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1.
Int J Syst Evol Microbiol ; 63(Pt 1): 208-211, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22389284

RESUMO

Phylogenetic analysis of the 16S rRNA gene sequences of strains TE26(T) and K6 belonging to Wautersia numazuensis Kageyama et al. 2005 showed the strains to be deeply intermingled among the species of the genus Cupriavidus. The comparison showed that strain TE26(T) was closely related to the type strains of Cupriavidus pinatubonensis (99.1 % 16S rRNA gene sequence similarity), C. basilensis (98.7 %), C. necator (98.7 %) and C. gilardii (98.0 %). However, DNA-DNA hybridization experiments (less than 20 % relatedness) demonstrated that strain TE26(T) is different from these Cupriavidus species. A comparative phenotypic and chemotaxonomic analysis (based on fatty acid profiles) in combination with the 16S rRNA gene sequence phylogenetic analysis and the DNA-DNA hybridization results supported the incorporation of Wautersia numazuensis into the genus Cupriavidus as Cupriavidus numazuensis comb. nov.; the type strain is TE26(T) (=LMG 26411(T) =DSM 15562(T) = CIP 108892(T)).


Assuntos
Cupriavidus/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Cupriavidus/genética , DNA Bacteriano/genética , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
2.
Appl Microbiol Biotechnol ; 97(10): 4639-49, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-22805783

RESUMO

Azospirillum are prominent plant growth-promoting rhizobacteria (PGPR) extensively used as phytostimulatory crop inoculants, but only few studies are dealing with Azospirillum-containing mixed inocula involving more than two microorganisms. We compared here three prominent Azospirillum strains as part of three-component consortia including also the PGPR Pseudomonas fluorescens F113 and a mycorrhizal inoculant mix composed of three Glomus strains. Inoculant colonization of maize was assessed by quantitative PCR, transcription of auxin synthesis gene ipdC (involved in phytostimulation) in Azospirillum by RT-PCR, and effects on maize by secondary metabolic profiling and shoot biomass measurements. Results showed that phytostimulation by all the three-component consortia was comparable, despite contrasted survival of the Azospirillum strains and different secondary metabolic responses of maize to inoculation. Unexpectedly, the presence of Azospirillum in the inoculum resulted in lower phytostimulation in comparison with the Pseudomonas-Glomus two-component consortium, but this effect was transient. Azospirillum's ipdC gene was transcribed in all treatments, especially with three-component consortia, but not with all plants and samplings. Inoculation had no negative impact on the prevalence of mycorrhizal taxa in roots. In conclusion, this study brought new insights in the functioning of microbial consortia and showed that Azospirillum-Pseudomonas-Glomus three-component inoculants may be useful in environmental biotechnology for maize growth promotion.


Assuntos
Azospirillum/fisiologia , Glomeromycota/fisiologia , Pseudomonas/fisiologia , Zea mays/crescimento & desenvolvimento , Azospirillum/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Zea mays/microbiologia
3.
Antonie Van Leeuwenhoek ; 104(6): 1063-71, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24057982

RESUMO

During a survey of Burkholderia species with potential use in agrobiotechnology, a group of 12 strains was isolated from the rhizosphere and rhizoplane of tomato plants growing in Mexico (Nepantla, Mexico State). A phylogenetic analysis of 16S rRNA gene sequences showed that the strains are related to Burkholderia kururiensis and Burkholderia mimosarum (97.4 and 97.1 %, respectively). However, they induced effective nitrogen-fixing nodules on roots of Phaseolus vulgaris. Based on polyphasic taxonomy, the group of strains represents a novel species for which the name Burkholderia caballeronis sp. nov. is proposed. The type species is TNe-841(T) (= LMG 26416(T) = CIP 110324(T)).


