Abstract
Isolation of bacteroids from effective (Fix+) and ineffective (Fix−) pea nodules, inoculated withRhizobium leguminosarum K, were performed by a density gradient centrifugation method using silica sol (Percoll). Only one zone (ρ=1.064–1.072; n-zone) was recognized in the Fix+ nodule which contained typical Y-shaped bacteroids while two zones (n-zone and ρ=1.125–1.145; n'-zone) were obtained from the Fix− nodule. The cells in the n'-zone, which are long rods differed morphologically from free-living cells at any growth phase (ρ=1.108–1.125; f-zone and ρ=1.074–1.078; f'-zone), and differed from Y-shaped bacteroids by cell density. The esterase isozyme pattern of bacteroids in the n-and n'-zones also showed clear differences from that of f-and f'-zone of free-living cells.
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References
Bergersen F J 1974 Formation and function of bacteroids.In The Biology of Nitrogen Fixation. Ed. A Quispel. Elsevier-North Holland Publishing Co., Amsterdam, pp. 471–498.
Blevins D G, Barnett N M and Bottino P J 1977 The effects of calcium and the ionophore A 23187 on nodulation, nitrogen fixation and growth of soybeans. Physiol. Plant. 41, 235–238.
Ching T M, Hedtke S and Newcomb W 1977 Isolation of bacteria, transforming bacteria, and bacteroids from soybean nodules. Plant Physiol. 60, 771–774.
Davis B B 1964 Disc electrophoresis. II. Method and application to human serum proteins. Ann. N. Y. Acad. Sci. 121, 404–427.
Gmelig-Meyling F and Waldmann T A 1980 Separation of human blood monocytes and lymphocytes on a continuous Percoll gradient. J. Immunol. Methods 33, 1–9.
Higashi S and Abe M 1980 Scanning electron microscopy ofRhizobium trifolii infection sites on root hairs of white clover. Appl. Environ. Microbiol. 40, 1094–1099.
Jackson G, Dench D E, Hall D O and Moore L 1979 Separation of mitochondria from contaminating subcellular structures utilizing silica sol gradient centrifugation. Plant Physiol. 64, 150–153.
Johnston A W B, Beynon J L, Buchanan-Wollaston A V, Setchell S M, Hirsh P R and Beringer J E 1978 High frequency transfer of nodulating ability between strains and species of Rhizobium. Nature London 276, 634–636.
Keele B B, Hamilton Jr P B and Elkan G H 1969 Glucose catabolism inRhizobium japonicum. J. Bacteriol. 97, 1184–1191.
Meistrich M L, Longtin J L and Brock W A 1979 Further purification of rat spermatogenic cells by density centrifugation. J. Cell Biol. 83, 226a.
Moustafa E and Greenwood R H 1967 Esterase and phosphatase isozymes in Rhizobia and Rhizobial bacteroids in relation to strain effectiveness and proportion of nodule tissue in Lotus. N. Z. J. Sci. 10, 548–555.
Newcomb W 1979 A correlated light and electron microscopic study of symbiotic growth and differentiation inPisum sativum root nodules. Can. J. Bot. 54, 2163–2186.
Pertoft H, Laurent T C, Låås T and Kågedal L 1978 Density gradients prepared from colloidal silica particles coated by polyvinylpyrrolidone (Percoll). Anal. Biochem. 88, 271–282.
Reibach P H, Mask P L and Streeter J G 1981 A rapid one-step method for the isolation of bacteroids from root nodules of soybean plants, utilizing self-generating Percoll gradients. Can. J. Microbiol. 27, 491–495.
Rolfe B G, Djordjevic M, Scott K F, Hughes J E, Badenoch Jones J, Gresshoff P M, Cen Y, Dudman W F, Zurkowski W and Shine J 1980 Analysis of nodule forming ability of fast-growing Rhizobium strains.In Current perspectives in Nitrogen Fixation. Eds. A H Gibson and W E Newton. Australian Academy of Science, Canberra, pp. 142–145.
Sutton W and Mahoney P 1977 Preparation and fractionation of Rhizobium bacteroids by zone sedimentation through sucrose gradients. Plant Physiol. 60, 800–802.
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Higashi, S., Katahira, S. & Abe, M. Isolation of pea nodule bacteroids utilizing silica sol (Percoll) density gradient centrifugation. Plant Soil 81, 93–99 (1984). https://doi.org/10.1007/BF02206898
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DOI: https://doi.org/10.1007/BF02206898