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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.

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SUMMARYThe aim of this research is to isolate and identify fungi with high lignin-degrading abilities that are autochthonous to southern Serbian region. Two novel fungal isolates identified as Trametes hirsuta F13 and Stereum gausapatum F28 were selected to assess their ligninolytic enzyme activities and the efficiency of lignin removal from beech wood sawdust. Obtained results show that both isolates are good sources of industrially valuable enzymes with a potential for application in various biotechnological and industrial processes. Both isolates showed laccase, manganese-dependent peroxidase, and versatile peroxidase activities, while only S. gausapatum F28 had lignin peroxidase activity. This is the first record of the ability of S. gausapatum species to produce lignin peroxidase. T. hirsuta F13 showed higher laccase activity than S. gausapatum F28, while S. gausapatum F28 had higher manganese peroxidase activity. Also, T. hirsuta F13 exhibited much higher laccase activity under submerged cultivation conditions than solid-state cultivation conditions, which is rare for fungi. This is important for industrial processes since the submerged fermentation is a dominant technique in industry. The test of the efficiency of lignin removal showed that both isolates are efficient lignin decomposers. After five weeks of incubation on beech wood sawdust, the total lignin losses were 33.84% with T. hirsuta F13 and 28.8% with S. gausapatum F28.

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