scholarly journals First Report of Phytoplasma Infection in Hop Plants

Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 908-908 ◽  
Author(s):  
E. Solarska ◽  
M. Kamińska ◽  
H. Śliwa
Keyword(s):  
Natural Infection ◽  
Shoot Proliferation ◽  
Rflp Analysis ◽  
Enzymatic Digestion ◽  
Early Spring ◽  
Direct Pcr ◽  
Universal Primer ◽  
Disease Symptoms ◽  
Group Specific Primer ◽  
Reference Strains

Disease symptoms of severe shoot proliferation resembling phytoplasmal disease symptoms were observed in early spring of 2003 in hop (Humulus lupulus L.) plant cvs. Magnum and Marynka that were grown in a commercial farm in Poland. Proliferation of shoots has not been previously reported in hop plants. To detect the presence of phytoplasmas in hops, young shoots from four symptomatic (two cultivars) and two symptomless (‘Magnum’) plants were assayed for phytoplasma 16S rDNA using polymerase chain reaction (PCR). In addition, leaf samples from healthy Catharanthus roseus plants and plants experimentally infected with the reference strains of aster yellows phytoplasma (AY1, group 16SrI-B) or apple proliferation phytoplasma (AP, group 16SrX-A) were included for comparison. Amplifications were performed using the universal phytoplasma primer pair P1/P7 in an initial assay, and universal primer pairs fA/rA (1), Pc399/P1694, or R16F2n/R16R2 (2) and group specific primer pair R16(I)F1/R16(I)R1 (3) in a nested reaction. Specific products were obtained in direct PCR with the universal primer pairs P1/P7 only for the control samples of the reference strains AY and AP. No visible product was amplified by the direct PCR from samples obtained from hops and healthy periwinkle plants. However, in nested PCR with primer pairs P1/P7 followed by primer pairs fA/rA, R16F2n/R16R2, Pc399/P1694, or R16(I)F1/R16(I)R1, specific DNA bands were observed from naturally infected hop plants (both four symptomatic and two symptomless) tested. No amplification products were observed from healthy periwinkle plants. The specificity of PCR products (obtained with universal R16F2n/R16R2 primer pair) was confirmed by restriction fragment length polymorphism (RFLP) analysis using AluI, MseI, HhaI, and RsaI for enzymatic digestion. RFLP patterns of these rDNA fragments for samples of naturally infected hops and for AY1 reference strain were similar and were characteristic of phytoplasma 16SrI-B subgroup. To our knowledge, this is the first evidence that hop shoot proliferation disease is associated with natural infection by phytoplasmas. Furthermore, detection of phytoplasma in asymptomatic hops underscores the need to fully elucidate the etiological role of this pathogen in the disease. References: (1) U. Ahrens and E. Seemüller. Phytopathology 82:828, 1992. (2) I.-M. Lee et al. Phytopathology 83:834, 1993. (3) I.-M. Lee et al. Phytopathology 84:559, 1994.

scholarly journals Molecular characterization of stolbur phytoplasmas in pepper and tomato from Bulgaria

2020 ◽  
Vol 18 ◽  
pp. 00025
Author(s):  
Dimitriyka Sakalieva
Keyword(s):  
Molecular Characterization ◽  
Reference Strain ◽  
Rflp Analysis ◽  
Plant Diseases ◽  
Vegetable Crops ◽  
Disease Symptoms ◽  
Natural Plant ◽  
Conserved Genes ◽  
Reference Strains

Tomato and pepper are the main vegetable crops cultivated in Bulgaria. Phytoplasma diseases, mainly stolbur, are important plant diseases for these crops in Bulgaria. The goal of the present paper was to verify association of phytoplasmas with the observed disease symptoms in tomato and pepper and to identify the phytoplasmas detected using RFLP analysis of conserved genes and other uncharacterised phytoplasma chromosomal regions. The presence of phytoplasmas was confirmed in all the samples of tomato and pepper showing typical stolbur symptoms. A phytoplasm sample, which caused severe symptoms, showed the same pattern as the reference strain Mol, while all other phytoplasmic reference strains showed different polymorphisms. RFLP profiles were found useful in distinguishing phytoplasmas in stolbur subgroup (16SrXII-A) in natural plant hosts.

