Transcriptome Profiling and Weighted Gene Correlation Network Analysis Reveal Hub Genes and Pathways Involved in the Response to Polyethylene-Glycol-Induced Drought Stress of Two Citrus Rootstocks

Agriculture faces the dual challenge of increasing food production and safeguarding the environment. Climate change exacerbates this challenge, reducing crop yield and biomass due to drought stress, especially in semi-arid regions where Citrus plants are cultivated. Understanding the molecular mechanisms underlying drought tolerance in Citrus is crucial for developing adaptive strategies. Plants of two citrus rootstocks, Carrizo Citrange and Bitters (C22), were grown in aerated half-strength Hoagland's nutrient solution. Post-acclimation, the plants were exposed to a solution containing 0% (control) or 15% PEG-8000 for 10 days. Leaf malonyl dialdehyde (MDA) and hydrogen peroxide (H2O2) content were measured to assess the reached oxidative stress level. Total RNA was extracted, sequenced, and de novo-assembled. Weighted Gene Correlation Network Analysis (WGCNA) was conducted to examine the relationship between gene expression patterns and the levels of MDA and H2O2 used as oxidative stress indicators. Plant visual inspection and MDA and H2O2 contents clearly indicate that Bitters is more tolerant than Carrizo towards PEG-induced drought stress. RNA-Seq analysis revealed a significantly higher number of differentially expressed genes (DEGs) in Carrizo (6092) than in Bitters (320), with most being associated with drought sensing, ROS scavenging, osmolyte biosynthesis, and cell wall metabolism. Moreover, the WGCNA identified transcription factors significantly correlated with MDA and H2O2 levels, thus providing insights into drought-coping strategies and offering candidate genes for enhancing citrus drought tolerance.