The straw lignocellulose-degrading bacteria were isolated from soil samples collected at various sampling dates using the serial dilution plate method [25 (link)]. To initiate the isolation process, 1 g of soil obtained from the straw surface was thoroughly mixed and added to 9 mL of sterile water in a shaker set at 30 °C and 120 rpm for 20 min. Subsequently, the soil suspension was serially diluted and inoculated onto a straw agar medium consisting of 20 g/L of commercial straw powder (<1 mm) and 1 g/L of urea and adjusted to a pH of 7.0. Following a 7-day incubation period at 30 °C, single colonies were selected and cultured on the same medium to achieve purification. Genomic DNA extraction was performed employing a bacterial genomic DNA extraction kit (Tiangen Biochemical Technology Co. Ltd., Beijing, China). Bacteria and fungi were identified using the primers 27F/1492R and ITS1/ITS4, respectively. The obtained 16S rRNA and ITS gene sequences were compared using the Ezbiocloud and NCBI databases. After the removal of potential clonal duplicates, a total of 59 bacterial and 14 fungal strains were successfully isolated, and their glycerol stocks were prepared and stored at −80 °C.
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