scholarly journals Characterization of Phytoplasmas Related to Aster Yellows Group Infecting Annual Plants in Iran, Based on the Studies of 16S rRNA and RP Genes

2014 ◽  
Vol 54 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Fereshteh Vali Sichani ◽  
Masoud Bahar ◽  
Leila Zirak
Keyword(s):  
16S Rrna ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Rrna Gene ◽  
Annual Plants ◽  
Universal Primer ◽  
Aster Yellows ◽  
Central Regions ◽  
Group Specific Primer

Abstract Several annual field crops, vegetables, ornamentals, oilseed crops, and weeds showing phytoplasma diseases symptoms were collected to detect phytoplasmas related to ‘Candidatus Phytoplasma asteris’. The collecting was done in the central regions of Iran. For general detection of phytoplasmas, 16S rRNA gene fragments were amplified using phytoplasma universal primer pair P1/P7 in polymerase chain reaction (PCR) followed by primer pair R16F2n/R16R2 in nested PCR. Then, for finer detection of phytoplasmas related to ‘Ca. P. asteris’, DNA samples were used to extend the rp and tuf gene fragments by PCR using aster yellows group specific primer pairs rp(I)F1A/rp(I)R1A and fTufAy/rTufAy, respectively. Restriction fragment lenght polymorphism (RFLP) analysis of rp gene fragments using digestion with AluI, MseI, and Tsp509I restriction enzymes indicated that aster yellows group related phytoplasmas in these Iranian regions, belong to rpI-B subgroups. Sequence analysis of partial 16S rRNA and rp genes from representative phytoplasma isolates confirmed the RFLP results. This research is the first report of annual plants infected with phytoplasmas related to subgroup rpI-B in Iran.

Tomato big bud, legume little leaf, and lucerne witches' broom: three diseases associated with different mycoplasma-like organisms in Australia

1974 ◽  
Vol 25 (3) ◽  
pp. 449 ◽  
Author(s):  
JW Bowyer
Keyword(s):  
Type Strain ◽  
Shoot Proliferation ◽  
Datura Stramonium ◽  
Mild Form ◽  
Disease Symptoms ◽  
Reference Type ◽  
Incubation Periods ◽  
Broom Disease ◽  
Reference Strains ◽  
Type Strains

The pathogens (presumed to be mycoplasma-like organisms) causing tomato big bud, legume little leaf, and a 'mild' form of lucerne witches' broom disease could be distinguished by differential symptomatology and incubation periods in tomato (Lycopersicon esculentum) and Datura stramonium. They were regarded as reference 'type strains' of the Australian yellows disease agents. Incubation periods of the tomato big bud and legume little leaf agents in the leafhopper vector Orosius argentatus were similar, but the pathogen causing mild lucerne witches' broom was not transmissible by this insect. Six species of plants with yellows disease symptoms (shoot proliferation, small leaves, chlorosis, virescence) were collected from northern and southern Queensland. The pathogens were transmitted by O. argentatus to D. stramonium. From symptoms produced in this host, the various isolates were characterized as either the original big bud or little leaf reference strains. An isolate causing 'severe' lucerne witches' broom could not be distinguished from the legume little leaf type strain.

scholarly journals First Report of Phytoplasma Belonging to Apple Proliferation Group in Roses in Poland

Plant Disease ◽  
2004 ◽  
Vol 88 (11) ◽  
pp. 1283-1283 ◽  
Author(s):  
M. Kamińska ◽  
H. Śliwa
Keyword(s):  
Reference Strain ◽  
Rflp Analysis ◽  
The United States ◽  
Apple Proliferation ◽  
Specific Primer ◽  
Disease Symptoms ◽  
Aster Yellows ◽  
Pcr Products ◽  
Aster Yellows Phytoplasma ◽  
Reference Strains

