scholarly journals Changes in the Concentration of an Allexivirus During the Crop Cycle of Two Garlic Cultivars

Plant Disease ◽  
2006 ◽  
Vol 90 (10) ◽  
pp. 1293-1296 ◽  
Author(s):  
Eva E. Cafrune ◽  
Mónica Balzarini ◽  
Vilma C. Conci

Garlic can be infected by a number of viruses, including allexiviruses. The coat protein sequence of an Allexivirus was detected in Argentina and deposited in the EMBL database as Garlic mite-borne filamentous virus (accession number X98991); it has high homology with Garlic virus A (GarV-A). For reliable virus detection, plants should be sampled when virus titer is high to reduce the risk of identifying infected plants as healthy. The objective of this study was to describe fluctuations in the concentration of this Argentine isolate of GarV-A in two garlic cultivars, Morado-INTA and Nieve-INTA, throughout the crop cycle using the double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Over a 2-year period, for both cultivars, virus concentration was assessed in samples from the tips section of the youngest leaves of GarV-A-infected plants, and from basal sections of both dormant and devernalized cloves of stored bulbs of Morado-INTA. The concentration of GarV-A varied during the crop cycle, but peaked at the beginning and again at the end of the crop cycle. Virus concentration was slightly higher in devernalized cloves compared with dormant cloves of Morado-INTA. No correlation between virus concentration and mean air temperature was observed. The results of this study recommend sampling times at the beginning of the crop cycle at 64 to 81 days after planting, and towards the end of the crop cycle to evaluate for the presence of GarV-A by DAS-ELISA.

Plant Disease ◽  
2002 ◽  
Vol 86 (10) ◽  
pp. 1085-1088 ◽  
Author(s):  
Vilma C. Conci ◽  
Pablo Lunello ◽  
Diana Buraschi ◽  
Rusell R. Italia ◽  
Sergio F. Nome

The purpose of this work was to determine variations in titer of Leek yellow stripe virus (LYSV) throughout the crop cycle and bulb storage, and to evaluate the incidence of infected plants in the main garlic-production regions of Argentina. One hundred plants with LYSV from each of five cultivars were analyzed by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) in six different vegetative stages in White- and Red-type garlic cultivars and seven stages in cv. Rosado Paraguayo, throughout the year. In two White-type garlic cultivars, LYSV showed peaks of viral concentration in May, at the beginning of the crop cycle, and in November, just before harvest. In two Red-type garlic selections, an increase was detected in November (period of bulbing). The highest virus titers for these four garlic cultivars were detected in devernalized clove. In Rosado Paraguayo, the peak virus concentration occurred in September prior to harvesting. In a survey at 14 different localities in Argentina, 3,066 random samples were analyzed. LYSV was found in 80 to 98% of the plants from all regions, except in Santa Cruz, where 34% of plants were infected. The importance of this study is that it allows us to recommend the most suitable moment of the year to make the analysis with DAS-ELISA.


Viruses ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2403
Author(s):  
Wei Wang ◽  
Jizong Li ◽  
Baochao Fan ◽  
Xuehan Zhang ◽  
Rongli Guo ◽  
...  

Porcine deltacoronavirus (PDCoV) can cause diarrhea and dehydration in newborn piglets. Here, we developed a double antibody sandwich quantitative enzyme-linked immunosorbent assay (DAS-ELISA) for detection of PDCoV by using a specific monoclonal antibody against the PDCoV N protein and an anti-PDCoV rabbit polyclonal antibody. Using DAS-ELISA, the detection limit of recombinant PDCoV N protein and virus titer were approximately 0.5 ng/mL and 103.0 TCID50/mL, respectively. A total of 59 intestinal and 205 fecal samples were screened for the presence of PDCoV by using DAS-ELISA and reverse transcriptase real-time PCR (RT-qPCR). The coincidence rate of the DAS-ELISA and RT-qPCR was 89.8%. DAS-ELISA had a sensitivity of 80.8% and specificity of 95.6%. More importantly, the DAS-ELISA could detect the antigen of PDCoV inactivated virus, and the viral antigen concentrations remained unchanged in the inactivated virus. These results suggest that DAS-ELISA could be used for antigen detection of clinical samples and inactivated vaccines. It is a novel method for detecting PDCoV infections and evaluating the PDCoV vaccine.


