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1.
W. Jarausch J. L. Danet G. Labonne F. Dosba J. M. Broquaire C. Saillard M. Garnier 《Plant pathology》2001,50(6):782-790
An epidemiological study on European stone fruit yellows (ESFY) phytoplasmas infecting Prunus fruit trees was carried out from 1994 to 2000 in Languedoc-Roussillon (southern France). The spread of the disease was monitored for 7 years by visual observation of symptoms and by PCR detection of the phytoplasma in an experimental orchard planted with apricot hybrid seedlings. This indicated that aerial vectors were responsible for disease spread, and that transmission rates were low at the beginning of the spread. Seventy thousand homopteran insects were captured within and in the surroundings of highly ESFY-infected apricot orchards, of which about 10 000 were used in PCR and nested-PCR assays with universal ribosomal and ESFY-specific nonribosomal primers to detect ESFY phytoplasmas. The other insects were confined in cages for trials of transmission to test plants. ESFY phytoplasmas could not be detected by PCR in any of the leafhopper species captured but could be detected in the psyllid Cacopsylla pruni caught on Prunus domestica and Prunus cerasifera rootstock suckers of apricot trees and on Prunus spinosa . Nested PCR revealed ESFY phytoplasmas in one individual of the deltocephalid Synophropsis lauri captured on an apricot tree. Transmission trials confirmed the role of Cacopsylla pruni as the ESFY phytoplasma vector in France. When apricot seedlings were used as bait plants from April to November during two consecutive years, no natural transmission could be demonstrated. However, one out of 50 apricot seedlings left for the whole year in the orchard became infected. An early spring ESFY infection is in agreement with both the natural transmission results and the life cycle of Cacopsylla pruni . 相似文献
2.
W. Jarausch B. Jarausch-Wehrheim J.L. Danet J.M. Broquaire F. Dosba C. Saillard M. Garnier 《European journal of plant pathology / European Foundation for Plant Pathology》2001,107(2):209-217
Between 1994 and 1998 a field study was conducted to identify plant hosts of the European stone fruit yellows (ESFY) phytoplasma in two apricot growing regions in southern and southwestern France where the incidence of apricot chlorotic leaf roll was high. A total of 431 samples from 51 different plant species were tested for the presence of phytoplasmas by PCR using universal and ESFY-specific primers. ESFY phytoplasma was detected in six different wild growing Prunus species exhibiting typical ESFY symptoms as well as in symptomless dog rose bushes (Rosa canina), ash trees (Fraxinus excelsior) and a declining hackberry (Celtis australis). The possible role of these plant species in the spread of ESFY phytoplasma is discussed. PCR-RFLP analysis of ribosomal DNA amplified with the universal primers was carried out to characterize the other phytoplasmas found. Thus, elm yellows phytoplasma, alder yellows phytoplasma and rubus stunt phytoplasma were detected in declining European field elm trees (Ulmus carpinifolia Gled), in declining European alder trees (Alnus glutinosa) and in proliferating Rubus spp. respectively. The presence of rubus stunt phytoplasma in great mallow (Malva sylvestris) and dog rose was demonstrated for the first time. Furthermore, the stolbur phytoplasma was detected in proliferating field bindweed (Convolvulus arvensis) and a previously undescribed phytoplasma type was detected in red dogwood (Cornus sanguinea). According to the 16S rDNA-RFLP pattern this new phytoplasma belongs to the stolbur phytoplasmas group. 相似文献
3.
Candidatus Phytoplasma prunorum was detected for the first time in almond (Prunus dulcis Mill.) cv. ‘Abiod’ in Tunisia. Infected trees showed emergence of new growth during dormancy and leafed out before flowers
opened in addition to early defoliation in summer. Phytoplasma was detected by nested polymerase chain reaction (PCR) using
universal phytoplasma primer pairs P1/P7 and F2n/R2. A band with expected size was observed in samples collected from five
symptomatic, but not symptomless almond trees. PCR products (1.2 kbp) were used for restriction fragment length polymorphism
(RFLP) analysis after digestion with endonucleases RsaI and SspI. RFLP patterns obtained were similar to those reported previously for the European stone fruit yellows (ESFY, 16SrX-B).
