoculated cotyledons of cucumbers co-infected with CMV. 相似文献
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Shin-ichi Fuji Nanae Mochizuki Masashi Fujinaga Makoto Ikeda Kouichi Shinoda Seiji Uematsu Hiromitsu Furuya Hideki Naito Fumiyoshi Fukumoto 《Journal of General Plant Pathology》2007,73(3):216-221
Alstroemeria plants were surveyed for viruses in Japan from 2002 to 2004. Seventy-two Alstroemeria plants were collected from Aichi, Nagano, and Hokkaido prefectures and 54.2% were infected with some species of virus. The
predominant virus was Alstroemeria mosaic virus, followed by Tomato spotted wilt virus, Youcai mosaic virus (YoMV), Cucumber mosaic virus (CMV), Alstroemeria virus X and Broad bean wilt virus-2 (BBWV-2). On the basis of nucleotide sequence of the coat protein genes, all four CMV isolates belong to subgroup IA. CMV
isolates induced mosaic and/or necrosis on Alstroemeria. YoMV and BBWV-2 were newly identified by traits such as host range, particle morphology, and nucleotide sequence as viruses
infecting Alstroemeria. A BBWV-2 isolate also induced mosaic symptoms on Alstroemeria seedlings. 相似文献
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ABSTRACT Mixed infections of cucurbits by Cucumber mosaic virus (CMV) and potyviruses exhibit a synergistic interaction. Zucchini squash and melon plants coinfected by the potyvirus Zucchini yellow mosaic virus (ZYMV) and either Fny-CMV (subgroup IA) or LS-CMV (subgroup II) displayed strong synergistic pathological responses, eventually progressing to vascular wilt and plant death. Accumulation of Fny- or LS-CMV RNAs in a mixed infection with ZYMV in zucchini squash was slightly higher than infection with CMV strains alone. There was an increase in CMV (+) strand RNA levels, but no increase in CMV (-) RNA3 levels during mixed infection with ZYMV. Moreover, only the level of capsid protein from LS-CMV increased in mixed infection. ZYMV accumulated to similar levels in singly and mixed infected zucchini squash and melon plants. Coinfection of squash with the potyvirus Watermelon mosaic virus (WMV) and CMV strains increased both the Fny-CMV RNA levels and the LS-CMV RNA levels. However, CMV (-) strand RNA3 levels were increased little or not at all for CMV on coinfection with WMV. Infection of CMV strains (LS and Fny) containing satellite RNAs (WL47-sat RNA and B5*-sat RNA) reduced the accumulation of the helper virus RNA, except when B5*-sat RNA was mixed with LS- CMV. However, mixed infection containing ZYMV and the CMV strains with satellites reversed the suppression effect of satellite RNAs on helper virus accumulation and increased satellite RNA accumulation. The synergistic interaction between CMV and potyviruses in cucurbits exhibited different features from that documented in tobacco, indicating there are differences in the mechanisms of potyvirus synergistic phenomena. 相似文献
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H. Ben Tamarzizt J. Montarry G. Girardot H. Fakhfakh M. Tepfer M. Jacquemond 《Plant pathology》2013,62(6):1415-1428
Cucumber mosaic virus is one of the most prevalent viruses in Tunisian pepper crops, where it has been detected in 68% of plants developing mosaic symptoms, making it essential to characterize the molecular and biological properties of local CMV populations. Two hundred and seventy‐eight isolates collected in the late 1990s, 2006 and 2008–2010 were characterized genetically. Isolates belonging to the three phylogenetic subgroups of CMV (IA, IB and II) were detected, but surprisingly, 90% of the isolates were reassortants between subgroups IA and IB, with two predominant haplotypes, IB‐IA‐IA and IB‐IA‐IB (nomenclature according to the subgrouping of the three genomic RNAs). The IB‐IA‐IA haplotype was present in all regions surveyed, while IB‐IA‐IB was observed only in northern Tunisia. This situation was unexpected, because CMV reassortants were previously thought to be counterselected in nature, and this raises the questions of the origin of IB strains in Tunisia and of the widespread distribution of these two reassortant types. Phylogenetic studies revealed low diversity within haplotypes, whatever the locality or the year of sampling. However, analysis of haplotype frequencies revealed a high genetic differentiation between CMV populations, which was better explained by the localities of sampling than by years. Geographic distances affected the differentiation of CMV populations, mainly between north and central Tunisia. When tested against a polygenic resistance to CMV movement in pepper, 55 of 57 isolates tested were able to break the resistance, indicating that this resistance would not be useful for controlling CMV in Tunisian pepper fields. 相似文献
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Thiago M. Costa Alice K. Inoue-Nagata Andreza H. Vidal Simone da G. Ribeiro Tatsuya Nagata 《Plant pathology》2020,69(6):1042-1050
The severe yellowing disease (amarelão) on melon plants is a serious problem in Brazil, although the causative agent remained unknown for a long time. Recently, recombinant isolates of cucurbit aphid-borne yellows virus (CABYV) were reported as the possible causative agents of this disease on melon plants. Although aphids are known to be the vectors of the common type of CABYV isolates, almost no aphid colony was observed in the major melon fields in Brazil with high incidence of the severe yellowing disease. In contrast, whiteflies are often abundant. Based on this observation, the hypothesis of the transmission of recombinant CABYV by whiteflies was evaluated. After thorough transmission experiments, we found that this recombinant CABYV isolate was transmitted by the whitefly Bemisia tabaci MEAM1, but not by Aphis gossipii. Furthermore, the host response by whitefly-based inoculation in cucurbits and other indicator plants showed differences in host range when compared to the common type of CABYV. Due to its transmissibility by the whitefly and the distant relationship of the P3/P5 protein to CABYV, the name “cucurbit whitefly-borne yellows virus” is proposed for this recombinant CABYV. This is the second report of polerovirus transmission by the whitefly B. tabaci, following the report of pepper whitefly-borne vein yellows virus. 相似文献