Abstract
This study examines the relationship between needle age and rust resistance in Picea crassifolia, focusing on the needle morphology, including size, shape, and physiological traits. One-year-old spruce needles are more susceptible to rust, while two-year-old needles show effective resistance. Using RNA-seq on the Illumina HiSeq500 platform, we analyzed both healthy and diseased one-year-old needles (N and B), as well as healthy one-year-old and two-year-old needles (N and L). We applied a fold change (FC) threshold of ≥2 and a false discovery rate (FDR) of <0.01, alongside GO annotation and KEGG pathway enrichment, to identify differentially expressed genes (DEGs). In N vs. B, DEGs were significantly enriched in processes such as metabolism, cellular function, catalysis, binding, ribosomal function, plant-pathogen interactions, endoplasmic reticulum protein processing, and signal transduction, revealing a polygenic network regulating the rust response. Similarly, in N vs. L, electron microscopy highlighted morphological differences in the wax layers of needles, with subsequent transcriptome sequencing uncovering genes involved in the development of one-year-old and two-year-old needles. DEGs were primarily found in pathways related to cutin, suberin, wax biosynthesis, fatty acid metabolism, photosynthesis, and phenylalanine synthesis. Two-year-old needles displayed reduced stomatal density, higher lignin content, and a thicker wax layer compared to one-year-old needles. Validation of the RNA-seq data through RT-qPCR on 10 DEGs confirmed the consistency of gene expression trends, enhancing our understanding of Picea crassifolia's genetic response to rust and supporting future research into its disease resistance.