scholarly journals Response of Endophytic Bacterial Communities in Potato Plants to Infection with Erwinia carotovora subsp. atroseptica

2002 ◽  
Vol 68 (5) ◽  
pp. 2261-2268 ◽  
Author(s):  
Birgit Reiter ◽  
Ulrike Pfeifer ◽  
Helmut Schwab ◽  
Angela Sessitsch
Keyword(s):  
Plant Stress ◽  
Erwinia Carotovora ◽  
Community Analysis ◽  
Antagonistic Activity ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Polymorphism Analysis ◽  
Blackleg Disease ◽  
Potato Plants

ABSTRACT The term endophyte refers to interior colonization of plants by microorganisms that do not have pathogenic effects on their hosts, and various endophytes have been found to play important roles in plant vitality. In this study, cultivation-independent terminal restriction fragment length polymorphism analysis of 16S ribosomal DNA directly amplified from plant tissue DNA was used in combination with molecular characterization of isolates to examine the influence of plant stress, achieved by infection with the blackleg pathogen Erwinia carotovora subsp. atroseptica, on the endophytic population in two different potato varieties. Community analysis clearly demonstrated increased bacterial diversity in infected plants compared to that in control plants. The results also indicated that the pathogen stress had a greater impact on the bacteria population than the plant genotype had. Partial sequencing of the 16S rRNA genes of isolated endophytes revealed a broad phylogenetic spectrum of bacteria, including members of the α, β, and γ subgroups of the Proteobacteria, high- and low-G+C-content gram-positive organisms, and microbes belonging to the Flexibacter-Cytophaga-Bacteroides group. Screening of the isolates for antagonistic activity against E. carotovora subsp. atroseptica revealed that 38% of the endophytes protected tissue culture plants from blackleg disease.

scholarly journals Response of Nursery Pigs to a Synbiotic Preparation of Starch and an Anti-Escherichia coli K88 Probiotic

2010 ◽  
Vol 76 (24) ◽  
pp. 8192-8200 ◽  
Author(s):  
D. O. Krause ◽  
S. K. Bhandari ◽  
J. D. House ◽  
C. M. Nyachoti
Keyword(s):  
Escherichia Coli ◽  
Potato Starch ◽  
Control Diet ◽  
Community Analysis ◽  
Positive Control ◽  
Microbial Community Analysis ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Polymorphism Analysis ◽  
E Coli

ABSTRACT Postweaning diarrhea in pigs is frequently caused by enterotoxigenic Escherichia coli K88 (ETEC). The aim of this study was to test the efficacy of E. coli probiotics (PRO) in young pigs challenged with E. coli K88. We also tested the synbiotic interaction with raw potato starch (RPS), which can be used as a prebiotic. Forty 17-day-old weaned piglets were randomly assigned to four treatments: treatment 1, positive-control diet (C), no probiotics or RPS but containing in-feed antibiotics; treatment 2, probiotic (PRO), no feed antibiotics plus a 50:50 mixture of probiotic E. coli strains UM-2 and UM-7; treatment 3, 14% RPS, no antibiotics (RPS); treatment 4, 14% RPS plus a 50:50 mixture of probiotic E. coli strains UM-2 and UM-7, no antibiotics (PRO-RPS). The pigs were challenged with pathogenic E. coli K88 strains on day 7 of the experiment (24-day-old pigs) and euthanized on day 10 of the experiment (35-day-old pigs). Probiotic and pathogenic E. coli strains were enumerated by selective enrichment on antibiotics, and microbial community analysis was conducted using terminal restriction length polymorphism analysis (T-RFLP) of 16S rRNA genes. The combination of raw potato starch and the probiotic had a beneficial effect on piglet growth performance and resulted in a reduction of diarrhea and increased microbial diversity in the gut. We conclude that the use of E. coli probiotic strains against E. coli K88 in the presence of raw potato starch is effective in reducing the negative effects of ETEC in a piglet challenge model.

scholarly journals Development and Characterization of Stable Sediment-Free Anaerobic Bacterial Enrichment Cultures That Dechlorinate Aroclor 1260