Assuntos
Burkholderia/classificação , Burkholderia/fisiologia , Fixação de Nitrogênio , Phaseolus/microbiologia , Nodulação , Solanum lycopersicum/microbiologia , Técnicas de Tipagem Bacteriana , Burkholderia/genética , Burkholderia/isolamento & purificação , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , México , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
4.
Curr Microbiol ; 67(1): 51-60, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23404651

RESUMO

Burkholderia comprises more than 60 species of environmental, clinical, and agro-biotechnological relevance. Previous phylogenetic analyses of 16S rRNA, recA, gyrB, rpoB, and acdS gene sequences as well as genome sequence comparisons of different Burkholderia species have revealed two major species clusters. In this study, we undertook a multilocus sequence analysis of 77 type and reference strains of Burkholderia using atpD, gltB, lepA, and recA genes in combination with the 16S rRNA gene sequence and employed maximum likelihood and neighbor-joining criteria to test this further. The phylogenetic analysis revealed, with high supporting values, distinct lineages within the genus Burkholderia. The two large groups were named A and B, whereas the B. rhizoxinica/B. endofungorum, and B. andropogonis groups consisted of two and one species, respectively. The group A encompasses several plant-associated and saprophytic bacterial species. The group B comprises the B. cepacia complex (opportunistic human pathogens), the B. pseudomallei subgroup, which includes both human and animal pathogens, and an assemblage of plant pathogenic species. The distinct lineages present in Burkholderia suggest that each group might represent a different genus. However, it will be necessary to analyze the full set of Burkholderia species and explore whether enough phenotypic features exist among the different clusters to propose that these groups should be considered separate genera.


Assuntos
Burkholderia/classificação , Burkholderia/genética , DNA Bacteriano/genética , Variação Genética , Tipagem de Sequências Multilocus/métodos , Filogenia , Animais , Análise por Conglomerados , Genes Bacterianos , Genótipo , Humanos
5.
Microb Ecol ; 63(2): 249-66, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21850446

RESUMO

The genus Burkholderia comprises more than 60 species isolated from a wide range of niches. Although they have been shown to be diverse and ubiquitously distributed, most studies have thus far focused on the pathogenic species due to their clinical importance. However, the increasing number of recently described Burkholderia species associated with plants or with the environment has highlighted the division of the genus into two main clusters, as suggested by phylogenetical analyses. The first cluster includes human, animal, and plant pathogens, such as Burkholderia glumae, Burkholderia pseudomallei, and Burkholderia mallei, as well as the 17 defined species of the Burkholderia cepacia complex, while the other, more recently established cluster comprises more than 30 non-pathogenic species, which in most cases have been found to be associated with plants, and thus might be considered to be potentially beneficial. Several species from the latter group share characteristics that are of use when associating with plants, such as a quorum sensing system, the presence of nitrogen fixation and/or nodulation genes, and the ability to degrade aromatic compounds. This review examines the commonalities in this growing subgroup of Burkholderia species and discusses their prospective biotechnological applications.


Assuntos
Burkholderia/classificação , Burkholderia/fisiologia , Magnoliopsida/microbiologia , Simbiose , Burkholderia/genética , Fungos/fisiologia , Magnoliopsida/fisiologia , Filogenia , Especificidade da Espécie
6.
BMC Microbiol ; 11: 228, 2011 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-21995705

RESUMO

BACKGROUND: A close association between maize roots and Burkholderia cepacia complex (BCC) bacteria has been observed in different locations globally. In this study we investigated by MultiLocus Restriction Typing (MLRT) the genetic diversity and relationships among Burkholderia cenocepacia IIIB and BCC6 populations associated with roots of maize plants cultivated in geographically distant countries (Italy and Mexico), in order to provide new insights into their population structure, evolution and ecology. RESULTS: The 31 B. cenocepacia IIIB and 65 BCC6 isolates gave rise to 29 and 39 different restriction types (RTs), respectively. Two pairs of isolates of B. cenocepacia IIIB and BCC6, recovered from both Italian and Mexican maize rhizospheres, were found to share the same RT. The eBURST (Based Upon Related Sequence Types) analysis of MLRT data grouped all the B. cenocepacia IIIB isolates into four clonal complexes, with the RT-4-complex including the 42% of them, while the majority of the BCC6 isolates (94%) were grouped into the RT-104-complex. These two main clonal complexes included RTs shared by both Italian and Mexican maize rhizospheres and a clear relationship between grouping and maize variety was also found. Grouping established by eBURST correlated well with the assessment using unweighted-pair group method with arithmetic mean (UPGMA). The standardized index of association values obtained in both B. cenocepacia IIIB and BCC6 suggests an epidemic population structure in which occasional clones emerge and spread. CONCLUSIONS: Taken together our data demonstrate a wide dispersal of certain B. cenocepacia IIIB and BCC6 isolates in Mexican and Italian maize rhizospheres. Despite the clear relationship found between the geographic origin of isolates and grouping, identical RTs and closely related isolates were observed in geographically distant regions. Ecological factors and selective pressure may preferably promote some genotypes within each local microbial population, favouring the spread of a single clone above the rest of the recombinant population.


