MicroRNA166-REVOLUTA-auxin module affects tuber shape, color and productivity in potato

Tuber shape and size are important traits in potato. Although several factors governing tuberization have been well studied, the molecular mechanisms regulating the tuber shape and size are still elusive. Here, we demonstrate that suppression of miR166, which targets class III HD-ZIP transcription factors, alters tuber morphology and productivity in potato. Target mimicry of miR166 (MIM166) in Solanum tuberosum ssp. Andigena resulted in elongated, pigmented tubers with reduced yield under short-day conditions. Transcriptome profiling of the tuberizing stolons - swollen head vs. stalk revealed differential expression of auxin-, cytokinin-, and gibberellin-associated genes, consistent with altered hormone levels. The coloured tubers from MIM166 line exhibited differential accumulation of cyanidin 3-glucoside, pelargonidin 3-glucoside, and delphinidin 3-glucoside. Tuber productivity in these lines was reduced possibly due to the decreased expression of SWEET11b and distorted vascular structures. Further, the miR166 target-StREVOLUTA exhibited dynamic expression during stolon-to-tuber transition, and could modulate StYUCCA7 expression, an auxin biosynthesis gene. Increased pigmentation and auxin accumulation in MIM166, reduced expression of auxin biosynthesis genes in REVOLUTA antisense, and low tuber yield collectively suggest miR166-REV as a regulatory module of auxin homeostasis in differentiating stolons that influences tuber morphology. These results reveal a previously unrecognized miRNA-mediated pathway governing storage organ shape, and extends the functional scope of the miR166-HD-ZIP III module beyond organ polarity. Significance statementLimited studies describe the molecular mechanisms regulating tuber shape in storage organ development. Here, we show that the miR166-REVOLUTA-auxin network has significant regulatory roles in potato development, influencing tuber shape, pigmentation, and productivity. These findings provide insights into molecular control of tuber morphology, uncover hitherto unknown functions of the miR166-REVOLUTA module in tuber crops, and broadens our understanding of miR166-mediated developmental regulation in plants.