scholarly journals Serological and molecular detection of Prune dwarf virus infecting stone fruits of Charmahal-va-Bakhtiari province, a central region of Iran

2013 ◽  
Vol 4 (1) ◽  
pp. 4
Author(s):  
Nourolah Soltani ◽  
Jamshid Hayati ◽  
Ghobad Babaei ◽  
Maryam Ebrahim Qomi

<em>Prune dwarf</em> virus (PDV) is one of the major positive RNA viruses which cause economical damages in stone fruit trees. The symptoms of PDV vary between different stone fruits namely sour and sweet cherry, almond, peach, apricot and plum including leaf narrowing, leaf chlorosis, vein clearing, mosaic, leaf whitening, leathery leaf, bushy branches and stunt trees. During the years 2011 and 2012, 251 leaf samples were collected for detection of PDV in stone fruit orchards of Charmahal-va-Bakhtiari province. DAS-ELISA test proved PDV presence serologically. Then, total RNA were extracted and tested by two-step RT-PCR which replicated partial and full coat protein sequence of PDV. One hundred and eighty one out of total samples (251 samples) showed PDV infection using serological and two-step RT-PCR assays, hence, incidence of PDV in Charmahal-va-Bakhtiari province was confirmed. This is the first report of PDV in stone fruit orchards of Charmahal-va-Bakhtiari province and in Iran.

2010 ◽  
Vol 22 (2) ◽  
pp. 51-57 ◽  
Author(s):  
Mirosława Cieślińska ◽  
Halina Morgaś

Abstract A survey was carried out on 38 commercial and experimental stone fruit orchards located in major growing areas of stone fruit trees in Poland to determine the incidence of lesser known viruses and phytoplasmas. Leaf samples from 145 sweet cherry and 102 sour cherry trees were tested for Little cherry virus 1 (LChV-1), Little cherry virus 2 (LChV-2), Cherry green ring mottle virus (CGRMV), Cherry mottle leaf virus (CMLV), and Cherry necrotic rusty mottle virus (CNRMV) using RT-PCR. Sixty samples collected from peach and 20 apricot trees were also tested for CGRMV. Eleven out of 145 sweet cherry and three out of 102 sour cherry trees were infected by LChV-1. CGRMV was detected in 10 sweet cherry, four sour cherry, 14 peach and two apricot trees. No LChV-2, CMLV and CNRMV were detected in any of the tested trees. Phloem tissue from samples of shoots collected from 145 sweet cherry, 102 sour cherry, 128 peach, 37 apricot, five nectarine and 20 European as well as Japanese plum trees were tested for phytoplasmas. The nested PCR of the extracted DNA with universal and specific primer pairs showed the presence of phytoplasmas in six sweet cherry, three sour cherry, nine peach, four apricot, one nectarine and three Japanese plum trees. The RFLP patterns of 16S rDNA fragments after digestion with RsaI, MseI, AluI, and SspI endonucleases indicated that selected stone fruit trees were infected by two distinct phytoplasmas belonging to the apple proliferation group. The stone fruit trees infected by LChV-1, CGRMV and phytoplasmas were grown in orchards localised in all seven regions


Plant Disease ◽  
2011 ◽  
Vol 95 (11) ◽  
pp. 1483-1483 ◽  
Author(s):  
H. G. Cui ◽  
N. Hong ◽  
W. X. Xu ◽  
J. F. Zhou ◽  
G. P. Wang

