Biochar (BC) amendment has been proposed as a promising strategy for mitigating greenhouse gas (GHG) emissions, specifically carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O). Conducting a meta-analysis to evaluate the impact of biochar on microbial genetic profile, community structure, and phospholipid fatty acid (PLFA) contents can aid in identifying key microbial groups involved in GHG production and consumption, and assessing the overall effectiveness of biochar in reducing GHG emissions. The present meta-analysis revealed that the addition of biochar resulted in a 22 % and 41 % reduction in pmoA and mcrA genes of methanogenic microorganisms, respectively. The mcrA/pmoA ratio significantly increased by 81 %. Gene abundances exhibited a positive response to biochar amendment, with increases observed in nifH, nirK, nirS, nosZ, and nosZ (nirS + nirK) genes by 13 %, 32 %, 37 %, 42 %, and 79 %, respectively. Moreover, biochar amendment influenced the microbial community structure accordingly. The concentration of PLFAs increased in response to BC treatment in the following order: A-bacteria (+49 %) < Fungi (+30 %) < Gram-pb (+21 %) < G-bacteria (+17 %) < Gram-nb (+11 %). These findings indicate that biochar amendment shapes the microbial community structure, further emphasizing its significance in enhancing soil fertility.