Identification has been further confirmed by PCR using ESFY specific primer pairs (ECA1/ECA2). This is the first report of
Ca. Phytoplasma prunorum infecting almonds in Tunisia. 相似文献
4.
Barbara Jarausch Isabel Mühlenz Annette Fuchs Isabelle Lampe Uwe Harzer Wolfgang Jarausch 《Gesunde Pflanzen》2007,59(4):183-192
From 2003 to 2007 surveys have been conducted in different stone fruit growing regions in southwest Germany to detect European stone fruit yellows (ESFY) disease in Germany. Samplings have been done regularly in selected reference orchards in the regions Neuwieder Becken, Rheinhessen, Vorderpfalz and Südpfalz in summer on trees showing ESFY typical symptoms as well as on branches of trees with unspecific symptoms. All samples have been analysed by PCR for infection with Candidatus Phytoplasma prunorum. The phytoplasma could be detected in all investigated regions on the cultivated Prunus species P. armeniaca, P. persica and P. domestica. No infection was found in wild Prunus species. The main spread of the disease appeared on apricot while peach and European plum were less affected. A good correlation between symptoms and molecular detection of the pathogen could be shown for the typical symptoms in summer and winter for apricot as well as for peach. During regular psyllid captures in the reference orchards the population dynamics of Cacopsylla pruni could be described in southwest Germany for several years. By PCR-testing all collected insects individually a yearly natural infection rate of about 1–2% of all individuals of C. pruni could be calculated. 相似文献
5.
K. De Jonghe I. De Roo M. Maes 《European journal of plant pathology / European Foundation for Plant Pathology》2017,147(4):749-759
Over the years, real-time PCR outflanked endpoint PCR in phytopathogen diagnostics, mainly because of the increase in sensitivity and timesaving aspects of the technique. However, a time consuming 16S rRNA-based nested PCR method is still the gold standard for phytoplasma diagnosis. This is also the case for phytoplasma detection in Malus, Pyrus and Prunus, the three main host plants of apple proliferation (AP), pear decline (PD) and European stone fruit yellows (ESFY) phytoplasma, respectively. The last decade, loop-mediated isothermal amplification (LAMP) (Notomi et al. 2000) is gaining a lot in significance and is also for phytoplasmas expected to become a widely used reliable diagnostic tool. High specificity and sensitivity which also requires a less stringent need for DNA purification, and the short analysis time and the limited equipment requirements makes the LAMP method a fast and affordable alternative with great point-of-care diagnostic potential. In this paper, we present a LAMP primer set for the ribosomal group 16SrX, containing the important fruit tree phytoplasmas AP, PD and ESFY. The primers were developed and validated for fast and sensitive detection and general use for diagnosis. We foresee that the LAMP technique will also have its application in on-site diagnosis of the fruit tree phytoplasmas during inspections and surveys. 相似文献
6.
Hsiu-Lin Liu Ching-Chung Chen Chan-Pin Lin 《European journal of plant pathology / European Foundation for Plant Pathology》2007,117(3):281-291
Pear decline (PD) is an important phytoplasmal disease that occurs mainly in Europe and North America. In 1994, pear trees
exhibiting symptoms typical of PD disease were observed in orchards of central Taiwan. The sequence of 16S rDNA and 16S–23S
rDNA intergenic spacer region (ISR) of the causative agent of pear decline in Taiwan (PDTW) were amplified with polymerase
chain reaction (PCR) using a DNA template prepared from the diseased leaves. Sequence analysis of 16S rDNA revealed that the
PDTW agent was closely related to the phytoplasmas of the apple proliferation group that cause diseases in stone fruits, pear
and apple. Consistent with the result of 16S rDNA sequence analysis, sequence analysis of the 16S–23S rDNA ISR and putative
restriction site analyses of 16S rDNA and 16S–23S rDNA ISR sequences provided further support for the view that the PDTW phytoplasma
causing pear decline in Taiwan may represent a new subgroup of the apple proliferation group. According to the rDNA sequence
of PDTW phytoplasma, two specific PCR primer pairs, APf2/L1n and fPD1/rPDS1, were designed in this study for the detection
of the etiological agent in pear trees and insect vectors. Based on the sequence analyses of the PCR-amplified fragments,
two species of pear psyllas, Cacopsylla qianli and Cacopsylla chinensis, were found to carry PDTW phytoplasma. 相似文献
7.
Genetic comparison of the peach yellow leaf roll agent with European fruit tree phytoplasmas of the apple proliferation group 总被引：1，自引：0，他引：1
Restriction fragment length polymorphism (RFLP) analysis of PCR-amplified ribosomal DNA and Southern blot hybridization using cloned chromosomal DNA fragments from the apple proliferation (AP) phytoplasma as probes were used to investigate the genetic relationship of the California peach yellow leaf roll (PYLR) agent with phytoplasmas causing fruit tree diseases in Europe. This comparison showed that the California PYLR phytoplasma is closely related to apple proliferation (AP), pear decline, and European stone fruit yellows phytoplasmas and that it is a member of the phylogenetic AP group. The PYLR agent could clearly be distinguished from the AP and European stone fruit yellows phytoplasmas by Southern blot hybridization with DNA fragments from the AP phytoplasma and by RFLP analysis of ribosomal DNA employing Ssp I, Bsa AI, and Rsa I restriction endonucleases. However, the PYLR phytoplasma was indistinguishable from the pear decline agent by RFLP analysis of PCR-amplified ribosomal DNA. 相似文献
8.
In vitro grafting was tested as a technique for inoculating Prunus rootstock Prunus marianna GF 8-1 with European stone fruit yellows (ESFY) phytoplasmas and apple rootstock Malus pumila MM106 with apple proliferation (AP) phytoplasmas. In vitro shoot cultures of ESFY-infected Prunus marianna GF 8-1 and AP-infected Malus pumila MM106 were used as graft inoculum to transmit the phytoplasmas to the respective healthy rootstock. Phytoplasma transmission was assessed after a graft contact of 1, 2 or 3 months. Healthy autografts were used as controls to monitor parameters of in vitro grafting. Successful graft union formation ranged from 58 to 79% irrespective of the plant species and the sanitary state of the graft. Pathogen-specific polymerase chain reaction (PCR) was used to test the inoculated rootstocks for the presence of ESFY and AP phytoplasmas, respectively. The rate of ESFY phytoplasma transmission in successful Prunus -grafts increased from 69 to 94% with the time of contact. AP phytoplasma transmission to Malus occurred in 80 to 97% of successful grafts. To our knowledge this is the first report of phytoplasma transmission by grafting in vitro . The results provide a good basis for the establishment of a preliminary in vitro screening method for phytoplasma resistance in Prunus and Malus . 相似文献
9.
Thomas Thomidis 《Phytoparasitica》2001,29(1):47-49
The relative virulence ofPhytophthora cactorum andP. syringae originating from almond trees, and ofP. citrophthora originating from citrus, to apple, pear, peach, cherry and plum rootstocks, was studiedin vivo andin vitro. Results of the different experiments were in good agreement. All testedPhytophthora isolates showed little virulence to pear rootstocks-causing only minor crown rot symptoms - and no virulence at all to apple
rootstocks. In contrast, they were highly virulent to stone fruit rootstocks, causing crown rot disease. The non-pathogenicity
of these isolates to pome rootstocks could be interpreted as strict host specificity. 相似文献
10.
