Assessment of bacterial species present in Pasig River and Marikina River soil using 16S rDNA phylogenetic analysis

2015 ◽  
Vol 8 (2) ◽  
pp. 73-77
Author(s):  
Maria Constancia Carrillo ◽  
Paul Kenny Ko ◽  
Arvin Marasigan ◽  
Arlou Kristina Angeles
Keyword(s):  
Phylogenetic Analysis ◽  
16S Rdna ◽  
Bacterial Species

16S rDNA Sequence based Phylogenetic Analysis of Some Bacterial Phytoplasma

2012 ◽  
Vol 3 (3) ◽  
pp. 302-304
Author(s):  
G. D.Sharma G. D.Sharma ◽  
◽  
* Dhritiman Chanda ◽  
D.K. Jha D.K. Jha
Keyword(s):  
Phylogenetic Analysis ◽  
16S Rdna ◽  
Rdna Sequence ◽  
16S Rdna Sequence

IDENTIFICATION AND PHYLOGENETIC ANALYSIS OF SOUTHEASTERN USA EARTHWORM SPECIES USING 16S RDNA AND CO1 SEQUENCES

2018 ◽  
Vol 4 (3) ◽  
pp. 80-85
Author(s):  
YU.А. LEONTIEVA ◽  
◽  
A.G. NALIAN ◽  
G.A. DAMOFF ◽  
A.V. MARTYNOVA-VAN KLEY ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
16S Rdna ◽  
Southeastern Usa ◽  
Earthworm Species

scholarly journals Natural Merodiploidy of the lux-rib Operon of Photobacterium leiognathi from Coastal Waters of Honshu, Japan

2007 ◽  
Vol 189 (17) ◽  
pp. 6148-6158 ◽  
Author(s):  
Jennifer C. Ast ◽  
Henryk Urbanczyk ◽  
Paul V. Dunlap
Keyword(s):  
Phylogenetic Analysis ◽  
Sequence Analysis ◽  
Coastal Waters ◽  
Efflux Pump ◽  
Bacterial Species ◽  
Genomic Analysis ◽  
Multidrug Efflux Pump ◽  
Photobacterium Leiognathi ◽  
Content Organization ◽  
Luminous Bacteria

ABSTRACT Sequence analysis of the bacterial luminescence (lux) genes has proven effective in helping resolve evolutionary relationships among luminous bacteria. Phylogenetic analysis using lux genes, however, is based on the assumptions that the lux genes are present as single copies on the bacterial chromosome and are vertically inherited. We report here that certain strains of Photobacterium leiognathi carry multiple phylogenetically distinct copies of the entire operon that codes for luminescence and riboflavin synthesis genes, luxCDABEG-ribEBHA. Merodiploid lux-rib strains of P. leiognathi were detected during sequence analysis of luxA. To define the gene content, organization, and sequence of each lux-rib operon, we constructed a fosmid library of genomic DNA from a representative merodiploid strain, lnuch.13.1. Sequence analysis of fosmid clones and genomic analysis of lnuch.13.1 defined two complete, physically separate, and apparently functional operons, designated lux-rib 1 and lux-rib 2. P. leiognathi strains lelon.2.1 and lnuch.21.1 were also found to carry lux-rib 1 and lux-rib 2, whereas ATCC 25521T apparently carries only lux-rib 1. In lnuch.13.1, lelon.2.1, lnuch.21.1, and ATCC 25521T, lux-rib 1 is flanked upstream by lumQ and putA and downstream by a gene for a hypothetical multidrug efflux pump. In contrast, transposase genes flank lux-rib 2 of lnuch.13.1, and the chromosomal location of lux-rib 2 apparently differs in lnuch.13.1, lelon.2.1, and lnuch.21.1. Phylogenetic analysis demonstrated that lux-rib 1 and lux-rib 2 are more closely related to each other than either one is to the lux and rib genes of other bacterial species, which rules out interspecies lateral gene transfer as the origin of lux-rib 2 in P. leiognathi; lux-rib 2 apparently arose within a previously unsampled or extinct P. leiognathi lineage. Analysis of 170 additional strains of P. leiognathi, for a total of 174 strains examined from coastal waters of Japan, Taiwan, the Philippine Islands, and Thailand, identified 106 strains that carry only a single lux-rib operon and 68 that carry multiple lux-rib operons. Strains bearing a single lux-rib operon were obtained throughout the geographic sampling range, whereas lux-rib merodiploid strains were found only in coastal waters of central Honshu. This is the first report of merodiploidy of lux or rib genes in a luminous bacterium and the first indication that a natural merodiploid state in bacteria can correlate with geography.

