Development of transgenic watermelon resistant to Cucumber mosaic virus and Watermelon mosaic virus by using a single chimeric transgene construct

2011 ◽  
Vol 21 (5) ◽  
pp. 983-993 ◽  
Author(s):  
Ching-Yi Lin ◽  
Hsin-Mei Ku ◽  
Yi-Hua Chiang ◽  
Hsiu-Yin Ho ◽  
Tsong-Ann Yu ◽  
...  
Plant Disease ◽  
2003 ◽  
Vol 87 (4) ◽  
pp. 324-328 ◽  
Author(s):  
F. Fukumoto ◽  
Y. Masuda ◽  
K. Hanada

Necrotic diseases of the stems, petioles, and leaves of pea plants (Pisum sativumL.), leading to wilting and death, occur in the Wakayama and Mie Prefectures of Japan. Based on host range, symptomatology, electron microscopy, and serological relationships, Watermelon mosaic virus (WMV) and three Cucumber mosaic virus (CMV) isolates (PE2, PE3A, and PB1) were isolated from diseased plants in the Wakayama Prefecture. In the Mie Prefecture, CMV (PEAN) also was isolated from pea plants with similar symptoms. Single infection with CMV (PB1 or PEAN) caused stem necrosis and eventual death of pea plants. Similar symptoms developed after double infection with WMV and PE2 or PE3A, whereas single infection with PE2 and PE3A induced symptomless infection in pea plants. We concluded either CMV alone or synergistic effects of mixed infection with CMV and WMV induced pea plant stem necrosis.


2005 ◽  
Vol 2 (3) ◽  
pp. 179-185 ◽  
Author(s):  
Niu Sheng-Niao ◽  
Huang Xue-Sen ◽  
Wong Sek-Man ◽  
Yu Jia-Lin ◽  
Zhao Fu-Xing ◽  
...  

AbstractA trivalent vector containing genes of Watermelon mosaic virus (WMV) coat protein (CP), and replicase genes of Zucchini yellow mosaic virus (ZYMV) and Cucumber mosaic virus (CMV), was constructed for transformation of watermelon (Citrullus lanatus) plants, mediated by Agrobacterium tumefaciens. The integrated foreign genes were identified in the regenerated progenies by polymerase chain reaction (PCR) and Southern blots. The transformation efficiency was about 1.7‰. Resistance to virus infection was determined by mechanical inoculation in the greenhouse and by field trials. The transgenic watermelon lines showed different phenotypes of susceptible, resistant, immune or recovery from virus infections in the late growth stage. A relatively high level of resistance was shown by T3 plants of the line BH1-7. This result indicates the possibility of creating, by transgenic protocols, new varieties of watermelon resistant to viral infection.


HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 471B-471
Author(s):  
S. Alan Walters ◽  
Jeff D. Kindhart ◽  
Houston A. Hobbs ◽  
Darin M. Eastburn

Cucurbit viruses are a major hindrance to cucurbit production in southern Illinois, often rendering cucumber and summer squash fruit unmarketable. Specific viruses infecting cucurbits in the region need to be determined since this would enable growers to make better decisions on virus disease management. Leaf samples of various cucurbit vegetables that had symptoms of viral infection were collected from grower fields during the 1998 and 1999 growing seasons to determine the predominant cucurbit viruses present. Samples were assayed for the presence of five individual viruses: cucumber mosaic virus (CMV), papaya ringspot virus (PRSV, formerly watermelon mosaic virus-1), squash mosaic virus (SqMV), watermelon mosaic virus (WMV, formerly watermelon mosaic virus-2), and zucchini yellow mosaic virus (ZYMV). Results from the two years indicated that WMV was the most prevalent virus in southern Illinois. Cucumber mosaic virus was found both years, but only in a low percentage of samples collected each year. Two cucurbit viruses, PRSV and ZYMV, were each identified only in one sample during the 1998 growing season, and neither were found in any of the samples collected during 1999. Squash mosaic virus was not identified in any of the samples collected during 1998; however, for the 1999 growing season, SqMV was identified in 19% of the samples collected, primarily from those samples of transgenic squash that were showing symptoms of virus infection.


