Shading increases the susceptibility of alfalfa (Medicago sativa) to Pst. DC3000 by inhibiting the expression of MsIFS1.

Shade is a stressful factor for most plants, leading to both morphological and physiological changes, and often resulting in increased susceptibility to diseases and pathogen attacks. Our study revealed that the isoflavonoid synthesis pathway was inhibited in alfalfa under shade, resulting in a significant reduction in disease resistance. Overexpression of MsIFS1 , a switch regulator in isoflavonoid synthesis, led to a notable increase in endogenous isoflavonoids and enhanced resistance to Pseudomonas syringae pv. tomato DC3000 (Pst. DC3000). Conversely, MsIFS1 -RNAi had the opposite effect. Yeast one-hybrid (Y1H) assays revealed that the shade-responsive transcription factor MsWRKY41 could directly bind to the MsIFS1 promoter. This interaction was confirmed through Dual-Luciferase Reporter (Dual-LUC) and Chromatin Immunoprecipitation coupled with quantitative PCR (ChIP-qPCR) assays, both in vitro and in vivo. Overexpression of MsWRKY41 not only enhanced alfalfa's resistance to Pst. DC3000 but also promoted the accumulation of isoflavonoids. Additionally, yeast two-hybrid (Y2H) assays showed that neither MsWRKY41 nor MsIFS1 physically interacted with the Type III effector (T3SE) HopZ1 secreted by Pst. DC3000, suggesting that the MsWRKY41-MsIFS1 module is not a direct target of HopZ1. These findings provide valuable theoretical insights and genetic resources for the development of shade-tolerant alfalfa with enhanced disease resistance. • Shade leads to reduced disease resistance in alfalfa. • Isoflavonoid synthesis is regulated by light. • MsIFS1 affects disease resistance by influencing Isoflavonoid metabolism in alfalfa. [ABSTRACT FROM AUTHOR]