Abstract
Leptochloa fusca (L.) Kunth (kallar grass) has previously been found to exhibit high rates of nitrogen fixation. A series of experiments to determine the level of biological nitrogen fixation using N isotopic dilution were carried out in nutrient solution and saline soil. These studies indicated an agronomically significant amount of nitrogen being fixed in soil. Kallar grass has a similar growth habitat to rice. Therefore similar studies were carried out with rice after isolating various diazotrophs from the roots which were also screened for their ability to produce auxin (IAA). Five such strains namely Azospirillum lipoferum N-4, Azospirillum brasilense Wb-3, Azoarcus K-1, Pseudomonas 96-51, Zoogloea Ky-1 were selected for inoculating two rice varieties i.e. NIAB-6 and BAS-370 under aseptic laboratory conditions. The nitrogen fixed was quantified using the N isotopic dilution method. Variety BAS-370 had nearly 70% nitrogen derived from atmosphere (Ndfa) when inoculated with Azospirillum N-4. Similar studies with the mixed inoculum using N fertilizer in the micro plots indicated that nearly 29% of plant nitrogen was derived from the atmosphere.
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Baldani J I, Baldani V L D, Seldin L and Döbereiner J 1986 Characterization of Herbaspirillum seropedicae gen. nov., sp. nov., a root associated N2-fixing bacteria. Int. J. Syst. Bacteriol. 36, 86–93.
Bilal R 1988 Associative nitrogen fixation in plants growing in saline environments. Ph.D. Thesis, Punjab University Lahore, Pakistan. 248 p.
Bilal R and Malik K A 1987 Isolation and identification of a N2-fixing Zoogloea-forming bacterium fromkallar grass histoplane. J. Appl. Bacteriol. 62, 289–294.
Bilal R, Rasul G, Arshad M and Malik K A 1993 Attachment, colonization and proliferation of Azospirillum brasilense and Enterobacter spp. on root surface of grasses. World J.Microbiol. Biotechnol. 9, 63–69.
Bilal R, Rasul G, Qureshi J A and Malik K A 1990 Characterization of Azospirillum and related diazotrophs associated with roots of plants growing in saline soils. World J. Microbiol. Biotechnol. 6, 46–52.
Boddey R M and Victoria R L 1986 Estimation of biological nitrogen fixation associated with Brachiaria and Paspalum notatum using 15N labelled organic matter and fertilizer. Plant Soil 90, 265–292.
Boddey R M, Alves BJ R and Urquiaga S 1996 Evaluation of biological nitrogen fixation associated with non-legumes. In Nitrogen Fixation with Non-Legumes. Eds. K A Malik, M S Mirza and J K Ladha, Proceedings of 7th International Symposium on BNF with non-legumes. Oct. 16–21, 1996. Faisalabad, Pakistan. (In press).
Boddey R M, Chalk P M, Victoria R L and Matsui E 1983 The 15N isotope dilution technique applied to the estimation of biological nitrogen fixation associated with Paspalum notatum cv. batata in the field. Soil Biol. Biochem. 15, 25–32.
Bric J M, Bostock R M and Silverstone S E 1991 Rapid in-situ assay for indoleacetic acid production by bacteria immobilized on a nitrocellulose membrane. Appl. Environ. Microbiol. 57, 535–538.
Cannon F C, Dixon R A and Postgate J R 1974 Chromosomal integration of Klebsiella nitrogen fixation gene in Escherichia coli. J. Gen. Microbiol. 80, 227–239.
Chalk P M 1991 The contribution of associative and symbiotic nitrogen fixation to the nitrogen nutrition of non-legumes. Plant Soil 132, 29–39.
Cochran W G 1950 Estimation of bacterial densities by means of the "Most Probable Number". Biometrics 6, 105–116.
Day J M, Harris D, Dart P J and Van Berkum P 1975 The Broadbalk experiment. An investigation of nitrogen gains from nonsymbiotic fixation. In Nitrogen fixation by free-living Microorganisms. Ed. W D P Stewart Vol. 6, pp 71–84. International Biological programme Series, Cambridge Univ. Press, Cambridge.
Döbereiner J, Marriel E and Nery M 1976 Ecological distribution of Spirillum lipoferum Beijerinck. Can. J. Microbiol. 22, 1464– 1473.
Eskew D L, Eaglesham A R J and App A A 1981 Heterotrophic 15N-fixation and distribution of newly fixed nitrogen in a rice-flooded soil system. Plant Physiol. 68, 48–52.
Fried M and Broeshart H 1975 An independent measurement of the amount of the nitrogen fixed by a legume crop. Plant Soil 43, 707–711.
