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).
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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
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
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
Bloemberg GV, Lugtenberg BJJ (2001) Molecular basis of plant growth promotion and biocontrol by rhizobacteria. Curr Opin Plant Biol 4:343–350
Brown GD, Rovira AD (1999) The rhizosphere and its management to improve plant growth. Adv Agron 66:1–102
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
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
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
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
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
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
Glick BR (1995) The enhancement of plant growth by free living bacteria. Can J Microbiol 41:109–117
Glick BR, Bashan Y (1997) Genetic manipulations of plant-growth-promoting bacteria to enhance biocontrol of phytopathogens. Biotech Adv 15:353–378
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
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
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
King EO, Ward MK, Ranney DE (1954) Two simple media for the demonstration of pyocynin and fluorescin. J Lab Clin Med 44:301–307
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
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
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
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
Lynch JM, Whipps JM (1990) Sustainable flow in the rhizosphere. Plant Soil 129:1–10
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
Nautiyal CS (1997) A method for the selection and characterization of rhizosphere competent bacteria of chickpea. Curr Microbiol 34:12–17
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
Persello-Cartieaux F, Nussaume L, Robaglia C (2003) Tales from the underground: molecular plant- microbe interaction. Plant Cell Environ 26:189–199
Pikovskaya RI (1948) Mobilization of phosphorus in connection with the vital activity of some microbial species. Microbiologia 17:362–370
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
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
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
SAS Institute (2003) SAS/STAT guide for personal computers. SAS Institute, Cary
Schwyn B, Neilands JB (1987) Universal chemical assay for the detection and determination of siderophores. Anal Biochem 160:47–56
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
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
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
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
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
Turgeon N, Laclamme C, Ho J, Duchaine C (2006) Elaboration of an electroporation protocol for Bacillus cereus ATTC14579. J Microbiol Methods 67:543–548
Van Loon LC, Bakker PAHM (2003) Signaling in rhizobacteria-plant interaction. Ecol Studies 168:297–330
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
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
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.
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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
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DOI: https://doi.org/10.1007/s11274-010-0365-z