Reactive oxygen species (ROS) and salicylic acid (SA) are essential signaling molecules in plant cells that participate in responses to biotic and abiotic stresses. Changes in ROS and SA signals during interactions between pear and the pear scab pathogen Venturia nashicola remain unclear. Herein, we analyzed the roles of ROS in the signal transduction pathway of pear scab resistance using the highly resistant Huangguan and susceptible Xuehua cultivars of pear (Pyrus bretschneideri Rehd). Protoplasts, calluses, and leaves were obtained from 14-year-old pear trees and treated with V. nashicola for different periods. The results showed that ROS rapidly accumulated in protoplasts of both cultivars within a 120-min treatment period, but the fluorescence intensity of ROS differed between cultivars. The H₂O₂ content in fruit-derived calluses of Huangguan peaked at 48 h post-infection at levels 1.85 times higher than those in Xuehua. Induction of H₂O₂ by V. nashicola in Huangguan was more intense than in Xuehua over a 96-h treatment period. At 96 h post-infection, the malondialdehyde content in leaves of Huangguan was significantly lower than in Xuehua, while the activities of superoxide dismutase, peroxidase, and catalase, and the relative expression levels of PbMnSOD, PbPOD, and PbCAT genes were higher in Huangguan than Xuehua. V. nashicola infection also caused a continuous increase in the leaf SA content of Huangguan, which was 6.76 times higher than in Xuehua at 96 h post-infection, and V. nashicola exposure upregulated the expression of PbPAL, PbICS, PbPR1, and PbPR5. In summary, both ROS and SA participated in the responses of pear trees to V. nashicola infection and played vital roles in the signal transduction pathway of pear scab resistance.

Real-time quantitative PCR (RT-qPCR) has become a widely used method for exploring plant gene expression level. The method requires using some stably expressed genes as a reference to accurately normalize the RT-qPCR data. However, under various stresses and hormone treatments, the levels of most reference genes vary. Environmental variations also influence their expression levels. The lack of validated, stably expressed reference genes can mislead the study of gene function in pears. “Huangguan” pears have recently become the focus of research on stress resistance mechanisms, such as high resistance. Therefore, the aim of this study was to select the optimal reference genes in Huangguan pears, and we analyzed the expression of the genes EF1α, ACT, SKD1, YLS8, UBQ, GAPDH, TUB, and WDP in a series of pear leaf sets under various stresses and hormone treatments. Using different statistical algorithms, we found that under various treatments, the WDP gene had more stable expression, ACT was the most stable under MeJA treatment, YLS8 was the most valuable reference gene under ABA hormone and heat stress conditions, and GAPDH showed worst results compared to other housekeeping genes, except under heat stress. These results will supply valuable and updated information for the selection of housekeeping genes in pears under biotic and abiotic stresses in the future.

Lilium holds significant horticultural and ecological importance. Understanding the morpho-anatomical diversity of the stems can provide insights into taxonomy and breeding strategies. This study comprehensively examined the stem morpho-anatomy of 71 Lilium taxa to elucidate taxonomic and structural differences. For the first time, four distinct jigsaw-puzzle-shaped shapes of epidermal cells (Ep) in monocot stems, novel I-shaped and Co-xylem (O-, X-, W-, Q-shaped) vascular bundles (Vb) in Lilium stems, and quantitative characteristics (Vb density, xylem/phloem area ratio, etc.) were systematically discovered and analyzed. Asiatic (A) and Longiflorum × A (LA) hybrids displayed epidermal appendages, while Oritenal × Trumpet (OT) hybrids featured thicker sclerenchymatous rings (Sr). Collateral Vb in hybrids visually displayed bicollateral with degraded bundle sheaths (Bs), contrasting with intact circular Bs in wild species. Ward.D clustering categorized Lilium taxa into group A (Oritenal and OT hybrids) and B (A, LA, Trumpet, Longiflorum × Oriental hybrids and wild species), with Mantel’s test identified height, Ep shape, Ep length/width ratio, cortex/Sr thickness ratio and Bs integrity as key discriminators. Bending stems exhibited a higher Vb area. These findings establish a comprehensive pheno-anatomical framework for Lilium, which can guide future breeding programs and ecological studies.