Production of Monoclonal Antibodies against Apple Proliferation Phytoplasma and their Use in Serological Detection 总被引：2，自引：0，他引：2
Nazia Loi Paolo Ermacora Luigi Carraro Ruggero Osler Tseh An Chen 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(1):81-86
Two monoclonal antibodies were obtained against the apple proliferation phytoplasma that provide easy, rapid, specific and sensitive serological detection. They reacted specifically by using ELISA and immunofluorescence techniques with apple proliferation-infected periwinkles and apple trees from different regions in northern Italy and Slovenia, but not with several other phytoplasma isolates. We did not observe any monoclonal antibody reaction even using phytoplasmas belonging to the same phylogenetic group such as European stone fruit yellows and pear decline. Two serological techniques, immunofluorescence and ELISA, were compared with DAPI staining and PCR. From July until leaf fall ELISA was as sensitive as PCR but was more rapid and convenient than PCR; immunofluorescence was useful for specific detection of apple proliferation phytoplasma on roots throughout the year. Serological techniques could be conveniently applied in the roots, stems and leaves of apple trees depending on specific phenological stages of the plants. 相似文献
11.
M. Hassan G. Gomez V. Pallás A. Myrta P. Rysanek 《European journal of plant pathology / European Foundation for Plant Pathology》2009,124(2):363-368
In this paper, we report a large-scale survey for the incidence of Peach latent mosaic viroid (PLMVd) and Hop stunt viroid (HSVd) in stone fruit collections and commercial orchards in the Czech Republic. From the 645 samples analysed, PLMVd was
detected in 80 (26.6%) of peaches and the HSVd in 3 (1.3%) of apricot and 1 (0.33%) of peach trees. Sixty-seven accession
of peach (44.6%) from the Czech Clonal GeneBank were infected by PLMVd. In addition, we used naturally infected trees to standardise
the simultaneous detection of PLMVd and HSVd plus host mRNA as the control by means of one-step multiplex RTC-PCR. Eleven
PLMVd and two HSVd isolates were sequenced and analysed. All the PLMVd variants were highly homologous (97–100%) to previously
reported PLMVd variants from Tunisian peach and almond trees, and clustered together in the previously reported phylogenetic
group III. The HSVd variants obtained from apricot and peach trees were included in the previously proposed recombinant group
PH/cit3. 相似文献
12.
Several uncultivated trees of the species Prunus spinosa , P. cerasifera and P. domestica , sampled both adjacent to European stone fruit yellows (ESFY)-infected orchards and in isolation from cultivated stone fruit plants, were found to be infected by ESFY phytoplasma. These species were also colonized by Cacopsylla pruni , vector of the ESFY agent. In contrast, uncultivated species of Prunus avium , P. cerasus and P. mahaleb hosted neither the pathogen nor the vector. Insect- and graft-transmission trials of ESFY phytoplasma conducted under controlled conditions confirmed the data obtained in the field. The role played by the wild Prunus species is discussed and appears to be fundamental in the epidemic cycle of the disease. 相似文献
13.
In the Campania region of southern ltaly. commercial orchards of European hazel ( Corylus avellana ) are severely affected by yellowing and decline. To determine whether phytoplasmas are associated with the disorder, stem samples from diseased trees were examined using polymerase chain reaction assays. No visible products were obtained by amplification of sample DNA with universal and group-specific phytoplasma primers. However, when the products obtained with universal primers were re-amplified with nested primers that were specific for the fruit tree phytoplasmas of the apple proliferation group, most samples tested positively. Restriction site analysis revealed that the trees were infected with the apple proliferation, pear decline, and European stone fruit yellows phytoplasmas in about the same proportion. Some of the trees were doubly infected with one of the fruit tree phytoplasmas and the aster yellows agent. Most of the infected trees were also identified by hybridization of the products obtained in the initial amplification with suitable oligonucleotide probes. 相似文献
14.
