Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Animal Production Science Animal Production Science Society
Food, fibre and pharmaceuticals from animals
RESEARCH ARTICLE

Coinoculation of chickpea with Rhizobium isolates from roots and nodules and phytohormone-producing Enterobacter strains

Babur Saeed Mirza A , M. Sajjad Mirza B D , Asghari Bano A and Kauser A. Malik C
+ Author Affiliations
- Author Affiliations

A Department of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan.

B National Institute for Biotechnology and Genetic Engineering, PO Box 577, Jhang Road, Faisalabad, Pakistan.

C Pakistan Atomic Energy Commission, PO Box 1114, Islamabad, Pakistan.

D Corresponding author. Email: sajjad_mirza@yahoo.com

Australian Journal of Experimental Agriculture 47(8) 1008-1015 https://doi.org/10.1071/EA06151
Submitted: 1 July 2006  Accepted: 6 February 2007   Published: 16 July 2007

Abstract

The aim of the present study was to isolate plant-beneficial bacteria (both Rhizobium and plant growth promoting rhizobacteria) from roots and nodules of chickpea (Cicer arietinum L.) and to study the effect of coinoculations on growth of two cultivars of chickpea. Four Rhizobium strains were obtained from roots and four from the nodules of field-grown chickpea cv. Parbat and identified on the basis of morphological characteristics, and biochemical and infectivity tests on the host seedlings. Only one type of nitrogen and carbon source utilisation pattern and DNA banding pattern of random amplified polymorphic DNA was observed in all isolates (Rn1, Rn2, Rn3, Rn4) from nodules, while two types of such patterns were detected among the isolates from roots. The isolate Rr1 from roots also exhibited a pattern identical to those of the isolates from nodules, whereas the remaining three isolates (Rr2, Rr3 and Rr4) from roots showed a different pattern. Two strains of plant growth-promoting rhizobacteria belonging to genus Enterobacter were also isolated from chickpea roots. All the Rhizobium strains and Enterobacter strains produced the plant growth hormones indole acetic acid and gibberellic acid in the growth medium. Effects of the bacterial isolates as single- or double-strain inocula were studied on two chickpea cultivars (NIFA 88 and Parbat) grown in sterilised soil. In cultivar NIFA 88, coinoculation of Rhizobium strain Rn1 with Enterobacter strain B resulted in maximum increase in plant biomass and nodulation, as compared with the control treatment (non-inoculated as well as inoculated with Rhizobium strain Rn1 only), whereas the combination of Rhizobium Rn1 with Enterobacter A was more efficient in growth promotion of chickpea cv. Parbat. In non-sterilised soil, the same combinations of the Rhizobium strain Rn1 with Enterobacter strains A and B were found to be the most effective inoculants for cvv. Parbat and NIFA 88, respectively. However, some negative effects on plant growth were also noted in cv. Parbat coinoculated with Rhizobium strain Rr2 and Enterobacter strain B.


References


Andreeva IN, Red’kina TV, Ismallov SF (1993) The involvement of indoleacetic acid in the stimulation of Rhizobium-legume symbiosis by Azospirillum brasilense. Russian Journal of Plant Physiology: a Comprehensive Russian Journal on Modern Phytophysiology 40, 901–906. open url image1

Atzorn R, Crozier A, Wheeler CT, Sanabuerg G (1988) Production of gibberellins and indole-3-acetic acid by Rhizobium phaseoli in relation to nodulation of Phaseolus vulgaris roots. Planta 175, 532–538.
Crossref | GoogleScholarGoogle Scholar | open url image1

Azcon-Aguilar C, Barea JM (1978) Effects of interactions between different culture fractions of ‘phosphobacteria’ and Rhizobium on mycorrhizal infection, growth, and nodulation of Medicago sativa. Canadian Journal of Microbiology 24, 520–524.
PubMed |
open url image1

Baldani JI, Baldani VLD, Seldin L, Dobereiner J (1986) Characterization of Herbaspirillum seropedicae gen. nov., sp. Nov., a root-associated nitrogen fixing bacterium. International Journal of Systematic Bacteriology 36, 86–93. open url image1

Bally R, Thomas-Bauzon D, Heulin T, Balandreau J, Richard C, De Lay J (1983) Determination of the most frequent N2-fixing bacteria in a rice rhizosphere. Canadian Journal of Microbiology 29, 881–887. open url image1

Beck DP, Materon LA, Afandi F (1993) Practical Rhizobium-legume technology manual. International Center for Agricultural Research in the Dry Areas, Technical Manual No. 19, Aleppo, Syria.

