BOX-polymerase Chain Reaction Research Articles

Culturable Diversity of Endophytic and Rhizoplane Colonizing Bacteria of Indian Mustard (Brassica juncea (L.) Czern & Coss), Affected by Soil Types and Assessment of Plant Growth Promoting Attributes

Microbial community in the rhizosphere of plant significantly influences growth and development of a plant. As the soil vary in microbial composition due to interaction of soil with plant roots and its exudates, the role of soil in shaping microbial diversity of a crop rhizosphere is least studied. In present study, Mustard (Brassica juncea cv Pusa Mehak) was grown in three different soil viz. Vertisols, Inceptisols and Alfisols, and the bacteria were isolated form root tissues and rhizoplane to study their plant growth promoting (PGP) attributes and diversity using BOX-Polymerase chain reaction (PCR) fingerprinting technique, as influenced by soil types. Form threes soil orders, total 37 isolates were obtained of which, 19 could grow on nitrogen free medium, 6 solubilized P from tricalcium phosphate, 2 isolate solubilized potassium from glauconite and 8 had zinc solubilizing ability with the highest zinc solubilization index of 3.84 by isolate VRB-7. Based on the fingerprint analysis, differences occurred in the endophytic bacteria of same host grown in different soil types suggesting influence of soil types in colonization of microbes in rhizoplane and endorhizosphere. The common organisms obtained from host grown in different soil types indicating host genotype driven assembly of bacteria. Similarity in the isolates on rhizoplane and endorhizosphere indicated endophytic colonization of the isolates capable of firmly adhering to the rhizoplane. The findings of this study will help in formulation of crop specific bioinoculant for specific soil types and edaphic properties.

Detection of Diverse N-Acyl Homoserine Lactone Signalling Molecules Among Bacteria Associated with Rice Rhizosphere.

Bacterial communities communicate, regulate and coordinate their cooperative activities and physiological process by releasing, sensing and responding to small diffusible signal molecules such as acyl homoserine lactones (AHLs), auto-inducing oligo-peptides (AIPs) and autoinducer-2, a process referred to as Quorum sensing (QS). The QS mediated communication in rhizosphere associated bacterial communities significantly influence traits governing plant-microbe interactions. This study aimed to identify AHL-mediated QS signals in bacterial communities associated with rice rhizosphere using two AHL biosensors reporter strains Chromobacterium violaceum CV026 and Agrobacterium tumefaciens NTL4 (pZLR4). Approximately 375 bacterial isolates isolated from rice rhizosphere and screened using both the biosensors, detected 49 (13%) AHLpositive isolates. The BOX-Polymerase Chain reaction (BOX-PCR) fingerprinting profiles of the 49 AHL positive isolates represented 11 distinct cluster groups. Subsequent 16S rRNA gene sequence analysis identified 11 different species affiliated to two different phyla; predominantly γ-proteobacteria, representing 5 genera and 1 genus in α-proteobacteria. Thin-layer chromatography (TLC) analysis detected diverse AHL profiles among the 11 AHL positive isolates with both substituted and unsubstituted acyl side chains of C4, C6 and C8 carbon. Further, AHL production in Acinetobacter lactucae, Aeromonas popoffii, Serratia oryzae, and Rhizobium wuzhouense is being reported for the first time. Detection of diverse AHLs from different groups of rhizobacteria associated with rice indicates that these signalling molecules may be involved in the regulation of rhizobacterial behaviour and symbiotic plant-microbe interactions. Future research on the role of AHLs in trans-kingdom communication particularly plant-microbe interaction using synthetic microbial community will enable in evaluating and developing potential plant specific bioproducts.

