Occurrences and characterization of endophytic Gluconacetobacter isolated from sugarcane tissues of North Eastern region of India

DOI: 10.18805/ijare.v0iOF.9362    | Article Id: A-4501 | Page : 287-294
Citation :- Occurrences and characterization of endophytic Gluconacetobacter isolated from sugarcane tissues of North Eastern region of India .Indian Journal of Agricultural Research.2016.(50):287-294

Merasenla Ao1, Anjuma Gayan, Smirta Buragohain and Dhruba Jyoti Nath* 

Address :

Department of Soil Science, Assam Agricultural University, Jorhat-785 013, India.

Submitted Date : 16-09-2015
Accepted Date : 12-01-2016


Sugarcane plants including roots were collected from 21 sugarcane growing locations of Golaghat, Dimapur and Jorhat districts of North Eastern region for isolation of endophytic Gluconacetobacter. Based on the nitrogenase activity, only eight isolates were selected for further characterization of Gluconacetobacter. The nitrogenase activity of the isolates ranged in between 12.4 to 72.4 n mole C2H4 h-1 culture-1. The characteristic yellowish surface pellicles were formed on inoculating the selected isolates into semisolid LGI medium and acetic LGI medium. On enumeration in LGI plates, the frequencies of well recognized Gluconacetobacter like colonies ranged in between 0.75 and 7.68 x 105 (cfu g-1 fresh tissue). Colony morphology of the isolates was irregular flat, irregular raised or circular raised yellow colonies of 1-5 mm in diameter. Light brown to brown pigmentation was observed on potato agar plates and light brown pigmentation with clearing of plates was observed on GYC plates. All the isolates exhibited gram negative and shape of the cells were rods. Reactions to extracellular and intracellular enzymes as well as to the various carbon sources by the isolates explained the diversity among them. The IAR profiles of the eight isolates against twelve antibiotics exhibited wide variation in their response to various antibiotics. 


Carbon sources Enzyme activities Gluconacetobacter Pigmentation Sugarcane tissues.


