A Study of the Distribution of Apple stem pitting virus in Tissues of Pear Tree Using In Situ Hybridization and In Situ RT-PCR

2009 ◽  
Vol 8 (11) ◽  
pp. 1351-1359 ◽  
Author(s):  
Ying ZHAO ◽  
Na LIU ◽  
Jian-xin NIU
Keyword(s):  
In Situ Hybridization ◽  
Rt Pcr ◽  
Pear Tree ◽  
Apple Stem ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

scholarly journals The application of RT-PCR assay for the detection of Apple stem pitting virus and Apple stem grooving virus in four apple cultivars

2012 ◽  
Vol 38 (No. 1) ◽  
pp. 13-17 ◽  
Author(s):  
J.K. Kundu
Keyword(s):  
Mixed Infection ◽  
Rt Pcr ◽  
Pome Fruit ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Apple Cultivars ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

The reverse transcription polymerace chain reaction (RT-PCR) assay was successfully used for the detection of Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) in four apple cultivars of a 25 years old orchard. These two main pome fruit viruses were detected frequently in all tested apple cultivars. ASGV and ASPV occurred in as many as 16 trees (in the cultivar Spartan) and 13 trees (in the cultivar Idared) out of 20 tested trees, respectively. Mixed infection by ASGV and ASPV was found in all tested cultivars (as many as 9 out of 20 tested trees of the cultivar Spartan).

Evaluation of several RT-PCR primer pairs for the detection of Apple stem pitting virus

2010 ◽  
Vol 168 (1-2) ◽  
pp. 242-247 ◽  
Author(s):  
B. Komorowska ◽  
T. Malinowski ◽  
L. Michalczuk
Keyword(s):  
Rt Pcr ◽  
Apple Stem ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting ◽  
Pcr Primer

scholarly journals Identidade do Apple stem pitting virus em São Paulo, Brasil, confirmada por meio de RT-PCR

2004 ◽  
Vol 29 (6) ◽  
pp. 691-691
Author(s):  
Juliana Freitas-Astua ◽  
Quelmo S. de Novaes ◽  
Juarez A. Betti ◽  
Jorge A. M. Rezende ◽  
Hugo Kuniyuki ◽  
...  
Keyword(s):  
Sao Paulo ◽  
São Paulo ◽  
Rt Pcr ◽  
Apple Stem ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

scholarly journals Loquat (Eriobotrya japonica) Is a New Natural Host of Apple Stem Pitting Virus

Plants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1560
Author(s):  
Félix Morán ◽  
Celia Canales ◽  
Antonio Olmos ◽  
Ana Ruiz-García
Keyword(s):  
Sequence Analysis ◽  
Natural Host ◽  
Eriobotrya Japonica ◽  
Phylogenetic Study ◽  
Rt Pcr ◽  
Apple Stem ◽  
Close Phylogenetic Relationship ◽  
Apple Stem Pitting Virus ◽  
Woody Crop ◽  
Stem Pitting

Loquat (Eriobotrya japonica) is a minor but important woody crop cultivated in Asia and Europe. High-throughput sequencing (HTS) analysis of an asymptomatic loquat plant using RNAseq Illumina technology has allowed the detection for the first time of apple stem pitting virus (ASPV), the type species of the genus Foveavirus in the family Betaflexiviridae, infecting this crop. A nearly complete genome of 9303 nts (ASPV-SL61) reconstructed bioinformatically shows the typical genomic structure of this viral species and a highest nucleotide identity (85.9%) with the Chinese ASPV isolate YLX from pear. A close phylogenetic relationship between ASPV-SL61 and ASPV-YLX has been confirmed by the sequence analysis of full-length ASPV genomic sequences available in the databases. In fact, a phylogenetic study based on a partial CP N-terminal sequence previously proposed to be involved in host adaptation has shown that ASPV-SL61 loquat isolate is more closely related to ASPV pear isolates. The presence of ASPV in loquat has been further confirmed by RT-PCR and Sanger sequencing and DAS-ELISA. An incidence of 15% was determined in one of the loquat Spanish growing areas. The sequence analysis of the partial CP sequences amplified by RT-PCR has shown a high level of variability between loquat isolates. To our knowledge, this is the first record of loquat as a natural host of ASPV.

scholarly journals Diagnóstico biológico e molecular e análise da seqüência de nucleotídeos do gene da proteína capsidial de um isolado do Apple stem pitting virus

2006 ◽  
Vol 31 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Paula Radaelli ◽  
Osmar Nickel ◽  
Jurema Schons ◽  
Francisco J.L. Aragão ◽  
Thor V. M. Fajardo
Keyword(s):  
Pyrus Communis ◽  
Rt Pcr ◽  
Apple Stem ◽  
Pyrus Communis L ◽  
Apple Stem Pitting Virus ◽  
Dot Elisa ◽  
Stem Pitting

