Skip to main content

Advertisement

Log in

Root colonization and growth enhancement in wheat and tomato by rhizobacteria isolated from the rhizoplane of grasses

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Rhizobacteria isolated from the rhizoplane of grasses growing at the Nylsvlei Nature Reserve in South Africa were investigated for growth promotion and root colonization in wheat (Triticum aestivum L.) and tomato (Lycopersicon esculentum Mill.) under greenhouse and microplot field conditions. The identities of the isolates were determined by means of 16S rRNA gene sequencing as Bacillus simplex (KBS1F-3), Bacillus megaterium (NAS7-L), Bacillus cereus (KFP9-F) and Paenibacillus alvei (NAS6G-6). The three Bacillus strains were isolated from the perennial grass Themeda triandra while the Paenibacillus strain was isolated from another perennial grass Sporobolus fimbriatus. Enhanced plant shoot and root weight in wheat was achieved by single inoculation with three of the isolates whereas no significant increase was observed in root length. Combined inoculation of Paenibacillus alvei (NAS6G-6) and Bacillus cereus (KFP9-F) on wheat resulted in significant increase in these parameters. Single inoculations of Bacillus simplex (KBS1F-3) and Bacillus cereus (KFP9-F) resulted in significant increase in root and shoots fresh weight, root dry weight and total root length in tomatoes. Indoleacetic acid production, phosphate solubilization and siderophore secretion were studied as possible mechanisms by which the bacterial isolates enhanced plant growth. Root colonization was studied by means of spontaneous rifampicin resistant strains of the wild type isolates. Except for B. megaterium (NAS7-L), the rest of the isolates colonized the roots efficiently resulting in concentrations of 106–108 cfu g−1 root. The root colonization of Bacillus simplex (KBS1F-3) and Paenibacillus alvei (NAS6G-6) was visualized by confocal scanning laser microscope (CSLM) after successful transformation of the isolates with the pNF8 plasmid carrying the gene for the green fluorescent protein (gfp).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Ashghar HN, Zahir ZA, Arshad M (2004) Screening for improving the growth, yield and oil content of canola (Brassica napus L.). Aust J Agric Res 55:187–194

    Article  Google Scholar 

  • Barneix AJ, Saubidet MI, Fatta N, Kade M (2005) Effect of rhizobacteria on growth and grain protein in wheat. Agron Sustain Dev 25:505–511

    Article  Google Scholar 

  • Berge O, Guinebretiere MH, Achouak W, Normand P, Heilin T (2002) Paenibacillus graminis sp. Nov. and Paenibacillus odorifersp.nov. isolated from plant roots, soil and food. Int J Syst Evol Microbiol 32:607–616

    Google Scholar 

  • Bloemberg GV, Lugtenberg BJJ (2001) Molecular basis of plant growth promotion and biocontrol by rhizobacteria. Curr Opin Plant Biol 4:343–350

    Article  CAS  Google Scholar 

  • Brown GD, Rovira AD (1999) The rhizosphere and its management to improve plant growth. Adv Agron 66:1–102

    Article  Google Scholar 

  • Cakmakcl R, Erat M, Erdagan U, Figen-Donmez M (2007) The influence of plant-growth-promoting rhizobacteria on growth and enzyme activities in wheat and spinach plants. J Plant Nutr Soil Sci 170:288–295

    Article  Google Scholar 

  • Donate-Correa J, Leon-Barrios M, Perez-Galdona R (2004) Screening for plant-growth-promoting rhizobacteria in Chamaecytisus proliferus (tagasate), a forage shrub legume endemic to the Canary islands. Plant Soil 266:261–272

    Article  CAS  Google Scholar 

  • Egamberdiyeva D (2008) Plant-growth-promoting properties of rhizobacteria isolated from wheat and pea grown in loamy sand soil. Turk J Biol 32:9–15

    Google Scholar 

  • Egamberdiyeva D, Hoflich G (2002) Root colonization and growth promotion of winter wheat and pea by Cellulomonas spp. at different temperatures. Plant Growth Regul 38:219–224

    Article  CAS  Google Scholar 

  • Fortineau N, Trieu-Cuot P, Gaillot O, Pellegrini E, Berche P, Gaillard JL (2000) Optimization of green fluorescent protein expression vectors for in vitro and in vivo detection of Listeria monocytogens. Res Microbiol 151:353–360