Bilal R, Rasul G, Qureshi JA, Malik KA (1990) Characterization of Azospirillum and related diazotrophs associated with roots of plants growing in saline soils. World Journal of Microbiology & Biotechnology 6, 46–52.
Crossref | GoogleScholarGoogle Scholar | open url image1

Biswas JC, Ladha JK, Dazzo FB (2000) Rhizobia inoculation improves nutrient uptake and growth of low land rice. Soil Science Society of America Journal 64, 1644–1650. open url image1

Brenner DJ (1992) Introduction to the family Enterobacteriaceae. In ‘The prokaryotes’. (Eds A Baloues, HG Truper, M Dworkin, W Harder, KH Schleifer) pp. 2673–2695. (Springer: Berlin, Germany)

Burdsman S, Volpin H, Kiegel J, Kapulnik Y, Okon Y (1996) Promotion of nod-gene inducers and nodulation in common bean (Phaseolus vulgaris) roots inoculated with Azospirillum brasilense Cd. Applied and Environmental Microbiology 62, 3030–3033.
PubMed |
open url image1

Camacho M, Santamaria C, Temprano F, Rodriguez-Navarro DN, Daza A (2001) Co-inoculation with Bacillus sp. CECT 450 improves nodulation in Phaseolus vulgaris L. Canadian Journal of Microbiology 47, 1058–1062.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Chabot R, Antoun H, Kloepper JW, Beauchamp CJ (1996) Root colonization of maize and lettuce by bioluminescent Rhizobium leguminosarum biovar phaseoli. Applied and Environmental Microbiology 62, 2767–2772.
PubMed |
open url image1

Chabot R, Beauchamp CJ, Kloepper JW, Anoun H (1998) Effect of phosphorus on root colonization and growth promotion of maize by bioluminescent mutants of phosphate-solubilizing Rhizobium leguminosarum biovar phaseolii. Soil Biology & Biochemistry 30, 1615–1618.
Crossref | GoogleScholarGoogle Scholar | open url image1

Chebotar VK, Asis CA, Akao S (2001) Production of growth-promoting substances and high colonization ability of rhizobacteria enhance the nitrogen fixation of soybean when coinoculated with Bradyrhizobium japonicum. Biology and Fertility of Soils 34, 427–432. open url image1

Costacura A, Vanderleyden J (1995) Synthesis of phytohormones by plant associated bacteria. Critical Reviews in Microbiology 21, 1–18.
PubMed |
open url image1

Del-Gallo M, Fabbri P (1990) Inoculation of Azospirillum brasilense Cd on chickpea (Cicer arientinum). Symbiosis 9, 283–287. open url image1

Dobbelaere S, Vanderleyden J, Okon Y (2003) Plant growth-promoting effects of diazotrophs in the rhizosphere. Critical Reviews in Plant Sciences 22, 107–149.
Crossref |
open url image1

Dobereiner J, Day JM (1976) Associative symbiosis in tropical grasses: characterization of micro-organisms and dinitrogen fixing sites. In ‘Proceedings of the First International Symposium on Nitrogen Fixation. Vol. 2’. (Eds WE Newton, CJ Nyman) pp. 518–538. (Washington State University Press: Pullman, WA)

Fabbri P, Del-Gallo M (1995) ‘Specific interaction between chickpea (Cicer arientinum) and three chickpea-Rhizobium strains inoculated singularly and in combination with Azospirillum brasilense Cd. NATO ASI series G37.’ (Springer: Berlin, Germany)

Gillis M, Kresters K, Hoste B, Janssens D, Kropenstedt RM, Stephan MP, Teixeira KRS, Dobereiner J, De Lay J (1989) Acetobacter diazotrophicus sp. nov., a nitrogen-fixing acetic acid bacterium associated with sugarcane. International Journal of Systematic Bacteriology 39, 361–364. open url image1

Gull M, Hafeez FY, Saleem M, Malik KA (2004) Phosphorus uptake and growth promotion of chickpea by co-inoculation of mineral phosphate solubilising bacteria and a mixed rhizobial culture. Australian Journal of Experimental Agriculture 44, 623–628.
Crossref | GoogleScholarGoogle Scholar | open url image1