O Apple stem pitting virus (ASPV) foi detectado por RT-PCR em amostras de cultivares de pereiras européias (Pyrus communis L.) cvs. Starkrimson e Abate Fetel, e asiáticas (P. pyrifolia var. culta) cvs. Kousui e Housui coletadas no início do outono de 2003 em pomar da Estação Experimental da Embrapa Uva e Vinho, Vacaria, RS. Utilizando várias combinações de oligonucleotídeos, foram amplificados fragmentos de DNA de 269 e 1554 pb, este último contendo o gene completo (1131 nt) da proteína capsidial do ASPV. Outro fragmento amplificado de 291 pb compreende parte do gene da polimerase viral. Estes fragmentos constituem-se em um excelente instrumento de diagnóstico do ASPV em pereiras. A comparação das seqüências de nucleotídeos do gene da proteína capsidial do ASPV com seqüências do banco de dados GenBank, revelou identidades de 89% com seqüências de um isolado alemão de macieira e de 85 a 88% com isolados poloneses, de pereiras. A indicadora herbácea Nicotiana occidentalis cv. 37B, inoculada mecanicamente com extrato foliar da cv. Housui, apresentou lesões locais necróticas, necrose foliar marginal e das nervuras. O ASPV também foi detectado por dot-ELISA nas cvs. Abate Fetel e Kousui, na cv. Starkrimson por imunoblot, e em Pyronia veitchii (Trabut) Guill. por enxertia de borbulhas da cv. Abate Fetel infetada.

scholarly journals The occurrence of Apple stem pitting virus and Apple stem grooving virus within field-grown apple cultivars evaluated by RT-PCR

2011 ◽  
Vol 39 (No. 3) ◽  
pp. 88-92 ◽  
Author(s):  
J.K. Kundu
Keyword(s):  
Polymerase Chain Reaction ◽  
Mixed Infection ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Apple Cultivars ◽  
Polymerase Chain ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

The reverse transcription polymerase chain reaction (RT-PCR) was successfully used to determine the occurrence of Apple stem pitting virus (ASPV) and Apple stem grooving virus (ASGV) in field-grown apple cultivars. Both viruses were detected frequently in all 16 tested apple cultivars. As many as 27.86% ASPV-infected and 44% ASGV-infected trees were recorded among a total of 420 tested trees from 15 different orchards. Mixed infection with ASGV and ASPV was recorded in 16.7% of the trees.  

scholarly journals Rapid and Sensitive Detection of Apple stem pitting virus in Apple Trees Through RNA Amplification and Probing with Fluorescent Molecular Beacons

2001 ◽  
Vol 91 (11) ◽  
pp. 1085-1091 ◽  
Author(s):  
M. M. Klerks ◽  
G. Leone ◽  
J. L. Lindner ◽  
C. D. Schoen ◽  
J. F. J. M. van den Heuvel
Keyword(s):  
Field Testing ◽  
Molecular Beacons ◽  
Fluorescent Detection ◽  
Rna Amplification ◽  
Apple Trees ◽  
Apple Stem ◽  
Bark Tissue ◽  
Specific Amplification ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

Currently, detection of Apple stem pitting virus (ASPV; genus Foveavirus) in apple trees for certification purposes occurs by woody indexing. This method requires a minimum of 12 to 24 weeks in greenhouse testing to up to 2 years in field testing. In this paper, the development of a single tube AmpliDet RNA system for the rapid gel-free detection of ASPV in apple tree tissues is described. The system relies on the specific amplification of the viral RNA by nucleic acid sequence-based amplification and the simultaneous fluorescent detection of the amplification product through molecular beacons. A sensitivity of a minimum of 100 molecules of transcript RNA was obtained by the ASPV-specific AmpliDet RNA. All biologically characterized ASPV isolates from a field trial and 12 of 14 isolates from a plant virus collection were readily detected with this AmpliDet RNA system. In addition, the efficiency of this method for detecting ASPV in ‘Golden Delicious’ and ‘Gravenstein’ apple trees was compared throughout the year with mechanical inoculation onto Nicotiana occidentalis 37B, a candidate indicator for ASPV. This revealed that only AmpliDet RNA consistently detected the virus in bark tissue, irrespective of the season. Season-specific tissues such as buds, petals, and fruits, but not leaves, also were reliable sources for detection of ASPV by the AmpliDet RNA system.

scholarly journals Quantitative real-time RT-PCR and chromogenic in situ hybridization: precise methods to detect HER-2 status in breast carcinoma

BMC Cancer ◽  
2009 ◽  
Vol 9 (1) ◽  
Author(s):  
Fabíola E Rosa ◽  
Sara M Silveira ◽  
Cássia GT Silveira ◽  
Nádia A Bérgamo ◽  
Francisco A Moraes Neto ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Breast Carcinoma ◽  
Real Time ◽  
Rt Pcr ◽  
Her 2 ◽  
Chromogenic In Situ Hybridization

Comet and cup genes in Drosophila spermatogenesis: the first demonstration of post-meiotic transcription

2008 ◽  
Vol 36 (3) ◽  
pp. 540-542 ◽  
Author(s):  
Carine Barreau ◽  
Elizabeth Benson ◽  
Helen White-Cooper
Keyword(s):  
In Situ Hybridization ◽  
Expression Pattern ◽  
Reverse Transcription ◽  
Protein Product ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Single Cyst ◽  
Reverse Transcription Pcr

Post-meiotic transcription is widespread in mammalian spermatogenesis, but is generally believed to be absent from Drosophila spermatogenesis. Genes required during meiosis, in early spermatids or later in spermiogenesis are typically transcribed in primary spermatocytes in Drosophila. Their mRNAs are then stored in the cytoplasm until the protein product is needed. Recently, using in situ hybridization, we identified 17 Drosophila genes, collectively named ‘comets’ and ‘cups’, whose mRNAs are most abundant in, and localize to the distal ends of, elongating spermatids. Using a single-cyst quantitative RT–PCR (reverse transcription–PCR) assay, we confirmed this unusual expression pattern and conclusively demonstrate the existence of post-meiotic transcription in Drosophila spermatids. We found that transcription of comets and cups occurs just before protamines can be detected in spermatid nuclei.

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