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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   

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The phloem‐sucking psyllid Cacopsylla picta plays an important role in transmitting the bacterium ‘Candidatus Phytoplasma mali’, the agent associated with apple proliferation disease. The psyllid can ingest ‘Ca. Phytoplasma mali’ from infected apple trees and spread the bacterium by subsequently feeding on uninfected trees. Until now, this has been the most important method of ‘Ca. Phytoplasma mali’ transmission. The aim of this study was to investigate whether infected C. picta are able to transmit ‘Ca. Phytoplasma mali’ directly to their progeny. This method of transmission would allow the bacteria to bypass a time‐consuming reproductive cycle in the host plant. Furthermore, this would cause a high number of infected F1 individuals in the vector population. To address this question, eggs, nymphs and adults derived from infected overwintering adults of C. picta were reared on non‐infected apple saplings and subsequently tested for the presence of ‘Ca. Phytoplasma mali’. In this study it was shown for the first time that infected C. picta individuals transmit ‘Ca. Phytoplasma mali’ to their eggs, nymphs and F1 adults, thus providing the basis for a more detailed understanding of ‘Ca. Phytoplasma mali’ transmission by C. picta.  相似文献   

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Three experimental treatments consisting of inoculation with an arbuscular mycorrhizal fungus, application of a synthetic antimicrobial peptide or application of a resistance inducer, were evaluated in Madagascar periwinkle as control methods for rubus stunt and stolbur diseases caused by ‘Candidatus Phytoplasma rubi’ and ‘Candidatus Phytoplasma solani’, respectively. Two experiments were conducted under controlled environment conditions. In the first experiment, 4 months after graft‐inoculating the phytoplasmas, the root colonization achieved by Rhizophagus irregularis significantly reduced both disease symptoms and the frequency of detection of the pathogens by real‐time PCR. In the second experiment, the antimicrobial peptide BP100 totally prevented disease symptoms, despite the molecular detection of the phytoplasmas in 75% and 50% of the plants inoculated with ‘Ca. Phytoplasma rubi’ and ‘Ca. Phytoplasma solani’, respectively, and was more effective than benzothiadiazole (BTH) at increasing resistance against the pathogenic infections. A potential combination of early mycorrhizal inoculation and BP100 antimicrobial peptide application is envisaged as a future control strategy for phytoplasma diseases.  相似文献   

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Epidemiological aspects and transmission routes of bois noir (BN), a grapevine yellows disease induced by ‘Candidatus Phytoplasma solani’, have been exhaustively studied in the affected vineyards of continental Europe but not in the Mediterranean coastal zone. Because ‘Ca. Phytoplasma solani’ and its principal vector Hyalesthes obsoletus presumably originate from the Mediterranean, gaining knowledge of the epidemiological peculiarities of the disease in this area is essential for understanding its global spread and diversification, as well as for designing local management strategies. In this study, molecular epidemiology was applied to trace transmission pathways of ‘Ca. Phytoplasma solani’ in the Mediterranean vineyards of Montenegro, using multilocus sequence typing of tuf, vmp1 and stamp genes of the isolates associated with various hosts. Thus, ‘Ca. Phytoplasma solani’ was tracked from a tentative reservoir plant (inoculum source) through an associated vector population to the infected grapevine. Three pathways of transmission were documented, originating from Urtica dioica, Convolvulus arvensis and Vitex agnus‐castus; however, only the route originating from U. dioica was direct, whereas the latter two were overlapping and could be intermixed. Vitex agnus‐castus is a natural source of ‘Ca. Phytoplasma solani’, representing an important link in disease epidemiology in the Mediterranean and a possible origin of several genotypes occurring in central Europe. Experimental confirmation of the role of Vitex‐associated H. obsoletus in BN transmission in Montenegrin vineyards indicates its tentative role as a vector in the wide area of the Mediterranean, where some of the major wine‐producing regions are located.  相似文献   

