The peu-miR160a-PeARF17.1/PeARF17.2 module participates in the adventitious root development of poplar

Deep roots give rise to flourishing leaves, and the two complement each other. However, the genetic mechanisms underlying adventitious rooting for forest trees have remained elusive. In this study, we verified that peu-miR160a targets six poplar genes AUXIN RESPONSE FACTORS (ARFs), PeARF10.1, PeARF16.1, PeARF16.2, PeARF16.3, PeARF17.1 and PeARF17.2, using 5'RLM-RACE. Interaction experiments with peu-miR160a and PeARFs in poplar protoplasts further confirmed that peu-miR160a targets and negatively regulates the six PeARFs. Peu-miR160a and its target genes exhibited obvious temporal expression in different stages of adventitious root development, and they could also be induced by IAA and abscisic acid. Peu-miR160a-overexpressing lines exhibited a significant shortening of adventitious root length, an increase in the number of lateral roots, severe dwarfing and shortened internodes. In addition, the overexpression of PeARF17.1 or mPeARF17.2 (peu-miR160a-resistant version of PeARF17.2) significantly increased the number of adventitious roots. Furthermore, PeARF17.1-overexpressing lines had multiple branches with no visible trunk, although the adventitious root length of the PeARF17.1-overexpressing lines was significantly increased. Our findings reveal that the peu-miR160a - PeARF17.1/PeARF17.2 module is an important regulator involved in the development of the adventitious roots of poplar.