    Article  CAS  Google Scholar 

  • Gamalero EA, Trotta A, Massa N, Coppeta A, Martinotti GM, Berta G (2004) Impact of two fluorescent pseudomonads and an arbuscular mycorrhizal fungus on tomato plant growth, root architecture and P-acquisition. Mycorrhiza 14:185–192

    Article  Google Scholar 

  • Glick BR (1995) The enhancement of plant growth by free living bacteria. Can J Microbiol 41:109–117

    Article  CAS  Google Scholar 

  • Glick BR, Bashan Y (1997) Genetic manipulations of plant-growth-promoting bacteria to enhance biocontrol of phytopathogens. Biotech Adv 15:353–378

    Article  CAS  Google Scholar 

  • Idris A, Labuschagne N, Korsten L (2009) Efficacy of rhizobacteria for plant growth promotion in sorghum under greenhouse conditions and selected modes of action studies. J Agric Sci 147:17–30

    Article  CAS  Google Scholar 

  • Itzigsohn R, Burdman S, Okon Y (2000) Plant growth promotion in natural pastures by inoculation with Azosprillum brasilense under suboptimal growth conditions. Arid Soil Res Rehabil 13:151–158

    Google Scholar 

  • Khalid A, Arshad M, Zahir ZA (2004) Screening plant-growth-promoting rhizobacteria for improving growth and yield of wheat. J Appl Microbiol 96:473–480

    Article  CAS  Google Scholar 

  • King EO, Ward MK, Ranney DE (1954) Two simple media for the demonstration of pyocynin and fluorescin. J Lab Clin Med 44:301–307

    CAS  Google Scholar 

  • Kishore GK, Pande S, Podile AR (2005) Phyloplane bacteria increase seedling emergence, growth and yield of field grown ground nut (Arachis hypogaea L). Lett Appl Microbiol 40:260–268

    Article  CAS  Google Scholar 

  • Kloepper JW (1993) Plant-growth-promoting rhizobacteria as biological control agents. In: Metting FB Jr (ed) Soil microbial ecology: applications in agriculture and environmental management. Marcel Dekker Inc., New York, pp 255–274

    Google Scholar 

  • Kohler J, Caravaca F, Carrasco L, Roldan A (2006) Contribution of Pseudomonas mendosina and Glomus intraradices to aggregate stabilization and promotion of biological fertility in rhizosphere soil of lettuce plants under field conditions. Soil Use Manag 22:298–304

    Article  Google Scholar 

  • Lane DJ (1991) 16S/23S rRNA sequencing. In: Stackebrandt E, Goodfellow M (eds) Nucleic acid techniques in bacterial systematics. John Wiley, New York, pp 115–175

    Google Scholar 

  • Lynch JM, Whipps JM (1990) Sustainable flow in the rhizosphere. Plant Soil 129:1–10

    Article  CAS  Google Scholar 

  • Mena-Violante HG, Olalde-Portugal V (2007) Alteration of fruit quality by root inoculation with plant-growth-promoting rhizobacteria (PGPR) Bacillus subtilis BEB13bs. Sci Hortic 113:103–106

    Article  CAS  Google Scholar 

  • Nautiyal CS (1997) A method for the selection and characterization of rhizosphere competent bacteria of chickpea. Curr Microbiol 34:12–17

    Article  CAS  Google Scholar 

  • Nelson LM (2004) Plant-growth-promoting rhizobacteria (PGPR): prospects for new inoculants. Crop Manag. doi:10.1094/CM-2004-031-05-RV

  • Newman ET (1966) A method of estimating the total length of root in a sample. J Appl Ecol 3:139–145

    Article  Google Scholar 

  • Persello-Cartieaux F, Nussaume L, Robaglia C (2003) Tales from the underground: molecular plant- microbe interaction. Plant Cell Environ 26:189–199

    Article  CAS  Google Scholar 

  • Pikovskaya RI (1948) Mobilization of phosphorus in connection with the vital activity of some microbial species. Microbiologia 17:362–370

    CAS  Google Scholar 

  • Probanza A, Lucas Garcia JA, Ruiz-Palomino M, Ramos B, Gutierrez-Manero FJ (2002) Pinus pinea L. seedling growth and bacterial rhizosphere structure after inoculation with PGPR Bacillus (B. lechinofermis CECT5106 and B. pumilis CECT5105). Appl Soil Ecol 20:75–84