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The phytoplasmas of groups 16SrI (‘Candidatus Phytoplasma asteris’) and 16SrVII (‘Ca. Phytoplasma fraxini’) have been associated with phytoplasma diseases in several urban tree species in Bogotá, Colombia and surrounding areas. The insect vectors responsible for this phytoplasma transmission are unknown. The objectives of this study were to test for the presence of phytoplasmas in leafhopper species (Cicadellidae) collected in areas with diseased trees and to determine the phytoplasma transmission ability of two of these species. Leafhoppers of nine species were collected at two sampling sites and tested by nested or double nested PCR using primers for the 16S rRNA gene. The amplicons were subjected to RFLP and/or sequencing analysis. Phytoplasmas of group 16SrI were detected in morphospecies MF05 (Haldorus sp.), group 16SrVII in MF07 (Xestocephalus desertorum), MF08 (Empoasca sp.) and MF09 (Typhlocybinae), and both groups 16SrI and 16SrVII in MF01 (Empoasca sp.), MF02 (Typhlocybinae), MF03 (Scaphytopius sp.), MF04 (Amplicephalus funzaensis) and MF06 (Exitianus atratus). Transmission tests to uninfected bean plants (Phaseolus vulgaris) were performed using field collected A. funzaensis and E. atratus individuals in separate assays. After 5 weeks, the test plants exposed to individuals of both species of leafhoppers showed symptoms, suggesting phytoplasma infection. Phytoplasma groups 16SrI and 16SrVII were detected in the two groups of exposed plants, indicating that A. funzaensis and E. atratus were able to transmit both groups of phytoplasmas. This is the first report of insect vectors for phytoplasmas of group 16SrVII in the world and of 16SrI in South America.  相似文献   

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The presence of phytoplasmas in seven coniferous plant species (Abies procera, Pinus banksiana, P. mugo, P. nigra, P. sylvestris, P. tabuliformis and Tsuga canadensis) was demonstrated using nested PCR with the primer pairs P1/P7 followed by R16F2n/R16R2. The phytoplasmas were detected in pine trees with witches’ broom symptoms growing in natural forest ecosystems and also in plants propagated from witches’ brooms. Identification of phytoplasmas was done using restriction fragment length polymorphism analysis (RFLP) of the 16S rDNA gene fragment with AluI, MseI and RsaI endonucleases. All samples showed RFLP patterns similar to the theoretical pattern of ‘Candidatus Phytoplasma pini’, based on the sequence of the reference isolate Pin127S. Nested PCR‐amplified products, obtained with primers R16F2n/R16R2, were sequenced. Comparison of the 16S rDNAs obtained revealed high (99·8–100%) nucleotide sequence identity between the phytoplasma isolates. The isolates were also closely related to four other phytoplasma isolates found in pine trees previously. Based on the results of RFLP and sequence analyses, the phytoplasma isolates tested were classified as members of the ‘Candidatus Phytoplasma pini’, group 16SrXXI.  相似文献   

10.
Phytoplasma suspected symptoms of little leaf, flat stem, witches’ broom and leaf yellowing were recorded on the four legume species, cowpea (Vigna unguiculata (L.) Walp.), pigeon pea (Cajanus cajan (L.) Millsp.), lentil (Lens culinaris Medikus) and mung bean (Vigna radiata (L.) Wilczek) in the states of Delhi, Uttar Pradesh (UP) and Kerala from 2014 to 2016. DNA specific fragments of approximately 1.3 kb were amplified from symptomatic samples of cowpea, pigeon pea, lentil and mung bean in nested PCR assays by using two sets of universal phytoplasma nested specific primers P1/P7 followed by 3Far/3Rev. No DNA amplifications were observed in any of the non-symptomatic legume samples with same primer pairs. Pair wise sequence comparison, phylogeny and virtual RFLP analysis of 16S rDNA sequences of the four legume species confirmed the association of four different groups and subgroups of phytoplasmas in the present study. The mung bean witches’ broom at Delhi was identified to be associated with strain related to ‘Ca. P. aurantifolia’ (16SrII-D), pigeon pea little leaf at Faizabad, UP with strain related to ‘Ca. P. phoenicium’ (16SrIX-C), lentil witches’ broom at Faizabad, UP with ‘Ca. P. trifolii’ (16SrVI-D) and cow pea flat stem disease at Kerala with ‘Ca. P. cynodontis’ (16SrXIV-A). Association of ‘Ca. P. cynodontis' (16SrXIV-A) infecting cowpea, ‘Ca. P. trifolii’ (16SrVI-D) in lentil and phytoplasmas strain related to ‘Ca. P. phoenicium’ (16SrIX-C) infecting pigeon pea are the new reports to the world.  相似文献   

