Optimization of tissue and time for rapid serological and molecular detection of Apple stem pitting virus and Apple stem grooving virus in apple’

2018 ◽  
Vol 46 (5) ◽  
pp. 705-713 ◽  
Author(s):  
Sajad Un Nabi ◽  
Javid Iqbal Mir ◽  
Om Chand Sharma ◽  
Desh Beer Singh ◽  
Shafia Zaffer ◽  
...  
Keyword(s):  
Molecular Detection ◽  
Apple Stem Grooving Virus ◽  
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).

scholarly journals Gospodarski važni virusi jezgričavog voća

2019 ◽  
Vol 22 (3-4) ◽  
pp. 123-136
Author(s):  
Dario Ivić
Keyword(s):  
Mosaic Virus ◽  
Leaf Spot ◽  
Spot Virus ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Chlorotic Leaf ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

Virusi jabuke, kruške ili dunje relativno su slabo poznati stručnjacima i voćarima. Najvažnijim virusima koji se javljaju na jezgričavim voćnim vrstama smatraju se virus mozaika jabuke (Apple mosaic virus, ApMV), virus klorotične pjegavosti lista jabuke (Apple chlorotic leaf spot virus, ACLSV), virus brazdavosti debla jabuke (Apple stem grooving virus, ASGV) i virus jamičavosti debla jabuke (Apple stem pitting virus,ASPV). U radu je ukratko opisana njihova važnost, biologija i regulativni status, kao i osnovne mjere zaštite.

scholarly journals First Report of Apple Top Working Disease Caused by Viruses (Apple stem grooving virus, Apple chlorotic leaf spot virus, and Apple stem pitting virus) in Apple in India

Plant Disease ◽  
2013 ◽  
Vol 97 (7) ◽  
pp. 1001-1001 ◽  
Author(s):  
A. Brakta ◽  
P. D. Thakur ◽  
A. Handa
Keyword(s):  
Leaf Spot ◽  
Spot Virus ◽  
Sequence Identity ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Commercial Cultivars ◽  
Chlorotic Leaf ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting ◽  
Das Elisa

Top working of apple (Malus domestica Borkh.) trees of old, unproductive, and less preferred cultivars with the newly introduced spur type commercial cultivars has become a common practice with many growers in the northwestern Himalayan region of India. Typical viral symptoms of curling, puckering, and necrosis on leaves were observed with an incidence of 80% on Red Chief, Super Chief, Scarlet Spur, Schillet Spur, Washington Red Delicious, and many other newly introduced cultivars during surveys conducted in May and June 2009. Leaf samples from top worked trees were tested for the presence of Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), and Apple stem pitting virus (ASPV) by employing biological detection (herbaceous and woody indicators), double antibody sandwich (DAS)-ELISA), and reverse transcriptase (RT)-PCR based detection. Mechanical transmission to herbaceous indicators produced chlorotic lesions on Chenopodium quinoa and C. amaranticolor, whereas marginal necrosis was induced on Phaseolous vulgaris within 9 to 21 days after sap inoculations. All three viruses, i.e., ASGV, ASPV, and ACLSV, were detected from these herbaceous indicators in DAS-ELISA (BIOREBA AG, Switzerland). Furthermore, symptoms similar to those observed in orchards were produced when the test budwood was inoculated onto the woody indicator (M. pumila ‘Spy 227’) plant by double grafting, grafting cum budding, and double budding methods within time periods ranging from 4 months in double grafting, 5 months in double budding, to 1 year 4 months in the grafting cum budding method. The presence of all three viruses was confirmed by DAS-ELISA again in Spy 227 woody indicator. PCR detection was carried out by using the coat protein gene specific primers (ASGV5641 [forward], ASGV6396 [reverse]; ACLSV6784 [forward], ACLSV7365 [reverse] [2]; ASP-C [sense], ASP-A [anti-sense] [1]) of all the viruses detected through ELISA. The amplified products were cloned, sequenced, and deposited in NCBI (GenBank Accessions KC110892 for ASGV, KC154859 for ASPV, and KC154862 for ACLSV). BLASTn analysis showed the ASGV isolate had 97 to 98% sequence identity with Indian (FM204881) and Brazilian (AF438409) ASGV isolates. The ASPV and ACLSV isolates had 98% and 99% sequence identity with Chinese (JF895517) and Japanese (AB326230) isolates, respectively. To the best of our knowledge, this is the first report of apple top working disease associated with ASGV, ASPV, and ACLSV infection in commercial cultivars of apple from India and seems to be a serious threat for growing virus-free healthy stocks in orchards. Top working disease in apple associated with ASGV, ASPV, and ACLSV viruses has been reported from Japan (3,4). References: (1) J. K. Kundu et al. Plant Prot. Sci. 39:88, 2003. (2) O. Nickel et al. Fitopatol. Brasil. 26:655, 2001. (3) H. Yanase. Bull. Fruit Tree Res. Stn., Japan Ser. C 1:47, 1974. (4) H. Yanase et al. Acta Hortic. 44:221, 1975.

Droplet-vitrification cryotherapy for eradication of apple stem grooving virus and apple stem pitting virus from “Marubakaido” apple rootstock

2020 ◽  
Vol 45 (2) ◽  
pp. 148-152 ◽  
Author(s):  
Juliana A. Souza ◽  
Amauri Bogo ◽  
Jean C. Bettoni ◽  
Murilo Dalla Costa ◽  
Fabio N. da Silva ◽  
...  
Keyword(s):  
Apple Rootstock ◽  
Droplet Vitrification ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

scholarly journals Simultaneous detection of four viruses affecting apple and pear by molecular hybridization using a polyprobe

2014 ◽  
Vol 44 (10) ◽  
pp. 1711-1714 ◽  
Author(s):  
Thor Vinícius Martins Fajardo ◽  
Osmar Nickel
Keyword(s):  
Leaf Spot ◽  
Simultaneous Detection ◽  
Hybridization Method ◽  
Spot Virus ◽  
Apple Stem Grooving Virus ◽  
Apple Stem ◽  
Chlorotic Leaf ◽  
Viral Cdna ◽  
Apple Stem Pitting Virus ◽  
Stem Pitting

The viruses Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus (ACLSV), Apple stem pitting virus (ASPV) and Apple mosaic virus (ApMV) are common in apples and pears and main targets of detection in propagation materials. This study aimed at demonstrating the usefulness of the hybridization method with a non-radioactive probe for simultaneous detection of these four viruses. The sensitivity of this method was sufficiently high enabling the detection of ASGV, ACLSV, ASPV and ApMV in total RNA extracted from infected samples. The probe specificity was confirmed by reaction with homologous viral cDNA, individually cloned for each virus.

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.

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