2001 ◽  
Vol 26 (1) ◽  
pp. 90-92 ◽  
Author(s):  
MARIA C. C. L. MOURA ◽  
J. ALBERSIO A. LIMA ◽  
VANÚZIA B. OLIVEIRA ◽  
M. FÁTIMA B. GONÇALVES

Os vírus representam sérios obstáculos para o sucesso da olericultura no mundo inteiro, constituindo a identificação daqueles de maior incidência numa região, papel fundamental para o estabelecimento de estratégias de controle. Visitas de campo foram realizadas a plantios de espécies de cucurbitáceas em áreas produtoras do Maranhão e amostras foliares foram coletadas de 118 plantas com sintomas ou suspeita de sintomas de vírus, sendo 46 de abóbora (Cucurbita moschata), 30 de melancia (Citrullus lanatus), 23 de maxixe (Cucumis anguria), 13 de pepino (C. sativus) e seis de melão (C. melo). Todas as amostras foram testadas contra anti-soros específicos para os principais vírus das famílias Bromoviridae, Comoviridae e Potyviridae que infetam cucurbitáceas no Nordeste, mediante "enzyme-linked immunosorbent assay" (ELISA) indireto e dupla difusão em agar. Os resultados revelaram a identificação sorológica de Papaya ringspot vírus (PRSV) em 64,4% das amostras analisadas, seguido de Watermelon mosaic virus-2 (WMV-2) em 15,2%, Cucumber mosaic virus (CMV) em 6,8%, Squash mosaic virus (SqMV) em 3,4% e Zucchini yellow mosaic virus (ZYMV) em 3,4%. Este levantamento confirma a predominância do PRSV em espécies de cucurbitáceas cultivadas no estado do Maranhão.


2002 ◽  
Vol 92 (1) ◽  
pp. 51-58 ◽  
Author(s):  
Yongzeng Wang ◽  
Victor Gaba ◽  
Jie Yang ◽  
Peter Palukaitis ◽  
Amit Gal-On

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.


Author(s):  
M. G. F. O. Soares ◽  
J. A. Soares ◽  
M. A. Cezar ◽  
T. A. L. Cardoso ◽  
J. A. A. Lima

<p>Por constituírem uma importante fonte de alimento,<strong> </strong>plantações de melancia e abóbora são comumente cultivadas no sertão paraibano, porém pouco se sabe sobre a ocorrência de patógenos causadores de doenças, os quais limitam a sua produtividade e renda aos produtores. Visando obter informações sobre a ocorrência dos patógenos virais e fúngicos em cultivos de abóbora e melancia situados em municípios produtores no sertão da Paraíba, amostras coletadas com sintomas de mosaico e deformação foliar, típicos de doenças virais foram analisadas pela técnica sorológica “enzime linked immuno sorbentassay” (Elisa) indireto para <em>Papaya ring spot virus, </em>type watermelon (PRSV-W), <em>Watermelon mosaic virus </em>(WMV), <em>Zucchini yellow mosaic virus </em>(ZYMV) e <em>Cucumber mosaic virus </em>(CMV). O teste de dupla difusão em Agar foi utilizado para verificar a presença de <em>Squash mosaic virus </em>(SqMV). Em contrapartida, amostras coletadas com sintomas de doenças fúngicas foram analisadas pelo isolamento do patógeno e visualização de suas características morfológicas em microscópio óptico. Em abóbora, houve prevalência dos vírus ZYMV e PRSV-W em infecções simples e mistas, e maior incidência dos fungos <em>Cladosporium</em> spp., e <em>Alternaria</em> spp. Em melancia detectou-se<em> </em>infecção simples e mistas das espécies PRSV-W, WMV e ZYMV, e maior freqüência de <em>Fusarium</em> spp. e <em>Alternaria</em> spp.. Não foram detectados os vírus CMV e SqMV. Os resultados obtidos revelam a ocorrência de vários patógenos fúngicos e viróticos em cultivos de abóbora e melancia situados no sertão da Paraíba e ressaltam a importância da utilização de estratégias de manejo que reduzem os danos ocasionados por esses patógenos.</p><p align="center"><strong><em>Occurrence of pathogens in watermelon and pumpkin crops in the State of Paraiba</em></strong></p><p><strong>Abstract:</strong> By constitute an important food source, watermelon and pumpkin plantations are commonly grown on Paraíba<strong> </strong>backlands, but little is known about the occurrence of disease-causing pathogens, which limit their productivity and income to producers. With objective to get information about the viral and fungal pathogens occurrence in pumpkin and watermelon crops in producing counties located in the backlands of Paraiba, samples with symptoms of mosaic and leaf distortion, typical of viral etiology of disease were collected and analyzed by "Enzyme linked immune sorbentassay" (Elisa) Indirect for species <em>Papaya ring spot virus</em>, type watermelon (PRSV-W), <em>Watermelon mosaic virus </em>(WMV), <em>Zucchini yellow mosaic virus</em> ( ZYMV) and <em>Cucumber mosaic virus</em> (CMV). The double diffusion test in Agar was used to verify the presence of <em>Squash mosaic virus</em> (SqMV). In contrast, samples collected with fungal disease symptoms were analyzed based on the isolation of the pathogen and viewing their morphological characteristics under an optical microscope. In pumpkin, there was prevalence of ZYMV and PRSV-W in single and mixed infections, and higher incidence of <em>Cladosporium</em> spp. and <em>Alternaria</em> spp. On the other hand, in watermelon was detected single and mixed infections of PRSV-W, WMV and ZYMV species, and higher frequency of <em>Fusarium </em>spp. and <em>Alternaria</em> spp.. Were not detected the virus CMV and SqMV. The results show the occurrence of various fungal and viral pathogens in pumpkin and watermelon crops located in the backlands of Paraiba and emphasize the importance of using management strategies that reduce the damage caused by these pathogens.</p>


2011 ◽  
Vol 26 (4) ◽  
pp. 325-336 ◽  
Author(s):  
Ana Vucurovic ◽  
Aleksandra Bulajic ◽  
Ivana Stankovic ◽  
Danijela Ristic ◽  
Janos Berenji ◽  
...  

