Environment factors, especially temperature and moisture, have significant effects on microbial community associated with straw decomposition. This study investigated the differences in metabolic diversity of microbes during decomposition of straws under different temperatures (15℃, 25℃, 35℃) and moistures (40%, 70%, 90% water holding content) by BIOLOG analysis. Litterbag method was used to distinguish the straw-decomposing microbial community and soil microbial community. The results showed that the average well color development (AWCD) of microbial community responsible for straw decomposition declined with increasing incubation temperature, the decrease of AWCD became more significant with increasing incubation time. Besides, the richness index and dominance index decreased with the increasing temperature. The sugar and lipid were the most important sources used by microbes during the incubation. Principal component analysis showed that temperature rather than moisture significantly affected the metabolic diversity of microbes during the decomposition period.