Hafeez FY, Safdar ME, Chaudhry AU, Malik KA (2004) Rhizobial inoculation improves seedling emergence, nutrient uptake and growth of cotton. Australian Journal of Experimental Agriculture 44, 617–622.
Crossref | GoogleScholarGoogle Scholar | open url image1

Hamaoui B, Abbadi J, Burdman S, Rasid A, Sarig S, Okon Y (2001) Effects of inoculation with Azospirillum brasilense on chickpeas (Cicer arietinum) and faba beans (Vicia faba) under different growth conditions. Journal of Agronomy 21, 553–560.
Crossref | GoogleScholarGoogle Scholar | open url image1

Holt JG, Kreig NR, Sneath PHA (1994) ‘Bergey’s manual of determinative bacteriology.’ 9th edn. (Williams and Wilkins: Baltimore, MD)

Janzen AR, Rood BS, Dormaar FJ, Mcgill BW (1992) Azospirillum brasilense produces gibberellin in pure culture on chemically-defined medium and in co-culture on straw. Soil Biology & Biochemistry 24, 1061–1064.
Crossref | GoogleScholarGoogle Scholar | open url image1

Kennedy IR, Choudhury ATMA, Kecskes ML (2004) Non-symbiotic bacterial diazotrophs in crop-farming systems: can their potential for plant growth promotion be better exploited? Soil Biology & Biochemistry 36, 1229–1244.
Crossref | GoogleScholarGoogle Scholar | open url image1

Kloepper JW (1993) Plant growth-promoting rhizobacteria as biological control agents. In ‘Soil microbial ecology: applications in agricultural and environmental management’. (Ed. FB Meting) pp. 255–274. (Marcel Dekker: New York)

Laguerre G, Allard MR, Revoy F, Amarger N (1994) Rapid identification of Rhizobium by restriction fragment length polymorphism analysis of PCR amplified 16S rRNA genes. Applied and Environmental Microbiology 60, 56–63.
PubMed |
open url image1

Laguerre G, Mayingui P, Allard MR, Charnay MP, Louvrier P, Mazurier SI, Rigottier-Gois L, Amarger N (1996) Typing of Rhizobia by PCR DNA fingerprinting and PCR-reaction fragment length polymorphism analysis of chromosomal and symbiotic gene regions: application to Rhizobium leguminosarum and its different biovars. Applied and Environmental Microbiology 62, 2029–2036.
PubMed |
open url image1

Li DM, Alexander M (1988) Co-inoculation with antibiotic-producing bacteria to increase colonization and nodulation by rhizobacteria. Plant and Soil 108, 211–219.
Crossref | GoogleScholarGoogle Scholar | open url image1

Li DM, Alexander M (1990) Factors affecting co-inoculation with antibiotic producing bacteria to enhance rhizobial colonization and nodulation. Plant and Soil 129, 195–201. open url image1

Li JC, Shi J, Zhao XL, Wang G, Yu HF, Ren YJ (1994) Separation and determination of three kinds of plant hormone by high-performance liquid chromatography. Chinese Journal of Analytical Chemistry 22, 801–804. open url image1

MacFaddin JF (1980) ‘Biochemical tests for identification of medical bacteria.’ (Williams and Wilkins: Baltimore, MD)

Mehnaz S, Mirza MS, Haurat J, Bally R, Normand P, Bano A, Malik KA (2001) Isolation and 16S rRNA sequence analysis of the beneficial bacteria from the rhizosphere of rice. Canadian Journal of Microbiology 47, 110–117.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Miller JH (Ed.) (1972) ‘Experiments in molecular genetics.’ (Cold Spring Harbor Laboratory: Cold Spring Harbor, NY)

Mirza MS, Ahmad W, Latif F, Haurat J, Bally R, Normand P, Malik KA (2001) Isolation, partial characterization, and the effect of plant growth-promoting bacteria (PGPB) on micro-propagated sugarcane in vitro. Plant and Soil 237, 47–54.
Crossref | GoogleScholarGoogle Scholar | open url image1

Nishijima I, Evans WR, Vesper SJ (1988) Enhanced nodulation of soybean by Bradyrhizobium in the presence of Pseudomonas fluorescens. Plant and Soil 111, 149–150.
Crossref | GoogleScholarGoogle Scholar | open url image1

Noel TC, Sheng C, Yost CK, Pharis RP, Hynes MF (1996) Rhizobium leguminosarum as a plant growth promoting rhizobacterium: direct growth promotion of canola and lettuce. Canadian Journal of Microbiology 42, 279–283.
PubMed |
open url image1