The identification and genetic diversity of endophytic bacteria isolated from selected crops

AbstractA collection of 45 isolates was created based on bacteria isolated from maize, broad bean, wheat, rye and wild plants such as horsetail and burdock. The aim of the current study was to isolate the bacteria, and then identify and assess the degree of genomic diversity. The molecular identification of microsymbionts isolated from the endosphere (root and stem) of plants grown in agricultural soils was performed using 16S rRNA gene sequencing. To evaluate the genomic diversity between strains that occurred in multiple host plants, 18 bacterial isolates representing four species were subjected to denaturing gradient gel electrophoresis. The 16S rDNA analysis assigned all bacterial isolates to ten genera, from whichRhizobiumwas represented by 19 isolates,Delftiaby 11,Agrobacteriumby five,Stenotrophomonasby three,Brevundimonasby two andNovosphingobium,Variovorax, Collimonas, AchromobacterandComamonasby only one isolate. Furthermore, the genomic diversity of the 11 isolates ofDelftiasp. was assessed using the BOX – polymerase chain reaction (BOX-PCR) and enterobacterial repetitive intergenic consensus – PCR (ERIC-PCR) methods. Typing patterns and analysis using BOX-PCR and ERIC-PCR data demonstrated similarities among the tested isolates. In general, the results obtained with BOX-PCR and ERIC-PCR were in good agreement. However, a greater degree of differentiation patterns of the genomic DNA was obtained in the ERIC-PCR method.

Molecular Epidemiology of Pseudomonas syringae pv. syringae Causing Bacterial Leaf Spot of Watermelon and Squash in Florida.

From 2013 to 2014, bacterial leaf spot epidemics incited by Pseudomonas syringae pv. syringae affected an estimated 3,000 ha of watermelon and squash in Florida, and caused foliar blighting and transplant losses in severely affected fields. To investigate the diversity of the causal agent, we isolated 28 P. syringae strains from diseased plants grown in 10 Florida and Georgia counties over the course of 2 years. Strains were confirmed as P. syringae through sequence analysis of the 16S ribosomal RNA, phenotypic, and biochemical profiling; however, 20 displayed an atypical phenotype by exhibiting nonfluorescent activity on King's medium B agar and being negative for ice-nucleating activity. Multilocus sequence analysis and BOX polymerase chain reaction revealed the presence of two haplotypes among the collected strains that grouped into two distinct clades within P. syringae phylogroup 2. Pathogenicity testing showed that watermelon, cantaloupe, and squash seedlings were susceptible to a majority of these strains. Although both haplotypes were equally virulent on cantaloupe, they differed in virulence on watermelon and squash. The distribution of one haplotype in 9 of 10 Florida and Georgia counties sampled indicated that these epidemics were associated with the recent introduction of a novel clonal P. syringae lineage throughout major watermelon production areas in Florida.

Genetic Diversity of Nitrogen-Fixing and Plant Growth Promoting Pseudomonas Species Isolated from Sugarcane Rhizosphere.

The study was designed to isolate and characterize Pseudomonas spp. from sugarcane rhizosphere, and to evaluate their plant- growth- promoting (PGP) traits and nitrogenase activity. A biological nitrogen-fixing microbe has great potential to replace chemical fertilizers and be used as a targeted biofertilizer in a plant. A total of 100 isolates from sugarcane rhizosphere, belonging to different species, were isolated; from these, 30 isolates were selected on the basis of preliminary screening, for in vitro antagonistic activities against sugarcane pathogens and for various PGP traits, as well as nitrogenase activity. The production of IAA varied from 312.07 to 13.12 μg mL−1 in tryptophan supplemented medium, with higher production in AN15 and lower in CN20 strain. The estimation of ACC deaminase activity, strains CY4 and BA2 produced maximum and minimum activity of 77.0 and 15.13 μmoL mg−1 h−1. For nitrogenase activity among the studied strains, CoA6 fixed higher and AY1 fixed lower in amounts (108.30 and 6.16 μmoL C2H2 h−1 mL−1). All the strains were identified on the basis of 16S rRNA gene sequencing, and the phylogenetic diversity of the strains was analyzed. The results identified all strains as being similar to Pseudomonas spp. Polymerase chain reaction (PCR) amplification of nifH and antibiotic genes was suggestive that the amplified strains had the capability to fix nitrogen and possessed biocontrol activities. Genotypic comparisons of the strains were determined by BOX, ERIC, and REP PCR profile analysis. Out of all the screened isolates, CY4 (Pseudomonas koreensis) and CN11 (Pseudomonas entomophila) showed the most prominent PGP traits, as well as nitrogenase activity. Therefore, only these two strains were selected for further studies; Biolog profiling; colonization through green fluorescent protein (GFP)-tagged bacteria; and nifH gene expression using quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The Biolog phenotypic profiling, which comprised utilization of C and N sources, and tolerance to osmolytes and pH, revealed the metabolic versatility of the selected strains. The colonization ability of the selected strains was evaluated by genetically tagging them with a constitutively expressing GFP-pPROBE-pTetr-OT plasmid. qRT-PCR results showed that both strains had the ability to express the nifH gene at 90 and 120 days, as compared to a control, in both sugarcane varieties GT11 and GXB9. Therefore, our isolated strains, P. koreensis and P. entomophila may be used as inoculums or in biofertilizer production for enhancing growth and nutrients, as well as for improving nitrogen levels, in sugarcane and other crops. The present study, to the best of our knowledge, is the first report on the diversity of Pseudomonas spp. associated with sugarcane in Guangxi, China.

