Creation of trivalent transgenic watermelon resistant to virus infection

2005 ◽  
Vol 2 (3) ◽  
pp. 179-185 ◽  
Author(s):  
Niu Sheng-Niao ◽  
Huang Xue-Sen ◽  
Wong Sek-Man ◽  
Yu Jia-Lin ◽  
Zhao Fu-Xing ◽  
...  
Keyword(s):  
Virus Infection ◽  
Mosaic Virus ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
Field Trials ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Virus Infections ◽  
High Level ◽  
Transgenic Watermelon

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.

scholarly journals Occurrence and Distribution of Pumpkin and Squash Viruses in Illinois

Plant Disease ◽  
2008 ◽  
Vol 92 (1) ◽  
pp. 61-68 ◽  
Author(s):  
S. Jossey ◽  
M. Babadoost
Keyword(s):  
Virus Infection ◽  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Dual Infection ◽  
Ringspot Virus ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Watermelon Mosaic Virus ◽  
Virus Infections ◽  
Dark Green

Surveys were conducted during 2004 to 2006 to identify the viruses infecting pumpkin and squash in Illinois. In 2004, 16 jack-o-lantern pumpkin (Cucurbita pepo) samples and one squash (C. pepo) sample were collected from 11 counties. In 2005, 85 jack-o-lantern pumpkin, 12 processing pumpkin (Cucurbita moschata), 37 squash, and six gourd (C. pepo) samples were collected from 54 counties. In 2006, 85 jack-o-lantern pumpkin, 16 processing pumpkin, 51 squash, and 18 gourd samples were collected from 47 counties. Cucumber mosaic virus (CMV), Papaya ringspot virus (PRSV), Squash mosaic virus (SqMV), Tobacco ringspot virus (TRSV), Tomato ringspot virus (ToRSV), Watermelon mosaic virus (WMV), Zucchini yellow mosaic virus (ZYMV), and unknown potyviruses were detected in pumpkin, squash, and gourd fields during the surveys, using enzyme-linked immunosorbent assay (ELISA). Overall, 86, 11, 75, and 79% of jack-o-lantern pumpkin, processing pumpkin, squash, and gourds, respectively, were tested positive for virus infection during the survey. WMV was detected in 47, 46, and 52% of the samples in 2004, 2005, and 2006, respectively, and was the most prevalent virus throughout the state. SqMV was detected in more counties than any other virus because it was identified in 65 and 88% of the counties surveyed in 2005 and 2006, respectively. SqMV was detected in 6, 41, and 48% of the samples in 2004, 2005, and 2006, respectively. During the surveys, CMV was detected in 6, 4, and 3% of the samples; PRSV was detected in 6, 11, and 4% of the samples; and ZYMV was detected in 18, 4, and 4% of the samples tested in 2004, 2005, and 2006, respectively. TRSV was detected in 3% of the samples in 2005, for the first time on pumpkin in Illinois. Pathogenicity of the detected viruses was proved for CMV, PRSV, SqMV, WMV, and ZYMV on summer squash (‘Fortune’ and ‘Grey Zucchini’), jack-o-lantern pumpkin (‘Howden’), and processing pumpkin (‘Dickinson’). All of the viruses were present alone and mixed in the samples tested. Earlier in the growing seasons (July and early August), single-virus infections were detected. Mixed infections were more common from the second week of August until the end of the growing season in October. Dual infection of WMV and SqMV was the most prevalent mixed virus infection detected in Illinois. Most viruses infecting pumpkin and squash showed similar symptoms. The most common symptoms observed in the commercial fields and in the greenhouse studies were light- and dark-green mosaic, veinbanding, veinclearing, puckering, and deformation of leaves of pumpkin, squash, and gourds. Severe symptoms included fernleaf and shoestring on leaves and color breaking and deformation of fruit.

scholarly journals Evaluation of ‘AU-Performance’ Watermelon for Its Response to Virus Inoculation

