Plant and Soil

, Volume 336, Issue 1–2, pp 129–142 | Cite as

Enhancement of rice production using endophytic strains of Rhizobium leguminosarum bv. trifolii in extensive field inoculation trials within the Egypt Nile delta

  • Youssef G. Yanni
  • Frank B. Dazzo
Regular Article


This study assessed the ability of biofertilizer inoculants containing Rhizobium leguminosarum bv. trifolii to enhance production of rice (Oryza sativa L.) under actual agricultural conditions in the Nile delta. Large-scale field experiments evaluated 5 rice varieties inoculated with 7 endophytic rhizobial strains during 5 growing seasons, including at sites ranked as the world’s highest in rice production. Inoculation with single strains or multi-strain consortia significantly increased grain yield in 19 of the 24 trials. By combining superior rhizobial inoculants with agricultural extension training, grain yield increased up to 47% in farmers’ fields, with an average increase of 19.5%. Data on rice straw production, harvest index and the agronomic fertilizer N-use efficiency also indicated positive agronomic benefits of rhizobial inoculation. These results establish the merit of deploying our biofertilization strategy using selected rhizobial strains to promote rice production capacity while reducing the need for additional chemical N-fertilizer inputs to maintain agricultural sustainability and acceptable production economy. Technology transfer of this important translational research can significantly help to alleviate hunger and meet the nutritional needs of many people in developing countries.


Biofertilizer Consortia Endophyte Field inoculation Inoculant Rhizobium Rice 





colony forming units


Food and Agriculture Organization


International Rice Research Institute


plant growth-promoting rhizobacteria


yeast extract mannitol



This work was supported by projects BIO2-001-017 and BIO5-001-015 (Contract/Agreement Award No. 27 and 015, respectively) of the US-Egypt Science and Technology Joint Fund. We thank the numerous rice farmers listed in Table 2 and our field experimentation experts for their cooperation in this study.