Virus-like diseases of rose plants of uncertain aetiology have been widely distributed throughout the world (4). Symptoms include dieback, rose rosette, witches' broom, and bud proliferation. Research conducted in England (4) and the United States (1) could not reveal the etiology of the diseases. Disease symptoms including stunted growth, leaf and flower malformation, and shoot and flower proliferation were observed in rose plants in Poland (2). In this previous work, we reported cases of phytoplasma closely related to group 16SrI in rose plants with shoot proliferation as determined by nested polymerase chain reaction (PCR) with ribosomal primers R16F1/R0 followed by rA/fA or R16(I)F1/R1. In this study, we examined 48 symptomatic rose plants of 12 cultivars using nested PCR primed by P1/P7 and followed by universal primer pairs R16F2n/R2, fA/rA, or group 16SrI-specific R16(I)F1/R1. To detect potential mixed infection in roses, the group 16SrX-specific primer pairs fAT/rAS, fAT/rPRUS, and fPD/rAT were used for nested PCRs. Samples of rose plants with disease symptoms and nonsymptomatic, samples of Catharanthus roseus, healthy and inoculated by grafting with the reference strain of aster yellows phytoplasma (AY1, group 16SrI-B, kindly supplied by I.-M. Lee, Beltsville, MD) and the reference strain of apple proliferation phytoplasma (AP, group 16SrX-A, kindly supplied by A. Bertaccini, Bologna, Italy), were tested for the presence of phytoplasma rDNA by PCR. Phytoplasma identification was accompanied by digestion with restriction endonucleases, AluI, HhaI, HpaII, MseI, and RsaI, and restriction fragment length polymorphism (RFLP) analysis of a 1.2-kb fragment of rDNA (3). A DNA amplification product was observed in all nested PCRs containing template DNA of samples collected from diseased roses and the reference strains but not from control plants. On the basis of RFLP analysis of PCR products and comparison of the RFLP patterns with those of the reference strains, we demonstrated the presence of aster yellows phytoplasma belonging to phytoplasma group 16SrI-B in roses of 11 cultivars. RFLP profile of samples collected from rose cv. Red Champ was identical to those obtained for reference AP strain (group 16SrX-A). Mixed RFLP profiles were observed in samples collected from rose cv. Memory, which were doubly infected by phytoplasmas belonging to groups 16SrI-B or 16SrX-A. These results were confirmed by PCR with group 16SrX-specific primer pairs. The target DNA was amplified when amplifications were conducted with subgroup 16SrX-A-specific primer pair fAt/rAS, whereas no observable PCR products were obtained with subgroup 16SrX-B- (fAT/rPRUS) or 16SrX-C- (fPD/rAT) specific primer pairs. This report confirms infection of roses by aster yellows phytoplasma belonging to group 16SrI-B, and to our knowledge, records for the first time, infection by phytoplasma of group 16SrX-A. References: (1) A. H. Epstein and J. H. Hill. J. Phytopathol. 143:353, 1995. (2) M. Kami ska et al. J. Phytopathol. 149:3, 2001. (3) I.-B. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1968. (4) B. J. Thomas. Ann. Rep. Glasshouse Crops Res. Inst. 1979:178, 1981.

scholarly journals Detection and identification of a novel 16SrXIII subgroup phytoplasma associated with strawberry red leaf disease in Argentina

2015 ◽  
Vol 65 (Pt_8) ◽  
pp. 2741-2747 ◽  
Author(s):  
Franco D. Fernández ◽  
Natalia G. Meneguzzi ◽  
Fabiana A. Guzmán ◽  
Daniel S. Kirschbaum ◽  
Vilma C. Conci ◽  
...  
Keyword(s):  
Rflp Analysis ◽  
Rrna Gene ◽  
The Novel ◽  
Universal Primer ◽  
Mature Leaves ◽  
Leaf Disease ◽  
Detection And Identification ◽  
Rflp Profile ◽  
Young Leaves ◽  
Leaf Symptoms

