Molecular characterization of stolbur phytoplasmas in pepper and tomato from Bulgaria

Reference Strain ◽

Rflp Analysis ◽

Plant Diseases ◽

Vegetable Crops ◽

Disease Symptoms ◽

Natural Plant ◽

Conserved Genes ◽

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.

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

Plant Disease ◽

10.1094/pdis.2004.88.11.1283a ◽

2004 ◽

Vol 88 (11) ◽

pp. 1283-1283 ◽

Cited By ~ 9

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 ◽

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.

Molecular Characterization of Pathogenicity Locus (PaLoc) and tcdC Genetic Diversity Among tcdA+B+ Clostridioides Difficile Clinical Isolates in Tehran, Iran

10.21203/rs.3.rs-56651/v1 ◽

2020 ◽

Author(s):

Mansoor Kodori ◽

Zohreh Ghalavand ◽

Abbas Yadegar ◽

Gita Eslami ◽

Masoumeh Azimirad ◽

...

Keyword(s):

Genetic Diversity ◽

Disease Severity ◽

Genetic Variants ◽

Rflp Analysis ◽

Clostridioides Difficile ◽

Pcr Rflp ◽

The Common ◽

Healthcare Associated

Abstract Background: Clostridioides difficile is the main cause of healthcare-associated diarrhea worldwide. It is proposed that certain C. difficile toxinotypes with distinct pathogenicity locus (PaLoc) variants are associated with disease severity and outcomes. Additionally, few studies have described the common C. difficile toxinotypes, and also little is known about the tcdC variants in Iranian isolates. We characterized the toxinotypes and the tcdC genotypes from a collection of Iranian clinical C. difficile tcdA+B+ isolates with known ribotypes (RTs).Methods: Fifty C. difficile isolates with known RTs and carrying the tcdA and tcdB toxin genes were analyzed. Toxinotyping was carried out based on a PCR-RFLP analysis of a 19.6 kb region encompassing the PaLoc. Genetic diversity of the tcdC gene was determined by the sequencing of the gene.Results: Of the 50 C. difficile isolates investigated, five distinct toxinotypes were recognized. Toxinotypes 0 (33/50, 66%) and V (11/50, 22%) were the most frequently found. C. difficile isolates of the toxinotype 0 mostly belonged to RT 001 (12/33, 36.4%), whereas toxinotype V consisted of RT 126 (9/11, 81.8%). The tcdC sequencing showed six variants (35/50, 70%); tcdC-sc3 (24%), tcdC-A (22%), tcdC-sc9 (18%), tcdC-B (2%), tcdC-sc14 (2%), and tcdC-sc15 (2%). The remaining isolates were wild-types (15/50, 30%) in the tcdC gene.Conclusions: The present study demonstrates that the majority of clinical tcdA+B+ isolates of C. difficile frequently harbor tcdC genetic variants. We also found that the RT 001/ toxinotype 0 and the RT 126/ toxinotype V are the most common types among Iranian isolates. Further studies are needed to investigate the putative association of various tcdC genotypes with CDI severity and its recurrence.

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

Canadian Journal of Microbiology ◽

10.1139/w06-127 ◽

2007 ◽

Vol 53 (3) ◽

pp. 427-434 ◽

Cited By ~ 17

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 .

Phenotypic And Genotypic Characterization

Phenotypic and genotypic characterization of Xanthomonas campestris strains isolated from cabbage, kale and broccoli

Archives of Biological Sciences ◽

10.2298/abs1302585p ◽

2013 ◽

Vol 65 (2) ◽

pp. 585-593 ◽

Cited By ~ 4

Author(s):

Tatjana Popovic ◽

Dragana Josic ◽

Mira Starovic ◽

P. Milovanovic ◽

N. Dolovac ◽

...

Keyword(s):

