Concepción Jordá

Genetic Structure of the Population of Pepino mosaic virus Infecting Tomato Crops in Spain.

ABSTRACT The population structure of Pepino mosaic virus (PepMV), which has caused severe epidemics in tomato in Spain since 2000, was analyzed. Isolates were characterized by the nucleotide sequence of the triple gene block and coat protein gene and, for a subset of isolates, a part of the RNA-dependent RNA polymerase gene. The full-length sequence of the genomic RNA of a Solanum muricatum isolate from Peru also was determined. In spite of high symptom diversity, the Spanish population of PepMV mostly comprised highly similar isolates belonging to the strain reported in Europe (European tomato strain), which has been the most prevalent genotype in Spain. The Spanish PepMV population was not structured spatially or temporally. Also, isolates highly similar to those from nontomato hosts from Peru (Peruvian strain) or to isolate US2 from the United States (US2 strain) were detected at lower frequency relative to the European strain. These two strains were detected in peninsular Spain only in 2004, but the Peruvian strain has been detected in the Canary Islands since 2000. These results suggest that PepMV was introduced into Spain more than once. Isolates from the Peruvian and US2 strains always were found in mixed infections with the European tomato strain, and interstrain recombinants were detected. The presence of different strains of the virus, and of recombinant isolates, should be considered for the development of control strategies based on genetic resistance.

A RT-PCR assay combined with RFLP analysis for detection and differentiation of isolates of Pepino mosaic virus (PepMV) from tomato

The partial nucleotide sequence of the RNA polymerase gene from one isolate of Pepino mosaic virus (PepMV) was determined. Phylogenetic and distance analysis indicated that this isolate was related to other isolates of PepMV previously reported. To develop a method for detecting PepMV by reverse transcriptase–polymerase chain reaction (RT–PCR), a pair of primers was designated from RNA polymerase sequences. RT–PCR with RNA from a large number of tomato samples with PepMV symptoms, positive controls of PepMV, weed samples containing PepMV, and other potexviruses as negative controls confirmed the specificity of the primers. Restriction endonuclease digestion of the RT–PCR products distinguished three restriction fragment length polymorphism (RFLP) types. The majority of isolates were included within type P1, which correspond with the PepMV isolates found in Europe. This type and type P2, which corresponded with the original PepMV isolated from Solanum muricatum, appear more closely related to each other than to type P3. Type P3 had completely different RFLPs from the other two types studied. It may represent a further line within the PepMV virus. The RT–PCR–RFLP assay is proposed as a rapid and easy method to detect and identify new isolates of the PepMV virus.

Multiplex PCR assay for the simultaneous detection and differentiation of Olpidium bornovanus, O. brassicae, and O. virulentus

A multiplex PCR method has been developed to detect, differentiate, and confirm the morphological identification of three root infecting Olpidium spp.: O. bornovanus, O. brassicae, and O. virulentus. Of the 132 root samples examined, 101 samples were infected by Olpidium spp.. Based on the morphology of resting spores, the presence of O. bornovanus was confirmed in 20.5% of the samples, whereas species identity could not be determined for the remaining samples because they failed to reproduce sexually. With multiplex PCR, it was possible to determine the Olpidium identity of all the infected samples, even when resting spores were not formed. This method was also effective for detecting Olpidium spp. in water samples. In addition, the specificity and sensitivity of multiplex PCR were evaluated. The multiplex PCR method was validated with samples of 9 different crops from 11 countries of America, Europe, and Africa.

Viruses of cucurbits in Panama

The main areas of field-grown cucurbit production in Panama were surveyed for the occurrence and distribution of viruses in the growing seasons of 2006 and 2008. Notably, in 2006 only Melon necrotic spot virus (MNSV) was detected, while Papaya ringspot virus (PRSV) was the most frequent in 2008 in terms of number of fields and samples analysed, followed by Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV) and MNSV. Cucumber mosaic virus (CMV), Squash mosaic virus (SqMV), Cucumber green mottle mosaic virus (CGMMV), Tomato spotted wilt virus (TSWV), Cucumber leaf spot virus (CLSV), Cucurbit yellow stunting disorder virus (CYSDV), Cucurbit aphid-borne yellows virus (CABYV), Beet pseudoyellows virus (BPYV), Beet yellows virus (BYV), and Cucumber vein yellowing virus (CVYV) were not detected . Single and double (PRSV+WMV; PRSV+ZYMV; ZYMV+MNSV) infections were detected in 60% and 40% of the samples tested, respectively. The occurrence and distribution of each virus varied according to the region and cucurbit species.