Sugar transport across tonoplasts is essential for maintaining cellular sugar homeostasis and metabolic balance in plant cells. It remains unclear, however, how this process is regulated among different classes of sugar transporters. Here, we identified a tonoplast H+/glucose symporter, MdERDL6-1, from apples, which was highly expressed in fruits and exhibited expression patterns similar to those of the tonoplast H+/sugar antiporters MdTST1 and MdTST2. Overexpression ofMdERDL6-1unexpectedly increased not only glucose (Glc) concentration but also that of fructose (Fru) and sucrose (Suc) in transgenic apple and tomato leaves and fruits. RNA sequencing (RNA-seq) and expression analyses showed an up-regulation ofTST1andTST2in the transgenic apple and tomato lines overexpressingMdERDL6-1. Further studies established that the increased sugar concentration in the transgenic lines correlated with up-regulation ofTST1andTST2expression. Suppression or knockout ofSlTST1andSlTST2in theMdERDL6-1–overexpressed tomato background reduced or abolished the positive effect ofMdERDL6-1on sugar accumulation, respectively. The findings demonstrate a regulation ofTST1andTST2byMdERDL6-1, in which Glc exported by MdERDL6-1 from vacuole up-regulatesTST1andTST2to import sugars from cytosol to vacuole for accumulation to high concentrations. The results provide insight into the regulatory mechanism of sugar accumulation in vacuoles mediated by the coordinated action of two classes of tonoplast sugar transporters.

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.

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In this paper, the splitting of microdroplet in a closed electrowetting-based digital microfluidic system has been studied via a numerical model. The governing equations for the fluid flow are solved by a finite volume formulation with a two-step projection method on a fixed computational domain. The free surface of the liquid is tracked by a coupled level-set and volume-of-fluid method, with the surface tension at the free surface computed by continuum surface force scheme. Contact angle hysteresis is implemented as an essential component of electrowetting modeling, and a simplified viscous force model is adopted to evaluate the viscous stress based on the Hele-Shaw model. Excellent agreement has been achieved between the numerical and published experimental results. A parametric study has been performed in which the effects of viscous stress, channel height, static contact angles, contact angle hysteresis, and electrode size on the splitting process have been analyzed. Three distinct splitting modes, which are “splitting with satellite droplet,” “normal splitting,” and “splitting cessation,” have been discussed. Based on the competition between the curvature in the z-direction (κz) and that on the x-y plane (κxy), the physical mechanism that separates the splitting into these three modes has been revealed. More importantly, a dimensionless parameter κ̃ has been proposed, which can be used for (a) determining the splitting mode and (b) estimating satellite droplet volume for electrowetting-induced droplet splitting process.

Microdrop generation with excellent controllability and volume precision is of paramount significance for a large variety of microfluidic applications. In this work, we propose a new configuration comprising only stripped electrodes of rectangular shape for the closed electrowetting-on-dielectric digital microfluidic (EWOD DMF) system and investigate its parallel microdrop generation outcomes via a numerical approach. The microfluidic droplet motion is solved by a finite-volume scheme on a fixed computational domain. The numerical model is verified by an experimental study of microdrop production from an EWOD DMF device with three different electrode designs. After model verification, we examine the influences of the equilibrium contact angle and the spacing of the microchannel on stripped electrode based microdrop generation outcomes and discover five different regimes including the phenomena of satellite droplet formation and separation cessation. Despite the various generation outcomes, the daughter droplet size is found to vary linearly with a dimensionless EWOD parameter κ*. More importantly, for all successful generations, the deviation of the daughter droplet size from that of the stripped electrode is smaller than 3.5%, which even reaches zero in proper conditions. This new configuration can be utilized as a convenient alternative for electrowetting-induced parallel microdrop production with excellent precision and controllability.