scholarly journals A Multiplex-PCR Method for Strain Identification and Detailed Phylogenetic Analysis of AY-Group Phytoplasmas

Plant Disease ◽  
2014 ◽  
Vol 98 (3) ◽  
pp. 299-305 ◽  
Author(s):  
Shigeyuki Kakizawa ◽  
Yoichi Kamagata
Keyword(s):  
Phylogenetic Analysis ◽  
Multiplex Pcr ◽  
16S Rdna ◽  
Pathogenic Bacteria ◽  
Target Genes ◽  
Direct Sequencing ◽  
Pcr Amplification ◽  
Single Copy ◽  
Strain Identification ◽  
Specific Detection

Phytoplasmas are plant pathogenic bacteria that cause devastating losses in the yield of diverse crops worldwide. Specific detection and strain identification of phytoplasmas is important to prevent the spread of phytoplasma-induced diseases. Hence, methods to rapidly detect these organisms are important for pest control. Polymerase chain reaction (PCR) methods using phytoplasma-specific primers are widely used to detect phytoplasmas from infected plants and insects because they are highly sensitive, easily handled, and have a variety of analytical secondary applications. The phytoplasma 16S rDNA was widely used as a target of the PCR detection method; however, further target genes and more rapid methods have been required for more specific detection of phytoplasmas. Here, we developed a multiplex-PCR system to amplify several phytoplasma genes. We designed 36 primers, based on the genome sequence of ‘Candidatus Phytoplasma asteris’, to amplify 18 single-copy genes covering wide regions of the phytoplasma genome. Nine genes could be simultaneously amplified in a single PCR. This multiplex-PCR was applied to DNAs from 10 phytoplasma strains belonging to the AY-group, and different amplification patterns were obtained between strains, suggesting that this method would allow us to differentiate phytoplasmas at the strain level. Direct sequencing was also possible after the multiplex-PCR amplification by a modified sequencing method. Detailed phylogenetic analysis was performed using concatenated sequences, and evolutionary relationships among four Japanese isolates were revealed, where these strains could not be distinguished by their 16S rDNA. Thus, this multiplex-PCR system is useful for rapid strain identification and detailed phylogenetic analysis of phytoplasmas.

Comparative analysis of microbiome between accurately identified 16S rDNA and quantified bacteria in simulated samples

2014 ◽  
Vol 63 (3) ◽  
pp. 433-440 ◽  
Author(s):  
Haiyin Wang ◽  
Pengcheng Du ◽  
Juan Li ◽  
Yuanyuan Zhang ◽  
Wen Zhang ◽  
...  
Keyword(s):  
Microbial Communities ◽  
16S Rdna ◽  
Gold Standard ◽  
Clinical Utility ◽  
Bacterial Species ◽  
Relative Proportion ◽  
Rrna Gene ◽  
Low Concentrations ◽  
Rdna Sequencing ◽  
V3 Region

Although 16S rRNA gene (rDNA) sequencing is the gold standard for categorizing bacteria or characterizing microbial communities its clinical utility is limited by bias in metagenomic studies, in either the experiments or the data analyses. To evaluate the efficiency of current metagenomic methods, we sequenced seven simulated samples of ten bacterial species mixed at different concentrations. The V3 region of 16S rDNA was targeted and used to determine the distribution of bacterial species. The number of target sequences in individual simulated samples was in the range 1–1000 to provide a better reflection of natural microbial communities. However, for a given bacterial species present in the same proportion but at different concentrations, the observed percentage of 16S rDNAs was similar, except at very low concentrations that cannot be detected by real-time PCR. These results confirmed that the comparative microbiome in a sample characterized by 16S rDNA sequencing is sufficient to detect not only potential infectious pathogens, but also the relative proportion of 16S rDNA in the sample.

