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A simple medium for quick growth of Rhizobium japonicum
ZLJ.Uakt. ALt. II, Bd. J28, S. 23-24
Short Communication [From the Diyision of Mierohio]ogy, Indian AgTicultural Research Institute, New j)elhi-12 (India)]
A simple medium for quick growth of Rhizobium japonicum V. Jswaran, Ahhiswar Sen, alld H. Aple Sumll1ar~·
Iihi=olJilllll jlljJOlliclI1II is a slow-growing I\hizobium in ('oIllLllonly use,lllledia. A medium, gi,"ell in this cOllllllunication, enahles the speeies to gr'oll" lJuickly, as evidenced by the eounts. "'"'ious synthetic media al'e heing used I'or the growth of Rhizobium spp. (BERGENSEN I!)(il. FRED et a!.W:12, ISWARAN and JAUHRI 1!169, ISWARAN et a!. 1969, XORRIS 1959, \VEST 'l1ld WILSONI!l3U, WILSON and UMBREIT l(40). In the ~etherlands, VAN SCHREVEN (1958) ltsed calciulll glycerol'hospl,ate as a constituent of the medium in large-scale manufaeture for cnltlll·es. YINCENT, JANCEY and ROUGHLEY (per. eomlllunjcation) nsed eostly galactose or amilinlls(', sterilized hy filtl'ation, as a carbon sou['(~e [or slow-growing species. ~lost of the fastgr'OIl"ing strains of rhizobia can be grown in an excess of 250 X 106 viable cells/rn!' in 3 dnys nt 28°(: in the usnal medium (FRED, BALDWIN, and McCoy 1932), whilc slow-growing types take at !t'ast 7 days in this Illedinm to 1'ench the nbovenlunber. However, no cheap and effi,iient n,edium for quick gl'owth o[ strains of SIIlW-gl'Owillg ffhi=obillfn juponicLtfn is available. I n the present nn attempt has heen made inH'stigation in this direction.
Malcl'ial and Methods The me,liuIll elllployed consists of manIliLol 01' SllCl'OSC, iOg.; K 2 LI PO~, 0.5 g.; MgSOI" 0.2 g.: XaCl, 0.1 g.; ealcitllll gltH'O'H1tt', 1.5 g.; 1"eC13 , O,()\ g.; yeast extl'act, 2.0 g.; H2 0, 1 litre,
[JH 6.1'.
Foul' strains of Hhizobiufn japo/licwlt were grown in the above mcdilllll, and were continuously shaken in a I'otatol'y shaker for 96 hours at 28°C. Standard yeast extract mannitol medium was used 1'01' comparison. Plate connts of each species we,'c lIlade by stalldard ,tilutioll proced" ..es (FRED, BALDWIN, '\[eeoy 1932) ;n the yeast extl'al'l lIlannitol agar, eontaining Congo red (1:,,00).
Hesulls The r'('slIlls a['(' shown hdo\\" ill T;rl>le J. TaLle 1 Counts of diffcl'ent stwins of Hhi=obillrll j{/[JolliclIlli ill till' tll"O tcst media (average of triplicates) II L !16 !tours Yeast cxtr'act mallnitol Sll'iJill
111 ediulII
Yeast extract mannitol cal. glueonate medi Ulll
(X
(X 108)
I'SIl.\ I> 1:J(i
Hi
~anking
28 :12
SH-G SB-J(j
,,2
J()6)
Jj ')
7 ~
24
V. Iswaran, Abhiswal' Sen and R.Apte, A simple medium for quick growth ••.
The results show that in this medium the Australian acceptable minimum standard for broth inoculum (250 X 106 viable rhizobia/mI.) are obtained in 96 hours, thereby minimizing the usual 7 days' period in the yeast extract mannitol medium. This medium differs from the common medium in having calcium gluconate and ferric chloride with additional 1 g./I. yeast extract. This medium is being now for used slow-growing species, including cowpea groups. Zusammenfassung Rhizobium japonicum ist auf gewohnlichen Niihrmedien als ein langsam wachsendes Knollchenbakterium zu betrachten. Das in diesel' Mitteilung angegebene Medium el'moglicht ein schnelleres Wachstum diesel' Bakteriengattung.
Literature BERGENSEN, F. J.: Growth of Rhizobium in synthetic media. Austr. J. BioI. Sci. 14 (1961), 349. - FRED, E. B., BALDWIN, 1. 1., and McCoy, E.: Root nodule bacteria and leguminous plants. Univ. Wisconsin Press, Madison 1932. - ISWARAN, V., and JAUHRI, K. S.: Malt extract as a source of carbon for large scale production of Rhizobium inoculants. CUlT. Sci. 38 (1969), 67. - ISWARAN, V., GRAHAM, P. H., and JAUHRI, K. S.: Modified Burton's medium for large scale production of cultures of Rhizobium japonicum. Curl'. Sci. 38 (1969), :J4G. NORRIS, D. 0.: The role of calcium and magnesium in the nutrition of Rhizobium. Austr. J. agric. Res. 10 (1959), 651. - VAN SCHREVEN, D. A.: Methods used in the Netherlands for the production of legume inoculants. In Nutrition of the Legumes (ed. Hallsworth), Butterwol·ths. London 1958, pp. 328-333. - WEST, P. M., and WILSON, P. W.: Growth factor requirements of the root nodule bacteria. J. Bacteriol. 37 (1939),161. - WILSON, J. B., and UMllREIT, W. W.: Growth studies of Rhizobium japocnium. Proc. Soil Sci. Soc. ArneI'. V (1940), 262.
Authors' address: Dr. V. Iswaran, Dr. Abhiswar Sen and Sri R. Apte, Division of Microbiology, Indian Agricultural Research Institute, New Delhi-12 (India).