Cucumber mosaic virus (CMV) is considered one of the most economically important plant viruses and has a worldwide distribution and a very wide host range including plants from family Cucurbitaceae. In Serbia, on cucurbits CMV was detected in single and mixed infections with Zucchini yellow mosaic virus (ZYMV) and Watermelon mosaic virus (WMV). Viruses, including CMV, are constantly present in cucurbit crops, but their frequency changes by year and locality. Surveys and sample collections were conducted in cucurbit crops in the period from 2008 to 2009 at 15 localities in Vojvodina province, and sample testing was carried out using the DAS-ELISA method and commercially available antisera for six economically most important cucurbit viruses. In 2008, a total of 51 samples were collected from 13 cucurbit crops of oilseed pumpkin Olinka variety, squash, and bottle gourd and CMV was detected in a total of 55% of tested samples with symptoms of viral infection. The most common infectious type was mixed infection with ZYMV and WMV (35.3%), and then mixed infection with ZYMV (17.7%) and WMV (2%). A total of 599 symptomatic samples of oilseed pumpkin Olinka variety, zucchini squash varieties Beogradska and Tosca, squash, and winter squash were collected in 15 cucurbits crops in 2009. CMV was present in 4.4% of total collected samples, in single infections in 1.3%, and in mixed with WMV or ZYMV in 1.3%, and 1.8%. Five CMV isolates were obtained by mechanical inoculations of N. glutinosa and one of them was selected for further biological characterization. Test plants which were described to be hosts of CMV expressed symptoms characteristic for those caused by CMV after inoculations by isolate 115-08. CMV specific primers Au1u/Au2d were used to amplify an 850 bp fragment using RT-PCR method. Amplified fragment encodes the entire viral coat protein (CP) gene and partial 5? and 3? UTRs of two selected CMV isolates. Amplified fragments were sequenced and deposited in the NCBI, where they were assigned accession numbers, HM065510 (115-08) and HM065509 (151-08). The sequences of CMV isolates from Serbia shared the highest nucleotide and amino acid identity with isolates from subgroup IA, from 99.5 to 97.4% and 99.1 to 97.4%, and the lowest identities were with the subgroup II isolates from 66.9 to 64, 5%, from 75.8 to 74.1%.


2004 ◽  
Vol 29 (6) ◽  
pp. 687-689 ◽  
Author(s):  
Bernardo A. Halfeld-Vieira ◽  
Najara F. Ramos ◽  
Francisco A.C. Rabelo Filho ◽  
M. Fátima B. Gonçalves ◽  
Katia L. Nechet ◽  
...  

No período de maio de 2003 a março de 2004, foram coletadas amostras foliares de plantas de melancia (Citrullus lanatus) de 21 campos de cultivo de cucurbitáceas, no Estado de Roraima. As amostras exibiam diferentes sintomas de vírus e foram levadas para o Laboratório de Virologia Vegetal da Universidade Federal do Ceará para serem testadas por "enzyme linked immunosorbent assay" (Elisa)-indireto, contra anti-soros específicos para Cucumber mosaic virus (CMV), Papaya ringspot virus estirpe melancia (PRSV-W), Watermelon mosaic virus (WMV) e Zucchini yellow mosaic virus (ZYMV). Nos testes de Elisa, utilizou-se o conjugado universal, anti-imunoglobulina (IgG) de coelho produzida em cabra conjugada à enzima fosfatase alcalina. Todas as amostras foram testadas, também, por dupla difusão contra o anti-soro para Squash mosaic virus (SqMV). Os resultados indicaram a presença do PRSV-W em 84,2% das amostras coletadas em maio de 2003, em 7,1% das amostras coletadas em dezembro de 2003 e em 55,6% das amostras coletadas em março de 2004. A presença do ZYMV foi observada em 10,5% das amostras coletadas em maio de 2003, 21,4% das amostras coletadas em dezembro de 2003 e em 25,9% das amostras de março de 2004. O WMV foi detectado somente em oito das amostras coletadas em março de 2004 (29,6%). Os resultados desta pesquisa confirmam a ampla dispersão do PRSV-W em cultivos de cucurbitáceas no território brasileiro e a preocupante expansão do ZYMV em razão dos elevados prejuízos que o mesmo tem causado em outras partes do mundo.


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