Significance: Sugar transport across membranes is essential for maintaining cellular sugar homeostasis and metabolic balance in plant cells. However, it remains unclear how this process is regulated among different classes of sugar transporters. Here, we identified an apple tonoplast H ⁺ /glucose symporter, MdERDL6-1, that exports glucose to cytosols to up-regulate the expression of H ⁺ /sugar antiporter genes TST1 and TST2 to import sugars from cytosol to vacuole for accumulation to high concentrations in apples and tomatoes. The findings provide insights into the regulatory mechanism underlying sugar exchange between cytosol and vacuole.

Summary: Sugar transporters are necessary to transfer hexose from cell wall spaces into parenchyma cells to boost hexose accumulation to high concentrations in fruit. Here, we have identified an apple hexose transporter (HTs), MdHT2.2, located in the plasma membrane, which is highly expressed in mature fruit. In a yeast system, the MdHT2.2 protein exhibited high ¹⁴C‐fructose and ¹⁴C‐glucose transport activity. In transgenic tomato heterologously expressing MdHT2.2, the levels of both fructose and glucose increased significantly in mature fruit, with sugar being unloaded via the apoplastic pathway, but the level of sucrose decreased significantly. Analysis of enzyme activity and the expression of genes related to sugar metabolism and transport revealed greatly up‐regulated expression of SlLIN5, a key gene encoding cell wall invertase (CWINV), as well as increased CWINV activity in tomatoes transformed with MdHT2.2. Moreover, the levels of fructose, glucose and sucrose recovered nearly to those of the wild type in the sllin5‐edited mutant of the MdHT2.2‐expressing lines. However, the overexpression of MdHT2.2 decreased hexose levels and increased sucrose levels in mature leaves and young fruit, suggesting that the response pathway for the apoplastic hexose signal differs among tomato tissues. The present study identifies a new HTs in apple that is able to take up fructose and glucose into cells and confirms that the apoplastic hexose levels regulated by HT controls CWINV activity to alter carbohydrate partitioning and sugar content.

To comprehensively understand the responses of carbohydrate metabolism and transport to different levels of nitrogen supply in growing shoot tips of crabapple (Malus hupehensis Rehd), enzyme activities and related genes involved in the sugar metabolism pathway were investigated. The nitrogen and chlorophyll content of plants increased with increasing nitrogen supply. High nitrogen application increased the net photosynthesis rate and the growth rate of shoot tips but decreased the synthesis capability of sucrose and sorbitol in mature leaves. However, the shoot tips of plants under high-nitrogen treatment had higher contents of sucrose and sorbitol than did those under low-nitrogen treatment, while the activity of sucrose phosphate synthase and aldose-6-phosphate was increased and the transporters MdSOT and MdSUT were up-regulated. Moreover, the activities of enzymes involved in sucrose and hexose metabolism (including sucrose synthase, fructokinase, and hexokinase) were enhanced in the shoot tips of plants under high-nitrogen conditions, and the expression levels of MdSUSY3 and MdHK1 were significantly up-regulated. These findings indicate that a high nitrogen supply increases the metabolic capacity of assimilatory substances in shoot tips, accelerates the efficiency of sugar utilization and eventually leads to a rapid increase in the growth of shoot tips. Our results highlight that high nitrogen increases the capacity of sugar unloading and metabolic utilization in growing shoot tissues.

Melatonin has been reported to play roles in regulating carbohydrate levels and plant growth. However, little is known about the exact mechanism by which melatonin regulates sugar levels and growth in plants. In this study, it was found that high levels of melatonin inhibited the growth of wild-type (WT) apple plants and induced significant accumulations of fructose, glucose, and sucrose in apple leaves, while MdFRK2 expression was significantly downregulated. MdFRK2 promoter transiently expressed in tobacco leaves further supported that the expression of MdFRK2 could be inhibited by exogenous melatonin. After applying exogenous melatonin, the suppression of MdFRK2 expression was significantly rescued in transgenic apples overexpressing MdFRK2 via the 35S promoter. Fructose, glucose, and sucrose concentrations increased less as compared to WT apple plants. Wild-type plants showed a stunted phenotype 21 days after melatonin treatment, while MdFRK2-overexpressing plants exhibited slightly inhibited growth, indicating that the downregulated MdFRK2 expression in response to melatonin was involved in melatonin-mediated growth inhibition. Taken together, these results demonstrate the involvement of MdFRK2 in melatonin-induced sugar accumulation and growth inhibition. Our findings shed light on the roles played by MdFRK2 in connecting melatonin action and plant growth.