2009 ◽  
Vol 19 (3) ◽  
pp. 609-612 ◽  
Author(s):  
John F. Murphy ◽  
Fenny Dane
Keyword(s):  
Mosaic Virus ◽  
Fungal Pathogens ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
Systemic Infection ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Susceptible Parent ◽  
Resistant Parent ◽  
Systemic Symptoms

The watermelon (Citrullus lanatus var. lanatus) ‘AU-Performance’ was developed for resistance to multiple fungal pathogens and the plant virus, zucchini yellow mosaic virus (ZYMV). A greenhouse-based evaluation was carried out to determine the response of ‘AU-Performance’ to inoculation with three important cucurbit (Cucurbitaceae) viruses in the genus Potyvirus: papaya ringspot virus (PRSV), watermelon mosaic virus (WMV), and ZYMV. The evaluation included the resistant parent (PI595203), the susceptible parent (‘AU-Producer’), and varieties AU-Allsweet and Charleston Gray. Each of the three viruses systemically infected ‘AU-Performance’ with 100% infection and development of characteristic systemic symptoms. The susceptible parent (‘AU-Producer’), ‘AU-Allsweet’, and ‘Charleston Gray’ responded similarly with 100% infection and systemic symptoms. In contrast, the resistant parent (PI595203) was resistant to WMV and ZYMV; however, PRSV-inoculated plants developed a systemic infection with accompanied symptoms and high levels of PRSV accumulation in noninoculated leaves. PI595203 was shown in previous studies to be resistant to PRSV. We show in this report that under greenhouse conditions and application of virus by mechanical inoculation, ‘AU-Performance’ was not resistant to infection by the three potyviruses.

Some characteristics of Australian isolates of Zucchini Yellow Mosaic Virus

1988 ◽  
Vol 39 (6) ◽  
pp. 1085 ◽  
Author(s):  
RS Greber ◽  
DM Persley ◽  
ME Herrington
Keyword(s):  
Mosaic Virus ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
Resistance Breeding ◽  
Seed Transmission ◽  
Fruit Production ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Marketable Fruit ◽  
Triton X

Isolates of zucchini yellow mosaic virus (ZYMV) were obtained from six cucurbit species from four states of Australia and identified by host range, electron microscopy and serology. A survey of coastal Queensland in 1986-87 showed no ZYMV north of Townsville nor around Rockhampton, but major centres of ZYMV incidence with specific pathotypes, around Ayr, Bowen and in south-east Queensland. Pathotypes were distinguished using Cucumis melo differentials, Trichosanthes anguina and Phaseolus vulgaris. Three ZYMV isolates were compared with watermelon mosaic virus 2 and papaya ringspot virus, cucurbit strain, on 20 hosts from 5 families. The effect of ZYMV on 19 cvv. of pumpkin, squash, zucchini, cucumber, rockmelon and watermelon was examined in the field where marketable fruit production was most affected in Cucurbita maxima and C. pepo, moderately in C. melo and Citrullus lanatus and least in C. moschata and C. sativus. One instance of ZYMV seed transmission in C. pepo and transmission by Aphisgossypii was demonstrated. Resistance breeding progress with C. maxima using a C. ecuadorensis source and in C. lanatus using Citrullus sp. cv. Egusi is reported. Virus purification was achieved using carbon tetrachloride and Triton X-100 clarification with sucrose pad and gradient centrifugations. An antiserum was produced and reacted well in agarose gel-diffusion tests without prior virus particle disruption.

scholarly journals Frequency and molecular characterization of watermelon mosaic virus from Serbia

2010 ◽  
Vol 25 (3) ◽  
pp. 213-230
Author(s):  
Ana Vucurovic ◽  
Aleksandra Bulajic ◽  
Ivana Stankovic ◽  
Danijela Ristic ◽  
Janos Berenji ◽  
...  
Keyword(s):  
Virus Infection ◽  
Mosaic Virus ◽  
Cucurbita Pepo ◽  
Zucchini Yellow Mosaic Virus ◽  
Growing Season ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Mixed Infections ◽  
Mediterranean Regions

