scholarly journals Preliminary Results on Behavior of Transgenic and Conventional Plum to Natural PPV Infection in a New Field Trial

2016 ◽  
Vol 73 (2) ◽  
pp. 274
Author(s):  
Ioan Zagrai ◽  
Luminita Zagrai ◽  
Angela Festila
Keyword(s):  
Field Trial ◽  
Visual Observation ◽  
Capsid Gene ◽  
Plum Pox Virus ◽  
Experimental Plot ◽  
Rt Pcr ◽  
Inoculum Source ◽  
Preliminary Results ◽  
Das Elisa ◽  
Durability Of Resistance

To assess the durability of resistance to PPV of HoneySweet transgenic plum harbouring plum pox virus (PPV) capsid gene, a new field trial was recently established in Romania at Fruit Research & Development Station Bistrita. Stanley and Reine Claude d’Althan cvs. are used as control. PPV inoculum source is secured within the experimental plot. PPV monitoring was made by visual observation of symptoms development on leaves and by serological (DAS-ELISA) and molecular (IC/-RT-PCR) testings. Three years after planting PPV was observed and detected only on Stanley. 'HoneySweet' and  Reine Claude d’Althan cultivars remained PPV free.

scholarly journals Interactions of Plum pox virus strain Rec with Apple chlorotic leafspot virus and Prune dwarf viruses in field-grown transgenic plum Prunus domestica L., clone C5

2008 ◽  
Vol 44 (No. 1) ◽  
pp. 1-5 ◽  
Author(s):  
J. Polák ◽  
M. Ravelonandro ◽  
J. Kumar-Kundu ◽  
J. Pívalová ◽  
R. Scorza
Keyword(s):  
Sequence Analysis ◽  
Transgenic Plants ◽  
Open Field ◽  
Recombinant Strain ◽  
Dwarf Virus ◽  
Plum Pox Virus ◽  
Prunus Domestica ◽  
Rt Pcr ◽  
Antagonistic Effects ◽  
Das Elisa

Transgenic plums, <I>Prunus domestica</I> L. clone C5, were inoculated by bud grafting with <I>Plum pox virus</I> (PPV-Rec, recombinant strain originated from plum), PPV-Rec + <I>Apple chlorotic leafspot virus</I> (ACLSV), PPV-Rec + <I>Prune dwarf virus</I> (PDV), and PPV-Rec + ACLSV + PDV. Non-inoculated transgenic plums served as controls. Plants were grown in an open field for 5 years. They were evaluated by visible symptoms, by DAS-ELISA and RT-PCR. Mild PPV symptoms, diffuse spots or rings appeared two years after inoculation in some leaves of plants artificially inoculated with PPV-Rec, PPV-Rec + ACLSV, PPV-Rec + PDV, and PPV-Rec + ACLSV + PDV. Severe PPV symptoms appeared in leaves of shoots growing from infected buds used for inoculation. During the following three years, further weakening of PPV symptoms was observed in transgenic plants. In 2007, very mild PPV symptoms were found in only a few leaves, and over 60%, resp. 70% of the C5 trees showed no PPV symptoms. The presence of PPV was confirmed by ELISA, ISEM and RT-PCR. No difference in PPV symptoms was observed between PPV-Rec and combinations PPV-Rec + ACLSV, PPV-Rec + PDV, PPV-Rec + ACLSV + PDV. No symptoms of ACLSV appeared in combinations of ACLSV with PPV-Rec and PPV-Rec + PDV during 2004–2007, but the presence of ACLSV in leaves of transgenic plants clone C5 was proved by ELISA and RT-PCR. Neither synergistic nor antagonistic effects of ACLSV on PPV-Rec were observed. No symptoms of PDV appeared in combinations of viruses with PDV during 2004–2007. PDV was not detected by ELISA, and the presence of PDV was uncertain by RT-PCR in most of inoculated trees in 2006 and 2007. The results of RT-PCR will be further confirmed by sequence analysis and discussed. These results suggest a possible antagonistic interaction between PPV-Rec and PDV in plum clone C5.