Supplementary material


  1. Arora S (1969) The role of algae in the availability of phosphorus in paddy fields. Riso 18:135–138Google Scholar
  2. Black A, Evans DD, Ensmingers FE, White JI, Clark FE, Dinaver RC (1965) Methods of soil analysis. II- Chemical and microbiological properties. No.9, Series of Agronomy, American Society of Agronomy Inc., Madison, Wisconsin, USAGoogle Scholar
  3. Biswas JC, Ladha JK, Dazzo FB, Yanni YG, Rolfe BG (2000a) Rhizobial inoculation influences seedling vigor and yield of rice. Agron J 92:880–886CrossRefGoogle Scholar
  4. Biswas JC, Ladha JK, Dazzo FB (2000b) Rhizobia inoculation improves nutrient uptake and growth of lowland rice. Soil Sci Soc Amer J 64:1644–1650CrossRefGoogle Scholar
  5. Chaintreuil C, Giraud E, Prin Y, Lorquin J, Ba A, Gillis M, de Laudie P, Dreyfus B (2000) Photosynthetic bradyrhizobia are natural endophytes of the African wild rice Oryza breviligulata. Appl Environ Microbiol 66:5437–5447CrossRefPubMedGoogle Scholar
  6. Chi F, Shen SH, Cheng HP, Jing YX, Yanni YG, Dazzo FB (2005) Ascending migration of endophytic rhizobia, from roots to leaves, inside rice plants and assessment of benefits to rice growth physiology. Appl Environ Microbiol 71:7271–7278CrossRefPubMedGoogle Scholar
  7. Dazzo FB, Yanni YG (2006) The natural Rhizobium-cereal crop association as an example of plant-bacteria interaction. In: Uphoff N et al (eds) Biological approaches to sustainable soil systems. Taylor and Francis, Boca Raton, pp 109–127CrossRefGoogle Scholar
  8. Dazzo FB, Yanni YG, Rizk R, deBruijn F, Squartini A, Mateos P, Martinez-Molina E, Biswas J, Ladha JK, Weinman J, Rolfe B, Vega-Hernandez M, Hollingsworth RI (1999) The natural beneficial association between Rhizobium leguminosarum bv. trifolii and rice and its exploitation in sustainable agriculture. Proceedings of the "International Symposium on Nitrogen-Fixation and Crop Production", Cairo, Egypt, May 11–13, 99Google Scholar
  9. Dazzo FB, Yanni YG, Rizk R, de Bruijn FJ, Rademaker J, Squartini A, Corich V, Mateos P, Martínez-Molina E, Velázquez E, Biswas JC, Hernandez RJ, Ladha JK, Hill J, Weinman J, Rolfe BG, Vega-Hernández M, Bradford JJ, Hollingsworth RI, Ostrom P, Marshall E, Jain T, Orgambide G, Philip-Hollingsworth S, Triplett E, Malik KA, Maya-Flores J, Hartmann A, Umali-Garcia M, Izaguirre-Mayoral ML (2000) Progress in multinational collaborative studies on the beneficial association between Rhizobium leguminosarum bv. trifolii and rice. In: Ladha JK, Reddy PM (eds) The quest for nitrogen fixation in rice. IRRI, Los Banos, pp 167–189Google Scholar
  10. Gupta A, Sharma BM, Kerni S (1989) Comparative efficiency of Azospirillum, Azotobacter and blue-green algae in submerged rice culture. Res Develop Reporter 6:169–171Google Scholar
  11. Gutierrez-Zamora M, Martinez-Romero E (2001) Natural endophytic association between Rhizobium etli and maize (Zea mays). J Biotechnol 91:117–126CrossRefPubMedGoogle Scholar
  12. Hilali A, Prevost D, Broughton W, Hartmann A (2001) Effets de l’inoculation avec des souches de Rhizobium leguminosarum biovar trifolii sur la croissance der le dans deux sols der Maroc. Can J Microbiol 41:590–593CrossRefGoogle Scholar
  13. Jack V, Roger PA (1977) Diminution des fonts de semis dues ‘a la sulfatore’ duction, par un pre’traitement des grains de riz avec des cyanophyc’ees. Cabiers ORSTOM Serie Biologie 12:101–107Google Scholar
  14. Jackson ML (1967) Soil chemical analysis. Prentice-Hall, New DelhiGoogle Scholar
  15. Jha B, Thakur MC, Gentia I, Albrecht V, Stoffels M, Schmid M, Hartmann A (2009) Isolation, partial identification and application of diazotrophic rhizobacteria from traditional Indian rice cultivars. Eur J Soil Biol 45:62–72CrossRefGoogle Scholar
  16. Lupway N, Clayton G, Hanson K, Rice W, Bierderbeck V (2004) Endophytic rhizobia in barley, wheat, and canola roots. Can J Pl Sci 84:37–45Google Scholar
  17. Mano H, Morisaki H (2008) Endophytic bacteria in the rice plant. Microbes Environ 23:109–117CrossRefGoogle Scholar
  18. Matiru VN, Dakora FD (2004) Potential use of rhizobial bacteria as promoters of plant growth for increased yield in landraces of African cereal crops. African J Biotechnol 3:1–7Google Scholar
  19. Mishra RPN, Singh RK, Jaiswal HK, Singh MK, Yanni YG, Dazzo FB (2008) Rice-rhizobia association: evolution of an alternate niche of beneficial plant-bacteria association. In: Ahmad I, Pichtel J, Hayat S (eds) Plant-bacteria interactions: strategies and techniques to promote plant growth. Wiley-VCH Verlag GmbH, Weinheim, pp 165–194Google Scholar
