MeJA-responsive MYB41-MYC2 cascade integrates glucomannan biosynthesis and DoTIP1–1 activation during salinity tolerance in Dendrobium officinale

Abstract

Soil salinization limits the yield and quality of Dendrobium officinale. The molecular mechanisms linking methyl jasmonate (MeJA) signaling to the biosynthesis of glucomannans (GMs) under salinity remain unclear. In the WGCNA of MeJA- and salinity-treated D. officinale transcriptome, an R2R3-MYB transcription factor was identified, DoMYB41, which was highly expressed in stems and induced by both treatments. Overexpression of DoMYB41 in PLBs increased GMs and anthocyanin, enhanced the activities of antioxidant enzymes, reduced oxidative damage, and elevated relative water content, whereas CRISPR/Cas9 knockout lines showed opposite phenotypes. Transcriptomic analysis identified DoTIP1–1 as a key downstream target, and biochemical assays (dual-LUC, Y1H, EMSA) confirmed that DoMYB41 activates DoTIP1–1 transcription by directly targeting the MBS motif present in its promoter. Furthermore, DoMYB41 physically interacted with the bHLH transcription factor DoMYC2, which alone bound to and activated the promoters of both DoTIP1–1 and DoMYB41, forming a positive feedback loop. Intriguingly, DoMYB41 and DoMYC2 synergistically activated DoTIP1–1. These findings uncovered a MeJA-responsive DoMYB41-DoMYC2 cascade that iteratively activates DoTIP1–1 expression and promotes the biosynthesis of GMs, thereby enhancing salinity tolerance in D. officinale. This study also provides genetic targets for salinity tolerance in breeding programs for this horticultural orchid.