The heterologous expression of Arabidopsis PAP2 induces anthocyanin accumulation and inhibits plant growth in tomato

Anthocyanins are naturally occurring secondary metabolites, responsible for the color of many plants. The Arabidopsis thaliana MYB90/PAP2 (production of anthocyanin pigment 2) was introduced into tomato to study its effect on anthocyanin accumulation. The transgenic tomato displayed much greater anthocyanin accumulation than wild type in all plant organs, but the organs were not fully purple in color except for the stamen. The expression of anthocyanin biosynthetic genes and an anthocyanin-related basic helix-loop-helix (bHLH) gene SlAN1 was significantly increased in the transgenic line, suggesting that ectopic expression of AtPAP2 increases the expression of anthocyanin-related structural and regulatory genes to enhance anthocyanin content. Yeast two-hybrid assays revealed that the endogenous MYB protein SlAN2 interacted with two putative bHLH partners, SlAN1 and SlJAF13, while AtPAP2 only interacted with SlJAF13, which may be why AtPAP2 transgenic plants showed limited anthocyanin accumulation in fruits. In addition to anthocyanin accumulation, the transgenic tomato plants were significantly smaller in size, and the length of primary roots and number of lateral roots were obviously decreased. The expression of lignin biosynthetic genes was downregulated in transgenic tomato plants, which may be the reason for the inhibited growth. The lateral organ boundaries-domain (LBD) genes, which regulate lateral root organogenesis in the auxin signaling pathway, were downregulated in transgenic tomato roots, which may partly account for the disturbed lateral root formation in the transformants. Taken together, the results demonstrate that heterologous expression of transcription factor AtPAP2 not only resulted in anthocyanin accumulation but also inhibited plant growth in tomato.