scholarly journals Identification of a 16SrII-E Phytoplasma in Calendula arvensis, Solanum nigrum, and Chenopodium spp.

Plant Disease ◽  
2006 ◽  
Vol 90 (3) ◽  
pp. 325-330 ◽  
Author(s):  
G. Tolu ◽  
S. Botti ◽  
R. Garau ◽  
V. A. Prota ◽  
A. Sechi ◽  
...  
Keyword(s):  
Solanum Nigrum ◽  
Nested Polymerase Chain Reaction ◽  
Bois Noir ◽  
Gene Coding ◽  
Epidemiological Surveys ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Taxonomic Groups ◽  
Stolbur Phytoplasma

Epidemiological surveys were performed in Northern Sardinia (Italy) in a 10-year-old vineyard affected by “Bois noir” disease. Samples collected between May and October 2003 from chlorotic and stunted weeds belonging to 14 different taxonomic groups were indexed molecularly for detection of phytoplasmas. Nested polymerase chain reaction (PCR) assays using primers specific for the phytoplasma 16SrDNA gene showed three of six Calendula arvensis, one of two Solanum nigrum, and one of seven Chenopodium spp. assayed positive. Restriction fragment length polymorphism analyses and sequencing of amplified 16SrDNA fragments identified a putative phytoplasma in the ribosomal subgroup 16SrII-E. Further characterization of the rps3 gene, coding a ribosomal protein, confirmed the identification. However, the weeds and leafhop-per species collected in the vineyard tested negative by PCR assays for the Stolbur phytoplasma, the causal agent of “Bois noir”. This is the first report of a phytoplasma of the 16SrII-E subgroup infecting C. arvensis, S. nigrum, and Chenopodium spp.

Identification and characterization of a gene coding a novel isoform of DEAD-box protein

2002 ◽  
Vol 14 (3) ◽  
pp. 185 ◽  
Author(s):  
Lanlan Yin ◽  
JianMin Li ◽  
Hu Zhu ◽  
Min Lin ◽  
Lijun Cheng ◽  
...  
Keyword(s):  
Human Testis ◽  
Microarray Hybridization ◽  
Chain Reaction ◽  
Dead Box ◽  
Gene Coding ◽  
Testis Cdna Library ◽  
Testis Cdna ◽  
Polymerase Chain ◽  
Identification And Characterization

A gene coding a novel isoform of DEAD-box protein named testicular DEAD-box protein (tDbp), presumably involved in testicular function, was identified and characterized. Testicular DEAD-box protein was cloned from a human testis cDNA library. The cDNA microarray hybridization showed that it was expressed at a higher level in adult testis than in embryo testis. Reverse transcription-polymerase chain reaction indicated that tDbp was specifically expressed in testis, but not in some other tissues.

scholarly journals Characterization of phytoplasmas related to 'Candidatus Phytoplasma asteris' subgroup rpI-L in Iran

2014 ◽  
Vol 54 (2) ◽  
pp. 199-203 ◽  
Author(s):  
Fereshteh Vali-Sichani ◽  
Masoud Bahar ◽  
Leila Zirak
Keyword(s):  
Fragment Length Polymorphism ◽  
Specific Primer ◽  
Sequence Analyses ◽  
Chain Reaction ◽  
Universal Primer ◽  
Disease Symptoms ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Niger Seed

Abstract In two of Iran's central provinces, several herbaceous plants showing phytoplasma disease symptoms were collected to detect 'Canididatus Phytoplasma asteris'-related phytoplasmas. Confirmation of an association of phytoplasmas with diseased plants was done using polymerase chain reaction (PCR) assays having the phytoplasma universal primer pairs P1/P7 followed by R16F2n/ R16R2 in nested PCR. Then, for detection of 'Ca. P. asteris', DNA samples were subjected to amplification of rp and tuf genes using specific primer pairs rp(I)F1A/rp(I)R1A and fTufAy/rTufAy, respectively. Restriction fragment length polymorphism or RFLP analyses of rp gene fragments using Tsp509I restriction enzyme as well as sequence analyses indicated that 'Ca. P. asteris'-related phytoplasmas associated with carrot, niger seed and scallion plants in these regions, belong to the rpI-L subgroup. This research is the first report of carrot, niger seed, and scallion infection with phytoplasmas belonging to the rpI-L subgroup.

