Biology and Fertility of Soils

, Volume 41, Issue 5, pp 350–358 | Cite as

Pink-pigmented facultative methylotrophic bacteria accelerate germination, growth and yield of sugarcane clone Co86032 (Saccharum officinarum L.)

  • M. Madhaiyan
  • S. Poonguzhali
  • H. S. Lee
  • K. Hari
  • S. P. Sundaram
  • T. M. Sa
Original Paper


The existence of Methylobacterium as a symbiont with sugarcane and its influence on crop growth at various stages was examined. Pink-pigmented facultative methylotrophic bacteria (PPFMs) strains isolated from different parts of the sugarcane clone Co86032 showed growth on methanol, and were further confirmed based on the mxaF gene encoding the α-subunit of the methanol dehydrogenase by polymerase chain reaction amplification using specific primers. True seeds inoculated with PPFMs had a higher germination percent and rate of germination than the control. A combined treatment of seed imbibition, soil application and phyllosphere spray increased specific leaf area, plant height, number of internodes, and cane yield. Immunological determination of cytokinin in young and mature leaves significantly increased when the epiphytic population on the leaf surface increased. Trends in sugar qualities in the form of Pol (sucrose) % in cane, Brix % in cane, and commercial cane sugar were similar to that of cane yield. These effects might be mediated by the production or synthesis of plant hormones.


Pink-pigmented facultative methylotrophic bacteria Methylobacterium spp. Plant hormones True seed germination Specific leaf area 



We thank M. A. Holland (Department of Biology, Salisbury University, Salisbury, Maryland) and Joe C. Polacco (Department of Biochemistry, University of Missouri–Columbia, Columbia, Missouri), for their valuable suggestions and provision of strains. The authors thank the Korea Science and Engineering Foundation for financial assistance through the Research Centre for the Development of Advanced Horticultural Technology at Chungbuk National University.