Transmission of ‘Candidatus Phytoplasma pyri’ by naturally infected Cacopsylla pyri to peach,an approach to the epidemiology of peach yellow leaf roll (PYLR) in Spain 
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下载免费PDF全文 Peach orchards in the northeast of Spain were severely affected in 2012 by a previously unreported disease in this area. The symptoms included early reddening, leaf curling, decline, abnormal fruits, and in some cases death of the peach trees. All the infected peach samples were positive for ‘Candidatus Phytoplasma pyri’, but none were infected by the ‘Ca. Phytoplasma prunorum’. In this work, potential vectors able to transmit ‘Ca. Phytoplasma pyri’ from pear to peach and between peach trees were studied and their infective potential was analysed at different times of the year. Transmission trials of the phytoplasma with potential vectors to an artificial feeding medium for insects and to healthy peach trees were conducted. Additionally, isolated phytoplasmas were genetically characterized to determine which isolates were able to infect peach trees. Results showed that the only insect species captured inside peach plots that was a carrier of the ‘Ca. Phytoplasma pyri’ phytoplasma was Cacopsylla pyri. Other insect species captured and known to be phytoplasma transmitters were present in very low numbers, and were not infected with ‘Ca. Phytoplasma pyri’ phytoplasma. A total of 1928 individuals of C. pyri were captured in the peach orchards, of which around 49% were phytoplasma carriers. All the peach trees exposed to C. pyri in 2014, and 65% in 2015, were infected by ‘Ca. Phytoplasma pyri’ 1 year after exposure, showing that this species is able to transmit the phytoplasma to peach. Molecular characterization showed that some genotypes are preferentially determined in peach. 相似文献
15.
Franco Daniel Fernández Fabiana Aida Guzmán Viviana Curzel Noemí Bejarano Luis Rogelio Conci 《European journal of plant pathology / European Foundation for Plant Pathology》2013,135(4):627-631
Peach (Prunus persica L.) plants with symptoms of yellowing, reddening, curling and leaf necrosis, premature defoliation and internode shortening were observed in production fields in Jujuy province (Argentina). A phytoplasma was detected by PCR using the universal primer pairs P1/P7 and R16F2n/R16R2 in all the symptomatic samples analysed. The RFLP profile of PCR products, amplified with R16F2n/R16R2 primers, shows that this phytoplasma, named Argentinean Peach Yellows (ArPY), belongs to subgroup 16Sr III-B. The phylogenetic analysis of the 1244 bp 16S rDNA cloned sequence, grouped the ArPY phytoplasma into the X-disease group with a closer relationship with CFSD, PssWB and ChTDIII phytoplasmas. This is the first report of a phytoplasma infecting peach trees in Argentina. 相似文献
16.
Gulnara Balakishiyeva Madat Qurbanov Alamdar Mammadov Shaniyar Bayramov Jalal Aliyev Xavier Foissac 《European journal of plant pathology / European Foundation for Plant Pathology》2011,130(4):457-462
‘Candidatus Phytoplasma brasiliense’, a phytoplasma taxon associated with hibiscus witches’ broom disease was first described in 2001
in Brazil. In September 2007, a peach tree (Prunus persica) displaying yellowing symptoms reminiscent of phytoplasma infection was sampled in Guba region of Azerbaijan. A phytoplasma
was detected in the diseased peach tree by nested PCR amplification of its 16S rDNA with universal primers for phytoplasmas.
Phylogenetical analyses of the amplified 16S rDNA showed that the phytoplasma infecting the peach tree corresponded to ‘Ca. P. brasiliense’, a species never reported in Euro-Mediterranean area. To set up a detection assay, cloning of a ‘Ca. P. brasiliense’ DNA fragment was undertaken by comparative RAPD. The amplified dnaK-dnaJ genetic locus was used to design a nested PCR assay able to amplify all ‘Ca. P. brasiliense’ isolates of the subgroup 16SrXV-A without amplifying the related members of the group 16SrII. This assay
also allowed confirming the first detection of ‘Ca. P. brasiliense’ in diseased basil collected in south Lebanon. 相似文献
17.