Assuntos
Burkholderia cenocepacia/genética , Rizosfera , Microbiologia do Solo , Zea mays/microbiologia , Alelos , Burkholderia cenocepacia/isolamento & purificação , DNA Bacteriano/genética , Variação Genética , Itália , Desequilíbrio de Ligação , México , Raízes de Plantas/microbiologia , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA
7.
Appl Environ Microbiol ; 76(13): 4302-17, 2010 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-20435760

RESUMO

The genus Burkholderia includes over 60 species isolated from a wide range of environmental niches and can be tentatively divided into two major species clusters. The first cluster includes pathogens such as Burkholderia glumae, B. pseudomallei, and B. mallei and 17 well-studied species of the Burkholderia cepacia complex. The other recently established cluster comprises at least 29 nonpathogenic species, which in most cases have been found to be associated with plants. It was previously established that Burkholderia kururiensis, a member of the latter cluster, possesses an N-acyl homoserine lactone (AHL) quorum-sensing (QS) system designated "BraI/R," which is found in all species of the plant-associated cluster. In the present study, two other BraI/R-like systems were characterized in B. xenovorans and B. unamae and were designated the BraI/R(XEN) and BraI/R(UNA) systems, respectively. Several phenotypes were analyzed, and it was determined that exopolysaccharide was positively regulated by the BraIR-like system in the species B. kururiensis, B. unamae, and B. xenovorans, highlighting commonality in targets. However, the three BraIR-like systems also revealed differences in targets since biofilm formation and plant colonization were differentially regulated. In addition, a second AHL QS system designated XenI2/R2 and an unpaired LuxR solo protein designated BxeR solo were also identified and characterized in B. xenovorans LB400(T). The two AHL QS systems of B. xenovorans are not transcriptionally regulating each other, whereas BxeR solo negatively regulated xenI2. The XenI2/R2 and BxeR solo proteins are not widespread in the Burkholderia species cluster. In conclusion, the present study represents an extensive analysis of AHL QS in the Burkholderia plant-associated cluster demonstrating both commonalities and differences, probably reflecting environmental adaptations of the various species.


Assuntos
Acil-Butirolactonas , Proteínas de Bactérias , Burkholderia/genética , Regulação Bacteriana da Expressão Gênica , Oryza/microbiologia , Percepção de Quorum , Acil-Butirolactonas/metabolismo , Acil-Butirolactonas/farmacologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Burkholderia/classificação , Burkholderia/crescimento & desenvolvimento , Burkholderia/metabolismo , Dados de Sequência Molecular , Percepção de Quorum/fisiologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Análise de Sequência de DNA , Especificidade da Espécie
8.
Appl Environ Microbiol ; 75(6): 1782-5, 2009 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-19139238

RESUMO

Gluconacetobacter diazotrophicus utilizes plant sucrose with a constitutively expressed levansucrase (LsdA), producing extracellular levan, which may be degraded under energetically unfavored conditions. Reverse transcriptase-PCR analysis revealed that lsdA and the downstream exolevanase gene (lsdB) form an operon. lsdB transcription was induced during growth with low fructose concentrations (0.44 to 33 mM) and repressed by glucose. Transport of LsdB to the periplasm involved N-terminal signal peptide cleavage. Type II secretion mutants failed to transfer LsdB across the outer membrane, impeding levan hydrolysis.


