Improved detection of prunus necrotic ringspot virus by the polymerase chain reaction

1996 ◽  
Vol 102 (7) ◽  
pp. 681-685 ◽  
Author(s):  
S. Spiegel ◽  
S. W. Scott ◽  
V. Bowman-Vance ◽  
Y. Tam ◽  
N. N. Galiakparov ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Ringspot Virus ◽  
Chain Reaction ◽  
Prunus Necrotic Ringspot Virus ◽  
Polymerase Chain

scholarly journals Blueberry red ringspot virus eliminated from highbush blueberry by shoot tip culture

2014 ◽  
Vol 50 (No. 4) ◽  
pp. 174-178
Author(s):  
J. Špak ◽  
D. Pavingerová ◽  
J. Přibylová ◽  
V. Špaková ◽  
F. Paprštein ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Woody Plant ◽  
Shoot Tips ◽  
Ringspot Virus ◽  
Highbush Blueberry ◽  
Shoot Tip Culture ◽  
Chain Reaction ◽  
Rooting Medium ◽  
Woody Plant Medium ◽  
Polymerase Chain

In order to find an effective elimination method of Blueberry red ringspot virus (BRRV), shoot tips of 2–3 mm size originating from buds of highbush blueberry cv. Darrow naturally infected by BRRV and collected in winter months were regenerated and multiplied on woody-plant medium (WPM) with 1.5 mg/l zeatin and 20 g/l sucrose. Developed shoots tested negative by polymerase chain reaction (PCR) for BRRV were transferred onto a rooting medium consisting of WPM supplemented with 1 mg/l indole-3-butyric acid. Rooting plants were transferred into soil and tested twice by PCR 6 months apart. Ninety per cent eradication of BRRV was achieved while isolating shoot tips, multiplying them in one vegetative generation, and regenerating whole 50–60 cm high plants within 30 months. All plants developed rich fruits after their overwintering in a cold greenhouse and were tested BRRV negative in summer 2013.  

scholarly journals Molecular Identification, Reverse Transcription-Polymerase Chain Reaction Detection, Host Reactions, and Specific Cytopathology of Artichoke yellow ringspot virus Infecting Onion Crops

2006 ◽  
Vol 96 (6) ◽  
pp. 622-629 ◽  
Author(s):  
Varvara I. Maliogka ◽  
Chrisostomos I. Dovas ◽  
Dietrich E. Lesemann ◽  
Stephan Winter ◽  
Nikolaos I. Katis
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Transmission Rate ◽  
Seed Transmission ◽  
Ringspot Virus ◽  
Chain Reaction ◽  
Degenerate Primers ◽  
Cytoplasmic Inclusions ◽  
The Family ◽  
Polymerase Chain

An isometric virus ca. 25 nm in diameter with angular contour was isolated from onion plants showing yellow leaf striping and necrotic tips. The virus was mechanically transmitted onto 28 species of indicator plants belonging to five families, viz. Amaranthaceae, Chenopodiaceae, Cucurbitaceae, Leguminosae, and Solanaceae where it causes ring spots, malformations, and/or tip necrosis. Cytopathological studies in infected Nicotiana benthamiana tissues revealed cytoplasmic inclusions resembling those caused by Artichoke yellow ringspot virus (AYRSV), a member of the family Comoviridae. Host range and symptomatology of the onion virus were also similar to AYRSV. A high seed transmission rate (20%) was found in onion. Reverse transcription-polymerase chain reaction using degenerate primers specific for the family Comoviridae allowed amplification of RNA-dependent RNA polymerase sequences, which upon sequence analysis and comparison with AYRSV isolates from Cynara scolymus (AYRSV-AtG) and Vicia faba (AYRSV-F) were highly similar, thus providing evidence that the nepovirus AYRSV is infecting onion in the field.

scholarly journals Partial characterization of Prunus Necrotic Ringspot Virus on apple in EGYPT 

2021 ◽  
Author(s):  
Aly M. Abdel-Salam ◽  
Samah A. Mokbel
Keyword(s):  
Binding Assay ◽  
Ringspot Virus ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Prunus Necrotic Ringspot Virus ◽  
Polymerase Chain ◽  
Severe Isolate ◽  
Microscopy Examination

Abstract A severe isolate of Prunus necrotic ringspot virus (PNRSV) was isolated from apple orchards in the vicinity of Nubaria city, Beheira governorate, Egypt. Infected-apple trees showed chlorotic, necrotic ringspots, and shoot holes on leaves. Severely infected- trees withered, became useless, and were removed causing severe economic losses. Reverse transcriptase (RT) polymerase chain reaction (PCR), RT-PCR, using degenerate primer pair for the coat protein (CP) gene of Ilarvirus amplified products similar to those produced from peach and apricot isolates of PNRSV-infecting stone fruits). Dot blotting immuno-binding assay (DBIA showed a positive reaction between PNRSV-infected apple sap and an Egyptian antiserum for PNRSV. Purified preparation from infected leaves, using the electro-elution technique yielded nucleoprotein, which had Amax and Amin at 260 and 240 nm respectively. Electron microscopy examination showed spherical virions with ca. 26 nm in diameter.

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

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