0681 Evaluation of genetic diversity of Lactobacillus plantarum isolated from alfalfa silage using the BOX-polymerase chain reaction

0681 Evaluation of genetic diversity of Lactobacillus plantarum isolated from alfalfa silage using the BOX-polymerase chain reaction

Isolation and identification by 16S rRNA sequence analysis of plant growth-promoting azospirilla from the rhizosphere of wheat.

The main objective of the present study was to isolate phytohormone-producing, phosphate-solubilizing strains of Azospirillum from wheat to be used as inoculants for plant growth promotion. Five Azospirillum strains were isolated from the rhizosphere of field-grown wheat (Triticum aestivum L.), and it was confirmed by BOX-polymerase chain reaction (PCR) that the isolates were different and not re-isolates of the same strain. Sequence analysis of the PCR-amplified 16S rRNA gene indicated that four isolates showed maximum similarity to Azospirillum brasilense and one isolate showed maximum similarity to Azospirillum zeae. This is the first report indicating the presence of an A. zeae like isolate in the wheat rhizosphere in Pakistan. The bacterial isolates were characterized for their plant growth-promoting traits, phosphate solubilization, and indole-3-acetic acid (IAA) production. None of the isolates showed phosphate solubilization activity in the commonly used Pikovskaya medium. However, all strains (except AzoK4) exhibited ability to solubilize tricalcium phosphate (TCP) in modified Pikovskaya medium in which sucrose was replaced by Na-malate, as well as in TCP-supplemented Luria-Bertani (LB) medium. Organic acids, such as acetic, citric, lactic, malic, and succinic acids, were detected in culture supernatants of the tested Azospirillum strains. All strains exhibited ability to produce IAA in the growth medium, except Azospirillum sp. AzoK1. Among the strains tested, the maximum IAA production (30.49 ± 1.04 mg L−1) and phosphate solubilization (105.50 ± 4.93 mg L−1) were shown by a pure culture of Azospirillum sp. AzoK2. In pot experiments, single-strain inocula of Azospirillum sp. AzoK1 and AzoK2 improved wheat plant growth.

Maternal pneumococcal endometritis may cause early-onset neonatal sepsis

Streptococcus pneumoniae is a rarely recognized cause of neonatal sepsis. A case of early-onset neonatal sepsis due to penicillin susceptible pneumococci and maternal endometritis is herein described. A male infant was born at 33 weeks of gestation. The baby developed S. pneumoniae bacteraemia and recovered completely following treatment with antibiotics. Four days after partum, the mother was readmitted with fever. S. pneumoniae was recovered from an endometrial sample. Isolates from mother and child were both serogroup 1 and confirmed to be identical on the basis of BOX-polymerase chain reaction typing and pulse-field gel electrophoresis. Perinatal transmission of S. pneumoniae had probably occurred due to ascending infection from the maternal genital tract.