Strawberry red leaf phytoplasma was found in strawberry plants from production fields in Lules (Tucumán province) and Bella Vista (Corrientes province), Argentina. Characteristic strawberry red leaf symptoms were stunting, young leaves with yellowing at the edges, mature leaves which curled and were reddish at the abaxial face, flower and fruit deformation and death. The pathogen was detected with phytoplasma-universal primer pairs P1/P7 followed by R16F2n/R16R2 as nested primers in 13 diseased plants. Based on RFLP and sequence analysis of the amplified 16S rRNA gene, the phytoplasma was related to the 16SrXIII group (Mexican periwinkle virescence). In silico the RFLP profile of all the samples analysed revealed the presence of a unique pattern, showing that the novel phytoplasma is different from all the phytoplasmas currently composing the 16SrXIII group. The phylogenetic analysis was consistent with RFLP analysis as the strawberry red leaf phytoplasma was grouped within the 16SrXIII group, but formed a particular cluster. On this basis, the Strawberry red leaf phytoplasma associated with strawberry red leaf disease was assigned to a new subgroup, 16SrXIII-F.

Phenotypic and molecular characterization of chickpea rhizobia isolated from different areas of Tunisia

2007 ◽  
Vol 53 (3) ◽  
pp. 427-434 ◽  
Author(s):  
Boulbaba L’taief ◽  
Bouaziz Sifi ◽  
Maher Gtari ◽  
Mainassara Zaman-Allah ◽  
Mokhtar Lachaâl
Keyword(s):  
Rflp Analysis ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Phenotypic Characteristics ◽  
Cicer Arietinum L ◽  
Mesorhizobium Ciceri ◽  
Length Polymorphisms ◽  
Reference Strains ◽  
Tolerance To Salinity

Several phenotypic markers were used in this study to determine the biodiversity of rhizobial strains nodulating Cicer arietinum L. in various areas of Tunisia. They include symbiotic traits, the use of 21 biochemical substrates, and tolerance to salinity and pH. In addition, restriction fragment length polymorphisms (RFLPs) of PCR-amplified 16S rDNA were compared with those of reference strains. Numeric analysis of the phenotypic characteristics showed that the 48 strains studied fell into three distinct groups. This heterogeneity was highly supported by the RFLP analysis of 16S rRNA genes, and two ribotypes were identified. Chickpea rhizobia isolated from Tunisian soils are both phenotypically and genetically diverse. Results showed that 40 and 8 isolates were assigned, respectively, to Mesorhizobium ciceri and Mesorhizobium mediterraneum .

Molecular variation in plant cell populations evolving in vitro in different physiological contexts

Genome ◽  
2001 ◽  
Vol 44 (4) ◽  
pp. 549-558 ◽  
Author(s):  
Patrizia Bogani ◽  
Alessandra Simoni ◽  
Pietro Lio' ◽  
Angela Germinario ◽  
Marcello Buiatti
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Rflp Analysis ◽  
Enzymatic Digestion ◽  
Noncoding Dna ◽  
Noncoding Regions ◽  
Total Dna ◽  
Context Specific ◽  
Hormone Control ◽  
Selection Of

Previous work has shown the fixation of context-specific random amplified polymorphic DNA (RAPD) patterns in tomato cell cultures grown for 2 years in different hormonal contexts. In this work, RAPD sequences were characterised and RAPD-derived molecular markers used for a further study of variation between and within auto- and auxo-trophic tomato cultures grown in different hormonal equilibria. Results were then compared with those obtained using microsatellite markers located in noncoding regions of differentiation- and hormone-related genes and with those obtained with the external transcribed spacer (ETS) from tomato rDNA. Hybridisation of RAPDs on a tomato genomic DNA bank, or on total DNA after enzymatic digestion, suggested that the markers were repetitive in nature. Sequence analysis, however, showed that the homology between different fragments was due mainly to the presence of homo-AT nucleotide stretches. Moreover, a series of computational methods, such as an information-theory algorithm coupled with ΔG estimates, suggested that the RAPD fragments isolated in our experiments are noncoding. The amplification of SSR-containing RAPD-derived markers, and of other SSRs located in noncoding regions of tomato functional genes, consistently showed polymorphism between auxo- and auto-trophic somaclones (the latter being either habituated or transgenic for Agrobacterium tumefaciens oncogenes) but not within these same clones. Differences were also found between auxotrophic clones and the differentiated tissue. These findings were confirmed by restriction fragment length polymorphism (RFLP) analysis with the REII repetitive element of the ETS from tomato rDNA, which was isolated during this study. The results obtained suggest a possible role for physiological context in the selection of RAPD patterns during the evolution of tomato cells with different endogenous hormonal equilibria. The results are discussed in terms of a possible role for variation in noncoding regions of hormone-related genes in the adaptation to different physiological contexts.Key words: Lycopersicon esculentum, RAPD, SSR, genetic variation, noncoding DNA, hormone control.

