Complete genome sequence of the sugarcane nitrogen-fixing endophyte Gluconacetobacter diazotrophicus Pal5
- Marcelo Bertalan,
- Rodolpho Albano,
- Vânia de Pádua,
- Luc Rouws,
- Cristian Rojas,
- Adriana Hemerly,
- Kátia Teixeira,
- Stefan Schwab,
- Jean Araujo,
- André Oliveira,
- Leonardo França,
- Viviane Magalhães,
- Sylvia Alquéres,
- Alexander Cardoso,
- Welington Almeida,
- Marcio Martins Loureiro,
- Eduardo Nogueira,
- Daniela Cidade,
- Denise Oliveira,
- Tatiana Simão,
- Jacyara Macedo,
- Ana Valadão,
- Marcela Dreschsel,
- Flávia Freitas,
- Marcia Vidal,
- Helma Guedes,
- Elisete Rodrigues,
- Carlos Meneses,
- Paulo Brioso,
- Luciana Pozzer,
- Daniel Figueiredo,
- Helena Montano,
- Jadier Junior,
- Gonçalo de Souza Filho,
- Victor Martin Quintana Flores,
- Beatriz Ferreira,
- Alan Branco,
- Paula Gonzalez,
- Heloisa Guillobel,
- Melissa Lemos,
- Luiz Seibel,
- José Macedo,
- Marcio Alves-Ferreira,
- Gilberto Sachetto-Martins,
- Ana Coelho,
- Eidy Santos,
- Gilda Amaral,
- Anna Neves,
- Ana Beatriz Pacheco,
- Daniela Carvalho,
- Letícia Lery,
- Paulo Bisch,
- Shaila C Rössle,
- Turán Ürményi,
- Alessandra Rael Pereira,
- Rosane Silva,
- Edson Rondinelli,
- Wanda von Krüger,
- Orlando Martins,
- José Ivo Baldani,
- Paulo CG Ferreira
- … show all 61 hide
Abstract
Background
Gluconacetobacter diazotrophicus Pal5 is an endophytic diazotrophic bacterium that lives in association with sugarcane plants. It has important biotechnological features such as nitrogen fixation, plant growth promotion, sugar metabolism pathways, secretion of organic acids, synthesis of auxin and the occurrence of bacteriocins.
Results
Gluconacetobacter diazotrophicus Pal5 is the third diazotrophic endophytic bacterium to be completely sequenced. Its genome is composed of a 3.9 Mb chromosome and 2 plasmids of 16.6 and 38.8 kb, respectively. We annotated 3,938 coding sequences which reveal several characteristics related to the endophytic lifestyle such as nitrogen fixation, plant growth promotion, sugar metabolism, transport systems, synthesis of auxin and the occurrence of bacteriocins. Genomic analysis identified a core component of 894 genes shared with phylogenetically related bacteria. Gene clusters for gum-like polysaccharide biosynthesis, tad pilus, quorum sensing, for modulation of plant growth by indole acetic acid and mechanisms involved in tolerance to acidic conditions were identified and may be related to the sugarcane endophytic and plant-growth promoting traits of G. diazotrophicus. An accessory component of at least 851 genes distributed in genome islands was identified, and was most likely acquired by horizontal gene transfer. This portion of the genome has likely contributed to adaptation to the plant habitat.
Conclusion
The genome data offer an important resource of information that can be used to manipulate plant/bacterium interactions with the aim of improving sugarcane crop production and other biotechnological applications.