Phenotypic and molecular fingerprinting of fast growing rhizobia of field-grown pigeonpea from the eastern edge of the Brazilian Pantanal

The aim of this study was to evaluate the diversity of rhizobial isolates obtained from root nodules of pigeonpea plants grown at the eastern edge of the Brazilian Pantanal. The bacterial isolates were isolated from root nodules from field-growing pigeonpea grown in two rural settlements of the Aquidauana municipality. The bacterial isolates were characterized phenotypically by means of cultural characterization, intrinsic antibiotic resistance (IAR), salt and high incubation temperature tolerance, and amylolytic and cellulolytic activities. The molecular characterization of the bacterial isolates was carried out using amplified ribosomal DNA restriction analysis (ARDRA) and Box-polymerase chain reaction (PCR) techniques. In addition, the symbiotic performance of selected rhizobial isolates was evaluated in a greenhouse experiment using sterile substrate. The phenotypic characterization revealed that the bacterial strains obtained from pigeonpea root nodules presented characteristics that are uncommon among rhizobial isolates, indicating the presence of new species nodulating the pigeonpea plants in the Brazilian Pantanal. The molecular fingerprinting of these bacterial isolates also showed a highly diverse collection, with both techniques revealing less than 25% similarity among bacterial isolates. The evaluation of symbiotic performance also indicated the presence of microorganisms with high potential to increase the growth and nitrogen content at the shoots of pigeonpea plants. The results obtained in this study indicate the presence of a highly diversified rhizobial community nodulating the pigeonpea at the eastern edge of the Brazilian Pantanal.

Multifarious plant growth promoting characteristics of chickpea rhizosphere associated Bacilli help to suppress soil-borne pathogens

Wilt and root rot are the major constraints in chickpea production and very difficult to manage through agrochemicals. Hence, for an ecofriendly and biological management, 240 strains of Bacillus and Bacillus derived genera were isolated from chickpea rhizosphere, further narrowed down to 14 strains on the basis of in vitro production of indole acetic acid, siderophore, phosphate solubilization, hydrolytic enzymes and were evaluated for antagonism against chickpea pathogens (Fusarium oxysporum f. sp. ciceri race 1, F. solani and Macrophomina phaseolina). The strains were identified on the basis of physiological characters and 16S RNA gene sequencing. The genotypic comparisons of strains were determined by BOX-polymerase chain reaction profiles and amplified rDNA restriction analysis. These isolates were evaluated in greenhouse assay in which B. subtilis (B-CM191, B-CV235, B-CL-122) proved to be effective in reducing wilt incidence and significant enhancement in growth (root and shoot length) and dry matter of chickpea plants. PCR amplification of bacillomycin (bmyB) and β-glucanase genes suggests that amplified genes from the Bacillus could have a role to further define the diversity, ecology, and biocontrol activities in the suppression of soil-borne pathogens.

Isolation and Identification of Toluene-Metabolizing Bacteria from Rhizospheres of Two Indoor Plants

The role of the rhizosphere microbial community in removing volatile organic compounds has not been well investigated. In this study, two species of indoor foliage plants, Fittonia verschaffeltii var. argyroneura and Hoya carnosa, were primed with toluene exposure for 2 months, followed by isolation and identification of the rhizosphere bacteria that were demonstrated to metabolize toluene. A total of 42 bacterial isolates were obtained. The number of bacterial isolates was narrowed down to 23, which had banding pattern similarities of 80 % or less, using BOX-polymerase chain reaction (PCR) fingerprinting technique. The 23 isolates were further characterized by sequencing part of their 16S rDNA after PCR. Their identities were examined using Basic Local Alignment Search Tool (BLAST), resulting in the isolates having the highest sequence similarities (97–100 %) to eight known bacteria strains, none of which had been previously reported to be capable of degrading toluene. The bacterial isolates were positive for toluene monooxygenase gene, confirming their genetic potential to metabolize toluene. Five of the isolates were further tested with 14C-labeled toluene to directly show their ability to metabolize toluene. Isolate type did not significantly affect the percent of toluene mineralized over 2 weeks' time. However, the isolates had differing response to varying toluene concentrations. Under low (0.05) and high (0.2 μCi/mL) concentrations, they mineralized 43 and 49 % of toluene, respectively. The isolation and characterization of toluene-metabolizing bacteria corroborates previous speculation that the rhizosphere microbial community contributes to the phytoremediation potential of indoor foliage plants.