scholarly journals First Report of “Candidatus Liberibacter solanacearum” Associated with Psyllid-Affected Carrots in Norway

Plant Disease ◽  
2012 ◽  
Vol 96 (3) ◽  
pp. 454-454 ◽  
Author(s):  
J. E. Munyaneza ◽  
V. G. Sengoda ◽  
L. Sundheim ◽  
R. Meadow
Keyword(s):  
Ribosomal Protein ◽  
16S Rdna ◽  
Bacterial Species ◽  
Consensus Sequence ◽  
Economic Damage ◽  
Carrot Root ◽  
Candidatus Liberibacter Solanacearum ◽  
First Report ◽  
Candidatus Liberibacter ◽  
Root Tissues

Carrot (Daucus carota) plants with symptoms resembling those associated with the carrot psyllid Trioza apicalis and the bacterium “Candidatus Liberibacter solanacearum” (1–4) were observed in 70 to 80% of commercial fields and experimental plots in southeastern Norway from late July to mid-September of 2011; all cultivars grown were affected with approximately 10 to 100% symptomatic plants per field. T. apicalis, a pest of carrot in northern and central Europe, including Norway, can cause as much as 100% crop loss and is associated with “Ca. L. solanacearum” (1–4). Symptoms on affected plants include leaf curling, yellow and purple discoloration of leaves, stunted growth of shoots and roots, and proliferation of secondary roots. Carrot plant samples were collected from five T. apicalis-infested fields in Ostfold, Vestfold, Oppland, and Hedmark counties. Total DNA was extracted from petiole and root tissues of 54 plants, including 27 symptomatic plants and 27 asymptomatic plants from four cultivars (Namdal, Panther, Romance, and Yukon) with the cetyltrimethylammonium bromide (CTAB) buffer extraction method (2,3). DNA samples were tested by PCR assay using primer pairs OA2/OI2c and CL514F/R to amplify a portion of 16S rDNA and rplJ/rplL ribosomal protein genes, respectively, of “Ca. L. solanacearum” (2,3). A 1,168-bp 16S rDNA fragment was detected in the DNA from 22 (81.5%) symptomatic plants and a 668-bp rplJ/rplL fragment was amplified from the DNA of 26 (96.3%) symptomatic and 5 (18.5%) asymptomatic plants, indicating the presence of liberibacter. No liberibacter was detected in the asymptomatic carrot plants with the primer pair OA2/OI2c. Amplicons from the DNA of four carrot root samples with each primer pair were cloned (pCR2.1-TOPO; Invitrogen, Carlsbad, CA) and three clones of each of the eight amplicons were sequenced (MCLAB, San Francisco, CA). BLAST analysis of the 16S rDNA consensus sequence from the carrot root tissues (GenBank Accession No. JN863097) showed 100% identity to those of “Ca. L. solanacearum” previously amplified from carrot (GU373048 and GU373049) and T. apicalis (GU477254 and GU477255) in Finland (2,3). The rplJ/rplL consensus sequence from the carrots (GenBank Accession No. JN863098) was 99% identical to the sequences of rplJ/rplL “Ca. L. solanacearum” ribosomal protein gene from carrots in Finland (GU373050 and GU373051). To our knowledge, this is the first report of “Ca. L. solanacearum” associated with carrot in Norway. This bacterial species has caused millions of dollars in losses to potato and several other solanaceous crops in North and Central America and New Zealand (1). This plant pathogen has also been reported from carrots and T. apicalis in Finland, where it has caused significant economic damage to carrot crops (2–4). References: (1) J. E. Munyaneza. Southwest. Entomol. 35:471, 2010. (2) J. E. Munyaneza et al. Plant Dis. 94:639, 2010. (3) J. E. Munyaneza et al. J. Econ. Entomol. 103:1060, 2010. (4) A. Nissinen et al. Entomol. Exp. Appl. 125:277, 2007.

