Abstract
Drosera villosa var. villosa A. St.-Hil is a carnivorous plant that grows in Brazilian flooded soils very poor on nutrients, including low levels of N. Under these conditions, the plant shows vigorous growth, low root number, low number of captured prey (less than 50%) and a great assemblage of bacteria associated with the roots and leaves that grow in N-free medium. These preliminary results have led us to investigate the number of colony forming units (log CFU) in the roots (rhizosphere and endorhizosphere) and leaves (phyllosphere and endophyllosphere) of D. villosa var. villosa by the tenfold serial dilution technique in two N-free culture media. The results showed that the phyllosphere had 6.65 log CFU g dry leaf−1 and the rhizosphere 6.47 log CFU g dry soil−1, with the lowest value detected in the endophyllosphere (4.39 log CFU g dry leaf−1). Sixty-three different bacteria morphotypes were isolated from the surface and interior of the roots and leaves and the amplification of the DNA with specific primers detected the nifH gene in 34 of those strains. The DNAs of the 34 strains were compared by the BOX-PCR technique and a great diversity was observed, with the bacteria clustering at a final level of similarity of only 12%. The strains were also submitted to the partial sequencing of the 16S rRNA gene and several genera of N2-fixing bacteria were detected, including Bacillus, Burkholderia, Methylobacterium, Paenibacillus, Pseudomonas and Sphingomonas.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Achouak W, Normand P, Heulin T (1999) Comparative phylogeny of rrs and nifH genes in the Bacillaceae. Int J Syst Bacteriol 49:961–967
Adamec L (1997) Mineral nutrition of carnivorous plants: a review. Bot Rev 63:273–299
Adhikari TB, Joseph CM, Yang GP, Phillips DA, Nelson LM (2001) Evaluation of bacteria isolated from rice for plant growth promotion and biological control of seedling disease of rice. Can J Microbiol 47:916–924
Albino U (2004) Isolamento e caracterização de bactérias diazotróficas associadas à planta carnívora D. villosa var. villosa e seu potencial como inoculante de plantas arbóreas associadas a fungos micorrízicos arbusculares. Ph.D. thesis, Universidade Estadual de Londrina
Andrade G, Esteban E, Velasco L, Lorite MJ, Bedmar EJ (1997a) Isolation and identification of N2-fixing microorganisms from the rhizosphere of Capparis spinosa (L.). Plant Soil 197:19–23
Andrade G, Mihara KL, Linderman RG, Bethlenfalvay GJ (1997b) Bacteria from rhizosphere and hyphosphere soils of different arbuscular mycorrhizal fungi. Plant Soil 192:71–79
Balota EL, Lopes ES, Hungria M, Dobereiner J (1999) Occurrence of diazotrophic bacteria and arbuscular mycorrhizal fungi on the cassava crop. Pesqui Agropecu Bras 34:1265–1276
Barraquio WL, Revilla L, Ladha JK (1997) Isolation of endophytic diazotrophic bacteria from wetland rice. Plant Soil 194:15–24
Bending GD, Poole EJ, Whipps JM, Read DJ (2002) Characterisation of bacteria from Pinus sylvestris–Suillus luteus mycorrhizas and their effects on root–fungus interactions and plant growth. FEMS Microbiol Ecol 39:219–227
Boddey RM, Urquiaga S, Alves BJR, Reis V (2003) Endophytic nitrogen fixation in sugarcane, present knowledge and future applications. Plant Soil 252:139–149
Caballero-Mellado J, Lopez-Reyes L, Bustillos-Cristales R (1999) Presence of 16S rRNA genes in multiple replicons in Azospirillum brasilense. FEMS Microbiol Lett 178:283–288
Chaintreuil C, Giraud E, Prin Y, Lorquin J, Bâ A, Gillis M, de Lajudie P, Dreyfus B (2000) Photosynthetic bradyrhizobia are natural endophytes of the African wild rice Oryza breviligulata. Appl Environ Microbiol 66:5437–5447
Chen LS, Figueredo A, Pedrosa FO, Hungria M (2000) Genetic characterization of soybean rhizobia in Paraguay. Appl Environ Microbiol 66:5099–5103
Chua KL, Chan YY, Gan YH (2003) Flagella are virulence determinants of Burkholderia pseudomallei. Infect Immun 71:1622–1629
Cruz LM, De Souza EM, Weber OB, Baldani JI, Döbereiner J, Pedrosa FO (2001) 16S ribosomal DNA characterization of nitrogen-fixing bacteria isolated from banana (Musa spp.) and pineapple (Ananas comosus (L.) Merril). Appl Environ Microbiol 67:2375–2379