Diversity of native rhizobia isolated in south Brazil and their growth promotion effect on white clover (Trifolium repens) and rice (Oryza sativa) plants

In this study, rhizobia strains isolated from white clover (Trifolium repens) root nodules were evaluated in an effort to identify an efficient nitrogen-fixing rhizobia strain that can also improve the growth of rice plants (Oryza sativa). White clover plants were collected from seven sites in south Brazil, and 78 native rhizobia isolates were obtained. The genetic diversity analysis of those isolates was carried out by BOX-polymerase chain reaction. Overall, the native rhizobia isolated showed a high genetic diversity, but when the bacterial isolates from the same site were compared, the diversity was lower. One native rhizobia, POA3 (isolated from the Porto Alegre locality), was able to promote the growth of both plants and is therefore a good candidate for new inoculant formulation. Finally, we can conclude that the community of native rhizobia symbiont of white clover plants in southern Brazil is highly diverse and the growth promotion effect of rhizobia inoculation on rice plants was more pronounced in a poor nutrient substrate condition than in a rich nutrient substrate condition.

Identification, Virulence, and Distribution of Two Biovars of Pseudomonas syringae pv. actinidiae in New Zealand.

Pseudomonas syringae pv. actinidiae, the causal agent of bacterial canker of kiwifruit, was detected for the first time in New Zealand in November 2010. Only in Bay of Plenty, one of the four regions where this pathogen had been detected, did symptoms evolve beyond leaf spots, resulting in cane die-back, wilting of canes, and canker, sometimes leading to death of the vine. Molecular analysis (cts haplotype and BOX-polymerase chain reaction [PCR] electrophoretic pattern) of strains isolated from different regions of New Zealand revealed that two biovars could be distinguished. They have been called biovar 3 and biovar 4 to differentiate them from strains from Japan (biovar 1) or Korea (biovar 2), which have a different cts haplotype or a different BOX-PCR pattern. Biovars 3 and 4 displayed different degrees of virulence, as measured by their ability to cause leaf spots on young, potted kiwifruit plants. Biovar 3, which has also been present in Italy since 2008 and in France, was found in the Bay of Plenty, where cane diebacks were observed. In contrast, no symptoms other than leaf spots have been observed in orchards where strains of biovar 4 have been isolated. We report the distribution and the disease progression of biovars 3 and 4 in New Zealand.

Genetic Structure of Clavibacter michiganensis subsp. michiganensis Populations in Michigan Commercial Tomato Fields.

Clavibacter michiganensis subsp. michiganensis, causal agent of bacterial canker of tomato, is distinguished into four fingerprint types (A, B, C, and D) using BOX-polymerase chain reaction (PCR). To characterize the variation within the C. michiganensis subsp. michiganensis population in Michigan, 718 strains of C. michiganensis subsp. michiganensis were isolated from infected foliage and fruit collected from 14 and 9 Michigan commercial tomato fields in 1997 and 1998, respectively. The frequency of PCR types detected with BOX-PCR in all strains, and Bayesian cluster analysis, pairwise differentiation index comparisons, and genetic diversity estimates of 96 strains genotyped for six virulence-related genes revealed that C. michiganensis subsp. michiganensis populations in Michigan tomato fields are geographically structured. A multilocus haplotype cladogram was also consistent with geographic stratification in C. michiganensis subsp. Michiganensis populations. Some regions had strains predominantly of only one PCR type or belonging mostly to one genetic cluster, while other regions presented more diversity of occurrence of PCR types and genetic clusters. Results derived from this study provide information about the genetic structure of C. michiganensis subsp. michiganensis populations in Michigan and genetic diversity of C. michiganensis subsp. michiganensis inocula, which is key in developing disease management strategies.