  1. Avezoux A, Goodwin MG, Anthony C (1995) The role of the novel disulphide ring in the active site of the quinoprotein methanol dehydrogenase from Methylobacterium extorquens. Biochem J 307:735–741Google Scholar
  2. Basile DV, Slade LL, Corpe WA (1969) An association between a bacterium and a liverwort, Scapania nemorosa. Bull Torrey Bot Club 96:6711–6714Google Scholar
  3. Beattie GA, Lindow SE (1995) The secret life of foliar bacterial pathogens on leaves. Annu Rev Phytopathol 33:145–172Google Scholar
  4. Breuer U, Ackermann JU, Babel W (1995) Accumulation of poly (3-hydroxybutyric acid) and overproduction of exopolysaccharides in a mutant of methylotrophic bacterium. Can J Microbiol 41:55–59Google Scholar
  5. Cervantes-Martinez J, Lopez-Diaz S, Rodriguez-Garay B (2004) Detection of the effects of Methylobacterium in Agave tequilana Weber var. azul by laser-induced fluorescence. Plant Sci 166:889–892Google Scholar
  6. Chanprame S, Todd JJ, Widholm JM (1996) Prevention of pink pigmented methylotrophic bacteria (Methylobacteirum mesophilicum) contamination of plant tissue cultures. Plant Cell Rep 16:222–225Google Scholar
  7. Corpe WA (1985) A method for detecting methylotrophic bacteria on solid surfaces. J Microbiol Methods 3:215–221Google Scholar
  8. Corpe WA, Basile DV (1982) Methanol-utilizing bacteria associated with green plants. Dev Ind Microbiol 23:483–493Google Scholar
  9. Corpe WA, Rheem S (1989) Ecology of the methylotrophic bacteria on living leaf surfaces. Microb Ecol 62:243–248Google Scholar
  10. Dileepkumar BS, Dube HC (1992) Seed bacterization with fluorescent Pseudomonas for enhanced plant growth, yield and disease control. Soil Biol Biochem 24:539–542Google Scholar
  11. Dunleavy JM (1988) Curtobacterium plantarum sp. nov. is ubiquitous in plant leaves and is seed transmitted in soybean and corn. Int J Syst Bacteriol 39:240–249Google Scholar
  12. Engelke J (2002) Sugarcane: measuring commercial quality. Department of Agriculture 2002 farmnote. Department of Agriculture, Western AustraliaGoogle Scholar
  13. Freyermuth SK, Long RLG, Mathur S (1996) Metabolic aspects of plant interaction with commensal methylotrophs. In: Lidstrom ME, Tabita FR (eds) Microbial growth on C1 compounds. Kluwer, Dordrecht, pp 277–284Google Scholar
  14. Green PN (1992) The genus Methylobacterium. In: Baloes A, Truper HG, Dworkin M, Harder W, Schleifer KH (eds) The prokaryotes. Springer, Berlin Heidelberg New York, pp 2342–2349Google Scholar
  15. Green PN, Bousifield IJ (1982) A taxonomic study of some Gram-negative facultatively methylotrophic bacteria. J Gen Microbiol 128:623–638Google Scholar
  16. Green PN, Bousifield IJ (1983) Emendation of Methylobacterium Patt, Cole and Hanson 1976, Methylobacterium rhodinum (Heumann 1962) comb. Nov. corig; Methylobacterium radiotolerans (Ito and Iizuka 1971), Comb. nov. corrig., and Methylobacterium mesophilicum (Austin and Goodfellow 1979) comb. nov. Int J Syst Bacteriol 33:875–877Google Scholar
  17. Hirano SS, Upper CD (1992) Bacterial community dynamics. In: Andrews JH, Hirano SS (eds) Microbial ecology of leaves. Springer, Berlin Heidelberg New York, pp 271–294Google Scholar
  18. Hirano SS, Upper CD (2000) Bacteria in the leaf ecosystem with emphasis on Pseudomonas syringae—a pathogen, ice nucleus, and epiphyte. Microbiol Mol Biol Rev 64:624–653Google Scholar
  19. Hirano SS, Baker S, Upper CD (1996) Raindrop momentum triggers growth of leaf-associated populations of Pseudomonas syringae on field-grown snap bean plants. Appl Environ Microbiol 62:2560–2566Google Scholar
  20. Holland MA (1997) Occams razor applied to hormonology. Are cytokinins produced by plants? Plant Physiol 115:865–868Google Scholar
  21. Holland MA, Polacco JC (1992) Urease-null and hydrogenase-null phenotypes of a phylloplane bacterium reveal altered nickel metabolism in two soybean mutants. Plant Physiol 98:942–948Google Scholar
  22. Holland MA, Polacco JC (1994) PPFMs and other contaminants: is there more to plant physiology than just plant? Annu Rev Plant Physiol Plant Mol Biol 45:197–209Google Scholar
  23. Holland MA, Long RLG, Polacco JC (2002) Methylobacterium spp.: phylloplane bacteria involved in cross-talk with the plant host? In: Lindow SE, Hecht-Poinar EI, Elliot VJ (eds) Phyllosphere microbiology. APS, St Paul, Minn., pp 125–135Google Scholar
  24. Ivanova EG, Doronina NV, Shepelyakovskaya AO, Laman AG, Brovko FA, Trotsenko YA (2000) Facultative and obligate aerobic methylobacteria synthesize cytokinins. Mikrobiologiya 69:764–769Google Scholar
  25. Ivanova EG, Doronina NV, Trotsenko YA (2001) Aerobic methylobacteria are capable of synthesizing auxins. Microbiology 70:392–397Google Scholar
  26. Knani M, Corpe WA, Rohmer M (1994) Bacterial hoponoids from pink-pigmented facultative methylotrophs and from green plant surfaces. Microbiology 140:2755–2759Google Scholar
  27. Koenig RL, Morris RO, Polacco JC (2002) tRNA is the source of low-level trans-Zeatin production in Methylobacterium spp. J Bacteriol 184:1832–1842Google Scholar
  28. Lidstorm ME (1992) The aerobic methylotrophic bacteria. In: Balows A, Truper HG, Dworkin M, Harder W, Schleifer KH (eds) The prokaryotes. Springer, Berlin Heidelberg New York, pp 431–445Google Scholar