scholarly journals Design and Evaluation of PCR Primers for Analysis of Bacterial Populations in Wine by Denaturing Gradient Gel Electrophoresis

2003 ◽  
Vol 69 (11) ◽  
pp. 6801-6807 ◽  
Author(s):  
Isabel Lopez ◽  
Fernanda Ruiz-Larrea ◽  
Luca Cocolin ◽  
Erica Orr ◽  
Trevor Phister ◽  
...  
Keyword(s):  
Lactic Acid ◽  
16S Rdna ◽  
Gel Electrophoresis ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Bacterial Species ◽  
Pcr Primers ◽  
Fungal Species ◽  
Bacterial Populations ◽  
Gradient Gel Electrophoresis ◽  
Primer Sets

ABSTRACT Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified ribosomal DNA (rDNA) is routinely used to compare levels of diversity of microbial communities and to monitor population dynamics. While using PCR-DGGE to examine the bacteria in wine fermentations, we noted that several commonly used PCR primers for amplifying bacterial 16S rDNA also coamplified yeast, fungal, or plant DNA present in samples. Unfortunately, amplification of nonbacterial DNA can result in a masking of bacterial populations in DGGE profiles. To surmount this problem, we developed two new primer sets for specific amplification of bacterial 16S rDNA in wine fermentation samples without amplification of eukaryotic DNA. One primer set, termed WLAB1 and WLAB2, amplified lactic acid bacteria, while another, termed WBAC1 and WBAC2, amplified both lactic acid bacterial and acetic acid bacterial populations found in wine. Primer specificity and efficacy were examined with DNA isolated from numerous bacterial, yeast, and fungal species commonly found in wine and must samples. Importantly, both primer sets effectively distinguished bacterial species in wine containing mixtures of yeast and bacteria.

Rapid identification of Medicago nodulating strains by using two oligonucleotide probes complementary to 16S rDNA sequences

1997 ◽  
Vol 43 (9) ◽  
pp. 854-861 ◽  
Author(s):  
Sophie Rome ◽  
Jean-Claude Cleyet-Marel ◽  
Luis A. Materon ◽  
Philippe Normand ◽  
Brigitte Brunel
Keyword(s):  
16S Rdna ◽  
Sinorhizobium Meliloti ◽  
Bacterial Species ◽  
Blot Hybridization ◽  
Dot Blot ◽  
Plant Host ◽  
Rdna Sequences ◽  
Sinorhizobium Medicae ◽  
16S Rdna Sequences ◽  
Annual Medicago

Symbiotic bacteria associated with the Medicago genus are separated into two closely related species named Sinorhizobium meliloti and Sinorhizobium medicae. To discriminate rapidly between these two bacterial species, two 15-base DNA probes, 16Smfs and 16Smed, were designed from the alignment of 16S rDNA sequences to differentiate S. meliloti from S. medicae. Their specificities were evaluated by dot-blot hybridization experiments on 25 reference strains representing 13 species of Rhizobium and Sinorhizobium, and by comparison with all 16S rDNA sequences available in the GenBank data base. No cross-reaction was found with 16Smed, which was thus considered species specific for S. medicae. By contrast, as expected according to the 16S rDNA sequence alignment, the labeled 16Smfs probe cross-hybridized with the DNAs of S. meliloti, Sinorhizobium fredii, and Sinorhizobium saheli but not with the DNA of S. medicae. Since S. saheli and S. fredii do not nodulate Medicago, 16Smed and 16Smfs can be routinely used to characterize the two Sinorhizobium species nodulating Medicago from pure cultures or from Medicago root nodules. Fifty strains isolated from eight annual Medicago species were then characterized by using colony hybridizations. Sinorhizobium meliloti was more frequently obtained (>80% isolates) than was S. medicae. Both Sinorhizobium species seemed to be trapped by annual Medicago and no plant-host specificity was detected.Key words: Sinorhizobium meliloti, Sinorhizobium medicae, Medicago, oligonucleotide probe, 16S rDNA gene.

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