A series of field experiments were carried out to study the effects of different applying modes of conditioners on selenium (Se) availability of highly acidic Se-rich soil in tea gardens. And for reaching this goal, 3 alkaline conditioners (straw charcoal, lime and calcium-magnesium phosphate) and “BIO” bio-organic fertilizer were used. The results showed that the endogenous regulations to tea garden soil were able to improve Se bioavailability on the premise of not adding exogenous Se. 3 treatments that combined straw charcoal, lime and calcium-magnesium phosphate with bio-organic fertilizer respectively at intervals worked most effectively, which increased the content of soil available Se by 105.98, 114.09 and 146.71 μg/kg respectively, increased the content of Se in tea by 0.96, 1.14 and 1.24 μg/g correspondingly, and these enhancements all reached the extremely significant level (P <0.01).