Carotenoids not only play important roles in light harvesting and photoprotection against excess light, but also serve as precursors for apocaroteniod hormones such as abscisic acid (ABA) and strigolactones (SLs). Although light- and ABA-associated phenotypes of the carotenoid biosynthesis mutants such as albino, leaf variegation and preharvest sprouting have been studied extensively, the SLs-related branching phenotype is rarely explored. Here we characterized four allelic rice mutants named mit3, which exhibited moderately increased tiller number, semi-dwarfism and leaf variegation. Map-based cloning revealed that MIT3 encodes a carotenoid isomerase (CRTISO), the key enzyme catalyzing the conversion from prolycopene to all-trans-lycopene in carotenoid biosynthesis. Prolycopene was accumulated while all-trans-lycopene was barely detectable in the dark-grown mit3 seedlings. Accordingly, content of lutein and β-carotene, the two most abundant carotenoids, was significantly reduced. Furthermore, content of epi-5DS, a native SL, was significantly reduced in mit3. Exogenously applied GR24, a synthetic SL, could rescue the tillering phenotype of mit3. Double mutant analysis of mit3 with the SLs biosynthesis mutant d17 revealed that MIT3 controls tiller development upstream of the SLs biosynthesis pathway. Our results reveal that the tillering phenotype of mit3 is due to SL deficiency and directly link carotenoid deficiency with SL-regulated rice tillering.