  20. Omar MNA, Hussein KRF, Zaher EA, Abou-Zeid MY (1993) Effect of inoculation with Azospirillum brasilense on yield and some chemical constituents of rice grain. In: Hegazi NA, Fayez M, Monib M (eds) Nitrogen fixation with non-legumes. American University in Cairo Press, Cairo, pp 493–494Google Scholar
  21. Peng G, Yuan Q, Zhang W, Tan Z (2008) Rhizobium oryzae sp. nov., isolated from the wild rice Oryza alta. Int J System Evol Microbiol 58:2158–2163CrossRefGoogle Scholar
  22. Perrine FM, Prayitno J, Weinman J, Dazzo FB, Rolfe BG (2001) Rhizobium plasmids are involved in the inhibition or stimulation of rice growth and development. Austr J Plant Physiol 28:923–937Google Scholar
  23. Prayitno J, Stefaniak J, McIver J, Weinman JJ, Dazzo FB, Ladha JK, Barraquio W, Yanni YG, Rolfe BG (1999) Interactions of rice seedlings with bacteria isolated from rice roots. Austr J Plant Physiol 26:521–535CrossRefGoogle Scholar
  24. Rajarmamohan RV, Charyulu PBBN, Nayas DN, Ramakrishna C (1978) Nitrogen fixation by free-living organisms in tropical rice soil. In: Dobereiner J, Burris RH, Hollaender A, Franco AA, Scott DB (eds) Limitation and potential for biological nitrogen fixation in the tropics. Plenum, New York, p 354Google Scholar
  25. Rao DLN, Sharma PC (1995) Alleviation of salinity stress in chickpea by Rhizobium inoculation or nitrate supply. Biologia Plantarum 37:405–410CrossRefGoogle Scholar
  26. Reddy PM, Ladha JK, So R, Hernandez R, Dazzo FB, Angeles OR, Ramos MC, de Bruijn FJ (1997) Rhizobial communication with rice: induction of phenotypic changes, mode of invasion and extent of colonization. Plant Soil 194:81–98CrossRefGoogle Scholar
  27. Roger PA, Kulasooriya SA (1980) Blue-green Algae and Rice. International Rice Research Institute, PhilippinesGoogle Scholar
  28. Shahaby AF, Amin G, Khalafalla GM (1993) Response of rice and tomato seedlings to inoculation with diazotrophs and their culture filtrates. In: Hegazi NA, Fayez M, Monib M (eds) Nitrogen fixation with non-legumes. American University in Cairo, Cairo, pp 375–376Google Scholar
  29. Singh RK, Mishra RPN, Jaiswal HK, Kumar V, Pandey SP, Rao SB, Annapurna K (2006) Isolation and identification of natural endophytic rhizobia from rice (Oryza sativa L.) through rDNA PCR-RFLP and sequence analysis. Curr Microbiol 52:117–122CrossRefPubMedGoogle Scholar
  30. Singh MK, Kushwaha C, Singh RK (2009) Studies on endophytic colonization ability of two upland rice endophytes, Rhizobium sp. and Burkholderia sp. using green fluorescent protein reporter. Springer Science and Business Media, LLC 2009, published online 30 May 2009Google Scholar
  31. Subbarao GV, Johansen C, Kumar Rao JVDK, Jana MK (1990) Response of the pigeonpea-Rhizobium symbiosis to salinity stress: variation among Rhizobium strains in symbiotic activity. Biol Fert Soils 9:49–53CrossRefGoogle Scholar
  32. Subrahmanyan R, Relwani LL, Manna GB (1965) Fertility build-up of rice field soils by blue-green algae. Proc Indian Acad Sci Series B 62:252–277Google Scholar
  33. Sun L, Qiu F, Zhang X, Dai X, Dong X, Song W (2008) Endophytic bacterial diversity in rice (Oryza sativa L.) roots estimated by 16S rDNA sequence analysis. Microbial Ecol 55:415–424CrossRefGoogle Scholar
  34. Tan Z, Hurek T, Gyaneshwar P, Ladha JK, Reinhold-Hurek B (2001) Specific detection of Bradyrhizobium and Rhizobium strains colonizing rice (Oryza sativa) roots by 16S–23S ribosomal DNA intergenic spacer-targeted PCR. Appl Environ Microbiol 67:3655–3664CrossRefPubMedGoogle Scholar
  35. Venkataraman GS (1966) Algalization. Phykos 5:164–174Google Scholar
  36. Yanni YG, Abd El-Fattah FK (1999) Towards integrated biofertilization management with free living and associative dinitrogen fixers for enhancing rice performance in the Nile Delta. Symbiosis 27:319–331Google Scholar
  37. Yanni YG, Rizk RY, Corich V, Squartini A, Ninke K, Hollingsworth SP, Orgambide G, de Bruijn F, Stoltzfus R, Buckley D, Schmidt T, Mateos PF, Ladha JK, Dazzo FB (1997) Natural endophytic association between Rhizobium leguminosarum bv. trifolii and rice roots and assessment of its potential to promote rice growth. Plant Soil 194:99–114CrossRefGoogle Scholar
  38. Yanni YG, Rizk RY, Abd El-Fattah F, Squartini A, Corich V, Giacomini A, de Bruijn F, Rademaker J, Maya-Flores J, Ostrom P, Vega-Hernandez M, Hollingsworth RI, Martinez-Molina E, Mateos P, Velazquez E, Wopereis J, Triplett E, Umali-Garcia M, Anarna JA, Rolfe BG, Ladha JK, Hill J, Mujoo R, Ng PK, Dazzo FB (2001) The beneficial plant growth-promoting association of Rhizobium leguminosarum bv. trifolii with rice roots. Austr J Plant Physiol 62:845–870Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  1. 1.Sakha Agricultural Research StationKafr El-SheikhEgypt
  2. 2.Department of Microbiology and Molecular GeneticsMichigan State UniversityEast LansingUSA

Personalised recommendations