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Technical approaches to inoculate micropropagated sugar cane plants were Acetobacter diazotrophicus

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Abstract

Micropropagated plantlets of sugar cane were inoculated with the N2-fixing bacterium Acetobacter diazotrophicus. Various modifications on the basic plant culture medium MS were made for the plant/bacteria association. The protocol required the inoculation of the bacteria at the end of the rooting period in a medium without hormones or vitamins, and with the concentration of sugar and mineral nutrients reduced by a factor of 10. Individual plants were inoculated with A. diazotrophicus and maintained under the appropriate light and temperature condition used for micropropagation up to 7 days. The system favored the infection and the establishment of the bacteria within the plant tissue. Bacteria colonized the plant tissue and accumulated in inter-cellular cavities and the region of lateral root emergence and also colonizes the xylem vessels. The inoculated plantlets were subsequently transferred to the acclimatization phase and after 30 days it was possible to isolate the bacteria from plant tissue. This protocol permitted studies of infection and comparison among strains.

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References

  • Cavalcante V A and Döbereiner J 1988 A new acid tolerant nitrogen-fixing bacterium associated with sugarcane. Plant Soil 108, 23–31.

    Article  Google Scholar 

  • Cojho E M, Reis V M, Schenberg A C and Döbereiner J 1993 Interactions of Acetobacter diazotrophicus with an amylolytic yeast in nitrogen-free batch culture. FEMS Microbiol. Lett. 106, 341–346.

    CAS  Google Scholar 

  • Edelman J and Hanson A D 1971 Sucrose suppression of chlorophyll synthesis in carrot callus cultures. Planta 98, 150–156.

    Article  CAS  Google Scholar 

  • Hendre R R, Iyor R S, Kotwalm M, Kluspe S S and Mascarenhas A F 1983 Rapid multiplication of sugar cane by tissue culture. Sugar Cane 1, 5–8.

    Google Scholar 

  • James E K, Reis V M, Olivares F L, Baldani J I and Döbereiner J 1994 Infection of sugar cane by nitrogen fixing bacterium Acetobacter diazotrophicus. J. Exp. Bot. 45, 757–766.

    Article  CAS  Google Scholar 

  • Li R and Macrae I C 1992 Specific identification and enumeration of Acetobacter diazotrophicus in sugarcane. Soil Biol. Biochem. 24, 413–419.

    Article  Google Scholar 

  • Lima E, Boddey R m and Döbereiner J 1987 Quantification of biological nitrogen fixation associated with sugar cane using 15N aided nitrogen balance. Soil Biol. Biochem. 19, 165–170.

    Article  CAS  Google Scholar 

  • Murashigue T and Skoog F A 1962 Revised medium for rapid growth and bioassays with Tobacco tissue cultures. Physiol. Plant. 15, 473–497.

    Article  Google Scholar 

  • Paula M A, Reis V M and Döbereiner J 1991 Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of roots and tops of Sweet Potatoe (Ipomea batatas) sugar cane (Saccharum spp.) and Sweet Sorghum (Sorghum vulgare). Biol. Fertil. Soils 11, 111–115.

    Article  Google Scholar 

  • Reis V M 1991 Aspectos ecológicos e fisiológicos da bactéria fixadora de N2 Acetobacter diazotrophicus. M.S. Thesis, Universidade Federal Rural do Rio de Janeiro, Brazil. 119 p.

    Google Scholar 

  • Reis V M, Olivares F L and Döbereiner J 1994 Improved methodology for isolation of Acetobacter diazotrophicus and confirmation of its endophytic habitat. World J. Appl. Microbiol. Biotecnol. 10, 101–104.

    Google Scholar 

  • Reis V M, Zang Y and Burris R H 1990 Regulation of nitrogenase activity by ammonium and oxigen in Acetobacter diazotrophicus. An. Acad. Bras. Ciên. 62, 317.

    Google Scholar 

  • Rodrigues Neto J, Malavolta Jr V A and Victor O 1986 Meio simples para o isolamento e cultivo de Xanthomonas campestris pv. citri TIPO B. Suma Phytopathologica 12, 16.

    Google Scholar 

  • Stephan M P, Oliveira M, Teixeira K R S, Martinez-Drets G and Döbereiner J 1991 Physiology and dinitrogen fixation of Acetobacter diazotrophicus. FEMS Microbiol. Lett. 77, 67–72.

    Article  CAS  Google Scholar 

  • Urquiaga S, Cruz K H S and Boddey R M 1992 Contribution of nitrogen fixation to sugar cane: Nitrogen-15 and nitrogen balance estimates. Soil Sci. Soc. Am. J. 56, 105–114.

    Article  Google Scholar 

  • Yamada, Y and Sato, F 1978 The photoautotrophic culture of chlorophyllous cells. Plant Cell Physiol. 19, 691–699.

    CAS  Google Scholar 

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Reis, V.M., Olivares, F.L., de Oliveira, A.L.M. et al. Technical approaches to inoculate micropropagated sugar cane plants were Acetobacter diazotrophicus . Plant Soil 206, 205–211 (1999). https://doi.org/10.1023/A:1004436611397

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  • DOI: https://doi.org/10.1023/A:1004436611397

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