Development of liquid rhizobial inoculants and pre-inoculation of alfalfa seeds



rhizobia, pre-inoculation, alfalfa seeds, liquid inoculants, Sinorhizobium meliloti


Paper description:

  • Inoculation of legume seeds with symbiotic nitrogen-fixing bacteria, rhizobia, is an efficient mechanism for improving the nitrogen content and legume growth. The use of pre-inoculated seeds simplifies sowing in the field and reduces the cost of cultivation.
  • We used the following additives: agar, sodium-alginate, calcium chloride, glycerol and ferric chloride as supplements for alfalfa rhizobium (Sinorhizobium meliloti L3Si) survival in liquid inoculants as the final commercial products, and for pre-inoculation of seeds and one month storage.
  • The liquid rhizobial inoculants were successful for inoculation and pre-inoculation of alfalfa seeds and improved the yield and nitrogen content in alfalfa plants.

Abstract: Application of liquid microbial inoculants on legume seeds is a sustainable agricultural practice that can improve plant nutrient uptake and increase crop productivity. Inoculants should provide long-term survival of rhizobia in the final product and after application, to legume seeds. Ten different medium formulations of microbial inoculants were examined (yeast mannitol broth with the addition of agar, sodium-alginate, calcium chloride, glycerol or ferric chloride and combinations thereof) for the survival of the efficient nitrogen-fixing rhizobium, Sinorhizobium (Ensifer) meliloti L3Si strain. The most suitable liquid inoculant for survival of L3Si during a storage time of 150 days was the medium formulation containing glycerol in combination with agar or sodium-alginate. Alfalfa seeds were pre-inoculated with four formulations (yeast mannitol broth (YMB), YMB with agar (1 g L-1), YMB with 1 or 5 g L-1 sodium-alginate) for up to three months. Seeds pre-inoculated and stored for one month produced successful alfalfa plants. The nitrogen content in alfalfa obtained from pre-inoculated seeds one month before sowing was adequate and ranged from 3.72-4.19%. Using S. meliloti-based liquid inoculants for alfalfa and application of the pre-inoculation technique can increase the quality of alfalfa crops and reduce cultivation cost.

Received: October 8, 2018; Revised: December 3, 2018; Accepted: December 20, 2018; Published online: December 27, 2018

How to cite this article: Buntić AV, Stajković-Srbinović OS, Knežević MM, Kuzmanović ĐŽ, Rasulić NV, Delić DI. Development of liquid rhizobial inoculants and pre-inoculation of alfalfa seeds. Arch Biol Sci. 2019;71(2):379-87.


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Author Biography

Olivera S. Stajković-Srbinović, Institute of Soil Science, Department of Microbiology, University of Belgrade, Teodora Drajzera 7, 11000, Belgrade


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How to Cite

Buntić AV, Stajković-Srbinović OS, Knežević MM, Kuzmanović Đorđe Ž., Rasulić NV, Delić DI. Development of liquid rhizobial inoculants and pre-inoculation of alfalfa seeds. Arch Biol Sci [Internet]. 2019Jun.4 [cited 2022Jan.21];71(2):379-87. Available from: