Parasitism with Striga poses a major threat to global food production. Striga germination and growth rely on strigolactones (SLs) exuded by crop roots under phosphate (Pi)-deficient conditions, although the mechanism of this host-parasite interaction remains elusive. In this study, transcriptomic and functional analyses of sorghum treated with Pi deficiency or the SL GR245DS identify two ABC transporter G (ABCG) transporters of SL, Sorghum biocolor strigolactones transporter 1 (SbSLT1) and SbSLT2. Using AlphaFold2 and amino acid conversion mutants, we identify highly conserved amino acids in SL transport channels essential for transport function. Sorghum lines with single or double knockouts of these transporters exhibit significantly reduced SL secretion from roots, leading to decreased Striga germination and parasitism in field experiments and consequently reducing the grain loss under Striga infestation. This study thus describes the mechanism of SL exudation in monocots and defines conserved residues essential for SL transporter function, offering a potential strategy for enhancing crop resistance to Striga parasitism.