Biochar-activated periodate (PI) is a promising technology toward antibiotic wastewater purification. However, the mechanism of pyrolysis temperature on PI activation efficiency by biochar has not yet been revealed. Herein, this work selected water hyacinth stems as raw materials to prepare biochar with different pyrolysis temperatures (400, 500, 600, and 700 degrees C), and applied it to degrade tetracycline (TC) wastewater through PI activation. The results show that biochar with a pyrolysis temperature of 700 degrees C (BC-700) possesses the best TC degradation performance (similar to 100% within 30 min). Besides, the degradation of TC by BC-700 is less interfered by coexisting anions and water matrix, and exhibits good reusability. Quenching experiments and open circuit voltage tests verified that IO3 center dot, 1O(2), and reactive complex BC-PI* are active species involved in TC degradation. In addition, by constructing the relationship between biochar surface properties and degradation rate k(obs), it was revealed that the dominant role of pyridinic N in PI adsorption and formation of reactive complexes as well as the promotion of sp(2)-hybridized carbon in the electron transfer process. This work provides novel insights into the application of biochar in antibiotic wastewater treatment via PI activation.