Plant Disease ◽  
2010 ◽  
Vol 94 (10) ◽  
pp. 1188-1194 ◽  
Author(s):  
S. C. Chinestra ◽  
C. Facchinetti ◽  
N. R. Curvetto ◽  
P. A. Marinangeli

In a survey of lily growing fields in various regions of Argentina, three viruses, Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV), were found in Longiflorum, Asiatic, Oriental, Longiflorum × Asiatic (LA), and Oriental × Trumpet (OT) hybrids. The areas surveyed were between latitude 26° 56′ S and 43° 03′ S, and longitude 65° 21′ W and 71° 29′ W. Virus detection was performed by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) using polyclonal antiserum. In infected samples, viruses detected in decreasing order were LSV (60.5%), LMoV (51.0%), and CMV (28.7%) present in single or mixed infections. Virus infection varied among tested hybrids from 36.0% (Oriental Montecristo) to 94.7% (Lilium longiflorum Avita) in 2006 and from 38.9% (OT Yelloween) to 82.1% (LO Triumphator) in 2007, with an overall incidence of 64.1 and 70.7% in 2006 and 2007, respectively. A variation in virus incidence among localities was also observed. The highest virus incidence (89.6 and 87.6% in 2006 and 2007, respectively) was observed in Bahía Blanca (38° 44′ S, 62° 16′ W). The lowest virus incidences, detected in Trevellin (43° 03′ S, 71° 29′ W) and in Malargüe (35° 28′ S, 69° 35′ W), were 47.4 and 48.6% in 2006 and 2007, respectively. Moreover, a different distribution of each virus was observed between localities. The high occurrence of viruses infecting lily crops in Argentina could be due to both the use of infected bulbs for propagation and the lack of preventive virus vector control measures.


1985 ◽  
Vol 17 (10) ◽  
pp. 39-41 ◽  
Author(s):  
A. Schnattinger

Ten litres of tapwater were seeded with 200 µl (8×108 HAV particles) of a commercial (Organon Teknika) suspension of hepatitis A virus. Following WALTER and RÜDIGER (1981), the contaminated tapwater was treated with a two-stage technique for concentration of viruses from solutions with low virus titers. The two-stage technique consists of aluminium hydroxideflocculation (200 mg/l Al2(SO4)3. 18 H2O, pH 5,4-5,6) as first stage, the second stage of a lysis of aluminium hydroxidegel with citric acid/sodium citrate-buffer (pH 4,7; 1 ml/l sample), separation of viruses from the lysate by ultracentrifugation and suspension in 1 ml phosphate buffer solution (pH 7,2). A commercial solid phase enzyme-linked immunosorbent assay (ELISA) was used for the detection of HAV. HAV was detecterl in the 10.000:1 concentrates, but not in the seeded 101 samples. Approximately 4×108 of the inoculated 8×108 HAV particles were found in the 1 ml concentrates. The efficiency of detection is about 50%, the virus concentration 5000-fold. Although the percentage loss of HAV in comparison with concentration by means of membrane filtration is similar, the ultracentrifugation method yields a larger sample/concentrate ratio, so that smaller amounts of HAV can be detected more efficiently because of the smaller end-volume.


Author(s):  
Rami Obeid ◽  
Elias Wehbe ◽  
Mohamad Rima ◽  
Mohammad Kabara ◽  
Romeo Al Bersaoui ◽  
...  

Background: Tobacco mosaic virus (TMV) is the most known virus in the plant mosaic virus family and is able to infect a wide range of crops, in particularly tobacco, causing a production loss. Objectives: Herein, and for the first time in Lebanon, we investigated the presence of TMV infection in crops by analyzing 88 samples of tobacco, tomato, cucumber and pepper collected from different regions in North Lebanon. Methods: Double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), revealed a potential TMV infection of four tobacco samples out of 88 crops samples collected. However, no tomato, cucumber and pepper samples were infected. The TMV+ tobacco samples were then extensively analyzed by RT-PCR to detect viral RNA using different primers covering all the viral genome. Results and Discussion: PCR results confirmed those of DAS-ELISA showing TMV infection of four tobacco samples collected from three crop fields of North Lebanon. In only one of four TMV+ samples, we were able to amplify almost all the regions of viral genome, suggesting possible mutations in the virus genome or an infection with a new, not yet identified, TMV strain. Conclusion: Our study is the first in Lebanon revealing TMV infection in crop fields, and highlighting the danger that may affect the future of agriculture.