16S Rdna ◽

Xanthomonas Campestris ◽

Reference Strain ◽

Tween 80 ◽

Genotypic Characterization ◽

Hypodermic Syringe ◽

Xanthomonas Campestris Pv Campestris ◽

Reference Strains ◽

Thirty-six strains of Xanthomonas campestris pv. campestris (Xcc) isolated from cabbage, kale and broccoli were identified according to their pathogenicity, phenotypic and genotypic characterization. Pathogenicity was confirmed by the injection method with a hypodermic syringe into the mesophilic tissue of cabbage leaves. All strains were Gramnegative, aerobic, catalase-positive, oxidase-negative, grew at 35?C, produced levan, H2S and indole, did not reduce nitrate, hydrolyzed Tween 80, starch, gelatin and esculin and did not show tolerance to 0.1 and 0.02% TTC. The strains produced acid from d-arabinose, arginine, dulcitol, galactose, d-glucose, maltose, mannose, sorbitol, sucrose and xylose. The genetic characterization was based on the sequence analyses of 16S rDNA and ERIC and BOX PCR. Strains of different pathovars were also used to compare PCR resulting patterns. BOX-PCR of the strains from kale and broccoli, obtained using (GTG)5 primer, yielded patterns with a high similarity level to pathovar reference strain Xcc. The strains from cabbage yielded BOX and ERIC product patterns, distinguishing them from the other tested strains and reference strains. 16S rDNA of the representative strains was closely related to Xcc strain ATCC 33913. ERIC PCR and BOX using (GTG)5 primer generated different Xcc patterns and were effective in distinguishing strains from different plant hosts.

Characterization of Three Species of Thrips on Weeping Fig, Nutmeg, and Marine Seruni Plants Based on Mtcoi DNA Sequences

Biosaintifika Journal of Biology & Biology Education ◽

10.15294/biosaintifika.v8i2.5448 ◽

2016 ◽

Vol 8 (2) ◽

pp. 185

Author(s):

Nia Kurniawaty ◽

Purnama Hidayat ◽

Aunu Rauf

Keyword(s):

Pest Management ◽

Dna Sequences ◽

Biology Education ◽

Morphological Data ◽

Coi Gene ◽

Molecular Characteristics ◽

Vegetable Crops ◽

Total Extraction

Thrips are widely reported as pests in vegetable crops. However, the existence of Phlaeothripidae members has a less concern in Indonesia. Phlaeothripidae is the only family of Tubulifera Suborder and some reports suggested that they had potential to be pests in several crops due to their ability to roll up and to make galls on leaves. The first step in pest management attempt is to identify the pest accurately and quickly, so the pest management can be on target and more efficient. One of the identification methods is the molecular identification using DNA barcoding techniques. This study aimed to characterize and to compare species thrips in banyan, nutmeg, and marine seruni based on their molecular characteristics. This research was conducted in Bogor and Kuningan. The process of molecular characterization consisteds of four steps DNA total extraction, amplification by using PCR, COI gene sequence, and data analysis. PCR programme was succesfully to amplified mtCOI gene fragment at 710 bp. The length of mtCOI gene of Gynaikothrips uzeli, Haplothrips ganglbaueri, and Pseudophilothrips ichini were 704, 686, and 702 bp dominated by A and T bases with nucleotide variation value of 27.8%. This results confirmed that molecular characterization using mtCOI gene mitochondrial had successfully supported the morphological data. How to CiteKurniawaty, N., Hidayat, P. & Rauf, A. (2016). Characterization of Three Species of Thrips on Banyan, Nutmeg, and Marine Seruni Plants Based on Coi Gene. Biosaintifika: Journal of Biology & Biology Education, 8(2), 185-192.

First Report of Phytoplasma Infection in Hop Plants

Plant Disease ◽

10.1094/pdis.2004.88.8.908b ◽

2004 ◽

Vol 88 (8) ◽

pp. 908-908 ◽

Cited By ~ 1

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 ◽

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.

Isolation and characterization of Bacillus endophytic strains producers for non ribosomal lipopeptides NRLPs from tomato

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v9i1.1209 ◽

2018 ◽

Vol 9 (1) ◽

pp. 128

Author(s):

Walaa Hussein ◽

Ramadan WA ◽

Sameh Fahim

Keyword(s):

High Performance ◽

Plant Defence ◽

Plant Diseases ◽

Bacillus Species ◽

Vegetable Crops ◽

Defence Mechanisms ◽

Isolation And Characterization ◽

Genes Encoding ◽

Plant Defence Mechanisms

Tomato (Solanum lycopersicum) are considered one of the most important vegetable crops and infected by numbers of different diseases. Studying the use of biological alternatives, instead of chemical substances against plant diseases became necessary for the treatment by beneficial microorganisms endophytes, which can excrete natural products benefits to plant in reducing disease severity, promoting growth and inducing plant defence mechanisms. In this work, three endophytes strains were isolated from tomato stems and their 16srDNA have been found to belong to Bacillus species. The first strain was named BMG100, the second BMG101 and the third BMG102. Two Bacillus strains BMG100 and BMG101 have been found to harbour synthetases genes from three lipopeptides families; surfactin, plipastatin, and iturin (mycosubtilin) which can be detected by degenerated primers designed to detect the presence of synthetases genes encoding lipopeptides. The lipopeptides production was proved by their quantification using High Performance Liquid Chromatography (HPLC), whereas BMG100 produced 105, 178 and 293 mg/L of plipastatin, mycosubtilin and surfactin, respectively, BMG101 produced 385 mg/L of surfactin and 236 mg/L of mycosubtilin, while BMG102 showed no lipopeptides production. Keywords: Tomato; Endophytic bacteria; Lipopeptides; Bacillus species