    Article  Google Scholar 

  • RHIBAC (2007) Rhizobacteria for reduced fertilizer inputs in wheat (RHIBAC). Available on line at http://www.rhibac.org. Accessed 23 Jan 2009

  • Rodriguez H, Fraga R (1999) Phosphate solubilising bacteria and their role in plant growth promotion. Biotech Adv 17:319–339

    Article  CAS  Google Scholar 

  • Rosas SB, Avanzini G, Carlier E, Pasluosta C, Pastor N, Rovera M (2008) Root colonization and growth promotion of wheatand maize by Pseudomonas aurantiaca. Soil Biol Biochem. doi:10.1016/jsoilbio.2008.10.009

  • Saleem M, Arshad M, Hussain S, Bhatti AS (2007) Prospectives of plant-growth-promoting rhizobacteria (PGPR) containing ACC deaminase in stress agriculture. J Ind Microbiol Biotechnol 34:635–648

    Article  CAS  Google Scholar 

  • SAS Institute (2003) SAS/STAT guide for personal computers. SAS Institute, Cary

    Google Scholar 

  • Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56

    Article  CAS  Google Scholar 

  • Silva HAS, Romeiro RS, Mounter A (2003) Development of a root colonization bioassay for rapid screening of rhizobacteria for potential biocontrol agents. J Phytopathol 151:42–46

    Article  Google Scholar 

  • Singh BK, Munro S, Potts JM, Millard P (2007) Influence of grass species and soil type on rhizosphere microbial community structure in grassland soils. Appl Soil Ecol 36:147–155

    Article  Google Scholar 

  • Takagi H, Kagiyama S, Kadowiki K, Tsukagoshi N, Udaka S (1989) Genetic transformation of Bacillus brevis with plasmid DNA by electroporation. Agric Biol Chem 53:3099–3100

    CAS  Google Scholar 

  • Tejera N, Lluch C, Martinez-Toledo MV, Gonzalez-Lopez J (2005) Isolation and characterization of Azotobacter and Azosprillum strain from sugarcane rhizosphere. Plant Soil 270:223–232

    Article  CAS  Google Scholar 

  • Thakuria D, Talukdar NC, Goswami C, Hazarika S, Boro RC, Khan MR (2004) Characterization and screening of bacteria from rhizosphere of rice grown in acidic soils of Assam. Curr Sci 86:978–985

    Google Scholar 

  • Turgeon N, Laclamme C, Ho J, Duchaine C (2006) Elaboration of an electroporation protocol for Bacillus cereus ATTC14579. J Microbiol Methods 67:543–548

    Article  CAS  Google Scholar 

  • Van Loon LC, Bakker PAHM (2003) Signaling in rhizobacteria-plant interaction. Ecol Studies 168:297–330

    Google Scholar 

  • von der Weid I, Artusson V, Seldin L, Jansson JK (2005) Antifungal and root surface colonization properties of GFP-tagged Paenibacillus brasilensis PB177. World J Microbiol Biotech 12:1591–1597

    Article  Google Scholar 

  • Yasmin F, Othman R, Said-Saad M, Sijman K (2007) Screening for beneficial properties of rhizobacteria isolated from sweet potato rhizosphere. Biotechnol 6:49–52

    Article  Google Scholar 

Download references

Acknowledgments

We would like to acknowledge Dr. Nicolas Fortineau (Institut National de la Santé et de la Recherche Médicale, France) who kindly provided us with the E. coli JM109 which carries the plasmid used in the transformation of Bacillus strains. We would also like to thank Mr. Alan Hall of the microscopy division of the University of Pretoria, for the confocal scanning laser microscopy, Me Eva Arrebola-Diez for help with sequencing of the bacterial isolates and Prof. Fanus Venter of the Department of Microbiology and, Dr. Lyn Maree of the Department of Biochemistry, University of Pretoria for allowing us the use of their laboratories and facilities.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Ahmed Idris Hassen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hassen, A.I., Labuschagne, N. Root colonization and growth enhancement in wheat and tomato by rhizobacteria isolated from the rhizoplane of grasses. World J Microbiol Biotechnol 26, 1837–1846 (2010). https://doi.org/10.1007/s11274-010-0365-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-010-0365-z

Keywords

Navigation