Detection of Prunus necrotic ringspot virus in almond: effect of sampling time on the efficiency of serological and biological indexing methodologies

2002 ◽  
Vol 42 (2) ◽  
pp. 207 ◽  
Author(s):  
T. Bertozzi ◽  
E. Alberts ◽  
M. Sedgley
Keyword(s):  
Stem Elongation ◽  
Leaf Tissue ◽  
Ringspot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sample Collection ◽  
Local Conditions ◽  
Nursery Stock ◽  
Young Leaves ◽  
Almond Trees ◽  
Prunus Necrotic Ringspot Virus

The reliability of enzyme-linked immunosorbent assay (ELISA) and indexing methodologies for the detection of Prunus necrotic ringspot virus (PNRSV) in almond was assessed under local conditions. Thirteen field-grown almond trees were sampled fortnightly throughout the growing season. Petal and leaf homogenates were used for ELISA and to inoculate herbaceous indicator species, while buds collected from October onwards were budded to woody indicators. ELISA reliably detected PNRSV in petals and young leaves from bud-break until the cessation of stem elongation. While herbaceous indexing was not as reliable as ELISA, woody indicators could differentiate positive from negative samples reliably throughout the testing period. However, for mass screening of foundation plantings, nursery stock and elite germplasm, woody indexing is too costly and slow to give results. The use of ELISA can overcome these limitations but the timing of sample collection is critical. Petal or leaf tissue collected early in the season will yield the most reliable results.

scholarly journals Strawberry latent ringspot virus Infecting Roses in India

Plant Disease ◽  
2004 ◽  
Vol 88 (1) ◽  
pp. 86-86 ◽  
Author(s):  
S. Kulshrestha ◽  
V. Hallan ◽  
G. Raikhy ◽  
R. Ram ◽  
A. A. Zaidi
Keyword(s):  
Plant Viruses ◽  
Ringspot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Specific Primers ◽  
Chenopodium Amaranticolor ◽  
Elisa Kit ◽  
Local Lesions ◽  
Young Leaves ◽  
Polymerase Chain

Rose is an economically important crop of India and the world. A survey of rose plantations in and near the Kangra Valley of Himachal Pradesh, India, showed virus-like symptoms, including yellow flecking in young leaves and reduction in leaflet size, while some were symptomless. These symptoms are similar to those for Strawberry latent ringspot virus (SLRSV) (1). Sap inoculation from symptomatic and some symptomless leaves to Chenopodium amaranticolor resulted in chlorotic local lesions followed by systemic chlorosis. SLRSV was detected in this indicator host and six rose cultivars (Happiness, Iceberg, First Prize, Ganga, Pink Panther, and Oklahoma) showing characteristic symptoms of SLRSV using enzyme-linked immunosorbent assay (ELISA) with ELISA kit (DSMZ, Braunschweig, Germany). Reverse transcription-polymerase chain reaction was performed with SLRSV-specific primers (2), and a product of the expected size of ˜181 bp was amplified. The authenticity of the fragment was confirmed by sequencing. Isolated SLRSV was also inoculated to seed-grown rose seedlings and after 20 days postinoculation the same symptoms (yellow flecking in young leaves) were observed. These results established the identity of the virus that caused yellow flecking on rose leaves in India as SLRSV. To our knowledge, this is the first report of SLRSV infecting rose in India. References: (1) A. F. Murant. Strawberry latent ringspot virus. No. 126 in: Description of Plant Viruses, CMI/AAB, Surrey, U.K., 1974. (2) E. Bertolini et al. J. Virol. Methods 96:33, 2001.

scholarly journals Detection of Tomato Ringspot Virus in Nectarine Trees Using ELISA and Transcribed RNA Probes

HortScience ◽  
1991 ◽  
Vol 26 (10) ◽  
pp. 1290-1292 ◽  
Author(s):  
C.A. Powell ◽  
A. Hadidi ◽  
J.M. Halbrendt
Keyword(s):  
Nucleic Acid ◽  
Prunus Persica ◽  
Leaf Tissue ◽  
Ringspot Virus ◽  
Nucleic Acid Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tomato Ringspot Virus ◽  
Dot Hybridization ◽  
Bark Tissue ◽  
Stem Pitting

The ability of 32P-labeled transcribed cRNA probes to detect tomato ringspot virus (TmRSV) RNA in nucleic acid extracts from roots, bark, and leaves of nectarine (Prunus persica [L.] Batsch) trees with the Prunus stem-pitting disease was assessed and compared with detection of TmRSV antigen by enzyme-linked immunosorbent assay (ELISA) in the same tissues. Neither TmRSV-specific nucleic acid nor antigen was detected in nectarine leaf tissue. ELISA detected TmRSV antigen in root extracts from 71% of the diseased trees, while dot hybridization detected virus-specific nucleic acid in 18% of the same samples. However, ELISA detected TmRSV antigen in only 47% of bark extracts; whereas TmRSV-specific nucleic acid was detected in 100% of the bark extracts from samples collected at or near the soil line. When nucleic acid extracts from bark were prepared from various locations on diseased trees and tested for TmRSV-specific nucleic acid by dot hybridization, there was an almost perfect correlation between the presence of stem-pitting symptoms and the detection of TmRSV nucleic acid. Detection of TmRSV RNA from the bark tissue of rootstock suckers from TmRSV-infected `Delicious'/MM.lO6 apple (Malus × domestica Borkh.) trees was unsuccessful using dot hybridization. The viral RNA, however, was usually detected in either leaf or root tissue of these same trees.