scholarly journals First Report of Clover Proliferation Phytoplasma in Strawberry

Plant Disease ◽  
1999 ◽  
Vol 83 (10) ◽  
pp. 967-967 ◽  
Author(s):  
R. Jomantiene ◽  
J. L. Maas ◽  
E. L. Dally ◽  
R. E. Davis ◽  
J. D. Postman
Keyword(s):  
Dna Sequences ◽  
Rrna Gene ◽  
Universal Primer ◽  
Disease Symptoms ◽  
First Report ◽  
Chain Reactions ◽  
Polymerase Chain Reactions ◽  
Fragaria Virginiana ◽  
Phytoplasma Infection ◽  
Group 2

In 1996, diseased plants of Fragaria virginiana Duchesne were collected from a native population in Quebec, Canada, and sent to the National Clonal Germplasm Repository in Corvallis, OR, where grafting onto disease-free plants of F. chiloensis (L.) Duchesne (4) was performed. Plants of both species were sent to Beltsville, MD, for identification of a phytoplasma possibly associated with the disease symptoms of dwarfing and multibranching crowns. A phytoplasma was found in both species and characterized as the strawberry “multicipita” (SM) phytoplasma, which is representative of subgroup 16SrVI-B, a new subgroup of the clover proliferation (CP) group (2). In 1999, we observed commercial strawberry (Fragaria × ananassa Duchesne) plants collected in California and Maryland that were stunted and chlorotic or exhibited these symptoms in addition to small, distorted leaves. Infected F. × ananassa plants, as well as diseased F. virginiana and grafted F. chiloensis plants previously infected by the SM phytoplasma, were assessed for phytoplasma infection by nested polymerase chain reactions primed by phytoplasma universal primer pairs R16mF2/R1 and F2n/R2 (1) or P1/P7 (3) and F2n/R2 for amplification of phytoplasma 16S rDNA (16S rRNA gene) sequences. Phytoplasma-characteristic 1.2-kbp DNA sequences were amplified from all diseased plants. No DNA sequences were amplified from healthy plants. Restriction fragment length polymorphism patterns of rDNA digested with AluI, KpnI, HhaI, HaeIII, HinfI, HpaII, MseI, RsaI, and Sau3A1 endonucleases indicated that all plants were infected by a phytoplasma that belonged to subgroup 16SrVI-A (CP phytoplasma subgroup) and that diseased F. virginiana and grafted F. chiloensis plants were infected by both SM and CP. This is the first report of the CP phytoplasma, subgroup 16SrVI-A, infecting strawberry. This report also indicates that the occurrence of the CP phytoplasma in strawberry may be widespread in North America and that F. chiloensis, F. virginiana, and F. × ananassa plants are susceptible to infection by the CP phytoplasma. References: (1) D. E. Gunderson and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (2) R. Jomantiene et al. HortScience 33:1069, 1998. (3) R. Jomantiene et al. Int. J. Syst. Bacteriol. 48:269, 1998. (4) J. D. Postman et al. Acta Hortic. 471:25, 1998.