  29. Lilley AK, Hails RS, Cory JS, Bailey MS (1997) The dispersal and establishment of pseudomonad populations in the phyllosphere of sugar beet by phytophagous caterpillars. FEMS Microbiol Ecol 24:151–157Google Scholar
  30. Lindemann J, Upper CD (1985) Aerial dispersal of epiphytic bacteria over bean plants. Appl Environ Microbiol 50:1229–1232Google Scholar
  31. Long R, Morris R, Polacco J (1997) Cytokinin production by plant-associated methylotrophic bacteria. Plant Physiol Abst 1168.
  32. Machackova I, Krekule J, Eder J, Seidlova F, Strnad (1993) Cytokinins in photoperiodic induction of flowering in Chenopodium species. Physiol Plant 87:160–166Google Scholar
  33. Machlin SM, Hanson RS (1988) Nucleotide sequence and transcriptional start site of the Methylobacterium organophilum XX methanol dehydrogenase structural gene. J Bacteriol 170:4739–4747Google Scholar
  34. Madhaiyan M (2003) Molecular aspects, diversity and plant interaction of facultative methylotrophs occurring in tropical plants. PhD dissertation. Tamilnadu Agricultural University, Coimbatore, TamilnaduGoogle Scholar
  35. Madhaiyan M, Sundaram SP, Kannaiyan S (2002) Influence of pink pigmented facultative methylotrophic bacteria on the seedling vigour of maize (Zea mays L.). J Microb World 4:117–121Google Scholar
  36. Madhaiyan M, Poonguzhali S, Senthilkumar M, Seshadri S, Chung HK, Yang JC, Sundaram SP, Sa TM (2004) Growth promotion and induction of systemic resistance in rice cultivar Co-47 (Oryza sativa L.) by Methylobacterium spp. Bot Bull Acad Sin 45:315–324Google Scholar
  37. Mathur RBL (1961) Handbook of cane sugar technology, 2nd edn. Oxford University Press, IBH, New DelhiGoogle Scholar
  38. Matsuchke J, Machackova I (2002) Changes in the content of indole-3-acetic acid and cytokinins in spruce, fir and oak trees after herbicide treatment. Biol Plant 45:375–382Google Scholar
  39. McDonald IR, Murrell JC (1997) The methanol dehydrogenase structural gene mxaF and its use as a functional gene probe for methanotrophs and methylotrophs. Appl Environ Microbiol 63(8):3218–3224Google Scholar
  40. McDonald IR, Kenna EM, Murrell JC (1995) Detection of methanotrophic bacteria in environmental samples with the PCR. Appl Environ Microbiol 61:116–121Google Scholar
  41. Meade GP (1963) Spencer-Meade cane sugar handbook. Wiley, New YorkGoogle Scholar
  42. Meade GP, Chen JCP (1977) Cane sugar handbook, 10th edn. Wiley, New YorkGoogle Scholar
  43. Nemecek-Marshall M, MacDonald RC, Franzen JJ, Wojciechowski CL, Fall R (1995) Methanol emission from leaves: enzymatic detection of gas-phase methanol and relation of methanol fluxes to stomatal conductance and leaf development. Plant Physiol 108:1359–1368Google Scholar
  44. Omer ZS, Tombolini R, Broberg A, Gerhardson B (2004) Indole-3-acetic acid production by pink-pigmented facultative methylotrophic bacteria. Plant Growth Regul 43:93–96Google Scholar
  45. Oppong D, King VM, Zhou X, Bowen JA (2000) Cultural and biochemical diversity of pink-pigmented bacteria isolated from paper mill slimes. J Ind Microbiol Biotech 25:74–80Google Scholar
  46. Patt TE, Cole GC, Hanson RS (1976) Methylobacterium, a new genus of facultatively methylotrophic bacteria. Int J Syst Bacteriol 26:226–229Google Scholar
  47. Peter TY, Jimmy DM, Benjamin LL (1998) Preservation of Saccharum spontaneum germplasm in the world collection of sugarcane and related grasses through storage of true seed. TEKTRAN, United States Department of Agriculture, Agricultural Research Service.
  48. SAS Institute (2001) SAS user’s guide, version 8.2. SAS Institute, Cary, N.C.Google Scholar
  49. Shepelyakovskaya AO, Doronina NV, Laman AG, Brovko FA, Trotsenko Yu A (1999) New data on the ability of aerobic methylotrophic bacteria to synthesize cytokinins. Dokl Akad Nauk 368:555–557Google Scholar
  50. Sy A, Girud E, Jourand P, Garcia N, Willems A, De Lajudie P, Prin Y, Neyra M, Gills M, Catherine BM, Dreyful B (2001) Methylotrophic Methylobacterium bacteria nodulate and fix atmospheric nitrogen in symbiosis with legumes. J Bacteriol 183:214–220CrossRefPubMedGoogle Scholar
  51. Terauchi T, Matsuoka M (2001) Analysis of the slow growth of sugarcane at the early stage. Proc Int Soc Sugarcane Technol 24:149–151Google Scholar
  52. Terauchi T, Matsuoka M, Nakagawa H, Nakano H (2001) A breeding index for improving the early growth of sugarcane. JIRCAS Res Highlights 2001Google Scholar
  53. Walker JC, Patel PN (1964) Splash dispersal and wind as factors in epidemiology of haloblight of bean. Phytopathology 54:140–141Google Scholar
  54. Whittenbury R, Davies SL, Wilkinson JF (1970) Enrichment, isolation and some properties of methane-utilizing bacteria. J Gen Microbiol 61:205–218PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • M. Madhaiyan
    • 1
  • S. Poonguzhali
    • 1
  • H. S. Lee
    • 1
  • K. Hari
    • 2
  • S. P. Sundaram
    • 3
  • T. M. Sa
    • 1
  1. 1.Department of Agricultural ChemistryChungbuk National UniversityChungbukRepublic of Korea
  2. 2.Division of Crop Production, Sugarcane Breeding InstituteICARTamilnaduIndia
  3. 3.Department of Agricultural MicrobiologyTamilnadu Agricultural UniversityTamilnaduIndia

Personalised recommendations