2006 ◽  
Vol 72 (4) ◽  
pp. 2460-2470 ◽  
Author(s):  
Donna L. Bedard ◽  
Jessica J. Bailey ◽  
Brandon L. Reiss ◽  
Greta Van Slyke Jerzak
Keyword(s):  
Fragment Length Polymorphism ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Polymorphism Analysis ◽  
Contaminated Sediment ◽  
Intensive Study ◽  
Bacterial Enrichment ◽  
Enrichment Cultures ◽  
Pcb Dechlorination

ABSTRACT We have developed sediment-free anaerobic enrichment cultures that dechlorinate a broad spectrum of highly chlorinated polychlorinated biphenyls (PCBs). The cultures were developed from Aroclor 1260-contaminated sediment from the Housatonic River in Lenox, MA. Sediment slurries were primed with 2,6-dibromobiphenyl to stimulate Process N dechlorination (primarily meta dechlorination), and sediment was gradually removed by successive transfers (10%) to minimal medium. The cultures grow on pyruvate, butyrate, or acetate plus H2. Gas chromatography-electron capture detector analysis demonstrated that the cultures extensively dechlorinate 50 to 500 μg/ml of Aroclor 1260 at 22 to 24°C by Dechlorination Process N. Triplicate cultures of the eighth transfer without sediment dechlorinated 76% of the hexa- through nonachlorobiphenyls in Aroclor 1260 (250 μg/ml) to tri- through pentachlorobiphenyls in 110 days. At least 64 PCB congeners, all of which are chlorinated on both rings and 47 of which have six or more chlorines, were substrates for this dechlorination. To characterize the bacterial diversity in the enrichments, we used eubacterial primers to amplify and clone 16S rRNA genes from DNA extracted from cultures grown on acetate plus H2. Restriction fragment length polymorphism analysis of 107 clones demonstrated the presence of Thauera-like Betaproteobacteria, Geobacter-like Deltaproteobacteria, Pseudomonas species, various Clostridiales, Bacteroidetes, Dehalococcoides of the Chloroflexi group, and unclassified Eubacteria. Our development of highly enriched, robust, stable, sediment-free cultures that extensively dechlorinate a highly chlorinated commercial PCB mixture is a major and unprecedented breakthrough in the field. It will enable intensive study of the organisms and genes responsible for a major PCB dechlorination process that occurs in the environment and could also lead to effective remediation applications.

scholarly journals Unraveling the Etiology of North American Grapevine Yellows (NAGY): Novel NAGY Phytoplasma Sequevars Related to ‘Candidatus Phytoplasma pruni’

Plant Disease ◽  
2015 ◽  
Vol 99 (8) ◽  
pp. 1087-1097 ◽  
Author(s):  
Robert E. Davis ◽  
Ellen L. Dally ◽  
Yan Zhao ◽  
Ing-Ming Lee ◽  
Wei Wei ◽  
...  
Keyword(s):  
16S Rdna ◽  
North American ◽  
Phylogenetic Analyses ◽  
New York State ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Vitis Vinifera L ◽  
Polymorphism Analysis ◽  
Ribosomal Protein Genes ◽  
Grapevine Yellows

North American grapevine yellows (NAGY) disease has sometimes been attributed to infection of Vitis vinifera L. by Prunus X-disease phytoplasma (‘Candidatus Phytoplasma pruni’) but this attribution may not be fully adequate. In this study, phytoplasma strains related to ‘Ca. Phytoplasma pruni’ were found in NAGY-diseased grapevines in Maryland, Pennsylvania, Virginia, Ohio, Missouri, and New York State. Based on restriction fragment length polymorphism analysis of 16S ribosomal RNA gene (16S rDNA) sequences, the strains (termed NAGYIII strains) were classified in group 16SrIII (X-disease group) but they contained a recognition site for the restriction endonuclease MseI that is not present in the 16S rDNA of ‘Ca. Phytoplasma pruni’. The 16S rDNA of the strains differed by three or four nucleotides from that of ‘Ca. Phytoplasma pruni’, indicating that they belonged to two novel 16S rDNA sequevars, designated NAGYIIIα and NAGYIIIβ. Both sequevars differed from ‘Ca. Phytoplasma pruni’ by a single base in each of three regions corresponding to species-unique (signature) sequences described for ‘Ca. Phytoplasma pruni’. Phylogenetic analyses of 16S rRNA genes and SecY proteins, and single-nucleotide polymorphism analyses of secY and ribosomal protein genes, further distinguished the two grapevine sequevar lineages from one another and from ‘Ca. Phytoplasma pruni’. The NAGYIIIα and NAGYIIIβ sequevars also differed from ‘Ca. Phytoplasma pruni’ in regions of the folded SecY protein that are predicted to be near or exposed at the outer surface of the phytoplasma membrane. No evidence indicated that diseased grapevines contained any phytoplasma strain conforming to ‘Ca. Phytoplasma pruni’ sensu stricto. Because the NAGYIII sequevars have not been reported in X-disease, a question is raised as to whether NAGYIII and Prunus X-disease are caused by different phytoplasma genotypes.