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1471-2164-10-450-S1.PDF
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Additional file 1: Distribution of mobile elements in plant endophyte complete genomes. The percentage column: Percentage of total number of mobile elements from all CDS annotated on the endophyte complete genomes. (PDF 13 KB)
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Additional file 2: Predicted Highly Expressed (PHX) genes. The PHX and proteomic analysis was used to indicate potentially important genes in the GDI genome. (XLS 46 KB)
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Additional file 3: 16S phylogenetic tree from Alphaproteobacteria. The Neighbor joining phylogenetic tree of 16S from Alphaproteobacteria was done using ClustalX. In blue are the three completed genomes closest to G. diazotrophicus PAL5 available in GenBank. (EPS 57 KB)
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1471-2164-10-450-S4.PDF
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Additional file 4: The 28 genome islands (GI) identified by GC3 and IVOMs. The GI column has the ID for each genome island. The integrase column shows which kind of integrase was found in each genome island. The CDS column shows how many CDS are inside the genome island. The Alien+GC3 column show how many CDS in each genome island were identified as accessory by both methods. The Related column shows which kinds of genes were found in each genome island. (PDF 13 KB)
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1471-2164-10-450-S5.PDF
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Additional file 5: Variation in G. diazotrophicus strains. 20 different strains were tested for gene variation. 37 CDS were selected from 21 putative genome islands and 17 CDS were selected from putative core regions of the chromossome as control. (+): PCR positive. (-): PCR negative. (PDF 66 KB)
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Additional file 6: Presence of homologues of the trbE gene among G. diazotrophicus strains. Total DNA of 11 Gluconacetobacter strains was completely digested with restriction enzymes EcoRI (a.) or EcoRV (b.), seParated on agarose gel and submitted to Southern blot analysis using a fragment of CDS GDI0133 (trbE, part of type IV secretion system) as a probe. Numbers 1-10 represent G. diazotrophicus strains: Pal5 (1), 3R2 (2), URU (3), 38f2 (4), PRJ50(5), Pal3 (6), AF3 (7), PCRI (8), PPe4 (9), CNFe-550 (10). Number 11 represents G. johannae. In strain Pal5, only 3 bands are present, although the genome sequence indicates the presence of four copies of the trbE gene. However, the fourth trbE Paralog (GDI1016) is more dissimilar to the probe sequence then the other three (GDI0133, GDI2742 e GDI2911), which may have prevented hybridization. (JPEG 103 KB)
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Additional file 7: Distribution of percent ID from RBH results. In red all the RBH from Rhodospirillales order. In yellow all RBH from other Alphaproteobacteria class and in blue RBH from other genomes beside Alphaproteobacteria class. (EPS 759 KB)
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Additional file 8: Number of Reciprocal Best Hits (RBH) in accessory and core regions. The first column shows the number of RBH for each organism in parentheses. The RBH in columns show the total number of RBH for each organism. The RBH % by organism columns shows the percent of RBH in relation with the total number of RBH found in accessory and core regions. RBH result has 708 RBH in accessory regions and 2,258 in core regions. The RBH % by organism columns shows the percentage of RBH in accessory and core regions for each organism or group. GOX = Gluconobacter oxydans 621H, GBE = Granulibacter bethesdensis CGDNIH, ACR = Acidiphilium cryptum JF-5, Rhiz = All the complete genomes from Rhizobiales order, Other Alpha = All other complete genomes from Alphaproteobacteria class, Beta = All complete genomes from Betaproteobacteria class, Gamma = All complete genomes from Gammaproteobacteria class, Others = All other complete genomes. (PDF 9 KB)
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Additional file 9: Endophyte comparison gene list. Endophyte gene list. (XLS 37 KB)
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Additional file 10: Endophyte comparison. In gray, genes similar to all genomes (core + endophyte, see Methods). In blue, genes present in all endophyte but not in core genomes. In red, genes only similar to GDI and Azoarcus sp BH72. In purple, genes only similar to GDI and Methylobacterium populi BJ001 and in green genes that are only present in GDI and at least two other endophyte genome. (EPS 1 MB)
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1471-2164-10-450-S11.PDF
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Additional file 11: Comparison of main signalling protein categories. AT, Agrobacterium tumefaciens C58; BJ, Bradyrhizobium japonicum USDA110; ML, Mesorhizobium loti MAFF303099, SM, Sinorhizobium meliloti 1021, GO, Gluconobacter oxydans 621H; RP, Rickettsia prowazekii MadridE; AB, Azoarcus sp. BH72; AE, Azoarcus sp. EbN1; XF, Xylella fastidiosa 9a5c; EC, Escherichia coli K12-MG1655. (PDF 14 KB)
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Additional file 12: Comparison of main transport-related protein categories. AT, Agrobacterium tumefaciens C58; BJ, Bradyrhizobium japonicum USDA110; ML, Mesorhizobium loti MAFF303099, SM, Sinorhizobium meliloti 1021, GO, Gluconobacter oxydans 621H; RP, Rickettsia prowazekii MadridE; AB, Azoarcus sp. BH72; AE, Azoarcus sp. BH72, XF, Xylella fastidiosa 9a5c; EC, Escherichia coli K12-MG1655. (PDF 14 KB)
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Additional file 13: Comparison among the two Gluconacetobacter diazotrophicus Pal5 genomic sequences. GDI-BR, NCBI RefSeq NC_010125, GDI-US, NCBI RefSeq NC_011365, GIs, Genome Islands. (PDF 13 KB)
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Additional file 14: CDS list of the two Gluconacetobacter diazotrophicus Pal5 sequences. Sheet 1: Blast best hits list of CDS found in both genomes. Sheet 2: List of unique CDS found in chromosome from GeneBank file CP001189. Sheet 3: List of unique genes found in chromosome from GeneBank file AM889285 (this work). (XLS 505 KB)
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- Title
- Complete genome sequence of the sugarcane nitrogen-fixing endophyte Gluconacetobacter diazotrophicus Pal5
- Open Access
- Available under Open Access This content is freely available online to anyone, anywhere at any time.