Short Communication [From the Diyision of Mierohio]ogy, Indian AgTicultural Research Institute, New j)elhi-12 (India)]
A simple medium for quick growth of Rhizobium japonicum V. Jswaran, Ahhiswar Sen, alld H. Aple Sumll1ar~·
Iihi=olJilllll jlljJOlliclI1II is a slow-growing I\hizobium in ('oIllLllonly use,lllledia. A medium, gi,"ell in this cOllllllunication, enahles the speeies to gr'oll" lJuickly, as evidenced by the eounts. "'"'ious synthetic media al'e heing used I'or the growth of Rhizobium spp. (BERGENSEN I!)(il. FRED et a!.W:12, ISWARAN and JAUHRI 1!169, ISWARAN et a!. 1969, XORRIS 1959, \VEST 'l1ld WILSONI!l3U, WILSON and UMBREIT l(40). In the ~etherlands, VAN SCHREVEN (1958) ltsed calciulll glycerol'hospl,ate as a constituent of the medium in large-scale manufaeture for cnltlll·es. YINCENT, JANCEY and ROUGHLEY (per. eomlllunjcation) nsed eostly galactose or amilinlls(', sterilized hy filtl'ation, as a carbon sou['(~e [or slow-growing species. ~lost of the fastgr'OIl"ing strains of rhizobia can be grown in an excess of 250 X 106 viable cells/rn!' in 3 dnys nt 28°(: in the usnal medium (FRED, BALDWIN, and McCoy 1932), whilc slow-growing types take at !t'ast 7 days in this Illedinm to 1'ench the nbovenlunber. However, no cheap and effi,iient n,edium for quick gl'owth o[ strains of SIIlW-gl'Owillg ffhi=obillfn juponicLtfn is available. I n the present nn attempt has heen made inH'stigation in this direction.
Malcl'ial and Methods The me,liuIll elllployed consists of manIliLol 01' SllCl'OSC, iOg.; K 2 LI PO~, 0.5 g.; MgSOI" 0.2 g.: XaCl, 0.1 g.; ealcitllll gltH'O'H1tt', 1.5 g.; 1"eC13 , O,()\ g.; yeast extl'act, 2.0 g.; H2 0, 1 litre,
[JH 6.1'.
Foul' strains of Hhizobiufn japo/licwlt were grown in the above mcdilllll, and were continuously shaken in a I'otatol'y shaker for 96 hours at 28°C. Standard yeast extract mannitol medium was used 1'01' comparison. Plate connts of each species we,'c lIlade by stalldard ,tilutioll proced" ..es (FRED, BALDWIN, '\[eeoy 1932) ;n the yeast extl'al'l lIlannitol agar, eontaining Congo red (1:,,00).
Hesulls The r'('slIlls a['(' shown hdo\\" ill T;rl>le J. TaLle 1 Counts of diffcl'ent stwins of Hhi=obillrll j{/[JolliclIlli ill till' tll"O tcst media (average of triplicates) II L !16 !tours Yeast cxtr'act mallnitol Sll'iJill
111 ediulII
Yeast extract mannitol cal. glueonate medi Ulll
(X
(X 108)
I'SIl.\ I> 1:J(i
Hi
~anking
28 :12
SH-G SB-J(j
,,2
J()6)
Jj ')
7 ~
24
V. Iswaran, Abhiswal' Sen and R.Apte, A simple medium for quick growth ••.
The results show that in this medium the Australian acceptable minimum standard for broth inoculum (250 X 106 viable rhizobia/mI.) are obtained in 96 hours, thereby minimizing the usual 7 days' period in the yeast extract mannitol medium. This medium differs from the common medium in having calcium gluconate and ferric chloride with additional 1 g./I. yeast extract. This medium is being now for used slow-growing species, including cowpea groups. Zusammenfassung Rhizobium japonicum ist auf gewohnlichen Niihrmedien als ein langsam wachsendes Knollchenbakterium zu betrachten. Das in diesel' Mitteilung angegebene Medium el'moglicht ein schnelleres Wachstum diesel' Bakteriengattung.
Literature BERGENSEN, F. J.: Growth of Rhizobium in synthetic media. Austr. J. BioI. Sci. 14 (1961), 349. - FRED, E. B., BALDWIN, 1. 1., and McCoy, E.: Root nodule bacteria and leguminous plants. Univ. Wisconsin Press, Madison 1932. - ISWARAN, V., and JAUHRI, K. S.: Malt extract as a source of carbon for large scale production of Rhizobium inoculants. CUlT. Sci. 38 (1969), 67. - ISWARAN, V., GRAHAM, P. H., and JAUHRI, K. S.: Modified Burton's medium for large scale production of cultures of Rhizobium japonicum. Curl'. Sci. 38 (1969), :J4G. NORRIS, D. 0.: The role of calcium and magnesium in the nutrition of Rhizobium. Austr. J. agric. Res. 10 (1959), 651. - VAN SCHREVEN, D. A.: Methods used in the Netherlands for the production of legume inoculants. In Nutrition of the Legumes (ed. Hallsworth), Butterwol·ths. London 1958, pp. 328-333. - WEST, P. M., and WILSON, P. W.: Growth factor requirements of the root nodule bacteria. J. Bacteriol. 37 (1939),161. - WILSON, J. B., and UMllREIT, W. W.: Growth studies of Rhizobium japocnium. Proc. Soil Sci. Soc. ArneI'. V (1940), 262.
Authors' address: Dr. V. Iswaran, Dr. Abhiswar Sen and Sri R. Apte, Division of Microbiology, Indian Agricultural Research Institute, New Delhi-12 (India).