Assuntos
Proteínas de Bactérias/biossíntese , Regulação Bacteriana da Expressão Gênica , Gluconacetobacter/enzimologia , Gluconacetobacter/genética , Glicosídeo Hidrolases/biossíntese , Transcrição Gênica , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Sequência de Bases , Frutanos/metabolismo , Frutose/metabolismo , Ordem dos Genes , Glucose/metabolismo , Glicosídeo Hidrolases/genética , Dados de Sequência Molecular , Óperon , Periplasma/enzimologia , Sinais Direcionadores de Proteínas , Reação em Cadeia da Polimerase Via Transcriptase Reversa
9.
Appl Environ Microbiol ; 75(20): 6581-90, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19700546

RESUMO

The genus Burkholderia includes pathogens of plants and animals and some human opportunistic pathogens, such as the Burkholderia cepacia complex (Bcc), but most species are nonpathogenic, plant associated, and rhizospheric or endophytic. Since rhizobacteria expressing ACC (1-aminocyclopropane-1-carboxylate) deaminase may enhance plant growth by lowering plant ethylene levels, in this work we investigated the presence of ACC deaminase activity and the acdS gene in 45 strains, most of which are plant associated, representing 20 well-known Burkholderia species. The results demonstrated that ACC deaminase activity is a widespread feature in the genus Burkholderia, since 18 species exhibited ACC deaminase activities in the range from 2 to 15 mumol of alpha-ketobutyrate/h/mg protein, which suggests that these species may be able to modulate ethylene levels and enhance plant growth. In these 18 Burkholderia species the acdS gene sequences were highly conserved (76 to 99% identity). Phylogenetic analysis of acdS gene sequences in Burkholderia showed tight clustering of the Bcc species, which were clearly distinct from diazotrophic plant-associated Burkholderia species. In addition, an acdS knockout mutant of the N(2)-fixing bacterium Burkholderia unamae MTl-641(T) and a transcriptional acdSp-gusA fusion constructed in this strain showed that ACC deaminase could play an important role in promotion of the growth of tomato plants. The widespread ACC deaminase activity in Burkholderia species and the common association of these species with plants suggest that this genus could be a major contributor to plant growth under natural conditions.


Assuntos
Burkholderia/enzimologia , Carbono-Carbono Liases/metabolismo , Solanum lycopersicum/crescimento & desenvolvimento , Solanum lycopersicum/microbiologia , Aminoácidos Cíclicos/metabolismo , Animais , Sequência de Bases , Burkholderia/classificação , Burkholderia/genética , Carbono-Carbono Liases/genética , Primers do DNA/genética , DNA Bacteriano/genética , Genes Bacterianos , Teste de Complementação Genética , Humanos , Solanum lycopersicum/metabolismo , Dados de Sequência Molecular , Mutação , Nitrogênio/metabolismo , Filogenia , Reguladores de Crescimento de Plantas/genética , Reguladores de Crescimento de Plantas/metabolismo , Especificidade da Espécie , Simbiose
10.
Appl Environ Microbiol ; 74(14): 4574-9, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18502926

RESUMO

Pulsed-field gel electrophoresis and 16S rRNA hybridization experiments showed that multichromosome genome structures and very large genome sizes (6.46 to 8.73 Mb) are prevalent in novel plant-associated Burkholderia species. (15)N(2) isotope dilution assays revealed unambiguous diazotrophy in these novel species. nifH gene sequence analysis, often used to determine phylogenetic relatedness among diazotrophs, showed tight clusters of Burkholderia species, which were clearly distinct from those of other diazotrophs.


Assuntos
Burkholderia/classificação , Burkholderia/genética , Cromossomos Bacterianos/genética , Genoma Bacteriano , Fixação de Nitrogênio , Sequência de Bases , Burkholderia/enzimologia , DNA Bacteriano/isolamento & purificação , Eletroforese em Gel de Campo Pulsado , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Oxirredutases/genética , Filogenia , Plantas/microbiologia , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/isolamento & purificação , Análise de Sequência de DNA , Microbiologia do Solo
11.
FEMS Microbiol Ecol ; 54(1): 57-66, 2005 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-16329972