scholarly journals Molecular Characterization of Carbapenemase-Producing Escherichia coli and Salmonella in children with diarrhea in rural Burkina Faso

2020 ◽  
Author(s):  
Rene DEMBELE ◽  
Issiaka Soulama ◽  
Wendpoulomdé A. D. Kaboré ◽  
Ali Konaté ◽  
Assèta Kagambèga ◽  
...  
Keyword(s):  
Burkina Faso ◽  
Treatment Options ◽  
Diffusion Method ◽  
Chain Reaction ◽  
Disk Diffusion Method ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Resistance Rates ◽  
Encoding Genes

Abstract Background: In recent years, carbapenemase-producing Enterobacterales (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso. Methods: Salmonella isolates were serotyped according to the Kauffman White scheme. Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were carried out using oligonucleotides to detect the presence of genes of the blaKPC, blaVIM, blaIMP, blaTEM, blaSHV, blaOXA and blaCTX-M types in all E. coli and Salmonella strains.Results: The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E. coli were imipenem resistant with carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each in E. coli strains. However, no Salmonella carbapenemases blaKPC, blaVIM or blaIMP were detected.Conclusions: This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacterales in patients is crucial.

scholarly journals Endophytic Fungal Mutualists: Seed-Borne Stagonospora Spp. Enhance Reed Biomass Production in Axenic Microcosms

2003 ◽  
Vol 16 (7) ◽  
pp. 580-587 ◽  
Author(s):  
Michael Ernst ◽  
Kurt W. Mendgen ◽  
Stefan G. R. Wirsel
Keyword(s):  
Polymerase Chain Reaction ◽  
Phragmites Australis ◽  
Immunofluorescence Microscopy ◽  
Fungal Endophytes ◽  
Common Reed ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Surface Sterilization ◽  
Polymerase Chain ◽  
Pcr Assays

Fungal endophytes mainly belong to the phylum Ascomycota and colonize plants without producing symptoms. We report on the isolation of seed-borne fungal endophytes from Phragmites australis (common reed) that were ascribed to the genus Stagonospora. Nested polymerase chain reaction (PCR) assays revealed that a Stagonospora sp. regularly colonized reed as shown for a period of three years. In spring, it was only detected in roots, whereas in autumn, it could frequently be found in all organs, including seeds. Microcosm experiments revealed that seeds harbored viable propagules of the fungus that colonized the developing germling, indicating vertical transmission. Endophytic growth was confirmed by immunofluorescence microscopy, reisolation of the fungus after surface sterilization, and PCR. Aseptic microcosms were established for studying fungal contributions towards host vitality. Several Stagonospora isolates enhanced reed biomass. Seed-borne endophytic Stagonospora spp. thus can provide improved vigor to common reed, which could be most important when seed-derived germlings establish new reed stands.

scholarly journals Characterization of a Phytoplasma Associated with Frogskin Disease in Cassava

Plant Disease ◽  
2009 ◽  
Vol 93 (11) ◽  
pp. 1139-1145 ◽  
Author(s):  
Elizabeth Alvarez ◽  
Juan F. Mejía ◽  
Germán A. Llano ◽  
John B. Loke ◽  
Alberto Calari ◽  
...  
Keyword(s):  
Ribosomal Protein ◽  
Manihot Esculenta ◽  
Fragment Length Polymorphism ◽  
Rrna Gene ◽  
South American ◽  
Sequence Analyses ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain

Cassava frogskin disease (CFSD) is an economically important root disease of cassava (Manihot esculenta) in Colombia and other South American countries, including Brazil, Venezuela, Peru, Costa Rica, and Panama. The roots of severely affected plants are thin, making them unsuitable for consumption. In Colombia, phytoplasma infections were confirmed in 35 of 39 genotypes exhibiting mild or severe CFSD symptoms either by direct or nested polymerase chain reaction (PCR) assays employing ribosomal (r)RNA operon primer pairs. The CFSD-associated phytoplasmas were identified as group 16SrIII strains by restriction fragment length polymorphism (RFLP) and sequence analyses of amplified rDNA products, and results were corroborated by PCRs employing group 16SrIII-specific rRNA gene or ribosomal protein (rp) gene primers. Collectively, RFLP analyses indicated that CFSD strains differed from all phytoplasmas described previously in group 16SrIII and, on this basis, the strains were tentatively assigned to new ribosomal and ribosomal protein subgroups 16SrIII-L and rpIII-H, respectively. This is the first molecular identification of a phytoplasma associated with CFSD in cassava in Colombia.

scholarly journals A Phytoplasma Belonging to a 16SrIII-A Subgroup and dsRNA Virus Associated with Cassava Frogskin Disease in Brazil

Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 771-779 ◽  
Author(s):  
Adriana N. de Souza ◽  
Fábio N. da Silva ◽  
Ivan P. Bedendo ◽  
Claudine M. Carvalho
Keyword(s):  
Dsrna Virus ◽  
Nested Polymerase Chain Reaction ◽  
First Report ◽  
Reverse Transcription Pcr ◽  
Minas Gerais State ◽  
Bacillus Spp ◽  
Total Dna ◽  
Polymerase Chain ◽  
Rna Polymerase Gene

Cassava frogskin disease (CFSD) is a particular threat in cassava because symptoms remain hidden until harvest and losses can be total. The information related to the etiological agent of this disease is contradictory, because some authors believe it is caused by phytoplasmas while others believe that it is caused by a virus. In order to refine detection protocols and to characterize organisms associated with CFSD in Brazil, 32 symptomatic and 20 asymptomatic cassava plants were collected in Minas Gerais state. Total DNA was extracted and used for nested polymerase chain reaction (PCR) to detect phytoplasmas. Because endophytic Bacillus spp. led to false positives, primers were designed to facilitate the detection of phytoplasma in the presence of bacteria. In addition, double-stranded (ds)RNA was extracted from tubers and used in reverse-transcription PCR for the detection of the RNA-dependent RNA polymerase gene from Cassava frogskin virus segment 4. The detected phytoplasma was identified as belonging to the group 16SrIII-A by restriction fragment length polymorphism (RFLP), sequencing, and RFLP in silico. This is the first report of a phytoplasma belonging to the 16SrIII-A group associated with cassava plants, the first molecular characterization of a phytoplasma associated with CFSD in Brazil, and a first report of phytoplasma and a dsRNA virus (possible reovirus) co-infecting cassava plants with CFSD symptoms.

scholarly journals Partial Molecular Characterization of Dahlia mosaic virus and Its Detection by PCR

Plant Disease ◽  
2003 ◽  
Vol 87 (8) ◽  
pp. 945-948 ◽  
Author(s):  
M. Nicolaisen
Keyword(s):  
Nucleotide Sequence ◽  
Mosaic Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Open Reading Frames ◽  
Dahlia Mosaic Virus ◽  
Polymerase Chain ◽  
Pcr Assays

Dahlia mosaic virus (DMV) is the causal agent of one of the most important diseases of Dahlia pinnata. The nucleotide sequence of a 1,195-bp fragment of its genome was amplified and characterized. Based on this sequence, polymerase chain reaction (PCR) assays were developed for detection of DMV. The nucleotide sequence confirmed the classification of DMV as a member of genus Caulimovirus since it was similar to a region covering partly open reading frames (ORFs) IV and V found in caulimoviruses. The two most closely related viruses on the basis of comparison of ORF V fragments were shown to be Figwort mosaic virus and Mirabilis mosaic virus with 66.6 and 68.1% identity, respectively. Two PCR assays were developed using identical primer pairs: a real-time PCR based on SYBR green chemistry and a conventional PCR. Both methods clearly discriminated DMV-infected and healthy dahlia. The real-time PCR assay detected DMV-infected material that was diluted 105-fold in healthy material.

Sign in / Sign up

Export Citation Format

Share Document