  1. Agrawal N and Singh H P. (2002) Antibiotic resistance and inhibitory effect of Azotobacter on soil borne plant pathogens. Indian Jour. Microbiol. 42: 245-246.
  2. Baldani V L D, Alvarez B, Baldani J L and Dobereiner J. (1986) Establishment of inoculated Azospirillum spp. in the rhizosphere and in roots of field grown wheat and sorghum. Plant Soil. 90:35-46.
  3. Becking J H. (1974) Family 2, Azotobacteriaceae In: Bergey’s Manual of Determinative Bacteriology, [Edited by Buchanan, R.E. and Gibbons, N.E.] (8th edition). The Williams and Wilkins Co., Baltimore, pp. 253-261.
  4. Blazevic D J and Ederer G M. (1975) Principles of Biochemical Tests in Diagnostic Microbiology. Wiley and Company, New York, pp.13-45.
  5. Boddey R M, Urquiaga S, Reis V M, Dobereiner J. (1991) Biological nitrogen fixation associated with sugarcane. Plant Soil. 37:111–117.
  6. Cappuccino J G and Sherman N. (1992) In: Microbiology: A Laboratory Manual. The Bejamin/Cummings Publishing Company, Inc., California.
  7. Cappuccino J G and Sherman N. (1996) In: Microbiology: A Laboratory Manual. The Benjamin/Cunnings Publishing Company Inc. (4th edition), Menlopark, California.
  8. Carrizo de Bellone S and Bellone C H. (2006) Presence of endophytic diazotrophs in sugarcane juice. World Jour. Microbiol. Biotech. 22:1065–1068.
  9. Cavalcante V A and Dobereiner J. (1988) A new acid-tolerant nitrogen-fixing bacterium associated with sugarcane. Plant. Soil. 108: 23–31.
  10. Cojho E H, Reis V M, Schenberg A C G and Dobereiner J. (1993) Interactions of Acetobacter diazotrophicus with an amylolytic yeast in nitrogen-free batch culture. FEMS Microbiol. Lett. 106: 341.
  11. Doern G V. (1995) Susceptibility tests of fastidious bacteria. In: Manual of Clinical Microbiology (P.R, Murray, E.J.P. Baron, M.A. Faller, F.C. Tenover and R, Yolken Eds), American Society for Microbiology, Washington DC, PP 1342-1349..
  12. Dobereiner J and Baldani V L D. (1979) Selective infection of maize roots by streptomycin -resistant Azospirillum lipoferum and bacteria. Canadian Jour. Microbiol. 25:1264-1269.
  13. Dobbelaere S, Vanderleyden J and Okon Y. (2003) Plant growth promoting effects of diazotrophs in the rhizosphere. Critical Reviews in Plant Sciences 22: 107–149.
  14. Dutta D and Gachhui R. (2007) Nitrogen-fixing and cellulose producing Gluconacetobacter kombuchae sp. nov. isolated from Kombucha tea. Intl. Jour. Syst. Evol.Microbiol. 57:353–357.
  15. Fuentes-Ramirez L E, Bustillos-Cristales R, Tapia-Hernandez A, Jimenez-Salgado T, Wang E T, Martinez-Romero E and Caballero-Mellado J. (2001) Novel nitrogen-fixing acetic acid bacteria Gluconacetobacter johannae sp. nov. and Gluconacetobacter azotocaptans sp. nov., associated with coffee plants. Intl. Jour. Syst. Evol.Microbiol. 51: 1305–1314.
  16. Fuentes-Ramirez L E, Jimenez-Salgado T, Abarca-Ocampo I R and Caballero-Mellado J. (1993) Acetobacter diazotrophicus, an indole acetic acid producing bacterium isolated from sugarcane cultivars of Mexico. Plant Soil. 154:145–150.
  17. Gerhardt P, Murry R G E, Costilow R N, Nester E W, Wood W A, Kreig N R and Phillips G B. (1981) In: Manual of Methods of General Bacteriology. American Society for Microbiology, Washing DC.
  18. Gillis M, Kersters K, Hoste B, Janssens D, Kroppenstedt R M, Stephan M P, Teixeira K R S, Dobereiner J and De Ley J. (1989) Acetobacter diazotrophicus sp. nov., a nitrogen fixing acetic acid bacterium associated with sugarcane. Intl.Jour. Syst.Bacteriol. 39: 361–364.
  19. Hernandez L, Sotolongo M, Rosabal Y, Menendez C, Ramirez R And Caballero-Mellado J. (2000) Structural levansucrase gene (lsdA) constitutes a functional locus conserved in the species Gluconacetobacter diazotrophicus. Arch. Microbiol.174: 120–124.
  20. Jimenez-Salgado T, Fuentes-Ramirez L E, Tapia-Hernandez A, Mascarua M A, Martinez-Romero E and Caballero-Mellado J. (1997) Coffea arabica L., a new host plant for Acetobacter diazotrophicus, and isolation of other nitrogen fixing acetobacteria. App. Environ.Microbiol. 63: 3676–3683.
  21. Loganathan P and Nair S. ( 2003) Crop-specific endophytic colonization by a novel, salt-tolerant, Nitrogen fixing and phosphate-solubilizing Gluconacetobacter sp. from wild rice. Biotech. Lett. 25: 497-501.
  22. Loganathan P, Sunitha R, Parida A K and Nair S. (1999) Isolation and characterization of two genetically distant groups of Acetobacter diazotrophicus from a new host plant Eleusine coracana L. Jour. App. Microbiol. 87:167–172.
  23. Madhaiyan, Saravanan V S, Bhakiya Silba Sandal Jovi D, Hyoungseok Lee, Thenmozhi R, Hari K And Tongmin Sa. (2004) Occurrence of Gluconacetobacter diazotrophicus in tropical and subtropical plants of Western Ghats, India. Microbiol. Res.159: 233–243.
  24. Mehnaz S, Weselowski B and Lazarovits G. (2006) Isolation and identification of Gluconacetobacter azotocaptans from corn rhizosphere: Syst. App.Microbiol. 29: 496-501.
  25. Muthukumarasamy R, Cleenwerck I, Revathi G, Vadivelu M, Janssens D, Hoste B, Gum K U, Park K D, Son C Y, Sa T M and Caballero-Mellado J. (2005) Natural association of Gluconacetobacter diazotrophicus and diazotrophic Acetobacter peroxydans with wetland rice. Syst. App.Microbiol. 28:277–286.
  26. Muthukumarasamy R, Revathi G and Lakshminarasimhan C. (1999) Influence of N fertilization on the isolation of Acetobacter diazotrophicus and Herbaspirillum spp. from Indian sugarcane varieties. Biol. Fert. Soils. 29:157–164.
  27. Muthukumarasamy R, Revathi G and Vadivelu M. (2000) Antagonistic potential of N2 fixing Acetobacter diazotrophicus against Colletotrichum falcatum Went, a causal organism of red-rot of sugarcane. Curr. Sci. 78:1063–1065.
  28. Muthukumarasamy R, Revathi G, Seshadri S and Lakshminarasimhan C. (2002) Gluconacetobacter diazotrophicus (Syn. Acetobacter diazotrophicus), a promising diazotrophic endophyte in tropics. Curr. Sci. 83:137–145.
  29. Rajaram K and Suri R K. (2000) In: Science and Technology in India. Spectrum India, pp. 68–69.
  30. Rangaswami G and Bagyaraj D J. (1993) Microbial Biotechnology. In: Agricultural Microbiology. Prentice Hall of India Pvt. Ltd., New Delhi, pp.389-405.
  31. Reis V M, Olivares F L and Dobereiner J. (1994) Improved methodology for isolation of Acetobacter diazotrophicus and confirmation of its endophytic habitat. World Jour. Microbiol. Biotech. 10:401–405.
  32. Seeley H W and Vandemark P J. (1981) Microbes in Action – A Laboratory Manual of Microbiology. Freeman and Company, San Francisco, USA, pp. 388.
  33. Smibert R M and Kreig N R. (1981) In: Methodology for General Bacteriology, Edited by Gerhardt, P. Academic Publisher, New York, pp.400-450. 
  34. Somasegaran P and Hoben H J. (1994) Handbook for Rhizobia: Methods in Legume Rhizobium Technology. New York: Springer­Verlag.
  35. Stephan M P, Oliveira M, Teixeira K R S, Martinez-Drets G and Dobereiner J. (1991) Physiology and dinitrogen fixation of Acetobacter diazotrophicus. FEMS Microbiol. Lett. 77: 67–72. 
  36. Suman A, Shasany A K, Singh M, Shahi H N, Gaur A and Khanuja S P S. (2001) Molecular assessment of diversity among endophytic diazotrophs isolated from subtropical Indian sugarcane. World Jour. Microbiol. Biotech. 17: 39–45.
  37. Yamada Y, Hoshino K I and Ishikawa T. (1998) Gluconacetobacter nom.corrig. Gluconacetobacter (sic). In: validation of publication of new names and new combinations previously effectively published outside the IJSB. List no. 64. Intl. Jour. Syst.Bacteriol. 48: 327-328.

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