Differential expression analysis of sugar accumulation-related genes during chestnut nut development

The chestnuts of Castanea mollissima Bl. are an important food crop in China, and have high nutritional content. To understand the pattern of sugar accumulation during chestnut nut development, the related enzyme gene regulatory pathways, and the molecular regulatory mechanisms of chestnut sugar biosynthesis metabolism, two chestnut varieties with different sugar content, namely Chengbu Youzhu (hereinafter referred to as CBYZ) and AnYou No. 1 (hereinafter referred to as AY01), were selected for investigation. Total sugar and starch content, and the activity of enzymes related to sugar accumulation, were measured in the nuts of the two chestnut varieties 10 days after flowering (DAF), 20 DAF, 30 DAF, 40 DAF, 50 DAF, 60 DAF, 70 DAF, 80 DAF, and 90 DAF. Changes in starch, straight-chain starch, and branched-chain starch content, and sucrose-phosphate synthase, soluble starch synthase, and granule-bound starch synthase enzyme activities were consistent with one-another. A total of 24 differentially expressed genes between the two varieties were associated with sugar biosynthesis and metabolism at three key stages (30, 60, and 90 DAF) of sugar accumulation. Further analysis showing upregulation of the expression of starch-related genes, such as β-amylase, GYS, and INV indicated that these genes were not actively expressed in AY01, resulting in slow accumulation of starch and reduced sugar content. By contrast, the downregulation of the expression of genes, such as PGK and MDH1, indicated that these genes were actively expressed in low-sugar chestnuts, resulting in the rapid fermentation of sugars. A link between gene up- or down-regulation during different developmental stages of chestnut and the effect of their expression on sugar content were established by KEGG pathway enrichment analysis. These findings provide further insights into the mechanism of sugar biosynthesis in chestnuts.