Molecular characterization of monocrotophos and chlorpyrifos tolerant bacterial strain for enhancing seed germination of vegetable crops

Chemosphere ◽

10.1016/j.chemosphere.2019.02.053 ◽

2019 ◽

Vol 223 ◽

pp. 636-650 ◽

Cited By ~ 7

Author(s):

Durgesh Kumar Jaiswal ◽

Jay Prakash Verma ◽

Ram Krishna ◽

Anand Kumar Gaurav ◽

Janardan Yadav

Keyword(s):

Seed Germination ◽

Bacterial Strain ◽

Vegetable Crops

Application of Siderotyping for Characterization of Pseudomonas tolaasii and “Pseudomonas reactans” Isolates Associated with Brown Blotch Disease of Cultivated Mushrooms

Applied and Environmental Microbiology ◽

10.1128/aem.66.11.4834-4841.2000 ◽

2000 ◽

Vol 66 (11) ◽

pp. 4834-4841 ◽

Cited By ~ 38

Author(s):

Patricia Munsch ◽

Valerie A. Geoffroy ◽

Tapani Alatossava ◽

Jean-Marie Meyer

Keyword(s):

Isoelectric Focusing ◽

Agaricus Bisporus ◽

White Line ◽

Disease Symptoms ◽

Line Test ◽

Brown Blotch ◽

Uptake Studies ◽

Brown Blotch Disease ◽

ABSTRACT Pyoverdine isoelectric focusing analysis and pyoverdine-mediated iron uptake were used as siderotyping methods to analyze a collection of 57 northern and central European isolates of P. tolaasiiand “P. reactans.” The bacteria, isolated from cultivated Agaricus bisporus or Pleurotus ostreatus mushroom sporophores presenting brown blotch disease symptoms, were identified according to the white line test (W. C. Wong and T. F. Preece, J. Appl. Bacteriol. 47:401–407, 1979) and their pathogenicity towards A. bisporus and were grouped into siderovars according to the type of pyoverdine they produced. Seventeen P. tolaasii isolates were recognized, which divided into two siderovars, with the first one containing reference strains and isolates of various geographical origins while the second one contained Finnish isolates exclusively. The 40 “P. reactans” isolates divided into eight siderovars. Pyoverdine isoelectric focusing profiles and cross-uptake studies demonstrated an identity for some “P. reactans” isolates, with reference strains belonging to theP. fluorescens biovars II, III, or V. Thus, the easy and rapid methods of siderotyping proved to be reliable by supporting and strengthening previous taxonomical data. Moreover, two potentially novel pyoverdines characterizing one P. tolaasiisiderovar and one “P. reactans” siderovar were found.

Molecular characterization of Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes) isolates

Scientia Agricola ◽

10.1590/s0103-90162000000400021 ◽

2000 ◽

Vol 57 (4) ◽

pp. 729-732 ◽

Cited By ~ 16

Author(s):

Andréia Cristiane Souza Azevedo ◽

Márcia Cristina Furlaneto ◽

Daniel Ricardo Sosa-Gómez ◽

Maria Helena Pelegrinelli Fungaro

Keyword(s):

Genetic Similarity ◽

Paecilomyces Fumosoroseus ◽

Base Pairs ◽

Restriction Profiles ◽

ITS and RAPD analyses were used to investigate molecular variations within samples of Paecilomyces isolates and to resolve five morphologically atypical isolates resembling P. fumosorosus, obtained from whitefly in Northern Paraná State. The ITS4-ITS5 amplicon was 700 base pairs (bp) long in all isolates. The five isolates of Paecilomyces not assigned to species produced restriction profiles identical to all the reference strains of P. fumosoroseus. The extent of fingerprint variability observed by RAPD was sufficient to discriminate all the isolates. The genetic similarity among unidentified isolates and strains of P. fumosoroseus was even higher than that observed among reference strains of this species, allowing us to conclude that isolates CNPso-P77, CNPso-P78, CNPso-P80, CNPso-P85 and CNPso-P91 are P. fumosoroseus.