scholarly journals Occurrence of Stone Fruit Viruses in Plum Orchards in Latvia

2013 ◽  
Vol 67 (2) ◽  
pp. 116-123
Author(s):  
Alina Gospodaryk ◽  
Inga Moročko-Bičevska ◽  
Neda Pūpola ◽  
Anna Kāle
Keyword(s):  
Mosaic Virus ◽  
Large Scale ◽  
Ringspot Virus ◽  
Stone Fruit ◽  
Detection Methods ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Detection Rates ◽  
Arabis Mosaic Virus ◽  
Das Elisa

To evaluate the occurrence of nine viruses infecting Prunus a large-scale survey and sampling in Latvian plum orchards was carried out. Occurrence of Apple mosaic virus (ApMV), Prune dwarf virus (PDV), Prunus necrotic ringspot virus (PNRSV), Apple chlorotic leaf spot virus (ACLSV), and Plum pox virus (PPV) was investigated by RT-PCR and DAS ELISA detection methods. The detection rates of both methods were compared. Screening of occurrence of Strawberry latent ringspot virus (SLRSV), Arabis mosaic virus (ArMV), Tomato ringspot virus (ToRSV) and Petunia asteroid mosaic virus (PeAMV) was performed by DAS-ELISA. In total, 38% of the tested trees by RT-PCR were infected at least with one of the analysed viruses. Among those 30.7% were infected with PNRSV and 16.4% with PDV, while ApMV, ACLSV and PPV were detected in few samples. The most widespread mixed infection was the combination of PDV+PNRSV. Observed symptoms characteristic for PPV were confirmed with RT-PCR and D strain was detected. Comparative analyses showed that detection rates by RT-PCR and DAS ELISA in plums depended on the particular virus tested. The results obtained in this study revealed that commonly grown plum cultivars in Latvia are infected with economically important stone fruit viruses and highlight the need to implement a programme to produce and propagate virus-free planting material.

scholarly journals First Report of Sharka Disease Caused by Plum Pox Virus in Lithuania

Plant Disease ◽  
1998 ◽  
Vol 82 (12) ◽  
pp. 1405-1405 ◽  
Author(s):  
J. Staniulis ◽  
J. Stankiene ◽  
K. Sasnauskas ◽  
A. Dargeviciute
Keyword(s):  
Coat Protein ◽  
Plant Protection ◽  
Virus Disease ◽  
Pcr Amplification ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
First Report ◽  
Das Elisa ◽  
Sharka Disease

Plum pox (sharka) disease caused by plum pox potyvirus (PPV) is considered the most important virus disease of stone fruit trees in Europe and the Mediterranean region. Nearly all those countries that produce stone fruits are affected (3). The causal virus of the disease is a European Plant Protection Organization A2 quarantine pathogen. Symptoms of leaf mottling, diffuse chlorotic spots, rings, and vein banding of varied intensity characteristic for plum pox virus infection were observed in the plum (Prunus domestica) orchard tree collection of the Lithuanian Institute of Horticulture in Babtai in 1996. Presence of this virus in the diseased trees was confirmed by double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) with kits from BIOREBA (Reinach, Switzerland) and by polyclonal antibodies raised against a Moldavian isolate of PPV courtesy of T. D. Verderevskaya (Institute of Horticulture, Kishinev, Moldova). ELISAs with both sources of antiserum were positive for presence of PPV. Electron microscopy revealed the presence of potyvirus-like particles averaging 770 nm in extracts of mechanically inoculated plants of Chenopodium foetidum (chlorotic LL [local lesions]) and Pisum sativum cvs. Rainiai and Citron (mottling). For molecular diagnosis and characterization of this isolate, PPV-971, reverse transcription-polymerase chain reaction (RT-PCR) was employed. Total RNA from the leaves of infected pea was isolated as described (2). High molecular weight RNA selectively precipitated with 2 M lithium chloride was used for RT-PCR amplification of the coat protein encoding sequence by use of specific primers complementary to 5′ and 3′ parts of PPV coat protein L1 (GenBank accession no. X81081). Amino acid sequence comparison with GenBank data indicated 98.2% similarity with coat protein of PPV potyvirus isolated by E. Mais et al. (accession no. X81083) and 97.3% with PPV strain Rankovic (1).The specific DNA fragment, corresponding to predicted coat protein sequence size, was cloned into Escherichia coli pUC57 for DNA sequencing. Expression of the cloned sequence in bacteria and yeast expression systems is under investigation. The presence of PPV in plum trees in the 9-year-old collection at Babtai was confirmed by DAS-ELISA in 1997 and again in 1998. PPV was then detected in 20% of symptomatic trees of three cultivars. The Lithuanian PPV isolate reacted positively with “universal” Mab.5b and with a Mab (Mab.4DG5) specific for PPV-D. No reaction was observed with Mabs specific for PPV-M (Mab.AL), PPV-C (Mab.AC and Mab.TUV), and PPV-El Amar (Mab.EA24). PPV-971 seems to be a typical member of the less aggressive Dideron strain cluster of PPV (D. Boscia, personal communication). This is the first report of PPV in Lithuania and confirms the necessity for continuing the precautionary measures established in this country for indexing of nursery plum trees used for graft propagation. References: (1) S. Lain et al. Virus Res. 13:157, 1989. (2) J. Logemann et al. Anal. Biochem. 163:16, 1987. (3) M. Nemeth. OEPP/EPPO Bull. 24:525, 1994.