Plant Disease ◽  
2021 ◽  
Author(s):  
Dan Edward Veloso Villamor ◽  
Karen E Keller ◽  
Robert Martin ◽  
Ioannis Emmanouil Tzanetakis

A comprehensive study comparing virus detection between high throughput sequencing (HTS) and standard protocols in 30 berry selections (12 Fragaria, 10 Vaccinium and 8 Rubus) with known virus profiles was completed. The study examined temporal detection of viruses at four sampling times encompassing two growing seasons. Within the standard protocols, RT-PCR proved better than biological indexing. Detection of known viruses by HTS and RT-PCR nearly mirrored each other. HTS provided superior detection compared to RT-PCR on a wide spectrum of virus variants and discovery of novel viruses. More importantly, in most cases where the two protocols showed parallel virus detection, 11 viruses in 16 berry selections were not consistently detected by both methods at all sampling points. Based on these data we propose a four sampling times/two-year testing requirement for berry and potentially other crops to ensure that no virus remains undetected independent of titer, distribution or other virus/virus or virus/host interactions.


Plant Disease ◽  
2003 ◽  
Vol 87 (2) ◽  
pp. 201-201 ◽  
Author(s):  
William M. Wintermantel ◽  
Teresa Crook ◽  
Ralph Fogg

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV) and vectored by the soilborne fungus Polymyxa betae Keskin, is one of the most economically damaging diseases affecting sugar beet (Beta vulgaris L.). The virus likely originated in Europe and was first identified in California in 1983 (1). It has since spread among American sugar beet production regions in spite of vigorous sanitation efforts, quarantine, and disease monitoring (3). In the fall of 2002, mature sugar beet plants exhibiting typical rhizomania root symptoms, including proliferation of hairy roots, vascular discoloration, and some root constriction (2) were found in several fields scattered throughout central and eastern Michigan. Symptomatic beets were from numerous cultivars, all susceptible to rhizomania. Two to five sugar beet root samples were collected from each field and sent to the USDA-ARS in Salinas, CA for analysis. Hairy root tissue from symptomatic plants was used for mechanical inoculation of indicator plants. Mechanical inoculation produced necrotic lesions on Chenopodium quinoa and systemic infection of Beta vulgaris ssp. macrocarpa, both typical of BNYVV and identical to control inoculations with BNYVV. Symptomatic sugar beet roots were washed and tested using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) for the presence of BNYVV using standard procedures and antiserum specific for BNYVV (3). Sugar beet roots were tested individually, and samples were considered positive when absorbance values were at least three times those of greenhouse-grown healthy sugar beet controls. Samples were tested from 16 fields, with 10 confirmed positive for BNYVV. Positive samples had mean absorbance values ranging from 0.341 to 1.631 (A405nm) after 30 min. The mean healthy control value was 0.097. Fields were considered positive if one beet tested positive for BNYVV, but in most cases, all beets tested from a field were uniformly positive or uniformly negative. In addition, soil-baiting experiments were conducted on seven of the fields. Sugar beet seedlings were grown in soil mixed with equal parts of sand for 6 weeks and were subsequently tested using DAS-ELISA for BNYVV. Results matched those of the root sampling. Fields testing positive for BNYVV were widely dispersed within a 100 square mile (160 km2) area including portions of Gratiot, Saginaw, Tuscola, and Sanilac counties in the central and eastern portions of the Lower Peninsula of Michigan. The confirmation of rhizomania in sugar beet from the Great Lakes Region marks the last major American sugar beet production region to be diagnosed with rhizomania disease, nearly 20 years after its discovery in California (1). In 2002, there were approximately 185,000 acres (approximately 75,00 ha) of sugar beet grown in the Great Lakes Region, (Michigan, Ohio, and southern Ontario, Canada). The wide geographic distribution of infested fields within the Michigan growing area suggests the entire region should monitor for symptoms, increase rotation to nonhost crops, and consider planting rhizomania resistant sugar beet cultivars to infested fields. References:(1) J. E. Duffus et al. Plant Dis. 68:251, 1984. (2) J. E. Duffus. Rhizomania. Pages 29–30 in: Compendium of Beet Diseases and Insects, E. D. Whitney and J. E. Duffus eds. The American Phytopathological Society, St. Paul, MN, 1986. (3) G. C. Wisler et al. Plant Dis. 83:864, 1999.