RESUMO

In this study the antagonistic activity among 55 Gluconacetobacter diazotrophicus strains, belonging to 13 electrophoretic types (ETs), in culture media was analyzed. Antagonistic effects were seen only in strains belonging to two ETs named ET-1 and ET-3. Two out of 29 ET-1 strains, and 3 out of 7 ET-3 strains of G. diazotrophicus showed antagonistic effects against many other strains belonging to all the ETs of this species analyzed, and against closely related strains of Gluconacetobacter species, including Gluconacetobacter johannae, Gluconacetobacter azotocaptans and Gluconacetobacter liquefaciens but not against other phylogenetically distant bacterial species. Results showed that the substance responsible of such antagonistic activity is a low molecular mass molecule (approximately 3400 Da), stable from pH 3.5 to 8.5, and very stable at 4 degrees C for 10 months. This substance was sensitive to proteases, and the antagonistic activity was lost after 2 h at 95 degrees C. All of these features show that the substance is related to bacteriocin-like molecules. The antagonistic substance should be chromosomally encoded because ET-3 strains of G. diazotrophicus do not harbor any plasmids. The antagonistic ability of ET-3 strains of G. diazotrophicus could be an advantage for the natural colonization of the sugarcane environment, as was observed in experiments with micropropagated sterile sugarcane plantlets co-inoculated with a bacteriocin-producer strain and a bacteriocin-sensitive strain of G. diazotrophicus. In these experiments, both in the rhizosphere as well as inside the roots, the bacteriocin-sensitive population decreased drastically. In addition, this study shows that inside the plants there may exist antagonistic interactions among endophytic bacteria like to those described among the rhizospheric community.


Assuntos
Bacteriocinas/biossíntese , Gluconacetobacter/crescimento & desenvolvimento , Saccharum/microbiologia , Meios de Cultura , Gluconacetobacter/efeitos dos fármacos
12.
Syst Appl Microbiol ; 28(3): 277-86, 2005 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15900973

RESUMO

The family Acetobacteraceae currently includes three known nitrogen-fixing species, Gluconacetobacter diazotrophicus, G. johannae and G. azotocaptans. In the present study, acetic acid-producing nitrogen-fixing bacteria were isolated from four different wetland rice varieties cultivated in the state of Tamilnadu, India. Most of these isolates were identified as G. diazotrophicus on the basis of their phenotypic characteristics and PCR assays using specific primers for that species. Based on 16S rDNA partial sequence analysis and DNA: DNA reassociation experiments the remaining isolates were identified as Acetobacter peroxydans, another species of the Acetobacteraceae family, thus far never reported as diazotrophic. The presence of nifH genes in A. peroxydans was confirmed by PCR amplification with nifH specific primers. Scope for the findings: This is the first report of the occurrence and association of N2-fixing Gluconacetobacter diazotrophicus and Acetobacter peroxydans with wetland rice varieties. This is the first report of diazotrophic nature of A. peroxydans.


Assuntos
Acetobacter/isolamento & purificação , Gluconacetobacter/isolamento & purificação , Oryza/microbiologia , Acetobacter/classificação , Acetobacter/genética , Acetobacter/metabolismo , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Microbiologia Ambiental , Genes de RNAr , Gluconacetobacter/classificação , Gluconacetobacter/genética , Gluconacetobacter/metabolismo , Índia , Dados de Sequência Molecular , Fixação de Nitrogênio , Hibridização de Ácido Nucleico , Oxirredutases/genética , Filogenia , Reação em Cadeia da Polimerase , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
13.
PLoS One ; 9(1): e83779, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24416172

RESUMO

Burkholderia is a diverse and dynamic genus, containing pathogenic species as well as species that form complex interactions with plants. Pathogenic strains, such as B. pseudomallei and B. mallei, can cause serious disease in mammals, while other Burkholderia strains are opportunistic pathogens, infecting humans or animals with a compromised immune system. Although some of the opportunistic Burkholderia pathogens are known to promote plant growth and even fix nitrogen, the risk of infection to infants, the elderly, and people who are immunocompromised has not only resulted in a restriction on their use, but has also limited the application of non-pathogenic, symbiotic species, several of which nodulate legume roots or have positive effects on plant growth. However, recent phylogenetic analyses have demonstrated that Burkholderia species separate into distinct lineages, suggesting the possibility for safe use of certain symbiotic species in agricultural contexts. A number of environmental strains that promote plant growth or degrade xenobiotics are also included in the symbiotic lineage. Many of these species have the potential to enhance agriculture in areas where fertilizers are not readily available and may serve in the future as inocula for crops growing in soils impacted by climate change. Here we address the pathogenic potential of several of the symbiotic Burkholderia strains using bioinformatics and functional tests. A series of infection experiments using Caenorhabditis elegans and HeLa cells, as well as genomic characterization of pathogenic loci, show that the risk of opportunistic infection by symbiotic strains such as B. tuberum is extremely low.