11.
Elaeocarpus yellows” (ELY) is a widely reported phytoplasma disease of Elaeocarpus zollingeri trees in Japan. The phytoplasma associated with ELY (ELY phytoplasma) had not been identified at the species level because its 16S rRNA sequence had yet to be reported. Here, we report the results of a sequence analysis based on 16S rRNA and secA gene sequences, which showed that the ELY phytoplasma is related to ‘Candidatus Phytoplasma malaysianum’. To our knowledge, this is the first report showing the occurrence of ‘Ca. P. malaysianum’ outside Malaysia and the infection of E. zollingeri by the phytoplasma.  相似文献   

12.
Witches’ broom disease (WBD), caused by ‘Candidatus Phytoplasma aurantifolia’, is a serious disease of acid lime (Citrus aurantifolia) in Oman and the UAE. However, little is known about the distribution of phytoplasma and the expression of WBD symptoms in different geographical locations. A survey was carried out in 18 districts in Oman and the UAE covering 143 orchards and 5823 acid lime trees. ‘Candidatus Phytoplasma aurantifolia’ was detected in acid lime in all the 18 surveyed districts. However, the development of typical symptoms of WBD was only observed in 12 districts. Districts in which the phytoplasma was present but symptoms were not expressed were located either in desert areas or in areas characterized by semitropical conditions. Phylogenetic analysis of 16 phytoplasma isolates from trees developing WBD symptoms and six phytoplasma isolates from trees with no WBD symptoms showed that all isolates share an identical 16S rRNA sequence, belonging to subgroup II‐B. Quantitative PCR analysis showed that the concentration of phytoplasma is significantly higher (8800–801 000 copies) in leaves developing WBD symptoms compared to 2–268 copies in symptomless leaves from the same trees and 8–874 copies in acid lime trees from areas where disease symptoms were not expressed. The lack of expression of WBD symptoms under certain environmental conditions may suggest that symptom development and phytoplasma are affected by certain unfavourable environmental conditions. These findings could provide a basis for managing WBD through encouraging lime cultivation under climatic conditions less conducive to WBD symptom expression.  相似文献   