Composition of the archaeal community involved in methane production during the decomposition of Microcystis blooms in the laboratory

2012 ◽  
Vol 58 (10) ◽  
pp. 1153-1158 ◽  
Author(s):  
Peng Xing ◽  
Huabing Li ◽  
Qing Liu ◽  
Jiuwen Zheng
Keyword(s):  
Lake Taihu ◽  
Archaeal Community ◽  
Oxygen Depletion ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Polymorphism Analysis ◽  
Methanogenic Community ◽  
Controlled Systems ◽  
Microcystis Blooms ◽  
Different Temperatures

We investigated the microbial processes involved in methane (CH4) production from Microcystis bloom scums at different temperatures. A Microcystis slurry was collected from Lake Taihu and incubated in airtight bottles at 15, 25, and 35 °C. The production of CH4 was monitored, and the emission rate was calculated. The dynamics of the methanogenic community were analyzed by terminal restriction fragment length polymorphism analysis of archaeal 16S rRNA genes. Phylogenetic information for the methanogens was obtained by cloning and sequencing selected samples. Significant CH4 emission from the Microcystis scums was delayed by approximately 12 days by the natural oxygen depletion process, and CH4 production was enhanced at higher temperatures. Phylogenetic analysis indicated that the archaeal community was composed of Methanomicrobiales, Methanobacteriaceae, and a novel cluster of Archaea. An apparent succession of the methanogenic community was demonstrated, with a predominance of Methanobacteriaceae at higher temperatures. Higher temperatures enhanced the methanogenic transformation of the Microcystis biomass and the phylogenetic dominance of hydrogenotrophic methanogens, suggesting that H2 and CO2 might be the primary substrates for CH4 production during Microcystis decomposition without the participation of lake sediment. This work provides insight into the microbial components involved in Microcystis biomass fermentation in controlled systems.

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 .

PCR-Ribotyping of Xenorhabdus andPhotorhabdus Isolates from the Caribbean Region in Relation to the Taxonomy and Geographic Distribution of Their Nematode Hosts

1998 ◽  
Vol 64 (11) ◽  
pp. 4246-4254 ◽  
Author(s):  
Marion Fischer-Le Saux ◽  
Hervé Mauléon ◽  
Philippe Constant ◽  
Brigitte Brunel ◽  
Noël Boemare
Keyword(s):  
Entomopathogenic Nematodes ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Caribbean Region ◽  
Polymorphism Analysis ◽  
Caribbean Islands ◽  
Pcr Ribotyping ◽  
The Caribbean ◽  
And Cluster Analysis ◽  
Reference Strains

ABSTRACT The genetic diversity of symbiotic Xenorhabdus andPhotorhabdus bacteria associated with entomopathogenic nematodes was examined by a restriction fragment length polymorphism analysis of PCR-amplified 16S rRNA genes (rDNAs). A total of 117 strains were studied, most of which were isolated from the Caribbean basin after an exhaustive soil sampling. The collection consisted of 77 isolates recovered from entomopathogenic nematodes in 14 Caribbean islands and of 40 reference strains belonging toXenorhabdus and Photorhabdus spp. collected at various localities worldwide. Thirty distinctive 16S rDNA genotypes were identified, and cluster analysis was used to distinguish the genus Xenorhabdus from the genusPhotorhabdus. The genus Xenorhabdusappears more diverse than the genusPhotorhabdus, and for both genera the bacterial genotype diversity is in congruence with the host-nematode taxonomy. The occurrence of symbiotic bacterial genotypes was related to the ecological distribution of host nematodes.

scholarly journals Prey Range Characterization, Ribotyping, and Diversity of Soil and Rhizosphere Bdellovibriospp. Isolated on Phytopathogenic Bacteria