- Journal
-
BMC Genomics
10:450
- Online Date
- September 2009
- DOI
- 10.1186/1471-2164-10-450
- Online ISSN
- 1471-2164
- Publisher
- BioMed Central
- Additional Links
- Topics
- Industry Sectors
- Authors
-
-
Marcelo Bertalan
(1)
-
Rodolpho Albano
(2)
-
Vânia de Pádua
(3)
-
Luc Rouws
(4)
-
Cristian Rojas
(1)
-
Adriana Hemerly
(1)
(12)
-
Kátia Teixeira
(4)
-
Stefan Schwab
(4)
-
Jean Araujo
(4)
-
André Oliveira
(4)
-
Leonardo França
(1)
-
Viviane Magalhães
(1)
-
Sylvia Alquéres
(1)
-
Alexander Cardoso
(1)
-
Welington Almeida
(1)
-
Marcio Martins Loureiro
(1)
-
Eduardo Nogueira
(11)
(3)
-
Daniela Cidade
(2)
-
Denise Oliveira
(2)
-
Tatiana Simão
(2)
-
Jacyara Macedo
(2)
-
Ana Valadão
(2)
-
Marcela Dreschsel
(4)
-
Flávia Freitas
(2)
-
Marcia Vidal
(4)
-
Helma Guedes
(4)
-
Elisete Rodrigues
(4)
-
Carlos Meneses
(4)
-
Paulo Brioso
(5)
-
Luciana Pozzer
(5)
-
Daniel Figueiredo
(5)
-
Helena Montano
(5)
-
Jadier Junior
(5)
-
Gonçalo de Souza Filho
(6)
-
Victor Martin Quintana Flores
(6)
-
Beatriz Ferreira
(6)
-
Alan Branco
(6)
-
Paula Gonzalez
(7)
-
Heloisa Guillobel
(7)
-
Melissa Lemos
(8)
-
Luiz Seibel
(8)
-
José Macedo
(8)
-
Marcio Alves-Ferreira
(9)
-
Gilberto Sachetto-Martins
(9)
-
Ana Coelho
(9)
-
Eidy Santos
(9)
-
Gilda Amaral
(9)
-
Anna Neves
(9)
-
Ana Beatriz Pacheco
(10)
-
Daniela Carvalho
(10)
-
Letícia Lery
(10)
-
Paulo Bisch
(10)
-
Shaila C Rössle
(10)
-
Turán Ürményi
(10)
-
Alessandra Rael Pereira
(2)
-
Rosane Silva
(10)
-
Edson Rondinelli
(10)
-
Wanda von Krüger
(10)
-
Orlando Martins
(1)
-
José Ivo Baldani
(4)
-
Paulo CG Ferreira
(1)
(12)
-
Marcelo Bertalan
- Author Affiliations
-
- 1. UFRJ, CCS, Bloco D, Instituto de Bioquímica Médica, 21491-590, subssolo, Rio de Janeiro, Brazil
- 2. Departamento de Bioquímica,UERJ, Instituto de Biologia Roberto Alcântara Gomes, Blv 28 de Setembro, 87, fundos, 4 andar, RJ 20551-013, Vila Isabel, Rio de Janeiro, Brazil
- 3. Laboratório de Tecnologia em Bioquímica e Microscopia, Centro Universitário Estadual da Zona Oeste, 23070-200, Rio de Janeiro, Brazil
- 4. Embrapa Agrobiologia BR465, Km 07 Seropédica Rio de Janeiro, 23851-970, Brazil
- 12. Laboratório de Biologia Molecular de Plantas,Botânico do Rio de Janeiro,Rio de Janeiro, Instituto de Pesquisas do Jardim, 22460-030, RJ, Brazil
- 11. Laboratório de Biologia Molecular, Departamento de Genética e Biologia Molecular,Rio de Janeiro, Universidade Federal do Estado do Rio de Janeiro, 22290-240, RJ, Brazil
- 5. Instituto de Biologia, Departamento de Entomologia e Fitopatologia, Universidade Federal Rural do Rio de Janeiro, Cx Postal 74585/BR 465, KM 07, 23851-970, Seropédica, RJ
- 6. Lab. Biotecnologia-Alberto Lamego 2000 Campos dos Goytacazes, Centro de Biociências e Biotecnologia Universidade Estadual do Norte Fluminense- Av., 28013-620, RJ, Brazil
- 7. Departamento de Biofísica e Biometria,87, fundos, 4 andar,Rio de Janeiro, Instituto de Biologia Roberto Alcântara Gomes UERJ, Blv 28 de Setembro, 20551-013, Vila Isabel, RJ, Brazil
- 8. Departamento de Informática, 225, Pontifícia Universidade Católica do Rio de Janeiro Rua Marquês de S. Vicente, 22453-900, Rio de Janeiro, Brazil
- 9. Departamento de Genética, Instituto de Biologia,Rio de Janeiro, Universidade Federal do Rio de Janeiro, 68011, 21941-617, RJ, Brazil
- 10. Instituto de Biofísica Carlos Chagas Filho Universidade Federal do Rio de Janeiro, CCS, Cidade Universitária, RJ21.949-900, Rio de Janeiro, Brazil