Plant trichomes are a specialized cellular tissue that functions in resistance to biotic and abiotic stresses. In rice, three transcription-factor genes: OsWOX3B, HL6, and OsSPL10, have been found to control trichome development. Although studies have shown interactions between the three genes, their full relationship in trichome development is unclear. We found that the expression levels of OsWOX3B and HL6 were both reduced in OsSPL10-knockout plants but increased in OsSPL10-overexpression plants, suggesting that OsSPL10 positively regulates their expression. Physical interaction between OsSPL10 and OsWOX3B was found both in vivo and in vitro and attenuated their abilities to bind to the promoter of HL6 to activate its transcription. This mechanism may regulate trichome length by adjusting the expression of HL6. A rice gene network regulating trichome development is proposed.