Plum bark necrosis and stem pitting disease was first observed on a ‘Black Beaut’ plum (Prunus salicina Lindl.) in the United States in 1986 and later is several other countries. Plum bark necrosis stem pitting-associated virus (PBNSPaV; genus Ampelovirus, family Closteroviridae), the putative causal agent of the disease, infects many stone fruit species and causes decline, gummosis, flattening of scaffold branches, and stem necrotic pits in some diseased trees (1,3). An investigation of the incidence of PBNSPaV on stone fruit trees in China was conducted during 2009 and 2010. Leaf samples were collected from 47 trees, including peach (P. persica L. Batsch), nectarine (P. persica L. var. nucipersica Schneider), plum (P. domestica L.), ornamental plum (P. cerasifera Ehrb), sweet cherry (P. avium L.), and flowering cherry (P. serrulata L.), grown in Hubei, Henan, and Shandong provinces in central and northern China. Most of sampled trees showed trunk gummosis or stem pitting. The presence of PBNSPaV was tested by reverse transcription (RT)-PCR using primer set PBN195F/PBN195R (5′-CTGGTCTTCCTGCTACTCCTT-3′/5′-AAGCCCACAATCTCAGAGCG-3′) designed for the detection of the coat protein (CP) gene of the virus. Total RNA was extracted from leaves using a CTAB protocol reported by Li et al. (2). Products of the expected size of 190 bp were amplified from 20 samples, including seven cultivated peach, four ornamental peach, one nectarine, two plum, one ornamental plum, three sweet cherry, and two flowering cherry samples. All trees positive for PBNSPaV showed stem pitting symptoms on the base of the trunk. To further confirm these results, a 590-base region of the heat shock protein 70 homolog (HSP70h) gene was amplified by RT-PCR using primers HSP-P1/HSP-P2 (5′-GGAATTGACTTCGGTACAAC-3′/5′-TCGAAAGTACCACCACCGAA-3′). Amplicons of the expected size were cloned into the vector pMD18-T (TaKaRa, Dalian, China) and sequenced by Genscript Corp. (Nanjing, China). Sequences of 18 PBNSPaV isolates were deposited in GenBank with Accession Nos. JF810177–JF810194. Sequence comparisons showed that the partial HSP70h gene from the Chinese PBNSPaV isolates shared 82.2 to 100% nucleotide (nt) and 94.0 to 100% amino acid (aa) similarities between them and 83.6 to 99.1% nt and 94 to 100% aa similarities with the corresponding region of the other PBNSPaV isolates deposited in GenBank. In July 2010, peach GF305 seedlings were inoculated by side grafting with budwoods from two PBNSPaV-positive ornamental peach plants. In June 2011, grooving symptom was observed on the stems of the seedlings and the virus was detected by RT-PCR. The results further confirmed PBNSPaV infection in China. These results show that PBNSPaV and the associated disease occur in main cultivated and ornamental Prunus species in China. Given the importance and the devastating symptoms of the disease, it is important to prevent virus spread by using virus-tested propagation materials. References: (1) M. Al Rwahnih et al. Arch. Virol. 152:2197, 2007. (2) R. Li et al. J. Virol. Methods 154:48, 2008. (3) D. B. Marini et al. Plant Dis. 86:415, 2002.


Plant Disease ◽  
2013 ◽  
Vol 97 (2) ◽  
pp. 290-290 ◽  
Author(s):  
J. F. Zhou ◽  
G. P. Wang ◽  
L. N. Qu ◽  
C. L. Deng ◽  
Y. Wang ◽  
...  

During the growing seasons of 2010 through 2012, leaf tissues from 206 stone fruit trees, including one flowering cherry, three sour cherry, six nectarine (Prunus persica L. var. nucipersica Schneider), 14 apricot, 24 plum (P. domestica L.), 41 sweet cherry, and 117 peach [P. persica (L.) Batsch] trees, grown in six provinces of China, were randomly collected and tested for the CNRMV infection by RT-PCR. Out of those sampled trees, 37 showed shot holes and vein yellowing symptoms. Total RNA was extracted from leaves using the CTAB protocol reported by Li et al. (2). The primer pair CGRMV1/CGRMV2 (1) was used to amplify a fragment of 949 bp from CNRMV genome, which includes the CP gene (804 bp). PCR products with the expected size were detected in one sweet cherry, one apricot, one peach, one plum, and two sour cherry plants. However, no correlation between PCR data and symptom expression could be found. PCR products were cloned into the vector pMD18-T (TaKaRa, Dalian, China). Three independent clones from each isolate were sequenced by Genscript Corp., Nanjing, China, and sequences were deposited in the GenBank under accession nos. JX491635, JX491636, JX491637, JX648205, and JX648206. Results of sequence analysis showed that sequences of the five CNRMV isolates shared the highest nt (99.0 to 99.6%) and aa (98.9 to 100%) similarities with a cherry isolate from Germany (GenBank Accession No. AF237816). The sequence of one isolate from a peach tree (JX648205) was divergent and shared only 84.7 to 86.1% nt and 94.4 to 95.1% aa similarities with those cp sequences. Clones intra each isolate shared more than 99% nt similarities. To confirm CNRMV infection, seedlings of peach GF 305 were graft-inoculated with bud-woods from a peach and a sweet cherry tree, which was positive to CNRMV and also two other viruses: Cherry green ring mottle virus (CGRMV) and Plum bark necrosis stem pitting-associated virus (PBNSPaV), as tested by RT-PCR. Grafted seedlings were kept in an insectproof greenhouse and observed for symptom development. In May of the following year, some newly developed leaves of inoculated seedlings showed vein yellowing, ringspot, and shot hole symptoms. Results of Protein A sandwich (PAS)-ELISA using an antiserum raised against the recombinant CP of a CNRMV isolate (unpublished) and RT-PCR confirmed CNRMV infection in inoculated trees. In addition to CNRMV, tested seedlings were also found to be infected with CGRMV and PBNSPaV by RT-PCR. To our knowledge, this is the first report on the occurrence of CNRMV on stone fruit trees in China, and also the first record of the CNRMV infection in peach and plum plants. Given the economic importance of its hosts and the visible symptoms of the viral disease, it is important to prevent the virus spread by using virus-tested propagation materials. References: (1) R. Li and R. Mock. J. Virol. Methods 129:162, 2005. (2) R. Li et al. J. Virol. Methods 154:48, 2008.


