Screening and Identification of Cellulose Degrading Bacteria and Determination of Cellulase Activity in Low-temperature Areas
[Objective]The cellulose degrading bacteria in low-temperature areas were screened to provide the theoretical basis for the harmless treatment of livestock manure.[Method]The low-temperature and cellulose degrading bacteria were preliminary isolated and screened from the frozen soil and cow dung in Jilin low-temperature areas by temperature suitability test and Congo red staining method;The molecular biological identification of the strain was carried out through 16S rDNA sequencing;The cellulase activity was determined by plotting the glucose standard curve.[Result]Three low-temperature strains(D-3,D-7 and D-16)were screened from cow dung and frozen soil by temperature suitability test,and five strains with high cellulase activity(X-1,X-5,X-9,X-11 and X-18)were screened by Congo red staining and enzyme activity test,which all belonged to bacteria.After 16S rDNA sequencing and identification,the homology alignment of sequencing results was carried out with BLAST software,then based on the established gene phylogenetic tree,the three low-temperature bacteria were identified as Pseudomonas frederiksbergensis,Acinetobacter calcoaceticus and Pseudomonas synxantha,and five cellulose degrading strains were identified as Bacillus cereus,Bacillus subtilis,Bacillus amyloliquefaciens,Bacillus pumilus and Bacillus safensis.[Conclusion]The group composition and cellulose degradation ability of cellulose degrading bacteria can be cultivated in Jilin low-temperature areas.The obtained five cellulose degrading strains with high activity(X-1,X-5,X-9,X-11 and X-18)can be used as potential strains for effectively degrading cellulose,which applied in the development of manure compost fermentation agent.
cellulose degrading bacterialow temperatureisolation and screeningoptimizationcellulase
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