scholarly journals First Report of Prunus necrotic ringspot virus in Peach in Mexico

Plant Disease ◽  
2008 ◽  
Vol 92 (3) ◽  
pp. 482-482 ◽  
Author(s):  
R. De La Torre-Almaraz ◽  
J. V. Montoya-Piña ◽  
S. Alcacio-Rangel ◽  
G. Camarena-Gutiérrez ◽  
M. Salazar-Segura
Keyword(s):  
Prunus Persica ◽  
Rna Extraction ◽  
Ringspot Virus ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
First Report ◽  
Field Samples ◽  
Prunus Necrotic Ringspot Virus ◽  
San Martín ◽  
Das Elisa

Peach (Prunus persica (L.) Batsch) is one of the most important fruit crops in the temperate regions of Mexico. In 2006, during a survey conducted in commercial peach orchards in Puebla, Mexico for viral diseases, many trees were observed with foliar symptoms that included yellow mottle, ringspot, line patterns, and mosaic. Samples (flowers, young shoot tips, and leaves) were collected from 120 symptomatic trees in three locations (San Martin Texmelucan, Domingo Arenas, and Tepetzala). All samples were tested using double-antibody sandwich (DAS)-ELISA kits (Agdia, Inc., Elkhart, IN) for the presence of the following viruses: Apple mosaic virus, Plum pox virus, Prune dwarf virus, and Prunus necrotic ringspot virus (PNRSV). Sap extracts from young symptomatic leaves and shoots were used to mechanically inoculate Chenopodium quinoa, C. amaranticolor, Gomphrena globosa, Nicotiana tabacum cv. Xanthi, N. glutinosa, N. clevelandii, N. benthamiana, Datura stramonium, Capsicum annuum, and Solanum lycopersicum. Plants were kept in a greenhouse with approximate temperatures of 25 to 35°C, humidity of 70%, and 12 h of light. Sap extracts were also used for dsRNA extraction and analyses (2) and RNA extraction for use in reverse transcription (RT)-PCR with the Access RT-PCR system (Promega, Madison, WI) and primers that annealed to a conserved region in the PNRSV coat protein gene (1). The expected size amplicons of approximately 450 bp were generated from all field-collected samples. The PCR products from three geographically distinct PNRSV isolates (Domingo Arenas [Accession No. DQ979004], Tepetzala [Accession No. DQ979005], and San Martin Texmelucan [Accession No. EF456771]) were directly sequenced with a Genetic Analyzer 3100 (Applied Biosystems, Foster City, CA) and their nucleotide and deduced amino acids sequences were more than 93% identical to corresponding sequences of PNRSV available in the NCBI/GenBank database. PNRSV was the only virus detected by DAS-ELISA in flowers and young shoots from 60 of the symptomatic field samples tested from the three locations. DsRNA banding patterns were obtained from 40 field-collected symptomatic samples; all showed three bands of approximately 3.6, 2.5, and 1.8 kb, the expected sizes for RNAs 1, 2, and 3 of PNRSV, respectively. DsRNAs were not detected in asymptomatic plants. PNRSV transmission by mechanical inoculation induced mosaic symptoms in N. tabacum cv. Xanthi and necrotic local lesions in G. globosa. Although G. globosa is reported to be a systemic host of PNRSV and N. tabacum is not reported to be a host, symptomatic plants were positive for PNRSV in DAS-ELISA tests. It is possible that there was an additional virus not detected in our assays that was responsible for the unexpected reactions in the host range studies. To our knowledge, this is the first report of PNRSV in peach in Mexico. References: (1) D. J. MacKenzie et al. Plant Dis. 81:222, 1997. (2) R. A. Valverde et al. Plant Dis. 74:255,1990.