Patten CL, Glick BR (1996) Bacterial biosynthesis of indole-3-acetic acid: a review. Canadian Journal of Microbiology 42, 207–220.
PubMed |
open url image1

Rasul G, Mirza MS, Latif F, Malik KA (1998) Identification of plant growth hormones produced by bacterial isolates from rice, wheat and Kallar grass. In ‘Nitrogen fixation with non-legumes’. (Eds KA Malik, MS Mirza, JK Ladha) pp. 25–37 (Kluwer: Dordrecht, The Netherlands)

Reinhold-Hurek B, Hurek T, Gillis M, Hoste B, Vancanneyt M, Kersters K, de Ley J (1993) Azoarcus gen. nov., nitrogen-fixing proteobacteria associated with roots of Kallar grass (Leptochloa fusca (L.) Kunth), and description of two species, Azoarcus indigens sp. nov. and Azoarcus communis sp. nov. International Journal of Systematic Bacteriology 43, 574–584. open url image1

Rennie RJ (1981) A single medium for the isolation of acetylene reducing (dinitrogen-fixing) bacteria from soils. Canadian Journal of Microbiology 27, 8–14.
PubMed |
open url image1

Roumiantseva ML, Andronov EE, Sharypova LA, Dammann-Kalinowski T, Keller M, Young JPW, Simarov BV (2002) Diversity of Sinorhizobium meliloti from the Central Asian Alfalfa Gene Center. Applied and Environmental Microbiology 68, 4694–4697.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Samasegaran P, Hoben H, Halliday J (1982) ‘The Nif-TAL manual for methods in legume-Rhizobium technology.’ (U.S. Agency for International Development: University of Hawaii)

Sarwar M, Arshad M, Martens DA, Frankenberger WT (1992) Tryptophan-dependent biosynthesis of auxins in soil. Plant and Soil 147, 207–215.
Crossref | GoogleScholarGoogle Scholar | open url image1

Sattar MA, Quader MA, Danso SKA (1995) Nodulation, nitrogen fixation and yield of chickpea as influenced by host cultivar and Bradyrhizobium strain differences. Soil Biology & Biochemistry 27, 725–727.
Crossref | GoogleScholarGoogle Scholar | open url image1

Steel KJ (1961) The oxidase reaction as a toxic tool. Journal of General Microbiology 25, 297. open url image1

Tas E, Saano L, Leinonen P, Lindstrom K (1995) Identification of Rhizobium spp. in peat-based inoculants by DNA hybridization and PCR and its application in inoculant quality control. Applied and Environmental Microbiology 61, 1822–1827.
PubMed |
open url image1

Tchebotar VK, Kang UGS, Asis CA, Akao S (1998) The use of the GUS-reporter gene to study the effect of Azospirillum-Rhizobium co-inoculation on nodulation of white clover. Biology and Fertility of Soils 27, 349–352.
Crossref | GoogleScholarGoogle Scholar | open url image1

Tien TM, Gaskins MH, Hubbel DH (1979) Plant growth substances produced by Azospirillum brasilense and their effect on growth of pearl millet (Pennisetum americanum L.). Applied and Environmental Microbiology 37, 1016–1024.
PubMed |
open url image1

Volpin H, Burdman S, Castro-Sowzinski S, Kapulnik Y, Okon Y (1996) Inoculation with Azospirillum increased exudation of rhizobial nod-gene inducers by alfalfa roots. Molecular Plant-Microbe Interactions 9, 388–394. open url image1

Welsh J, McClelland M (1990) Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Research 18, 7213–7218.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Yahalom E, Okon Y, Dovrat A (1987) Azospirillum effects on susceptibility to Rhizobium nodulation and on nitrogen fixation of several forage legumes. Canadian Journal of Microbiology 33, 510–514. open url image1

Yanni YG, Rizk RY, Abd El-Fattah FK, Squartini A, Corich V , et al. (2001) The beneficial plant growth promoting association of Rhizobium leguminosarum bv. trifolii with rice roots. Australian Journal of Plant Physiology 28, 845–870. open url image1

Zhang F, Smith DL (1996) Inoculation of soybean (Glycine max (L.) Merr.) with genistein-preincubated Bradyrhizobium japonicum or genistein directly applied into soil increases soybean protein and dry matter yield under short season conditions. Plant and Soil 179, 233–241.
Crossref | GoogleScholarGoogle Scholar | open url image1