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‘Korla’ fragrant pear (Pyrus sinkiangensis T.T. Yu) variety has shown severe coarse skin in recent years. The intrinsic quality of its coarse fruit shows an increase in the number of stone cells and poor taste. In this study, stone cells and the cell wall of coarse pear (CP) and normal pear (NP) during various development stages were compared using paraffin-sectioning and transmission electron microscopy (TEM), and the relationships between lignin-related genes and stone cell formation and cell wall thickening were also analyzed. Our results show that giant stone cells are formed and distributed in the core of pear, whereas many of these crack 60 days after flowering (DAF). The period of stone cell fragmentation occurs later in CP fruits than in NP fruits. Parenchyma cell wall development in CP and NP fruits varies from 120 DAF to maturity. The parenchyma cell wall of CP fruits thickens, whereas that of NP fruits is thinner during the same period. The expression pattern of five genes (Pp4CL1-l, PpHCT-l, Pp4CL2-l, PpPOD4, and PpPOD25) coincides with changes in stone cell content in the pulp. Correlation analysis demonstrates a significant correlation between stone cell content and the expression level of the five genes (ρ < 0.05). In addition, the expression of those five genes and PpCCR1 genes in CP fruits significantly increases during maturation and is highly correlated with the thickness of the parenchyma cell wall. The aim of this work is to provide insights into the mechanism of stone cell and parenchyma cell wall development in pear fruits and identify important candidate genes to regulate the quality of fruit texture using bioengineering methods.

Quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) is a sensitive and widely used technique for gene expression analysis that depends on stability of the reference genes used for data normalization. Tree peony (Paeonia suffruticosa), known as one of the most famous traditional ornamental plants in China, is very popular in both domestic and international markets for its showy and colorful flowers. To date, no systematic studies on reference genes have been performed in tree peony with different flower colors. In this study, we evaluated the expression stability of 12 candidate reference genes in different tissues and five flower developmental stages of tree peony with six different colors by three algorithms: geNorm, NormFinder, and BestKeeper. The results showed that protein phosphatase 2A (PP2A), ubiquitin protein ligase (UPL), and ubiquitin (UBQ) were the most stable genes across all samples. Helicase, alpha-tubulin (TUA), and eukaryotic translation initiation factor 5A (EIF5A) also exhibited high expression stability in different tissues, in samples with different colors, and at different flower developmental stages. According to the geNorm analysis, the combination of two most stable reference genes was optimal for normalization in all tested sample sets in this study. To further validate the suitability of the reference genes identified in this study, the expression patterns of two putative homologs of chalcone synthase gene (PsCHS1) and chalcone isomerase gene (PsCHI1) were studied at different developmental stages of white flowers. The results provide information for transcriptional analyses in future studies of gene expression on tree peony flower development and pigmentation.