Drought stress severely compromises the physiological integrity and secondary metabolism of medicinal plants. This study integrated physiological, biochemical, and ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) analyses to investigate the effects of exogenous methyl jasmonate (MeJA) on drought-stressed Ilex rotunda seedlings. Drought reduced relative water content by 29% and chlorophyll by >50%, while elevating H₂O₂ (76%) and malondialdehyde (120%). MeJA application mitigated these impairments, reducing oxidative markers by 25% and enhancing non-enzymatic antioxidant capacity, as shown by a 74% increase in DPPH radical scavenging activity and a 141% rise in total phenolic content. Hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA), and pathway mapping confirmed a significant reprogramming of the phenolic metabolome, particularly within phenylpropanoid and benzoate biosynthesis pathways. Drought + MeJA-treated plants exhibited a distinct and enriched profile compared to both well-watered control and drought-stressed groups. This reprogramming specifically elevated key hydroxycinnamates, including verbascoside and neochlorogenic acid (increased by 50% and 52%, respectively), while suppressing alternative phenolic branches. These findings demonstrate that MeJA orchestrates a shift from enzymatic scavenging to a potent metabolite-based antioxidant system, positioning it as an effective elicitor for enhancing drought resilience and enriching the high-value phytochemicals in I. rotunda.

Drought stress imposes oxidative damage on plants, yet the tissue-specific roles of salicylic acid (SA) in modulating phenolic metabolism remain poorly understood in woody species. Using Ilex rotunda seedlings, we investigated whether exogenous SA (100 µM) mitigates drought-induced oxidative damage and reshapes phenolic profiles in different tissues. Drought alone increased leaf total phenolics by 32% but depleted root phenolics by 29%, whereas combined drought + SA (DSA) treatment partially restored root phenolic levels, coinciding with elevated malondialdehyde (MDA) (2.2-fold in leaves, 2.6-fold in roots) and H₂O₂. Leaf antioxidant capacity increased under drought (DPPH by 73%, •OH by 33%), whereas root DPPH declined by 27% despite a 26% rise in •OH scavenging. SA alone induced mild oxidative responses and selectively upregulated caffeoylquinic and galloyl derivatives, notably 1-Caffeoylquinic acid (log₂FC = 6.38) in leaves. DSA treatment mitigated oxidative damage—reducing leaf MDA by 44% and root H₂O₂ by 38%. Metabolomics revealed tissue-specific reprogramming leaves accumulated dicaffeoylshikimic acid (log₂FC = 10.66) and trilobatin D (log₂FC = 11.18) under DSA, whereas roots showed contrasting patterns with up-accumulation of vanillate (log₂FC = 5.77) and suppression of 3,5-dicaffeoylquinic acid (log₂FC = −7.21) under drought, with stronger metabolic reprogramming in leaves than roots. Our findings indicate that SA-mediated drought tolerance is associated with tissue-specific phenolic reprogramming, identifying candidate indicators that advance the mechanistic understanding of woody plant resilience to drought. These results provide a framework for translating metabolomic signatures into practical strategies for stress mitigation in medicinal perennials facing climate change.

Ilex verticillata (winterberry) is a valuable ornamental shrub increasingly threatened by leaf blight, a disease that compromises its aesthetic and economic value. While fungal pathogens like Alternaria alternata are known to cause leaf blight in horticultural crops, their role in I. verticillata and the host’s defense mechanisms have not been fully characterized. Our study investigated the pathogen-host interaction by identifying the causal agent and examining the physiological and molecular defense mechanisms of I. verticillata. Through morphological and multi-locus molecular analyses (ITS, TEF1-α, G3PDH, RPB2), A. alternata was confirmed as the primary pathogen, fulfilling Koch’s postulates. Pathogenicity assays revealed distinct disease progression stages, from necrotic lesions to tissue degradation. Transcriptomic profiling uncovered dynamic host responses, with early upregulation of pattern recognition receptors (PRRs) and transcripts encoding antioxidant enzymes (SOD, CAT), followed by downregulation of metabolic pathway genes. Phytohormone analysis highlighted intricate crosstalk, with salicylic acid (SA) peaking during mid-infection and jasmonic acid (JA) rebounding later, reflecting a coordinated defense strategy. Additionally, the oxidative stress marker malondialdehyde (MDA), an indicator of membrane lipid peroxidation, surged early, indicating membrane damage, while sustained induction of antioxidant enzymes suggested adaptive responses. The key finding was distinct phytohormone crosstalk, characterized by a mid-infection SA peak followed by a late JA rebound, alongside an early oxidative burst marked by MDA accumulation and sustained antioxidant enzyme activity. These findings provide a framework for understanding I. verticillata’s defense mechanisms and offer insights for developing targeted disease management strategies, such as resistant cultivar breeding or hormone-mediated interventions.