Assuntos
Infecções por Burkholderia/microbiologia , Burkholderia/fisiologia , Mamíferos/microbiologia , Plantas/microbiologia , Simbiose , Animais , Sistemas de Secreção Bacterianos/genética , Burkholderia/genética , Burkholderia/patogenicidade , Caenorhabditis elegans/microbiologia , Resistência Microbiana a Medicamentos/genética , Flagelos/genética , Genes Bacterianos/genética , Loci Gênicos , Células HeLa , Humanos , Família Multigênica , Filogenia , Simbiose/genética , Virulência/genética
14.
Bioengineered ; 4(4): 236-43, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23680857

RESUMO

It has been documented that bacteria from the Burkholderia genera produce different kinds of compounds that inhibit plant pathogens, however in Burkholderia tropica, an endophytic diazotrophic and phosphate-solubilizing bacterium isolated from a wide diversity of plants, the capacity to produce antifungal compounds has not been evaluated. In order to expand our knowledge about Burkholderia tropica as a potential biological control agent, we analyzed 15 different strains of this bacterium to evaluate their capacities to inhibit the growth of four phytopathogenic fungi, Colletotrichum gloeosporioides, Fusarium culmorum, Fusarium oxysporum and Sclerotium rolffsi. Diverse analytical techniques, including plant root protection and dish plate growth assays and gas chromatography-mass spectroscopy showed that the fungal growth inhibition was intimately associated with the volatile compounds produced by B. tropica and, in particular, two bacterial strains (MTo293 and TTe203) exhibited the highest radial mycelial growth inhibition. Morphological changes associated with these compounds, such as disruption of fungal hyphae, were identified by using photomicrographic analysis. By using gas chromatography-mass spectroscopy technique, 18 volatile compounds involved in the growth inhibition mechanism were identified, including α-pinene and limonene. In addition, we found a high proportion of bacterial strains that produced siderophores during growth with different carbon sources, such as alanine and glutamic acid; however, their roles in the antagonism mechanism remain unclear.


Assuntos
Burkholderia/química , Fungos/efeitos dos fármacos , Compostos Orgânicos Voláteis/química , Compostos Orgânicos Voláteis/farmacologia , Agentes de Controle Biológico , Colletotrichum/efeitos dos fármacos , Fusarium/efeitos dos fármacos
15.
Syst Appl Microbiol ; 35(5): 310-4, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22738764

RESUMO

A group of 20 bacterial strains was isolated from the rhizosphere of different agricultural plants growing in alkaline soils in the northeast of Mexico. The phylogenetic analysis of the 16S rRNA gene sequence from four strains showed that this novel group belonged to the Cupriavidus genus, with C. taiwanensis (∼98.9%) and C. necator (∼98.8%) as the closest species. However, DNA-DNA reassociation values were less than 20%. The novel group did not fix nitrogen and lacked nifH and nodA genes, unlike C. taiwanensis. Whole-cell protein patterns were highly similar among the 20 strains but different from the closest Cupriavidus species. BOX-PCR patterns were distinct among the 20 strains but also differed from other Cupriavidus type species. The major cellular fatty acids from strains ASC-732(T) and SLV-2362 were C(16:0), C(18:1) ω7c/12t/9t and C(16:1) ω7c and/or C(15:0) iso 2OH. The major polar lipids consisted of phosphatidylglycerol, cardiolipin, phosphatidylethanolamine, 2-hydroxylated-phosphatidylethanolamine and an unknown aminolipid. The DNA G+C content of strain ASC-732(T) was 66.8mol%. All 20 strains grew in the presence of 5-10mgmL(-1) arsenic, 1mgmL(-1) zinc, and 0.1mgmL(-1) copper. Consequently, the group of strains was considered to represent a novel species for which the name Cupriavidus alkaliphilus sp. nov. is proposed. The type strain is ASC-732(T) (=LMG 26294(T)=CIP 110330(T)).