scholarly journals Evaluation of Presence and Concentration of PPV in Rootstocks Derived from Prunus davidiana (Carr.) Franch

2019 ◽  
Vol 67 (1) ◽  
pp. 121-131
Author(s):  
Petra Pavelková ◽  
Tomáš Kiss ◽  
Tomáš Nečas
Keyword(s):  
Real Time ◽  
Growing Season ◽  
Elisa Test ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Serological Method ◽  
Low Concentrations ◽  
Symptom Intensity ◽  
Das Elisa ◽  
Prunus Davidiana

Evaluation of the presence and concentration of PPV (Plum pox virus) in selected rootstocks was carried out in 2016–2017. For the purpose of the experiment we used rootstocks derived from crossbreeding of Prunus davidiana (Carr.) Franch, such as Cadaman and Barrier, and also a P. davidiana seedling. Peach seedling rootstock GF‑305 was used as a control. The rootstocks were inoculated artificially with PPV strain M (Marcus). Both the rootstock and the inoculum were tested for presence of the virus by a serological method – semiquantitative DAS‑ELISA test and molecular methods – RT‑PCR, real‑time RT‑PCR and RT‑LAMP. During the growing season the plants were evaluated for symptom intensity by using a scoring scale. The results show interdependency between symptom intensity and the amount of PPV in leaves, with DAS‑ELISA test giving less positive samples than RT‑PCR. The RT‑LAMP and real‑time RT‑PCR methods were capable of revealing low concentrations of the virus even in symptom‑free plants. The lowest PPV concentrations of all the four rootstocks were detected by real‑time RT‑PCR in P. davidiana. The highest PPV concentrations were detected in Barrier rootstock. In inocula, the lowest concentration was found in the inocula on Cadaman rootstock, whereas the highest PPV concentration was detected in the inocula inoculated on Barrier rootstock.

scholarly journals Experimental evaluation of apricot genotypes for resistance to Plum pox virus

2000 ◽  
Vol 36 (No. 4) ◽  
pp. 123-127 ◽  
Author(s):  
M. Glasa ◽  
D. Benediková ◽  
t.. Glasová ◽  
I. Hričovský ◽  
O. Kůdela
Keyword(s):  
Experimental Evaluation ◽  
Visual Inspection ◽  
Artificial Inoculation ◽  
First Year ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
Pcr Assay ◽  
The Third ◽  
Breeding Station ◽  
Das Elisa

The reaction of 19 Slovak apricot cultivars and hybrids (breeding program of Research Breeding Station at Vesele) to infection by M isolates of plum pox virus (PPV-M) was evaluated. The genotypes were inoculated by grafting to naturally infected plum trees in the field and by chip-budding in the glasshouse. Monitoring of PPV infection was done over a 3 year period by visual inspection and DAS-ELISA. In the third year of evaluation the RT-PCR assay was also applied. The tested apricot genotypes differed in their reaction to PPV infection. Most of them developed mild or severe symptoms on leaves in the first year and/or next two consecutive years after artificial inoculation. The four Slovak apricot genotypes Veharda, Vemina, VS 15811 and VS 157/8 were resistant to PPV-M infection.

scholarly journals Gene flow was not detected from a field trial of transgenic plum cv. HoneySweet – Short Communication

2019 ◽  
Vol 55 (No. 2) ◽  
pp. 90-92
Author(s):  
Petr Komínek ◽  
Jaroslav Polák ◽  
Marcela Komínková ◽  
Ralph Scorza
Keyword(s):  
Czech Republic ◽  
Gene Flow ◽  
Field Test ◽  
Field Trial ◽  
Short Communication ◽  
Test Site ◽  
Plum Pox Virus ◽  
Experimental Plot ◽  
The Czech Republic ◽  
Crop Research