13.
Kerala wilt disease of coconut palm is a major threat of coconut production in Kerala caused by phytoplasma. The genomic DNA purified from the insect tissues of Proutista moesta (PM) and Stephanitis typica (ST) was subjected to PCR assay using the primer combination P1/P6, P1/P7 and P4/P7. The amplified products resolved a prominent band of 650 bp for the universal primer P4/P7 and no bands were noticed for the primer pairs P1/P6 and P1/P7 combination. Since P4/P7 amplifies the 16S–23S intergenic spacer region of 16SrRNA gene, the PCR product 650 bp of the insect PM indicate the phytoplasma DNA. The presence of 650 bp for the primer P4/P7 in the genomic DNA isolated from P. moesta indicates the vectoral ability of the insect. No sign of amplification was noticed in the case of ST for the three sets of primers suggesting the inability of this insect as vector. The amplified product 650 bp from the genomic DNA of KWD palms as well as the insect tissues of P. moesta was gel purified and sequenced. The sequential similarity of 650 bp of both KWD phytoplasma and the insect phytoplasma supports the transmission of phytoplasma through the vector PM. Moreover, the sequence of 650 bp was compared with other sequences of 26 coconut phytoplasmas so far reported internationally and a cladogram was prepared for determining the phylogenetic status. It is obvious from the cladogram that the KWD disease phytoplasma is evolutionarily closest to coconut phytoplasma of coconut lethal yellowing of Mexican palms within the group 16SrIV. Phylogenetically, KWD phytoplasma is grouped in the new subgroup 16SrIV-C subsequent to the groups 16SrIV-A and 16SrIV-B for Mexican coconut lethal yellowing and Tanzanian coconut lethal decline, respectively. The restriction enzyme analysis of the PCR product 650 bp using the enzymes AluI, BclI, HindIII and RsaI further supports the phytoplasmic nature of DNA. This data records the first finding of the vector of Kerala wilt disease by detecting KWD phytoplasma in insect tissue of PM by PCR based methods. Moreover, the study reveals the phylogenetic status of KWD phytoplasma compared to other coconut phytoplasmas internationally.  相似文献   

14.
Phytoplasma-induced leaf yellowing, stunting and rosetting were observed on Japanese spurge (Pachysandra terminalis) in Jung-gu of Daegu city, Korea. Molecular analyses based on PCR–RFLP and 16S rDNA sequences indicated that the phytoplasma associated with Japanese spurge yellows disease is a new strain of Candidatus Phytoplasma asteris. This report is the first on the occurrence of phytoplasma disease on Japanese spurge in Korea.  相似文献   

15.
A screening system for apple proliferation resistance was developed, based on in vitro graft‐inoculation with the causal agent ‘Candidatus Phytoplasma mali’. For this, in vitro cultures of the field‐resistant apomictic genotypes Malus sieboldii, H0909, D2212 and the susceptible Malus × domestica genotypes Golden Delicious and rootstock M9 were established, as well as in vitro cultures of Rubinette and Golden Delicious infected with ‘Ca. P. mali’ strains PM4 and PM6, respectively. Healthy in vitro shoots were inoculated by micrografting with infected shoots used as graft tip. After 6 weeks graft contact no significant differences for graft quality were observed between healthy and infected grafts. Mortality of grafts and transmission rates were not significantly different among the different genotypes. The phytoplasma concentration in inoculated shoots was determined at different times post‐inoculation (p.i.) by quantitative real‐time PCR. Infected M. sieboldii and D2212 had lower phytoplasma concentration than the susceptible controls and showed no symptoms. H0909 showed an intermediate behaviour exhibiting lower phytoplasma concentrations with strain PM4 but growth was affected. The dynamics of phytoplasma concentration reached a maximum at 6–8 months p.i. for all genotypes but the values for 3–5 and 10–12 months p.i. were similar. The resistance of M. sieboldii and D2212 was confirmed in vitro. A significant difference in phytoplasma concentration was observed between strains PM4 and PM6.  相似文献   

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Symptoms of shoot proliferation characteristic of phytoplasma diseases were observed on nectarine (Prunus persica var. nucipersica) and peach (P. persica) trees in the Sarada plain, south of Lebanon. The presence of phytoplasmas in the two orchards visited was confirmed by nested polymerase chain reaction using universal primers. The amplified DNA fragments were cloned and sequenced. Blast analysis of over 1000 nucleotides demonstrated the presence of ‘Candidatus Phytoplasma phoenicium’ which is considered to be the causal agent of Almond witches’ broom. This phytoplasma which belongs to the pigeon pea witches’ broom group (16SrIX) can be devastating since Almond witches’ broom has killed thousands of almond trees in Lebanon and Iran. Previous reports indicated that Almond witches’ broom may be transmitted by grafting to peach and nectarine under experimental conditions. This is the first report of a natural and epidemic spread of ‘Ca. Phytoplasma phoenicium’ in peach and nectarine. Farmers in the region were advised to eradicate the infected trees immediately. Further studies on the epidemiology of ‘Ca. Phytoplasma phoenicium’ and its vector(s) are recommended in order to develop successful eradication or disease management programmes.  相似文献   