Assuntos
Cupriavidus/classificação , Cupriavidus/isolamento & purificação , Rizosfera , Agricultura , Composição de Bases , Cupriavidus/genética , Concentração de Íons de Hidrogênio , México , Filogenia , Solo/química , Microbiologia do Solo
16.
Environ Microbiol Rep ; 3(3): 383-9, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23761284

RESUMO

Although some sugarcane cultivars may benefit substantially from biological nitrogen fixation (BNF), the responsible bacteria have been not identified yet. Here, we examined the active diazotrophic bacterial community in sugarcane roots from Africa and America by reverse transcription (RT)-PCR using broad-range nifH-specific primers. Denaturing gradient gel electrophoresis (DGGE) profiles obtained from sugarcane showed a low diversity at all sample locations with one phylotype amounting up to 100% of the nifH transcripts. This major phylotype has 93.9-99.6% DNA identity to the partial nifH sequence from a strain affiliated with Rhizobium rosettiformans. In addition, nifH transcripts of this phylotype were also detected in spruce roots sampled in Germany, where they made up 91% of nifH transcripts detected. In contrast, in control soil or shoot samples two distinct nifH transcript sequences distantly related to nifH from Sulfurospirillum multivorans or Bradyrhizobium elkanii, respectively, were predominant. These results suggest that R. rosettiformans is involved in root-associated nitrogen fixation with sugarcane and spruce, plants that do not form root-nodule symbioses.

17.
Syst Appl Microbiol ; 34(7): 531-5, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21592712

RESUMO

The aim of this work is the taxonomic characterization of three biosurfactant-producing bacterial isolates from the Churince system at Cuatro Ciénegas Basin (CCB) in the Mexican State of Coahuila, and the study of the possible role of biosurfactant production in their ecology and evolution. We determined that these isolates belong to a Pseudomonas koreensis lineage endemic to CCB, using standard taxonomical techniques, phylogenetic analysis of three chromosomal loci and phenotypic characterization. This new lineage has the distinct capacity to produce a biosurfactant when compared with previously reported P. koreensis isolates recovered from agricultural soils in Korea. We present evidence suggesting that the biosurfactant secreted by CCB P. koreensis strains is involved in their ability to compete with a CCB Exiguobacterium aurantiacum strain (m5-66) used as a model organism in competition experiments. Furthermore, the ethyl acetate extract of culture supernatant of CCB P. koreensis strains results in growth inhibition not only of E. aurantiacum m5-66, but also of a Bacillus subtilis type strain (ATCC6633). Based on these results we propose that the production of biosurfactant could be of ecological importance and could play a role in the separation of the P. koreensis CCB lineage.


Assuntos
Pseudomonas/classificação , Tensoativos/química , Antibacterianos/química , Antibacterianos/farmacologia , Bacillaceae/efeitos dos fármacos , Bacillaceae/crescimento & desenvolvimento , Proteínas de Bactérias/química , Proteínas de Bactérias/farmacologia , Técnicas de Tipagem Bacteriana , Cromossomos/genética , DNA Girase/genética , DNA Ribossômico/genética , Ecologia , Evolução Molecular , Genes Bacterianos , Loci Gênicos , México , Testes de Sensibilidade Microbiana , Fenótipo , Filogenia , Pseudomonas/química , Pseudomonas/genética , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/isolamento & purificação , Recombinases Rec A/genética , Ribossomos/genética , Tensoativos/farmacologia , Microbiologia da Água
18.
J Microbiol ; 49(6): 867-76, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22203548

RESUMO

The presence of Burkholderia, Cupriavidus, and Ralstonia species in northeastern Mexico was investigated. An analysis of the root surrounding soil from different agricultural plants led to the isolation of Burkholderia and Cupriavidus species but no Ralstonia strains. Most Cupriavidus species were unknown and grouped into two clusters according to ARDRA profiles. The 16S rRNA sequence analysis showed that the Cupriavidus isolates were highly related among them and with different Cupriavidus species with validated names. However, SDS-PAGE profiles were distinct among the different ARDRA profiles and to other Cupriavidus species examined, suggesting new species in the genus. This shows that Cupriavidus is more widely associated with plants than previously appreciated. The BCC isolate was 99% similar to B. cenocepacia by recA sequence analysis. Additionally, most Cupriavidus strains from the two largest groups grew on media containing up to 0.1 mg/ml of copper, 10.0 mg/ml arsenic and 1.0 mg/ml zinc. Burkholderia strains grew on media containing up to 10.0 mg/ml zinc, 5.0 mg/ml arsenic and 0.1 mg/ml copper.