A field trial with plum cv. HoneySweet was established in 2003 in an experimental plot near Crop Research Institute in Prague-Ruzyně, Czech Republic, on the basis of a permission issued by the Ministry of the Environment of the Czech Republic. In addition to the evaluation of resistance of plum cv. HoneySweet to Plum pox virus, the field trial was used to evaluate gene flow of the inserted transgene. Sampling of blackthorn and myrobalan trees outside the field test site occurred at distances ranging from 544 m to 845 m from the test site and showed no gene flow, testing both plants and seeds collected from blackthorns and myrobalans. Similarly, seeds from plums cv. Jojo growing directly at the field test place did not show any presence of the transgene after seven years of evaluation. 

scholarly journals Distribution of Plum pox virus strains in natural sources in the Czech Republic

2009 ◽  
Vol 45 (No. 4) ◽  
pp. 144-147 ◽  
Author(s):  
J. Polák ◽  
P. Komínek
Keyword(s):  
Monoclonal Antibodies ◽  
Czech Republic ◽  
Sweet Cherry ◽  
Polyclonal Antibodies ◽  
Sour Cherry ◽  
Plum Pox Virus ◽  
Rt Pcr ◽  
The Czech Republic ◽  
Natural Sources ◽  
Das Elisa

The distribution of <i>Plum pox virus</i> (PPV) strains, PPV-D, PPV-M and PPV-Rec, was investigated in the Czech Republic in 2005–2008. Fifty-two to ninety-four samples of flowers or leaves of plum, myrobalan and blackthorn trees from different regions were tested in individual years. The presence of PPV was detected by DAS-ELISA with serotype-specific polyclonal antibodies. PPV-M was proved by DASI-ELISA with serotype-specific monoclonal antibodies; PPV-D, PPV-M and PPV-Rec were detected by RT-PCR in leaf samples from PPV infected trees. The presence of PPV-D ranged from 94.6% to 100%, the presence of PPV-M from 0.0% to 2.3% and the presence of PPV-Rec from 0.0% to 3.1% during 2005–2008. More than 95% of analysed samples of PPV were infected with PPV-D and less than 2.5% of analysed samples of PPV were infected with PPV-M or PPV-Rec. The presence of PPV-C was not proved in sweet cherry and sour cherry trees. The presence of PPV-EA was not proved in apricot trees.

scholarly journals Reaction of transgenic plum cv. HoneySweet to the Plum pox virus after a severe infection of Monilinia sp. – short communication

2018 ◽  
Vol 55 (No. 1) ◽  
pp. 8-10 ◽  
Author(s):  
Polák Jaroslav ◽  
Neubauerová Tereza ◽  
Komínek Petr ◽  
Kundu Jiban Kumar
Keyword(s):  
Czech Republic ◽  
Short Communication ◽  
Severe Infection ◽  
Pcr Detection ◽  
Plum Pox Virus ◽  
Prunus Domestica ◽  
Rt Pcr ◽  
The Czech Republic ◽  
Das Elisa ◽  
Year 2013

Resistance to Plum pox virus (PPV) in transgenic Prunus domestica L., clone C5 (cv. HoneySweet) was evaluated in a regulated field in the Czech Republic for fifteen years (2002–2016). PPV mild symptoms appeared in C5 trees only in several leaves situated close to the point of inoculum grafting up to 2010. No symptoms of PPV were observed in the years 2011–2013 and results of ELISA and RT-PCR detection tests were negative. In the twelfth year (2013), there was a severe unusual natural attack of plum trees by Monilinia sp. This Monilinia sp. attack occurred only one time – in 2013. There was no Monilinia sp. infection in 2002–2012 and in 2014–2016. Mild PPV symptoms reappeared in several leaves of transgenic plum trees in the next two years (2014–2015) and the presence of PPV was proved by DAS-ELISA and confirmed by RT-PCR.

P01.86 Preliminary results from the international personality disorders examination (IPDE) field trial in Greece

2000 ◽  
Vol 15 (S2) ◽  
pp. 343s-343s
Author(s):  
K.N. Fountoulakis ◽  
A. Iacovides ◽  
C. loannidou ◽  
F. Bassala ◽  
G. Kaprinis
Keyword(s):  
Personality Disorders ◽  
Field Trial ◽  
Preliminary Results
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