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2020 ◽  
Vol 3 (1) ◽  
pp. 81
Author(s):  
Algirdas Ivanauskas ◽  
Jolanta Rimsaite ◽  
Jurij Danilov ◽  
Guy Soderman ◽  
Donatas Sneideris ◽  
...  

Mountain pine (Pinus mugo Turra) is a coniferous native to the highlands of central Europe. Our previous study revealed that mountain pine proliferation decline (MPPD) disease in the Curonian Spit of Lithuania is caused by a ‘Candidatus Phytoplasma pini’-related strain (16SrXXI-A). However, the insect vector of MPPD has not been identified. In this study, we conducted a survey to determine potential insect vectors of MPPD phytoplasma for three consecutive years (2016–2019). More than 1000 insect samples were collected from four locations in the Curonian Spit. These insects were identified as belonging to six families and ten genera. The presence of phytoplasma in insect samples was examined by nested polymerase chain reaction (PCR) using phytoplasma-specific primers (P1A/16S-SR and R16F2n/R16R2n). Phytoplasmas were detected in Cinara (Cinara) pini (Scots pine aphid), Cinara (Cinara) piniphila and Cinara (Schizolachnus) pineti (waxy grey pine needle aphid) insect samples. Subsequent restriction fragment length polymorphism (RFLP) analysis showed that the PCR-RFLP profile of these positive insect samples was consistent with that of the MPPD of diseased pine trees. These results suggest that C. (C.) pini, C. (C.) piniphila and C. (S.) pineti may be potential insect vectors of MPPD phytoplasma. The findings from this survey will provide useful information for the management of MPPD disease.


2020 ◽  
Vol 59 (1) ◽  
pp. 55-61
Author(s):  
Ghobad BABAEI ◽  
Seyyed Alireza ESMAEILZADEH-HOSSEINI ◽  
Mahbobeh ZANDIAN ◽  
Vahid NIKBAKHT

Phytoplasma symptoms, including proliferation, witches’ broom, leaf rolling and yellowing, were observed in jujube (Ziziphus jujube) nurseries in the East of Iran. Total nucleic acid was extracted from symptomatic and symptomless plants, and was tested for phytoplasma presence using nested PCR. Amplicons of about 1.8 kb (primer pair P1/P7) and 1.25 kb (R16F2n/R16R2) were obtained from all symptomatic plants but not from symptomless plants. Restriction fragment length polymorphism (RFLP) analysis of R16F2n/R2 amplicons using KpnI, HaeIII, RsaI, AluI, HpaII, HhaI, TaqI, MseI, BfaI and ThaI restriction enzymes showed two RFLP patterns referable to 16SrI and 16SrVI phytoplasma groups. The consensus sequences of Z. jujube yellowing and witches’ broom of six samples correspond to ‘Candidatus Phytoplasma asteris’ and ‘Candidatus Phytoplasma trifolii’-related strains. Two R16F2n/R16R2 16S rDNA sequences representative of each RFLP profile, one each from witches’ broom (accession number MK379605) and yellowing (MK379604) host symptoms, were submitted to the GenBank. Phylogenetic analysis confirmed that the phytoplasma strains associated with jujube yellowing clustered within the 16SrI phytoplasma clade, and those associated with witches’ broom clustered within the 16SrVI clade. Restriction analysis confirmed that virtual RFLP patterns of the jujube yellowing and witches’ broom phytoplasma strains were identical to the reference pattern of 16SrI-B and 16SrVI-A. This is the first report of these phytoplasma strains associations with witches’ broom and yellowing in jujube plants.


Author(s):  
Hossein Meghdadi ◽  
Azar Dokht Khosravi ◽  
Ahmad Farajzadeh Sheikh ◽  
Ameneh Alami ◽  
Nerssy Nassirabady

Background and Objectives: Due to the widespread distribution of Listeria monocytogenes in environmental and animal sources and serious clinical complications in human, this study was aimed to isolate L. monocytogenes from water and clin- ical specimens by culture and PCR methods and to investigate the presence of hlyA and inlA virulence genes. Materials and Methods: Water and clinical samples of vaginal and fecal were screened for the presence of L. monocyto- genes by phenotypic and standard biochemical tests. PCR amplification was performed on extracted DNA using primers based on the hlyA and inlA genes. A 733-bp fragment of inlA gene was used for investigation of polymorphism using RFLP analysis. Results: In total, 45 phenotypically and molecularly confirmed L. monocytogenes strains were isolated from different sourc- es including 30 (16.7%) from water, 9 (11.3%) from vaginal swabs and 6 (7.5%) from fecal samples. RFLP analysis of PCR products using AluI and Tsp509I restriction enzymes, generated two profiles with 8 to 10 bands ranging in size from 15 to 210 bp. The majority of water and clinical isolates were classified in profile 2. Conclusion: We demonstrated 45 L. monocytogenes isolates from tested water and clinical samples by phenotypic and mo- lecular tests. The majority of the isolates were classified in the same RFLP profile, showing the water as a potential source of clinical complications in patients in the region of study.


Nova Hedwigia ◽  
2016 ◽  
Vol 103 (3) ◽  
pp. 475-490
Author(s):  
Yeny Meza-Meneses ◽  
Gema Galindo Flores ◽  
José Luis Martínez y Pérez ◽  
Arturo Estrada-Torres ◽  
Laura V. Hernández-Cuevas ◽  
...  

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 ◽  
...  

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.