Hassan U, Mirza M S, Mehnaz S, Rasul G and Malik K A 1996 Isolation and identification of diazotrophic bacteria from rice, wheat and kallar grass. In Nitrogen Fixation with Non-Legumes. Eds. K A Malik, M S Mirza and J K Ladha, Proceedings of 7th International Symposium on BNF with non-legumes. Oct. 16– 21, 1996. Faisalabad, Pakistan. (In press).
Hegazi N A, Amer H A and Monib M 1979 Enumeration ofN2-fixing Spirilla. Soil Biol. Biochem. 11, 437–438.
Hurek T, Reinhold-Hurek B, van Montagu M and Kellenberger E 1991 Infection of intact roots of kallar grass and rice seedlings by "Azoarcus". In Nitrogen Fixation. Proceedings of the 5th International Symposium on Nitrogen Fixation with Non-Legumes. Eds. M Polsinelli, R Materassi and M Vincenzeni. Kluwer Academic Publishers, Dordrecht.
Kreig N R and Holt J G (Eds.) 1984 The genus Zoogloea. In Shorter Bergey's Manual of Determinative Bacteriology, 8th Edition, pp 83–84. Williams & Wilkins Co., Baltimore, USA.
Ladha JK, So R B and Watanabe I 1987 Composition of Azospirillum species associated with wetland rice plant grown in different soils. Plant Soil 102, 127–129.
Lethbridge G and Davidson M S 1983 Root-associated nitrogen-fixing bacteria and their role in the nitrogen nutrition of wheat estimated by 15N isotope dilution. Soil Biol. Biochem. 15, 365– 374.
Lifshitz R, Kloepper J W, Kozlowski M, Simonson C, Carlson J, Tipping E M and Zaleska I 1987 Growth promotion of canola (rapeseed) seedlings by a strain of Pseudomonas putida under gnotobiotic conditions. Can. J. Microbiol. 33, 390–395.
Malik K A, Aslam Z and Naqvi S H M 1986 Kallar grass: a plant for saline land. Nuclear Institute for Agriculture and Biology (NIAB), Faisalabad, Pakistan.
Malik K A and Bilal R 1989 Survival and colonization of inoculated bacteria in kallar grass rhizosphere and quantification of N2-fixation. In Nitrogen Fixation with Non-Legumes. Eds. F A Skinner, R M Boddey and I Fendrik. pp 301–310. Kluwer Academic Publishers, Dordrecht.
Malik K A, Bilal R, Azam F and Sajjad M I 1988 Quantification of N2-fixation and survival of inoculated diazotrophs associated with roots of Kallar grass. Plant Soil 108, 43–51.
Malik K A, Bilal R, Rasul G, Mahmood K and Sajjad M I 1991 N2-fixation in plants growing in saline sodic soils and its relative quantification based on 15N natural abundance. Plant Soil 137, 67–74.
Malik K A, Rasul G, Hassan U, Mehnaz S and Ashraf M 1993 Role of N2-fixing and growth hormones producing bacteria in improving growth of wheat and rice. In Nitrogen Fixation with non-legumes. Eds. N A Hegazi, M Fayez and M Monib. pp 409–422. The Am. Univ. in Cairo Press, Cairo.
Malik K A and Zafar Y 1985 Quantification of root associated nitrogen fixation in kallar grass as estimated by 15N isotope dilution. In Nitrogen and the Environment. Eds. K A Malik, S H M Naqvi, M I H Aleem. pp 161–171. NIAB, Faisalabad, Pakistan.
Malik K A, Zafar Y and Hussain A 1980 Nitrogenase activity in the rhizosphere of kallar grass (Diplachne fusca Linn Beauv). Biologia 26, 107–112.
Malik K A, Zafar Y and Hussain A 1982 Associative dinitrogen fixation in Diplachne fusca (kallar grass). In Biological Nitrogen Fixation Technology for Tropical Agriculture. Eds. P H Graham and S C Harris. pp 503–507. CIAT, Cali, Columbia.
Malik K A, Zafar Y, Bilal R and Azam F 1987 Use of 15N isotope dilution for quantification of N2-fixation associated with roots of kallar grass (Leptochloa fusca (L.) Biol. Fertil. Soils 4, 103–108.
Mehnaz S, Mirza M S, Hassan U and Malik K A 1996 Detection of inoculated plant growth promoting rhizobacteria in the rhizosphere of rice. In Nitrogen Fixation with Non-Legumes. Eds. K A Malik, M S Mirza and J K Ladha, Proceedings of 7th International Symposium on BNF with non-legumes. Oct. 16–21, 1996. Faisalabad, Pakistan (In press).