Characterisation and identification of carbofuran-utilising bacteria isolated from agricultural soil

A total of 96 bacterial cultures were isolated from soil. Seventeen bacterial isolates were selected following their cultivation on solid media containing 100 mg · L−1 carbofuran as the sole source of carbon and nitrogen. Of the 17 isolates, 10F, 11M, 17N, 23B and 26M were specifically chosen because of their relatively higher growth efficiency and genetic diversity based on Box-polymerase chain reaction analysis. These bacterial cultures had 16S rRNA gene sequences that were most similar to Acinetobacter baumannii (10F), Agrobacterium tumefaciens (11M), Ochrobactrum anthropi (17N), Escherichia coli (23B) and Agrobacterium tumefaciens (26M) with 97, 95, 93, 95 and 94% similarity in their 16S rDNA gene sequence, respectively. Degradation rates of carbofuran in soil inoculated with these isolates were 1.9, 1.5, 1.6, 1.7 and 1.6 times, respectively, faster in comparison with uninoculated soil after 10 days of incubation. The maximum degradation rates of carbofuran (45 and 91%) were detected in soil inoculated with A. baumannii (10F) after 10 and 20 days’ incubation, respectively. These data indicate that these isolates may have the potential for use in bioremediation of pesticide contaminated soil.

Analysis of plant growth promoting rhizobacteria population in apple rhizosphere soils

利用选择性培养基, 对多年生苹果树根际与连作幼树根际促生细菌进行了分离和测数, 并采用BOX-PCR技术进行聚类分析。结果表明: 多年生苹果树根际细菌总量及固氮细菌、解磷细菌、硅酸盐细菌、拮抗细菌4类根际促生细菌的数量均高于连作幼树根际。在多年生苹果树根际, 硅酸盐细菌的数量最大, 解磷细菌和固氮细菌的数量次之, 拮抗细菌的数量最小; 在连作幼树根际, 解磷细菌的数量最大, 硅酸盐细菌和固氮细菌的数量次之, 拮抗细菌的数量最小。从两种土壤中获得的促生细菌分离株的BOX-PCR图谱最大的相异百分数都在1.25以上, 说明这些细菌分离株的遗传进化距离比较接近。在细菌BOX-PCR图谱相异百分数为0.25的水平上, 多年生苹果树根际促生细菌分为79个聚类群, 其中固氮细菌18个聚类群, 解磷细菌29个聚类群, 硅酸盐细菌19个聚类群, 拮抗细菌18个聚类群; 连作幼树根际促生细菌分为46个聚类群, 其中固氮细菌15个聚类群, 解磷细菌19个聚类群, 硅酸盐细菌8个聚类群, 拮抗细菌9个聚类群。多年生苹果树4类根际促生细菌的多样性、丰富度和均匀度指数均高于连作幼树根际, 而优势度指数低于连作幼树根际。与连作幼树相比, 多年生苹果树根际促生细菌具有丰富的种属多样性。

Dissemination of an Erythromycin-Resistant Penicillin-NonsusceptibleStreptococcus pneumoniaePoland6B-20 Clone in Argentina

Prevalence of serotype 6B penicillin (PEN)-nonsusceptible Streptococcus pneumoniae significantly increased from 15.8% (1993-1997) to 67.3% (1998-2002) (p<0.001) in Argentina. Serogroup 6 ranks fourth among different capsular types within invasive isolates from Argentinean patients <6 years of age. To evaluate whether the increase in PEN resistance in serotype 6B pneumococci was due to the dissemination of one or more clones, the genetic diversity of 93 S. pneumoniae serotype 6B isolates was analyzed. Five BOX-polymerase chain reaction types were obtained (65.5% isolates) and a group of 15 isolates, representing 41.6% of those having a decreased susceptibility to PEN, were further characterized. The antibiotype of these isolates showed their multiresistance, with 100% of the isolates being resistant to erythromycin, 80% to tetracycline, and 73.3% to trimethoprim-sulfamethoxazole. Of the 15 isolates, 13 belonged to the same pulsed-field gel electrophoresis type and galU cluster and were members of the same clone. The identity of the clone was confirmed in four isolates by multilocus sequence typing. The sequence type found (ST315) corresponds to the Poland(6B)-20 clone. In summary, BOX-polymerase chain reaction, pulsed-field gel electrophoresis, and galU polymorphism were useful tools to detect the presence of a clone whose identity was confirmed by multilocus sequence typing. The isolates belonging to Poland(6B)-20 found in this work are described for the first time in Latin America.