Watermelon mosaic virus (WMV) is widespread in cucurbit crops, most commonly occuring in temperate and Mediterranean regions. In Serbia WMV has been detected in single and mixed infections with Zucchini yellow mosaic virus and Cucumber mosaic virus in field-grown pumpkin and squash crops. Among pumpkin-affecting viruses WMV is the most frequent one, both by the number of localities and its incidence at each location. During the growing season of 2009, samples from 583 plants of Cucurbita pepo cvs. Olinka, Belgrade zucchini and Tosca (Zucchini group), as well as from C. maxima and C. moschata showing symptoms of virus infection were collected from 12 commercial fields at eight localities and analyzed by DAS-ELI

scholarly journals First Report of Zucchini yellow mosaic virus in Cucumber (Cucumis sativus) in India

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 906-906 ◽  
Author(s):  
Raj Verma ◽  
Satya Prakash ◽  
S. P. S. Tomer
Keyword(s):  
Mosaic Virus ◽  
Cucumis Sativus ◽  
Cucurbita Pepo ◽  
Citrullus Lanatus ◽  
Natural Infection ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Cucumis Sativus L ◽  
First Report

In August of 2002, cucumber (Cucumis sativus L. cv. Himangi) plants grown in commercial fields in Pune, western Maharashtra, India, exhibited chlorotic spots, veinal chlorosis, mosaic, blister formation and shoestring symptoms on leaves, stunted growth, and distortion of fruits. Incidence of virus infection in the fields varied from 25 to 38%. Crude sap extracted from infected cucumber leaf samples was inoculated mechanically onto cucumber and indicator host plants. The inoculated glasshouse-grown cucumber plants showed virus symptoms similar to those observed in the field. The virus produced chlorotic local lesions on Chenopodium amaranticolor and chlorotic spots followed by veinal chlorosis, mosaic, vein banding, and leaf distortion on Citrullus lanatus, C. melo, C. sativus, Cucurbita maxima, Cucurbita pepo, Luffa acutangula, and Trichosanthes anguina. The virus did not infect Nicotiana benthamiana, N. glutinosa, and N. tabacum cv. White Burley (1,2). Cucumber and indicator plants were tested using direct antigen coating enzyme-linked immunosorbent assays. A positive reaction was obtained with monoclonal antiserum to Zucchini yellow mosaic virus (ZYMV) but not with antisera to Papaya ringspot virus-P, Cucumber mosaic virus, and Watermelon mosaic virus (Agdia, Inc., Elkhart, IN). The disease was observed again in the field during July to September 2003. Natural infection of zucchini (Cucurbita pepo) by ZYMV has been reported previously from India (3). To our knowledge, this is the first report of ZYMV in cucumber in India. References: (1) H. Pospieszny et al. Plant Dis. 87:1399, 2003. (2) R. Provvidenti et al. Plant Dis. 68:443, 1984. (3) S. J. Singh et al. Indian Phytopathol. 56:174, 2003.

scholarly journals Identificação sorológica de espécies de vírus que infetam cucurbitáceas em áreas produtoras do Maranhão

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
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Watermelon Mosaic Virus ◽  
Papaya Ringspot Virus ◽  
Cucurbita Moschata

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.

scholarly journals Distribution of Viruses Infecting Cucurbit Crops and Isolation of Potential New Virus-Like Sequences from Weeds in Oklahoma

Plant Disease ◽  
2012 ◽  
Vol 96 (2) ◽  
pp. 243-248 ◽  
Author(s):  
Akhtar Ali ◽  
Osama Mohammad ◽  
Abeer Khattab
Keyword(s):  
Mosaic Virus ◽  
Zucchini Yellow Mosaic Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Virus Infections ◽  
Rt Pcr ◽  
Weed Species ◽  
Field Surveys ◽  
Weed Host ◽  
Symptomatic Leaf

