Abstract
The codling moth (Cydia pomonella) has a major effect on the quality and yield of walnut fruit. Plant defences respond to insect infestation by activating hormonal signalling and the flavonoid biosynthetic pathway. However, little is known about the role of walnut husk hormones and flavonoid biosynthesis in response to C. pomonella infestation. The phytohormone content assay revealed that the contents of salicylic acid (SA), abscisic acid (ABA), jasmonic acid (JA), jasmonic acid-isoleucine conjugate (JA-ILE), jasmonic acid-valine (JA-Val) and methyl jasmonate (MeJA) increased after feeding at different time points (0, 12, 24, 36, 48, and 72 h) of walnut husk. RNA-seq analysis of walnut husks following C. pomonella feeding revealed a temporal pattern in differentially expressed genes (DEGs), with the number increasing from 3,988 at 12 h to 5,929 at 72 h postfeeding compared with the control at 0 h postfeeding. Walnut husks exhibited significant upregulation of genes involved in various defence pathways, including flavonoid biosynthesis (PAL, CYP73A, 4CL, CHS, CHI, F3H, ANS, and LAR), SA (PAL), ABA (ZEP and ABA2), and JA (AOS, AOC, OPR, JAZ, and MYC2) pathways. Three gene coexpression networks that had a significant positive association with these hormonal changes were constructed based on the basis of weighted gene coexpression network analysis (WGCNA). We identified several hub transcription factors, including the turquoise module (AIL6, MYB4, PRE6, WRKY71, WRKY31, ERF003, and WRKY75), the green module (bHLH79, PCL1, APRR5, ABI5, and ILR3), and the magenta module (ERF27, bHLH35, bHLH18, TIFY5A, WRKY31, and MYB44). Taken together, these findings provide useful genetic resources for exploring the defence response mediated by phytohormones in walnut husks.