Döbereiner J, Day JM (1976) Associative symbiosis in tropical grasses, characterization of microorganisms and dinitrogen fixing sites. In: Proceedings of the 1st international symposium of nitrogen fixation. Washington State University Press, Pullman
Elbeltagy A, Nishioka K, Suzuki H, Sato T, Sato YI, Morisaki H, Mitsui H, Minamisawa K (2000) Isolation and characterization of endophytic bacteria from wild and traditionally cultivated rice varieties. Soil Sci Plant Nutr 46:617–629
Estrada-De Los Santos P, Bustillos-Cristales R, Caballero-Mellado J (2001) Burkholderia, a genus rich in plant-associated nitrogen fixers with wide environmental and geographic distribution. Appl Environ Microbiol 67:2790–2798
Fernandes MF, Fernandes RPM, Rodrigues LD (2001) Diazotrophic bacteria associated to coconut palms in a coastal lowland region in Sergipe State, Brazil. Pesqui Agropecu Bras 36:1509–1517
Gough C, Vasse J, Galera C, Webster G, Cocking E, Dénarié J (1997) Interactions between bacterial diazotrophs and non-legume dicots: Arabidopsis thaliana as a model plant. Plant Soil 194:123–130
Gyaneshwar P, James EK, Mathan N, Reddy PM, Renhold-Hurek B, Ladha JK (2001) Endophytic colonization of rice by a diazotrophic strain of Serratia marcescens. J Bacteriol 183:2634–2645
Kaschuk G, Hungria M, Santos JCP, Berton-Junior JF (2006) Differences in common bean rhizobial populations associated with soil tillage management in southern Brazil. Soil Till Res 87:205–217
Menna P, Hungria M, Barcellos FG, Bangel EV, Hess PN, Martínez-Romero E (2006) Molecular phylogeny based on the 16S rRNA gene of elite rhizobial strains used in Brazilian commercial inoculants. Syst Appl Microbiol 29:315–332
Minerdi D, Fani R, Gallo R, Boarino A, Bonfante P (2001) Nitrogen fixation genes in an endosymbiotic Burkholderia strain. Appl Environ Microbiol 67:725–732
O’Callaghan KJ, Stone PJ, Hu X, Griffiths DW, Davey MR, Cocking EC (2000) Effects of glucosinolates and flavonoids on colonization of the roots of Brassica napus by Azorhizobium caulinodans ORS571. Appl Environ Microbiol 66:2185–2191
Peix A, Mateos PF, Rodriguez-Barrueco C, Martinez-Molina E, Velásquez E (2001) Growth promotion of common bean (Phaseolus vulgaris L.) by a strain of Burkholderia cepacia under growth chamber conditions. Soil Biol Biochem 33:1927–1935
Perica MC, Berljak J (1996) In vitro growth and regeneration of D. spatulata Labill. on various media. Hortic Sci 31:1033–1034
Rosado AS, Duarte GF, Seldin L, Van Elsas JD (1998) Genetic diversity of nifH gene sequences in Paenibacillus azotofixans strains and soil samples analyzed by denaturing gradient gel electrophoresis of PCR-amplified gene fragments. Appl Environ Microbiol 64:2770–2779
Róźycki H, Dahm H, Strzelczyk E, Li CY (1999) Diazotrophic bacteria in root-free soil in the root of pine (Pinus sylvestris L.) and oak (Quercus robur L.). Appl Soil Ecol 12:239–250
Sy A, Giraud E, Samba R, de Lajudie P, Gillis M, Dreyfus B (2001) Certain legumes of the genus Crotalaria are specifically nodulated by a new species of Methylobacterium. Can J Microbiol 47:503–508
Ueda T, Suga Y, Yahiro N, Matsuguchi T (1995) Remarkable N2-fixing bacterial diversity detected in rice roots by molecular evolutionary analysis of nifH gene sequences. J Bacteriol 177:1414–1417
Wilson PW, Knight SC (1952) Experiments in bacterial physiology. Burguess, Minneapolis, pp 49
Yoon JH, Seo WT, Shin YK, Kho YH, Kang KH, Park YH (2002) Paenibacillus chinjuensis sp nov., anovel exopolysaccharide-producing bacterium. Int J Syst Evol Microbiol 52:415–421
Acknowledgments
We thank to the National Council of Scientific and Technological Development (CNPq) who enabled the execution of this study by conceding PIBIC, M.Sc., Ph.D. and Productivity in Research grants and to Ligia Maria de O. Chueire for valuable collaboration in the DNA sample sequencing. PRONEX-CNPq partially funded the study.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Albino, U., Saridakis, D.P., Ferreira, M.C. et al. High diversity of diazotrophic bacteria associated with the carnivorous plant Drosera villosa var. villosa growing in oligotrophic habitats in Brazil. Plant Soil 287, 199–207 (2006). https://doi.org/10.1007/s11104-006-9066-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11104-006-9066-7