Xylogenesis and phloemogenesis in the flesh of sweet cherry fruit are limited to early-stage development.

Water inflows into sweet cherry fruit occur via the xylem and the phloem vasculatures of the pedicel. The rates of these inflows are subject to marked changes during fruit development. The objective was to establish if, and when, xylogenesis and phloemogenesis occur in the fruit flesh (mesocarp) during fruit development. Fruit were cut in half and the median and the lateral bundles inspected by light microscopy. Fruit mass increased with time in a double sigmoid pattern. Xylogenesis and phloemogenesis were both limited to early fruit development (stage I). There were no consistent changes in the areas of either xylem or phloem after stage I until maturity (i.e., during stages II and III). The cross-sectional areas of xylem and of phloem in a bundle were both linearly related to total bundle area. Most of the increases (stage I) in bundle area (62%, r2 = 0.99***) were accounted for by increases in phloem area and about 35% (r2 = 0.97***) by increases in xylem area. A small percentage of the xylem area increase (about 4% of the total area of the bundle; r2 = 0.48***) was contributed by the appearance of intercellular spaces within the xylem. Our results suggest, that new xylem and phloem tissues are differentiated only during early development.

Prediction and characterization of the stability enhancing effect of the Cherry-Tag™ in highly concentrated protein solutions by complex rheological measurements and MD simulations.

Solution stability attributes are one of the key parameters within the production and launching phase of new biopharmaceuticals. Instabilities of active biological compounds can reduce the yield of biopharmaceutical productions, and may induce undesired reactions in patients, such as immunogenic rejections. Protein solution stability thus needs to be engineered and monitored throughout production and storage. In contrast to the gold standard of long-term storage experiments applied in industry, novel experimental and in silico molecular dynamics tools for predicting protein solution stability can be applied within several minutes or hours. Here, a rheological approach in combination with molecular dynamics simulations are presented, for determining and predicting long-term phase behavior of highly concentrated protein solutions. A diversity of liquid phase conditions, including salt type, ionic strength, pH and protein concentration are tested in a Glutathione-S-Transferase (GST) case study, in combination with the enzyme with and without solubility-enhancing Cherry-Tag™. The rheological characterization of GST and Cherry-GST solutions enabled a fast and efficient prediction of protein instabilities without the need of long-term protein phase diagrams. Finally, the strong solubility enhancing properties of the Cherry-Tag™ were revealed by investigating protein surface properties in MD simulations. The tag highly altered the overall surface charge and hydrophobicity of GST, making it less accessible to alteration by the chemical surrounding.