scholarly journals Occurrence of Prunus necrotic ringspot virus and Arabis mosaic virus on Rose in Iran

Plant Disease ◽  
2006 ◽  
Vol 90 (7) ◽  
pp. 975-975 ◽  
Author(s):  
F. Rakhshandehroo ◽  
H. R. Zamani Zadeh ◽  
A. Modarresi ◽  
S. Hajmansoor
Keyword(s):  
Mosaic Virus ◽  
French Bean ◽  
Ringspot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rosa Damascena ◽  
Natural Occurrence ◽  
Arabis Mosaic Virus ◽  
Local Lesions ◽  
Prunus Necrotic Ringspot Virus ◽  
Das Elisa

Rose is an economically important crop for Iran and the world. A survey was carried out from March 2005 to January 2006 to identify viruses infecting rose plants (Rosa × damascena, R. chinensis, R. canina, R. indica, and R. multiflora) in five plantations (Damavand, Tehran, Karaj, Shahre-Rey, and Varamin) in and near the Tehran Province of Iran. Samples (526) from eight rose-growing plantations were collected. All samples were tested for Prunus necrotic ringspot virus (PNRSV), Arabis mosaic virus (ArMV), and Cucumber mosaic virus (CMV) using the dot-immunobinding assay (1) and double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) according to the manufacturer's instructions (Bioreba, Basel, Switzerland). Among the samples tested, PNRSV and ArMV were found in 23.1 and 18.8% of the collection, respectively. No CMV was detected in any of the samples. The presence of ArMV and PNRSV was verified in samples by transmission to indicator test plants, cucumber (Cucumis sativus), French bean (Phaseolus vulgaris), and Cowpea (Vigna unguiculata). Inoculation with extracts from PNRSV-positive plants produced systemic mosaic, stunting, and vein banding on cucumber, and necrotic local lesions on cowpea. No symptoms were observed in French bean. Inoculation with extracts from ArMV-positive plants produced systemic vein banding on cucumber, chlorotic local lesions on French bean, and systemic mosaic on cowpea. These symptoms were similar to those that were described previously for these viruses (2,4). The symptoms observed on indicator plants for each virus corresponded to the results of DAS-ELISA. Examination of crude sap prepared from ArMV- and PNRSV-infected cucumber leaves using immu-nosorbent electron microscopy (IEM) revealed the presence of isometric virus particles with a diameter of approximately 30 and 25 nm, respectively. Frequencies of occurrence of these two viruses as determined by serological detection showed ArMV to be the most prevalent virus in high altitudes (1,700 to 1,900 m above sea level) compared with the lowland regions. Serological tests also indicate that PNRSV is mostly distributed through the red rose varieties (Rosa × damascena, R. chinensis, R. canina, and R. multiflora) and ArMV is within the white varieties (R. canina, R. indica, and R. multiflora). However, mixed infections of PNRSV and ArMV were detected in all rose samples tested. An infection by PNRSV and ArMV either singly or in combination is usually responsible for rose mosaic disease. PNRSV has been isolated in many rose-growing regions worldwide. ArMV alone or in complexes with ilarviruses infect garden and greenhouse rose in Europe and India (3). Mosaic is probably the most commonly found virus on roses. To our knowledge, this is the first report of a natural occurrence of ArMV and PNRSV on rose in Iran. References: (1) E. E. Banttari and P. H. Goodwin. Plant Dis. 69:202, 1985. (2) M. Boulila and M. Marrakchi. Phytopathol. Mediterr. 40:125, 2001. (3) S. Kulshrestha et al. Curr. Sci. 89:1759, 2005. (4) N. Salem et al. Plant Pathol. 86:85, 2004.

scholarly journals Genetic Diversity of Prunus necrotic ringspot virus Isolates Within a Cherry Orchard in New York

Plant Disease ◽  
2009 ◽  
Vol 93 (6) ◽  
pp. 599-606 ◽  
Author(s):  
J. E. Oliver ◽  
J. Freer ◽  
R. L. Andersen ◽  
K. D. Cox ◽  
T. L. Robinson ◽  
...  
Keyword(s):  
New York ◽  
Prunus Avium ◽  
Phylogenetic Analyses ◽  
Ringspot Virus ◽  
Spatial Proximity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Leaf ◽  
Cp Gene ◽  
Prunus Necrotic Ringspot Virus ◽  
Cherry Trees