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A survey for phytoplasma diseases in tomato and pepper fields in Lebanon was conducted during 2003 and 2004. Tomato plants with stunting, yellowing or purplish leaves, proliferation of laterals buds, hypertrophic calyxes and virescent flowers were found in 25% of the tomato fields surveyed, where they represented 2–8% of the plants. Pepper plants displaying stunting and yellowing of leaves, were found in 27% of the fields and 1–4% of the plants were affected. Phytoplasmas infecting tomato and pepper had identical 16S-rDNA RFLP profiles and sequences. A phytoplasma isolate named PTL was transmitted by dodder from a diseased tomato plant to a periwinkle (Catharanthus roseus) plant in which it induced leaf yellowing, virescence and phyllody. 16S-rDNA phylogenetic analysis classified PTL as a strain of ‘Candidatus Phytoplasma trifolii’.  相似文献   

18.
《EPPO Bulletin》2017,47(2):146-163

Specific scope

This Standard describes a diagnostic protocol for ‘Candidatus Phytoplasma mali’, ‘Ca. P. pyri’ and ‘Ca. P. prunorum’. This Standard should be used in conjunction with PM 7/76 Use of EPPO diagnostic protocols

Specific approval and amendment

Approved as PM 7/62 Candidatus Phytoplasma mali and PM 7/63 Ca. P. pyri in 2006. Revised in 2017‐02 as a single Standard as PM 7/62 (2) with the addition of ‘Ca. P. prunorum’.  相似文献   

19.
This test performance study (TPS) was carried out on DNA samples from grapevine, clematis, fungi and bacteria to compare and validate loop‐mediated isothermal amplification (LAMP) tests for detection of Grapevine flavescence dorée phytoplasma and ‘Candidatus Phytoplasma solani’ (Grapevine Bois noir phytoplasma). Two LAMP tests, for Grapevine flavescence dorée phytoplasma and ‘Candidatus Phytoplasma solani’ (as developed by Kogov?ek and colleagues), with proven applicability for rapid laboratory or on‐site detection were included in this study. They were performed in 10 laboratories. In addition, the commercial Qualiplante/Hyris isothermal amplification test for Grapevine flavescence dorée phytoplasma was performed in three laboratories. The accuracy of the three tests was shown to be over 98%. Moreover, the high accuracy of these tests, which used different devices across different laboratories, confirmed their reproducibility.  相似文献   

20.
Candidatus Phytoplasma prunorum’ is the causal agent of the European stone fruit yellows (ESFY) disease. This phytoplasma affects wild and cultivated species of Prunus to different degrees, depending on their susceptibility. ‘Candidatus Phytoplasma prunorum’ is present in the four regions of Spain surveyed in this study (Aragon, Catalonia, Extremadura and Valencia) with a variable incidence. Results showed that ‘Ca. Phytoplasma prunorum’ was detected in all of the cultivated Prunus species studied, except P. avium and P. dulcis, and was widespread in Spain. The most affected species was P. salicina, with symptoms including early bud break and blooming, leaf curling and yellowing, collapse, and a major decrease in production. In some plots in the Baix Llobregat area of Barcelona province (Catalonia), the incidence of ESFY on P. salicina was as high as 80%. The insect vector, Cacopsylla pruni, was present in all four of the regions studied, with the highest captures in yellow sticky traps in Catalonia on P. mahaleb and in Extremadura in peach orchards. In Baix Llobregat, large populations of C. pruni were present on infected P. mahaleb bushes, and with high infection rates. This was a key factor in the local pathogenic cycle that caused a major ESFY outbreak in the nearby P. salicina orchards. In the Ebro valley (Lleida and Aragon) and Valencia, the surveys showed very low incidences of the disease and low C. pruni populations.  相似文献   

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