Menéndez C, Trujillo LE, Ramírez R, González-Peña D, Espinosa D, Enriquez GA, Hernández L

Language: Spanish
References: 13
Page: 111-115
PDF size: 445.59 Kb.

ABSTRACT

Symbiotic nitrogen fixation is critical for high soybean (Glycine max L. Merril) yields with protection to the environment. This paper describes a procedure for massive production of Bradyrhizobium japonicum in an orbital multi-platform shaker and the use of the bacterial culture for seed inoculation in soybean areas under mechanized sowing. The B. japonicum strain was selected based on its highly efficient symbiotic interaction with major soybean varieties cropped in Cuba. Optimization of the growth medium and the use of serial and scaled inoculations allowed maintaining the cultures in continuous logarithmic phase while increasing its volume 100-fold in 7 days. The final culture having average cell viability of 5 x 10¹⁰ c.f.u./mL and high exopolysaccharides content does not require additives and can be stably stored for at least 8 months at 4 ºC. Soybean seeds were sprayed with the inoculant at equivalent dose of 150 mL/ha and sowed mechanically in production fields of Ciego de Avila province during the campaigns 2010-2013. The plants showed abundant nodulation and a proper nutritional state during the whole cycle. The bacterium strain was reisolated from root nodules of inoculated plants and its identity was confirmed by phenotype characterization. The high-cell density liquid inoculant rich in polysaccharides produced in this work is compatible with mechanized seed sowing, favors the bacterium survival in soil, and promotes the establishment of an efficient symbiotic interaction with the host plant.

REFERENCES

1. Producción de determinados productos básicos agrícolas, Grupo I (2004). In: FAO. Anuario estadístico de la FAO. 2005-2006. Roma: FAO; 2004. p. 79-82.

2. Clive J. ISAAA Brief 42. Global Status of Commercialized Biotech/GM Crops: 2010. New York: International Service for de Acquisition of Agri-Biotech Applications (ISAAA); 2010.

3. Singh MS. Effect of Bradyrhizobium inoculation on growth, nodulation and yield attributes of soybean - A Review. Agric Rev. 2005;26(4):305-8.

4. Appelbaum, E. The Rhizobium/Bradyrhizobium- legume symbiosis. In: Gresshoff PM, editor. The molecular biology of symbiotic nitrogen fixation. Boca Raton: CRC Press; 1989. p. 131-58.

5. Hollis AB, Kloos WE, Elkan GE. DNA: DNA hybridization studies of Rhizobium japonicum and related Rhizobiacea. J Gen Microbiol. 1981;123(2):215-22.

6. Alberton O, Kaschuk G, Hungria M. Sampling effects on the assessment of genetic diversity of rhizobia associated with soybean and common bean. Soil Biol Biochem. 2006;38(6):1298-307.

7. Soto N, Ferreira A, Delgado C, Enríquez GA. Regeneración in vitro de plantas de soya de la variedad cubana IncaSoy-36. Biotecnol Apl. 2013;30(1):29-33.

8. Romero-Arias A, Peña-MolinaL, González-Cruz M, González M. Evaluación de cultivares de soya (Glycine max) en las condiciones edafoclimáticas del municipio Majibacoa, Las Tunas. Innov Tecnol. 2013;19:1-9.

9. Díaz MF, Padilla C, Torres V, González A, Curbelo F. Caracterización bromofológica de variedades de soya (Glycine max) en producción de forrajes, forrajes integrales y granos de siembras de verano. Rev Cubana Cienc Agríc. 2003;37(3):311-7.

10. Somasegaran P, Hoben HJ. Handbook for rhizobia. Methods in legume-Rhizobium technology. New York: Springer-Verlag New York, Inc.; 1994.

11. Subba Rao NS. Soil microorganisms and plant growth. New Delhi: Oxford and IBH Publishing Co.; 1977.

12. Louch HA, Miller KJ. Synthesis of a low-molecular-weight form of exopolysaccharide by Bradyrhizobium japonicum USDA 110. Appl Environ Microbiol. 2001; 67(2):1011-4.

13. Maurice S, Beauclair P, Giraud J, Sommer G, Hartmann A. Survival and change in physiological state of Bradyrhizobium japonicum in soybean (Glycine max L. Merril) liquid inoculants after longterm storage. World J Microbiol Biotech. 2001; 17(6):635-43.