2000 ◽  
Vol 66 (6) ◽  
pp. 2365-2371 ◽  
Author(s):  
E. Jurkevitch ◽  
D. Minz ◽  
Barak Ramati ◽  
Gili Barel
Keyword(s):  
16S Rrna ◽  
Common Bean ◽  
Pcr Amplification ◽  
Erwinia Carotovora ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Range Analysis ◽  
Pcr Product ◽  
Utilization Patterns ◽  
Range Characterization

ABSTRACT Thirty new Bdellovibrio strains were isolated from an agricultural soil and from the rhizosphere of plants grown in that soil. Using a combined molecular and culture-based approach, we found that the soil bdellovibrios included subpopulations of organisms that differed from rhizosphere bdellovibrios. Thirteen soil and seven common bean rhizosphere Bdellovibrio strains were isolated when Pseudomonas corrugata was used as prey; seven and two soil strains were isolated when Erwinia carotovora subsp.carotovora and Agrobacterium tumefaciens, respectively, were used as prey; and one tomato rhizosphere strain was isolated when A. tumefaciens was used as prey. In soil and in the rhizosphere, depending on the prey cells used, the concentrations of bdellovibrios were between 3 × 102 to 6 × 103 and 2.8 × 102 to 2.3 × 104 PFU g−1. A prey range analysis of five soil and rhizosphereBdellovibrio isolates performed with 22 substrate species, most of which were plant-pathogenic and plant growth-enhancing bacteria, revealed unique utilization patterns and differences between closely related prey cells. An approximately 830-bp fragment of the 16S rRNA genes of all of theBdellovibrio strains used was obtained by PCR amplification by using a Bdellovibrio-specific primer combination. Soil and common bean rhizosphere strains produced two and one restriction patterns for this PCR product, respectively. The 16S rRNA genes of three soil isolates and three root-associated isolates were sequenced. One soil isolate belonged to theBdellovibrio stolpii-Bdellovibrio starrii clade, while all of the other isolates clustered withBdellovibrio bacteriovorus and formed two distantly related, heterogeneous groups.

scholarly journals Ammonium Concentrations in Produced Waters from a Mesothermic Oil Field Subjected to Nitrate Injection Decrease through Formation of Denitrifying Biomass and Anammox Activity

2010 ◽  
Vol 76 (15) ◽  
pp. 4977-4987 ◽  
Author(s):  
Sabrina L. Cornish Shartau ◽  
Marcy Yurkiw ◽  
Shiping Lin ◽  
Aleksandr A. Grigoryan ◽  
Adewale Lambo ◽  
...  
Keyword(s):  
Nitrate Reduction ◽  
Produced Water ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Community Analysis ◽  
Oil Field ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Ammonium Concentration ◽  
Anammox Activity ◽  
Produced Waters

ABSTRACT Community analysis of a mesothermic oil field, subjected to continuous field-wide injection of nitrate to remove sulfide, with denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes indicated the presence of heterotrophic and sulfide-oxidizing, nitrate-reducing bacteria (hNRB and soNRB). These reduce nitrate by dissimilatory nitrate reduction to ammonium (e.g., Sulfurospirillum and Denitrovibrio) or by denitrification (e.g., Sulfurimonas, Arcobacter, and Thauera). Monitoring of ammonium concentrations in producing wells (PWs) indicated that denitrification was the main pathway for nitrate reduction in the field: breakthrough of nitrate and nitrite in two PWs was not associated with an increase in the ammonium concentration, and no increase in the ammonium concentration was seen in any of 11 producing wells during periods of increased nitrate injection. Instead, ammonium concentrations in produced waters decreased on average from 0.3 to 0.2 mM during 2 years of nitrate injection. Physiological studies with produced water-derived hNRB microcosms indicated increased biomass formation associated with denitrification as a possible cause for decreasing ammonium concentrations. Use of anammox-specific primers and cloning of the resulting PCR product gave clones affiliated with the known anammox genera “Candidatus Brocadia” and “Candidatus Kuenenia,” indicating that the anammox reaction may also contribute to declining ammonium concentrations. Overall, the results indicate the following: (i) that nitrate injected into an oil field to oxidize sulfide is primarily reduced by denitrifying bacteria, of which many genera have been identified by DGGE, and (ii) that perhaps counterintuitively, nitrate injection leads to decreasing ammonium concentrations in produced waters.

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