Qureshi J A, Zafar Y and Malik K A 1988 Klebsiella sp. NIAB-1: A new diazotroph, associated with the roots of kallar grass from saline sodic soils. Plant Soil 110, 219–224.
Reinhold B, Hurek T and Fendrik I 1987 Cross-reaction of predominant nitrogen-fixing bacteria with enveloped, round bodies in the root interior of kallar grass. Appl. Environ. Microbiol. 53, 889–891.
Reinhold B, Hurek T, Fendrik I, Pot B, Gillis M, Kersters K, Thielemans S and De Ley J 1987a Azospirillum halopraeferens sp. Nov., a nitrogen-fixing organism associated with the roots of kallar grass (Leptochloa fusca (L.) Kunth). Int. J. Syst. Bacteriol. 37, 43–51.
Reinhold B, Hurek T, Niemann E G and Fendrik I 1986 Close association of Azospirillum and diazotrophic rods with different root zones of kallar grass. Appl. Environ. Microbiol. 52, 520–526.
Reinhold-Hurek B, Hurek T, Gillis M, Hoste B, Vancanneyt M, Kersters K and 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. Int. J. Syst. Bacteriol. 43, 574–584.
Rennie R J 1980 Isotope dilution as a measure of dinitrogen fixation by Azospirillum brasilense associated with maize. Can. J. Bot. 58, 21–24.
Rennie R J 1981 A single medium for the isolation of acetylene reducing (dinitrogen fixing) bacteria from soil. Can. J. Microbiol. 27, 8–14.
Rennie R J and Larson R I 1979 Dinitrogen fixation with disomic chromosome substitution lines of spring wheat. Can. J. Bot. 57, 2771–2775.
Rennie R J, de Freitas J R, Ruschel A P and Vose P B 1983 15N dilution to quantify dinitrogen (N2)-fixation associated with Canadian and Brazilian wheat. Can. J. Bot. 61, 1667–1671.
Roger P A and Ladha J K 1992 Biological N2 fixation in wetland rice fields: estimation and contribution to nitrogen balance. Plant Soil 141, 41–55.
Ruschel A, Henis Y and Salati E 1975 Nitrogen 15N tracing of N2-fixation with soil grown sugarcane seedlings. Soil Biol. Biochem. 5, 83–89.
Sandhu G R and Malik K A 1975 Plant succession – A key to utilization of saline soils. Nucleus 12, 35–38.
Shrestha R K and Ladha J K 1996 Genotypic variation in promotion of rice dinitrogen fixation as determined by nitrogen-15 dilution. Soil Sci. Soc. Am. J. 60, 1815–1821.
Sundaram S, Arunkumari A and Klucas R V 1988 Characterization of azospirilla isolated from seeds and roots of turf grass. Can. J. Microbiol. 34, 212–217.
Tarrand J J, Krieg N R and Döbereiner J 1978 A taxonomic study of Spirillum lipoferum group with descriptions of a new genus, Azospirillum gen. nov. and two species, Azospirillum lipoferum (Beijerinck) comb. nov. and Azospirillum brasilense sp. nov. Can. J. Microbiol. 24, 967–980.
Tien T M, Gaskins M H and Hubbell D H 1979 Plant growth substances produced by Azospirillum brasilense and their effect on the growth of pearl millet (Pennisetum americanum L.) Appl. Environ. Microbiol. 37, 1016–1024.
Van Berkum P 1980 Evaluation of acetylene reduction by excised roots for the determination of nitrogen fixation in grasses. Soil Biol. Biochem. 12, 141–145.
Ventura W and Watanabe I 1983 15N dilution of accessing the contribution of nitrogen fixation to rice plant. Soil Sci. Plant Nutr. 29, 123–131.
Witty J F 1979 Acetylene reduction assay can overestimate nitrogen fixation in soil. Soil Biol. Biochem. 11, 209–210.
Zafar Y, Ashraf M and Malik K A 1986 Nitrogen fixation associated with the roots of kallar grass (Leptochloa fusca (L.) Kunth). Plant Soil 90, 93–106.
Zafar Y, Malik K A and Niemann E G 1987 Studies on N2-fixing bacteria associated with salt tolerant grass Leptochloa fusca (L.) Kunth. Mircen J. Appl. Microbiol. 3, 45–56.
Zimmer W, Roeben K and Boothe H (1988) An alternative explanation for plant growth promotion by bacteria of the genus Azospirillum. Planta 176, 333–342.
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Malik, K., Bilal, R., Mehnaz, S. et al. Association of nitrogen-fixing, plant-growth-promoting rhizobacteria (PGPR) with kallar grass and rice. Plant and Soil 194, 37–44 (1997). https://doi.org/10.1023/A:1004295714181
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DOI: https://doi.org/10.1023/A:1004295714181