Plant Disease ◽  
1998 ◽  
Vol 82 (11) ◽  
pp. 1282-1282 ◽  
Author(s):  
H. R. Pappu ◽  
J. W. Todd ◽  
A. K. Culbreath ◽  
M. D. Bandla ◽  
J. L. Sherwood

In Georgia, tomato spotted wilt tospovirus (TSWV) causes significant losses in peanut, tobacco, tomato, and pepper. Transmission of TSWV in Georgia primarily is by tobacco thrips (TT), Frankliniella fusca (Hinds), and western flower thrips, F. occidentalis (Pergande), with TT being the predominant vector species in peanut (2). TSWV must be acquired at the larval stage for the adult to transmit the virus. Detection of NSs (a non-structural TSWV protein present only following virus replication) in thrips by enzyme-linked immunosorbent assay (ELISA) is a reliable indicator that the virus had multiplied in the vector and thus the vector is competent to transmit TSWV. While this has been accomplished with F. occidentalis (1), information is lacking for F. fusca, the predominant vector in Georgia and other states in the Southeast. Thus, the nature of the TSWV-TT association was investigated and the proportion of transmitters in a field population determined in 1,436 individual adult TT collected from sticky cards positioned in selected peanut fields in south Georgia. Additionally, 650 larvae collected from volunteer peanut plants were reared to adults in the laboratory and the resulting 295 adult TT were individually evaluated by ELISA. Of those collected from the sticky cards, NSs was detected in 8% of the adult insects, indicating that the virus had multiplied in TT. NSs was not detected in control TT that had no access to the virus. Of the adult TT that emerged from larvae collected from volunteer peanuts, 6.1% were positive for NSs. Our study provides the first immunological evidence that TSWV multiplies in TT. References: (1) M. D. Bandla et al. Phytopathology 84:1427, 1994. (2) J. R. Chamberlin et al. J. Econ. Entomol. 86:40, 1993.


Plant Disease ◽  
2006 ◽  
Vol 90 (4) ◽  
pp. 526-526 ◽  
Author(s):  
K. Bananej ◽  
C. Desbiez ◽  
C. Wipf-Scheibel ◽  
I. Vahdat ◽  
A. Kheyr-Pour ◽  
...  

A survey was conducted from 2001 to 2004 in the major cucurbit-growing areas in Iran to reassess the relative incidence of cucurbit viruses. Severe yellowing symptoms were observed frequently on older leaves of cucurbit plants in various regions in outdoor crops, suggesting the presence of Cucurbit aphid-borne yellows virus (CABYV, genus Polerovirus, family Luteoviridae) (1,2). Leaf samples (n = 1019) were collected from plants of melon (Cucumis melo L.), cucumber (C. sativus L.), squash (Cucurbita sp.), and watermelon (Citrullus lanatus L.) showing various virus-like symptoms (mosaic, leaf deformation, yellowing). All samples, collected from 15 provinces, were screened for the presence of CABYV by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with IgGs and alkaline phosphatase-conjugated IgGs against a CABYV reference isolate (1). Of the 1,019 samples tested, 471 were positive for CABYV using DAS-ELISA. Some of the positive samples had typical severe yellowing symptoms while symptoms in other samples were masked by mosaic or leaf deformations caused by other viruses frequently found in mixed infections (data not shown). During the entire survey, CABYV was detected by DAS-ELISA in 201 of 503 melon samples, 72 of 129 cucumber samples, 158 of 249 squash samples, and 40 of 138 watermelon samples. These results indicate that CABYV is widely distributed on four cucurbit species in the major growing areas of Iran. In order to confirm CABYV identification, total RNA extracts (TRI-Reagent, Sigma Chemical, St Louis, MO) were obtained from 25 samples that were positive using DAS-ELISA originating from Khorasan (n = 4), Esfahan (n = 6), Teheran (n = 3), Hormozgan (n = 4), Azerbaiejan-E-Sharqi (n = 4), and Kerman (n = 4). Reverse transcription-polymerase chain reactions (RT-PCR) were carried out using forward (5′-CGCGTGGTTGTGG-TCAACCC-3′) and reverse (5′-CCYGCAACCGAGGAAGATCC-3′) primers designed in conserved regions of the coat protein gene according to the sequence of a CABYV reference isolate (3) and three other unpublished CABYV sequences. RT-PCR experiments yielded an expected 479-bp product similar to the fragment amplified with extracts from the reference isolate. No amplification of the product occurred from healthy plant extracts. To our knowledge, this is the first report of the occurrence of CABYV in Iran on various cucurbit species. The high frequency (46.2%) with which CABYV was detected in the samples assayed indicates that this virus is one of the most common virus infecting cucurbits in Iran. References: (1) H. Lecoq et al. Plant Pathol. 41:749, 1992 (2) M. A. Mayo and C. J. D'Arcy. Page 15 in: The Luteoviridae. H. G. Smith and H. Barker, eds. CAB International Mycological Institute, Wallingford, UK, 1999. (3) H. Guilley et al. Virology 202:1012, 1994.


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