scholarly journals A Novel Source of Resistance to Tomato Yellow Leaf Curl Virus Exhibiting a Symptomless Reaction to Viral Infection

1998 ◽  
Vol 123 (6) ◽  
pp. 1004-1007 ◽  
Author(s):  
M. Friedmann ◽  
M. Lapidot ◽  
S. Cohen ◽  
M. Pilowsky
Keyword(s):  
Natural Infection ◽  
Tomato Yellow Leaf ◽  
Yellow Leaf ◽  
Susceptible Parent ◽  
Partial Dominance ◽  
Disease Symptoms ◽  
Leaf Curl Virus ◽  
Very High ◽  
Different Levels ◽  
Leaf Curl

Tomato yellow leaf curl virus (TYLCV), transmitted by the tobacco whitefy (Bemisia tabaci Genn.), can be devastating to tomato (Lycopersicon esculentum L.) crops in tropical and subtropical regions. The development of resistant cultivars is the best option for control of TYLCV. However, all the available resistant commercial cultivars tested at the Volcani Center, when inoculated with TYLCV, developed different levels of disease symptoms. In this study, we report the development of a breeding line, TY172, which is a symptomless carrier of TYLCV. Line TY172, whether infected in the greenhouse with viruliferous whiteflies, or when grown in the field under natural infection, showed no symptoms of the disease. Viral DNA was detected in infected TY172 plants, albeit at much lower levels than a susceptible infected control. In addition, grafting experiments using infected susceptible scions grafted onto TY172 stocks, showed that even when exposed continuously to very high levels of virus, line TY172 did not develop disease symptoms, nor did it accumulate high levels of the virus. When TY172 was crossed with susceptible lines, the hybrids exhibited milder symptoms and lower viral content than the susceptible parent, yet higher than that of TY172, suggesting a partial dominance for the TY172 resistance. Upon inoculation of F2 populations, the amount of symptomless individuals appeared in a ratio of≈7:64. This suggests that at least three genes may account for the resistance.

scholarly journals Detection, in silico analysis and molecular diversity of phytoplasmas from solanaceous crops in Turkey

2021 ◽  
Vol 58 (No. 1) ◽  
pp. 31-39
Author(s):  
Mustafa Usta ◽  
Abdullah Güller ◽  
Hikmet Murat Sipahioglu
Keyword(s):  
Geographical Area ◽  
Solanum Melongena ◽  
In Silico Analysis ◽  
Rflp Analysis ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Natural Occurrence ◽  
Similarity Coefficients ◽  
Universal Primer ◽  
Virtual Rflp

Phytoplasma-like symptoms of leaf yellowing and calyx malformation were observed in eggplant (Solanum melongena L.), upward leaves and fruit malformation in pepper (Capsicum annuum L.), and aerial tuber formation in potato (S. tuberosum L.) during the survey performed in the late season (August to September) of 2015 and 2016 in Van province (Turkey). A total of 100 samples were tested by nested-PCR using universal primer pairs to assess the sanitary status of the solanaceous crops and to characterise the phytoplasma isolates. Among them, seven samples resulted in a 1.25 kb DNA fragment, and five (two eggplants, two peppers, and one potato) were molecularly characterised (Accession No.: KY579357, KT595210, MF564267, MF564266, and MH683601). BLAST and the virtual restriction fragment length polymorphism (RFLP) analysis of 16S rRNA genes revealed the presence of two distinct phytoplasma infections in solanaceous crops: ‘Candidatus Phytoplasma trifolii’ a member of the clover proliferation group (16SrVI) and subgroup A and ‘Candidatus P. solani’ a member of the stolbur group (16SrXII) and subgroup A. The virtual RFLP analysis and calculated coefficients of RFLP pattern similarities further revealed a remarkable genetic diversity among the ‘Candidatus P. solani’ isolates infecting pepper (similarity coefficient of 0.90) and eggplant (similarity coefficients of 0.98 and 1.00) at the same geographical area. This is the first report of the natural occurrence of ‘Candidadtus P. trifolii’ in potato from the Eastern Anatolia region, Turkey.

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