Transcription factor RhRAP2.4L orchestrates cell proliferation and expansion to control petal size in rose

Abstract Maintaining proper flower size is vital for plant reproduction and adaption to the environment. Petal size is determined by spatiotemporally regulated cell proliferation and expansion. However, the mechanisms underlying the orchestration of cell proliferation and expansion during petal growth remains elusive. Here, we determined that the transition from cell proliferation to expansion involves a series of distinct and overlapping processes during rose (Rosa hybrida) petal growth. Changes in cytokinin content were associated with the transition from cell proliferation to expansion during petal growth. RNA sequencing identified the AP2/ERF transcription factor gene RELATED TO AP2 4-LIKE (RhRAP2.4L), whose expression pattern positively associated with cytokinin levels during rose petal development. Silencing RhRAP2.4L promoted the transition from cell proliferation to expansion and decreased petal size. RhRAP2.4L regulates cell proliferation by directly repressing the expression of KIP RELATED PROTEIN 2 (RhKRP2), encoding a cell cycle inhibitor. In addition, we also identified BIG PETALub (RhBPEub) as another direct target gene of RhRAP2.4L. Silencing RhBPEub decreased cell size, leading to reduced petal size. Furthermore, the cytokinin signaling protein ARABIDOPSIS RESPONSE REGULATOR 14 (RhARR14) activated RhRAP2.4L expression to inhibit the transition from cell proliferation to expansion, thereby regulating petal size. Our results demonstrate that RhRAP2.4L performs dual functions in orchestrating cell proliferation and expansion during petal growth.