Field surveys were conducted from 2008 to 2010 to detect and determine the incidence of viruses in the major cucurbit-growing areas of Oklahoma. In total, 1,049 symptomatic leaf samples (890 from cucurbits, 109 from weed species, and 50 from crop plants [agricultural crops]) were collected from 90 fields in four counties (Atoka, Blaine, Jefferson, and Tulsa) of Oklahoma. Samples were tested against seven viruses, including Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV), Melon necrotic spot virus (MNSV), Papaya ringspot virus-watermelon strain (PRSV-W, formerly known as Watermelon mosaic virus-1), Squash mosaic virus (SqMV), Watermelon mosaic virus-2 (WMV-2), and Zucchini yellow mosaic virus (ZYMV), using dot-immunobinding assay (DIBA). Results showed the highest incidence for PRSV (51%), followed by WMV-2 (14%) and ZYMV (10%) among the collected samples. SqMV, MNSV, and CMV were detected in 3.8, 3.3, and 1.1% of the samples, respectively. None of the samples collected during surveys reacted positively against the antiserum of CGMMV. Mixed virus infections were common involving two (5.18%) or three (4.61%) viruses in various combinations. New weed host species were found to be infected with PRSV when confirmed by both DIBA and reverse-transcription polymerase chain reaction (RT-PCR). Some weed species contained possible new viruses when analyzed by random RT-PCR, followed by cloning, sequencing, and BLAST analysis with sequences in GenBank.

scholarly journals First Report of Zucchini yellow mosaic virus Associated with Leaf Crinkle and Yellow Mosaic Diseases of Cucurbit Plants in Mali

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 923-923 ◽  
Author(s):  
W. S. Tsai ◽  
I. K. Abdourhamane ◽  
D. Knierim ◽  
J. T. Wang ◽  
L. Kenyon
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
The United States ◽  
Yellow Mosaic Virus ◽  
Nucleotide Identity ◽  
Watermelon Mosaic Virus ◽  
Virus Species ◽  
First Report

The aphid-transmitted Zucchini yellow mosaic virus (ZYMV; genus Potyvirus, family Potyviridae) has been reported to cause severe epidemics and yield losses in cucurbit crops worldwide (1). In Africa, ZYMV has been detected in Algeria, Egypt, Madagascar, Mauritius, Mayotte, Morocco, Nigeria, Reunion, South Africa, Sudan, Swaziland, and Tunisia (1). In April 2009, leaf yellowing, mosaic, crinkling, and curling were common on cucurbit plants in fields in Mali. Symptomatic leaf samples were collected from five cucumber (Cucumis sativus) plants in Kati, two watermelon (Citrullus lanatus) plants in Samanko, and one weedy melon (Cucumis sp.) plant in Baguineda. All samples tested positive for ZYMV and were negative for Cucumber mosaic virus (CMV), Cucumber green mottle mosaic virus (CGMMV), Papaya ringspot virus type W (PRSV-W), Watermelon mosaic virus (WMV), and Watermelon silver mottle virus (WSMoV) by double-antibody sandwich (DAS)-ELISA. They also tested negative for Melon yellow spot virus (MYSV) by indirect ELISA. Antibodies against ZYMV and WMV were obtained from DSMZ, Braunschweig, Germany, and those against CGMMV, MYSV, PRSV-W, and WSMoV were provided by Shyi-Dong Yeh, National Chung Hsing University, Taichung, Taiwan. Six ZYMV ELISA-positive samples (three cucumber, two watermelon, and the weedy melon sample) were also tested by reverse transcription (RT)-PCR using the potyvirus universal primer pair Sprimer1/Oligo(dT) (2). The expected 1.6-kb viral cDNA was amplified from all six samples and each was sequenced. All sequences obtained from cucumber (GenBank Accession Nos. HM005307, HM005308, and HM005309), watermelon (GenBank Accession Nos. HM005311 and HM005312), and weedy melon (GenBank Accession No. HM005310) isolates were 1,684 nucleotides (nt) long excluding the 3′ poly-A tails. They comprised the 3′-terminal of the NIb region (1 to 633 nt), the coat protein region (634 to 1473 nt), and the 3′-untranslated region (1,474 to 1,684 nt). Because the sequences shared high nucleotide identity (98.3 to 99.7%), these isolates were considered to be the same virus species. When the sequences were compared by BLASTn searching in GenBank and analyzed by DNAMAN Sequence Analysis Software (Lynnon Corporation, St-Louis, Pointe-Claire, Quebec, Canada), they were found to have the greatest nucleotide identity (97.4 to 98.0%) with the Connecticut strain of ZYMV (ZYMV-Connecticut; GenBank Accession No. D00692), within a clade of isolates from China, Italy, Japan, and the United States. When assessed separately, their coat protein (97.7 to 98.3% nucleotide and 98.9 to 99.6% amino acid identity) and 3′-untranslated regions (96.7 to 97.2% identity) also had greatest homology with ZYMV-Connecticut. To our knowledge, this is the first report of ZYMV infecting cucurbit plants in Mali. ZYMV should be taken into consideration when breeding cucurbit crops for this region, and managing viral diseases. References: (1) C. Desbiez et al. Plant Pathol. 46:809, 1997. (2) W. S. Tsai et al. Plant Dis. 94:378, 2010.