Pcr Assay for specific detection of European stone fruit yellows phytoplasmas and its use for epidemiological studies in France 总被引：2，自引：0，他引：2
W. Jarausch M. Lansac C. Saillard J.M. Broquaire F. Dosba 《European journal of plant pathology / European Foundation for Plant Pathology》1998,104(1):17-27
DNA amplification by polymerase chain reaction was used to specifically detect phytoplasmas associated with severe decline diseases of European stone fruits. PCR primers were designed according to the partial sequence of a nonribosomal genomic fragment of European stone fruit yellows phytoplasmas obtained by direct sequencing of a specific PCR product. A PCR assay was developed which resulted in specific amplification of a 237 bp-DNA fragment from total DNA extracts derived from over 300 stone fruit samples. No PCR product was obtained with DNA from healthy controls or plants diseased with various other phytoplasmas, e.g. the closely related apple proliferation and pear decline phytoplasmas. Phytoplasma infection was checked in all samples by PCR amplification with universal ribosomal primers. Detection rate with specific and universal primers was correlated by 97%. European stone fruit yellows phytoplasmas were detected in samples of 114 out of 139 examined orchards which represent the major stone fruit growing regions of France. Typical symptoms like chlorotic leaf roll in summer and off-season growth in winter were correlated by 95% to the presence of phytoplasmas. However, phytoplasmas were also detected in 51% of samples derived from trees showing non-specific symptoms. A comparison study including 201 samples showed that 81% of the PCR-positive samples were also tested positive using fluorescence microscopy with DAPI staining. 相似文献
18.
Viruses and viroids of stone fruits in Syria 总被引：1，自引：0，他引：1
F. Ismaeil A. Myrta N. Abou Ghanem-Sabanadzovic S. Al Chaabi V. Savino 《EPPO Bulletin》2002,32(3):485-488
Field surveys were carried out in the main stone fruit-growing areas of Syria to evaluate the sanitary status of mother blocks, varietal collections and commercial orchards. The presence of virus and virus-like diseases was checked by enzyme-linked immunosorbent assay (ELISA), sap transmission to herbaceous hosts, testing on the woody indicators Prunus persica cv. GF 305 and Prunus serrulata cv. Kwanzan and dot-blot hybridization tests. A total of 1337 samples was tested by ELISA (444 apricot, 283 peach, 246 cherry, 222 almond and 142 plum). The overall mean infection rate was 13%, and the percentage infection level of single species was: peach 24%, cherry 16%, almond 13.5%, apricot 6%, plum 5%. The following viruses and viroids were detected: PNRSV, PDV, ACLSV, PPV, ApMV, PLMVd and HSVd 1 . 相似文献
19.
Sharka disease has a limited distribution in Turkey and does not present a problem for stone fruit production. However, sharka
is the most common virus disease of apricots, plums and peaches in Ankara, although it is not a common disease in other cities
in Turkey. In different parts of Ankara, 212 private gardens in 21 locations were surveyed forPlum pox virus (PPV) incidence. All together 935 trees of apricot, plum, peach, sweet cherry and sour cherry were sampled and tested for the
presence ofPPV by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA).PPV was identified in numerous trees,viz. 286 apricots, 172 plums and 65 peaches. Strain differentiation ofPPV was accomplished using double-antibody sandwich indirect enzyme-linked immunosorbent assay (DASI-ELISA). These assays confirmed
the presence of isolates belonging toPPV-M, PPV-D and mixed infections ofPPV-M+D. This is the first report of the presence ofPPV-D and mixed infections ofPPV-M+D in Turkey.
http://www.phytoparasitica.org posting July 14, 2004. 相似文献
20.
Hosts and symptoms of Plum pox virus: fruiting Prunus species 总被引：1，自引：0，他引：1