2014 ◽  
Vol 2014 ◽  
pp. 1-10 ◽  
Author(s):  
Reza Shafiei ◽  
Bahador Sarkari ◽  
Seyed Mahmuod Sadjjadi ◽  
Gholam Reza Mowlavi ◽  
Abdolali Moshfe

The current study aimed to find out the morphometric and genotypic divergences of the flukes isolated from different hosts in a newly emerging focus of human fascioliasis in Iran. AdultFasciolaspp. were collected from 34 cattle, 13 sheep, and 11 goats from Kohgiluyeh and Boyer-Ahmad province, southwest of Iran. Genomic DNA was extracted from the flukes and PCR-RFLP was used to characterize the isolates. The ITS1, ITS2, and mitochondrial genes (mtDNA) of NDI and COI from individual liver flukes were amplified and the amplicons were sequenced. Genetic variation within and between the species was evaluated by comparing the sequences. Moreover, morphometric characteristics of flukes were measured through a computer image analysis system. Based on RFLP profile, from the total of 58 isolates, 41 isolates (from cattle, sheep, and goat) were identified asFasciola hepatica, while 17 isolates from cattle were identified asFasciola gigantica. Comparison of the ITS1 and ITS2 sequences showed six and seven single-base substitutions, resulting in segregation of the specimens into two different genotypes. The sequences of COI markers showed seven DNA polymorphic sites forF. hepaticaand 35 DNA polymorphic sites forF. gigantica. Morphological diversity of the two species was observed in linear, ratios, and areas measurements. The findings have implications for studying the population genetics, epidemiology, and control of the disease.


Author(s):  
Vartika Chandra ◽  
B. B. Bhanderi ◽  
R. A. Mathakiya ◽  
M. K. Jhala
Keyword(s):  
Rt Pcr ◽  

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Bijan Esmaeilnejad ◽  
Mousa Tavassoli ◽  
Siamak Asri-Rezaei ◽  
Bahram Dalir-Naghadeh ◽  
Karim Mardani ◽  
...  

This study aimed to assess the prevalence ofBabesia ovisinfection in adultRhipicephalus bursaand small ruminants in West Azerbaijan province, Iran. Blood samples were collected from 280 sheep and 122 goats of forty randomly selected flocks. SpecificB. ovisfragment was detected in 67 animals (16.7%), of which 52 animals (18.6%) were sheep and 15 animals (12.2%) goats (P<0.05). Of the 848R. bursacollected from naturally infested small ruminants and farm dogs,Babesia oviswas detected by PCR in salivary glands of 94 adult ticks. The frequency ofB. ovisinfection was higher in flocks with tick in comparison with animals without tick (P<0.05). Positive amplification from blood of ruminants, ticks, oviposition ticks, eggs, and larvae was subjected to restriction digestion withHphI. One RFLP profile was produced. The PCR-RFLP results indicated that one strain ofB. ovisexists in this area. The results showed that the PCR was useful method to investigate the epidemiology of small ruminants’ babesiosis. Furthermore,R. Bursa, which can transovarially transmitB. ovisand as well as being widely distributed in West Azerbaijan province, Iran, might play an important role in the field as a natural vector ofB. ovis.


Plant Disease ◽  
2013 ◽  
Vol 97 (6) ◽  
pp. 835-835 ◽  
Author(s):  
M. Catal ◽  
C. Ikten ◽  
E. Yol ◽  
R. Üstün ◽  
B. Uzun

Sesame (Sesamum indicum L.) is an important oilseed crops widely grown in the southern regions of Turkey. Sesame seeds are primarily used in production of tahini as well as a garnish on sweets and bakery products in the country. Sesame plants with phyllody disease symptoms have increasingly been observed in the fields of Antalya province since 2007. The disease incidence in these fields was found to range from 37 to 62% (2). Infected plants display a variety of the disease symptoms such as virescence, asymptomatic shoot proliferation, infertile flower formation, reduced leaf size, and thin and weak capsule development. Total genomic DNA was extracted from samples collected from symptomatic (10 plants) and asymptomatic healthy-looking plants (10 plants) using a CTAB method and amplified with universal primers P1/P7 and R16F2n/R16R2 in direct and nested PCR, respectively (1,3). Amplifications of the DNA from the symptomatic plants yielded a product of 1.8 kb in direct and 1.2 kb in nested PCR assays. No amplification was observed in symptomless plants of the same age and collected from the same fields. Amplicons were purified, cloned in a pTZ57R/T Vector, and sequenced using a Beckman Coulter 8000 CEQ Genetic Analysis System. Four aligned 16S rDNA sequences (1,845 bp) were found to be all identical and belonging to one species. One sequence was deposited in GenBank under the accession number KC139791. A BLAST similarity search revealed that the sequence shared 99% homology with the sequences of the members of 16SrIX group phytoplasmas, ‘Brassica rapa’ phyllody phytoplasma (HM559246.1) and Iranian Almond witches'-broom phytoplasma (DQ195209.1) available in GenBank. In addition, iPhyClassifier software (4) was employed to create a virtual RFLP profile. The analysis showed that the RFLP profile of the sesame phytoplasma 16S rDNA sequence is identical (a similarity coefficient of 1.00) to the profile of the 16Sr group IX phytoplasma reference sequence (Y16389). A phylogenetic tree was also constructed using the neighbor joining plot option of the Clustal X program. The sequence clustered together with 16SrIX group phytoplasmas. To our knowledge, this is the first report of a natural infection of sesame by a new phytoplasma species from the 16SrIX group in Turkey. References: (1) D. E. Gundersen and I.-M. Lee. Phytopathol. Mediterr. 35:144, 1996. (2) C. Ikten et al. Phytopathogenic Mollicutes 1:101, 2011. (3) C. D. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996. (4) Y. Zhao et al. Int. J. Syst. Evol. Microbiol. 59:2582, 2009.


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