A survey for Prunus necrotic ringspot virus (PNRSV) in an orchard of Prunus cerasus cv. Montmorency and Prunus avium cv. Hedelfingen in New York by enzyme-linked immunosorbent assay indicated an eightfold higher infection rate in sour cherry (33%, 32 of 96) than in sweet cherry (4%, 6 of 136) trees. The presence of PNRSV was confirmed by reverse transcription-polymerase chain reaction and amplification of the coat protein (CP) gene in total RNA from infected leaf tissue. Latent infection was prevalent in the majority of trees infected (87%, 33 of 38), while a few of them exhibited shock symptoms or had severely reduced growth (13%, 5 of 38). Asymptomatic PNRSV-infected trees clustered in spatial proximity to symptomatic trees. Sequence analysis of the CP gene (675 bp) indicated a population structure consisting of one predominant molecular variant for 10 isolates and six minor molecular variants for seven isolates. A high sequence identity was found between the CP gene of PNRSV isolates from cherry trees and other isolates from diverse hosts and various geographic origins at the nucleotide and amino acid levels (88 to 100%). Phylogenetic analyses showed a clustering of PNRSV isolates from cherry trees in New York in the predominant group PV-96.

Viruses infecting hop, Humulus lupulus, in Australia

1987 ◽  
Vol 38 (1) ◽  
pp. 83 ◽  
Author(s):  
D Munro
Keyword(s):  
Heat Treatment ◽  
Mosaic Virus ◽  
Immunosorbent Assay ◽  
Latent Virus ◽  
Ringspot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Meristem Culture ◽  
Arabis Mosaic Virus ◽  
Foliar Symptoms ◽  
Prunus Necrotic Ringspot Virus

In 1981 a survey of the main commercial hop cultivar in Australia, Pride of Ringwood, showed that foliar symptoms were associated with the presence of carlavirus particles. The incidence of plants containing particles varied between fields from 0 to 58%. In 1982 a survey based on enzyme-linked immunosorbent assay showed that the carlaviruses were hop mosaic and hop latent viruses and that they infected 1-68% and 1-58% of plants respectively. Prunus necrotic ringspot virus infected 4-9% of plants. Arabis mosaic virus was detected in a cultivar of English origin and American hop latent virus in cultivars of recent American origin, but neither of these viruses was detected in Australian-bred hops. Virus-free plants suitable for propagation were located in cultivars Pride of Ringwood and B23, and were produced by heat treatment and meristem culture of cultivar Southern Cross.

scholarly journals Fluctuations of Prunus Necrotic Ringspot Virus (PNRSV) at Various Phenological Stages in Peach Cultivars

Plant Disease ◽  
1999 ◽  
Vol 83 (11) ◽  
pp. 1055-1057 ◽  
Author(s):  
A. Dal Zotto ◽  
S. F. Nome ◽  
J. A. Di Rienzo ◽  
D. M. Docampo
Keyword(s):  
Ringspot Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Steady Increase ◽  
Virus Concentration ◽  
Mature Leaves ◽  
Low Profile ◽  
Yearly Average ◽  
Prunus Necrotic Ringspot Virus ◽  
The Difference ◽  
Peach Trees

Fluctuations in Prunus necrotic ringspot virus (PNRSV) concentration were researched in single plants of six peach (Prunus persicae) cultivars—Kurakata, Red Haven, Nectar Red, Start Delicious, Meadowlark, and Loadel—by double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) of dormant buds (May, June), flowers (September), new sprouts (November), and mature leaves (January) (Southern Hemisphere). The optimum extract dilution (sample weight per buffer volume) to detect the virus was also quantified. The average absorbance patterns of the six cultivars show a steady increase in virus concentration, ranging from A 405nm 0.61 in May to A 405nm 0.86 in July for dormant buds, to A 405nm 1.22 in September in flowers, to 1.53 in November in new sprouts, where the highest concentration was found. Virus concentrations in mature leaves drop to values similar to those of noninfected plants in January ( A405nm 0.12). The yearly average (six noninfected peach trees) ranged from A405nm 0.04 to A405nm 0.08. This drop coincides with an increase in summer temperature and attenuates foliation symptoms caused by PNRSV. Analysis of dormants buds, flowers, or new sprouts with 5-cm-long leaves was reliable to differentiate infected from noninfected plants. Cluster analysis of absorbance profiles for single plants of cvs. Loadel and Meadowlark, however, showed a comparatively low profile, with a drop at flowering time (A405nm 0.20 in September) close to the average of healthy controls. The difference between infected and healthy plants did not become apparent in all cultivars from the analysis of plants at a given phenological stage, for example by the analysis of flower only, the material most preferred to diagnose the virus. Therefore, plants should be analyzed during flowering and sprouting or flowering and dormancy (dormant buds).

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