2019 ◽  
pp. 121-127
Author(s):  
Jakub Pečenka ◽  
Eliška Peňázová ◽  
Dorota Tekielska ◽  
Ivo Ondrášek ◽  
Tomáš Nečas ◽  
...  

This study was focused on detection of the spectrum of fungi in the wood of stone fruits using molecular genetic methods. Samples were obtained from apricots, plums and sweet cherry trees from region of Moravia, one sample was obtained from Myjava (Slovakia). Segments of symptomatic wood were obtained from dying stone fruit trees with very significant symptoms. This study describes detection of the fungi in the wood of 11 trees in general in 5 localities. The cultivation of the fungi from symptomatic wood and sequencing of ITS was carried out. Eleven fungal genera were determined in the stone fruits wood, particularly Irpex lacteus, Fomes fomentarius, Neofabraea corticola, Calosphaeria pulchella, Cytospora leucostoma, Phellinus tuberculosus, Stereum hirsutum, Collophora sp., Pithomyces chartarum, Aureobasidium pullulans,Fusarium sp. The results of this study demonstrate that the reason of declining of stone fruit trees in Moravia is caused probably by trunk pathogens.


Author(s):  
V. P. Hayova

Abstract A description is provided for Leucostoma cinctum. Information is included on the disease caused by the organism, its transmission, geographical distribution, and hosts. DISEASE: Leucostoma cinctum, especially in its conidial state, is a well-known pathogen of stone-fruit trees causing necrosis of twigs, perennial Cytospora-canker. The fungus penetrates mainly through the scars, and may result in dieback of branches or even whole trees. Tree susceptibility to L. cinctum is influenced by lesions (Stanova, 1990). Comparative anatomy and host response of peach cultivars inoculated with L. cinctum was studied by Biggs (1986). Resistance of different cultivars of stone-fruit trees to L cinctum has been investigated by many authors (Cociu et al., 1991; Miles et al., 1989; Pedryc & Rozsnyai, 1991). HOSTS: On dead or dying, attached or fallen twigs of the Rosaceae, mainly Prunoideae (Amygdalus, Armeniaca, Cerasus, Persica, Prunus) and rarely other subfamilies of the Rosaceae, including genera such as Cotoneaster, Crataegus, Malus and Pyrus. GEOGRAPHICAL DISTRIBUTION: Asia: Armenia, Republic of Georgia, Iran, Kazakhstan, Russia, Turkmenistan, Uzbekistan. Australasia: Australia. Europe: Czech Republic, France, Germany, Hungary, Italy, Moldova, Rumania, Russia, Slovakia, Spain, Switzerland, Sweden, Turkey, UK, Ukraine, former Yugoslavia. North America: Canada, USA (Idaho, Michigan, New-Jersey, Oregon). TRANSMISSION: Both conidia and ascospores are air-borne, especially under humid conditions. Orange or reddish droplets or tendrils of conidia extruded from conidiomata can be often seen after rain. It is also known that arthropods can carry propagules in stone-fruit orchards (Helton et al., 1988).


2010 ◽  
pp. 34-41
Author(s):  
Gábor Tarcali ◽  
Emese Kiss ◽  
György J. Kövics ◽  
Sándor Süle ◽  
László Irinyi ◽  
...  

Plant diseases caused by phytoplasmas have increasing importance in all over the world for fruit growers. Lately, phytoplasma diseases occur on many fruit varieties and responsible for serious losses both in quality and quantity of fruit production. In the long-run these diseases cause destruction of fruit trees. The apricot phytoplasma disease (Ca. Phytoplasma prunorum) was first reported in Europe in 1924 from France. In 1992 the disease has also been identified in Hungary. On the base of growers' signals serious damages of "Candidatus Phytoplasma prunorum" Seemüller and Schneider, 2004 (formerly: European stone fruit yellows phytoplasma) could be observed in different stone fruit plantations in the famous apricot-growing area nearby Gönc town, Northern-Hungary. Field examinations have been begun in 2009 in several stone fruit plantations in Borsod-Abaúj-Zemplén County mainly in Gönc region which is one of the most important apricot growing regions in Hungary, named “Gönc Apricot Growing Area”. Our goals were to diagnose the occurrence of Ca. Phytoplasma prunorum on stone fruits (especially on apricot) in the North-Hungarian growing areas by visual diagnostics and confirm data by laboratory PCR-based examinations. All the 28 collected samples were tested in laboratory trials and at 13 samples from apricot, peach, sour cherry and wild plum were confirmed the presence of phytoplasma (ESFY). On the base of observations it seems evident that the notable losses caused by "Ca. Phytoplasma prunorum" is a new plant health problem to manage for fruit growers, especially apricot producers in Hungary. 


Author(s):  
Alina Gospodaryk ◽  
Inga Moročko-Bičevska ◽  
Neda Pūpola ◽  
Anna Kāle

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.


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