Assuntos
Burkholderia/isolamento & purificação , Produtos Agrícolas/microbiologia , Cupriavidus/isolamento & purificação , Microbiologia do Solo , Solo/análise , Burkholderia/classificação , Burkholderia/genética , Produtos Agrícolas/crescimento & desenvolvimento , Cupriavidus/classificação , Cupriavidus/genética , DNA Bacteriano/genética , México , Dados de Sequência Molecular , Filogenia , Raízes de Plantas/microbiologia , RNA Ribossômico 16S/genética
19.
Syst Appl Microbiol ; 33(1): 35-43, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19945811

RESUMO

Several novel N(2)-fixing Burkholderia species associated with plants, including legume-nodulating species, have recently been discovered. Presently, considerable interest exists in studying the diazotrophic Burkholderia species, both for their ecology and their great potential for agro-biotechnological applications. However, the available methods used in the identification of these Burkholderia species are time-consuming and expensive. In this study, PCR species-specific primers based on the 16S rRNA gene were designed, which allowed rapid, easy, and correct identification of most known N(2)-fixing Burkholderia. With this approach, type and reference strains of Burkholderia kururiensis, B. unamae, B. xenovorans, B. tropica, and B. silvatlantica, as well as the legume-nodulating B. phymatum, B. tuberum, B. mimosarum, and B. nodosa, were unambiguously identified. In addition, the PCR species-specific primers allowed the diversity of the diazotrophic Burkholderia associated with field-grown tomato and sorghum plants to be determined. B. tropica and B. xenovorans were the predominant species found in association with tomato, but the occurrence of B. tropica with sorghum plants was practically exclusive. The efficiency of the species-specific primers was validated with the detection of B. tropica and B. xenovorans from DNA directly recovered from tomato rhizosphere soil samples. Additionally, using PCR species-specific primers, all of the legume-nodulating Burkholderia were correctly identified, even from single nodules collected from inoculated common bean plants. These primers could contribute to rapid identification of the diazotrophic and nodulating Burkholderia species associated with important crop plants and legumes, as well as revealing their environmental distribution.


Assuntos
Burkholderia/isolamento & purificação , Fabaceae/microbiologia , Fixação de Nitrogênio , Nodulação , Reação em Cadeia da Polimerase/métodos , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Biodiversidade , Burkholderia/classificação , Burkholderia/genética , Burkholderia/fisiologia , Primers do DNA/genética , Sensibilidade e Especificidade
20.
FEMS Microbiol Lett ; 296(1): 52-9, 2009 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-19459961

RESUMO

Bacteria of the genus Azospirillum increase the grain yield of several grass crops. In this work the effect of inoculating maize plants with genetically engineered Azospirillum brasilense for trehalose biosynthesis was determined. Transformed bacteria with a plasmid harboring a trehalose biosynthesis gene-fusion from Saccharomyces cerevisiae were able to grow up to 0.5 M NaCl and to accumulate trehalose, whereas wild-type A. brasilense did not tolerate osmotic stress or accumulate significant levels of the disaccharide. Moreover, 85% of maize plants inoculated with transformed A. brasilense survived drought stress, in contrast with only 55% of plants inoculated with the wild-type strain. A 73% increase in biomass of maize plants inoculated with transformed A. brasilense compared with inoculation with the wild-type strain was found. In addition, there was a significant increase of leaf and root length in maize plants inoculated with transformed A. brasilense. Therefore, inoculation of maize plants with A. brasilense containing higher levels of trehalose confers drought tolerance and a significant increase in leaf and root biomass. This work opens the possibility that A. brasilense modified with a chimeric trehalose biosynthetic gene from yeast could increase the biomass, grain yield and stress tolerance in other relevant crops.


Assuntos
Azospirillum brasilense/metabolismo , Estresse Fisiológico , Trealose/metabolismo , Zea mays/microbiologia , Zea mays/fisiologia , Biomassa , Vias Biossintéticas/genética , Dessecação , Folhas de Planta/crescimento & desenvolvimento , Raízes de Plantas/crescimento & desenvolvimento , Plasmídeos , Saccharomyces cerevisiae/genética , Análise de Sobrevida
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