Non-oral bifidobacteria and the aciduric microbiota of the denture plaque biofilm

The microbiota of the denture plaque biofilm colonizing the fitting surface of dentures in edentulous subjects with healthy palates (n = 20) and in edentulous subjects with denture stomatitis (n = 20) was studied. The numbers of bacteria colonizing the dentures of healthy subjects was significantly less than the numbers colonizing the dentures of stomatitis subjects. The proportions and frequency of isolation of mutans streptococci, lactobacilli, bifidobacteria and yeasts were significantly (P < 0.05) greater in the subjects with denture stomatitis. The proportions of these organisms in the denture plaque biofilm of the subjects with denture stomatitis were similar to those found in carious lesions, indicating that the site is a low pH environment. The predominant bifidobacterial species in the mouths of dentate subjects is Bifidobacterium dentium but in the edentulous subjects wearing dentures B. dentium was isolated from only one of the 20 subjects with denture stomatitis and from none of the 20 subjects with healthy palates. Instead, Bifidobacterium breve, Bifidobacterium scardovii and Bifidobacterium longum subsp. longum were isolated. Only a single non-oral bifidobacterial species was isolated from each individual and repetitive extragenic palindromic- and BOX-polymerase chain reaction typing methods indicated that the same genotypes were shared between subjects. Using deferred antagonism spot plate assays, interspecies inhibition was demonstrated between oral isolates of B. dentium, B. breve, B. scardovii and B. longum subsp. longum. Here we have shown that bifidobacteria and caries-associated microbiota are present in denture plaque at levels similar to those of carious lesions and B. dentium cannot be maintained in an edentulous mouth.

Isolation and characterization of endophytic plant growth-promoting (PGPB) or stress homeostasis-regulating (PSHB) bacteria associated to the halophyte Prosopis strombulifera

This study was designed to isolate and characterize endophytic bacteria from halophyte Prosopis strombulifera grown under extreme salinity and to evaluate in vitro the bacterial mechanisms related to plant growth promotion or stress homeostasis regulation. Isolates obtained from P. strombulifera were compared genotypically by BOX-polymerase chain reaction, grouped according to similarity, and identified by amplification and partial sequences of 16S DNAr. Isolates were grown until exponential growth phase to evaluate the atmospheric nitrogen fixation, phosphate solubilization, siderophores, and phytohormones, such as indole-3-acetic acid, zeatin, gibberellic acid and abscisic acid production, as well as antifungal, protease, and 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. A total of 29 endophytic strains were grouped into seven according to similarity. All bacteria were able to grow and to produce some phytohormone in chemically defined medium with or without addition of a nitrogen source. Only one was able to produce siderophores, and none of them solubilized phosphate. ACC deaminase activity was positive for six strains. Antifungal and protease activity were confirmed for two of them. In this work, we discuss the possible implications of these bacterial mechanisms on the plant growth promotion or homeostasis regulation in natural conditions.

Macrolide Resistance Determinants in Erythromycin-Resistant &lt;i&gt;Streptococcus pneumoniae&lt;/i&gt; in Turkey

To determine the major molecular mechanisms of macrolide resistance among Streptococcus pneumoniae isolates in Turkey, we examined a total of 151 isolates collected from different regions of Turkey. Overall, 40 (26.4%) isolates were resistant to erythromycin. The most common mechanism (38/40) was target site modification due to erm (B) genes. Only two isolates harbored the mef (A)/(E) efflux gene. A clonal spread of resistant strains could not be demonstrated by BOX-polymerase chain reaction. The results from this study have shown that the erm (B) gene is predominant in Turkish S. pneumoniae isolates, as in isolates from the rest of the world, and a clonal dissemination is not responsible for this resistance profile.