scholarly journals Identificação sorológica de espécies de potyvirus em melancia, no estado de Roraima

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 ◽  
...  
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
Ringspot Virus ◽  
Yellow Mosaic Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Watermelon Mosaic Virus ◽  
Papaya Ringspot Virus

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.

scholarly journals Fontes de resistência em melancia aos principais potyvírus isolados de cucurbitáceas no Nordeste brasileiro

2002 ◽  
Vol 20 (4) ◽  
pp. 589-592 ◽  
Author(s):  
Vanuzia B. de Oliveira ◽  
Manoel Abilio de Queiroz ◽  
Albersio A. Lima
Keyword(s):  
Mosaic Virus ◽  
Virus Type ◽  
Citrullus Lanatus ◽  
Zucchini Yellow Mosaic Virus ◽  
Ringspot Virus ◽  
Yellow Mosaic Virus ◽  
Watermelon Mosaic Virus ◽  
Papaya Ringspot Virus

A melancia, Citrullus spp., pode ser infetada por várias espécies de vírus, dentre as quais se destacam as pertencentes à família Potyviridae. Visando identificar fontes de resistência a potyvírus, foi iniciado na UFC um programa de indexação do banco de germoplasma de melancia da Embrapa Semi-Árido. Foram utilizadas as espécies de potyvírus, isoladas de cucurbitáceas no Nordeste: Papaya ringspot virus type W (PRSV-W); Watermelon mosaic virus-2 (WMV-2) e Zucchini yellow mosaic virus (ZYMV). Dos 50 acessos de melancia avaliados, 37 pertencem a Citrullus lanatus, 9 a C. lanatus var. citroides e 4 a C. colocynthis. Em casa-de-vegetação foram inoculadas 4 plantas envasadas de cada acesso com PRSV-W, 4 com WMV-2 e 4 com ZYMV, ficando 4 sem inoculação. As inoculações foram efetuadas 8 dias após o plantio, e 15 dias depois. As plantas assintomáticas foram reinoculadas. As plantas que persistiram sem sintomas, após um período de 25 dias após a primeira inoculação, foram testadas por ELISA contra os anti-soros correspondentes. Dentre os acessos avaliados, 5 mostraram-se resistentes aos 3 vírus, um resistente somente ao PRSV-W, 21 resistentes somente ao ZYMV e nenhum com resistência isolada ao WMV-2. No entanto, 45 acessos apresentaram resistência ao ZYMV isoladamente ou em combinação com outro potyvírus. Os acessos suscetíveis, apresentaram sintomas 10 dias após a primeira inoculação enquanto que os resistentes permaneceram sem sintomas após a segunda inoculação e com resultados negativos em ELISA. Alguns acessos poderão ser cruzados visando obter populações segregantes para a seleção de indivíduos homozigotos recessivos e, juntamente com os acessos com resistência tripla identificados, ampliar a variabilidade genética de fontes quanto a características de planta e fruto.

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