Swelling of cell walls in mature sweet cherry fruit: factors and mechanisms.

MAIN CONCLUSION: Swelling of sweet cherry cell walls is a physical process counterbalanced by turgor. Cell turgor prevents swelling in intact cells, whereas loss of turgor allows cell walls to swell. Swelling of epidermal cell walls precedes skin failure in sweet cherry (Prunus avium) cracking. Swollen cell walls lead to diminished cell:cell adhesions. We identify the mechanism of cell wall swelling. Swelling was quantified microscopically on epidermal sections following freeze/thaw treatment or by determining swelling pressure or swelling capacity of cell wall extracts. Releasing turgor by a freeze/thaw treatment increased cell wall thickness 1.6-fold within 2 h. Pressurizing cell wall extracts at > 12 kPa prevented swelling in water, while releasing the pressure increased swelling. The effect was fully reversible. Across cultivars, cell wall thickness before and after turgor release in two subsequent seasons was significantly correlated (before release of turgor: r = 0.71**, n = 14; after release of turgor: r = 0.73**, n = 14) as was the swelling of cell walls upon turgor release (r = 0.71**, n = 14). Close relationships were also identified for cell wall thickness of fruit of the same cultivars grown in the greenhouse and the field (before release of turgor: r = 0.60, n = 10; after release of turgor: r = 0.78**, n = 10). Release of turgor by heating, plasmolysis, incubation in solvents or surfactants resulted in similar swelling (range 2.0-3.1 µm). Cell wall swelling increased from 1.4 to 3.0 µm as pH increased from pH 2.0 to 5.0 but remained nearly constant between pH 5.0 and 8.0. Increasing ethanol concentration decreased swelling. Swelling of sweet cherry cell walls is a physical process counterbalanced by turgor.

Combining Histochemical Staining and Image Analysis to Quantify Starch in the Ovary Primordia of Sweet Cherry during Winter Dormancy.

Changes in starch in small structures are associated with key events during several plant developmental processes, including the reproductive phase from pollination to fertilization and the onset of fruiting. However, variations in starch during flower differentiation are not completely known, mainly due to the difficulty of quantifying the starch content in the particularly small structures of the flower primordia. Here, we describe a method for the quantification of starch in the ovary primordia of sweet cherry (Prunus avium L.) by using an image analysis system attached to the microscope, which allows relating the changes in starch content with the different phases of dormancy from autumn to spring. For this purpose, the dormancy status of flower buds is determined by evaluating the bud growth of shoots transferred to controlled conditions at different moments in winter time. For the quantification of starch in the ovary primordia, flower buds are sequentially collected, fixed, embedded in paraffin wax, sectioned, and stained with I2Kl (potassium iodide-iodine). Preparations are observed under the microscope and analyzed by an image analyzer that clearly distinguishes starch from the background. Starch content values are obtained by measuring the optical density of the image that corresponds to the stained starch, considering the sum of the optical density of each pixel as an estimation of the starch content of the frame studied.

Anther and pollen development in sweet cherry (Prunus avium L.) in relation to winter dormancy.

Anther and pollen development is a highly conserved process in angiosperms, but while pollen formation in annual plants occurs in a few days, in temperate woody perennials, it requires several months. How anther and pollen development is framed in terms of seasonality plays a clear part in reproductive success. In this study, seasonal anther and pollen development is characterized in two sweet cherry cultivars over 2 years, paying special attention to the period of dormancy and unveiling the role of starch in this process. We evaluated starch content from the autumn until bud burst with the help of an image analysis system fitted to a light microscope. Microscope observations allowed the temporal relationship of pollen development to the phenological stages of flower and bud development to be determined. In both cultivars and years, anther and pollen development followed the same pattern. Development was halted by dormancy, when the anthers showed no morphological changes until several weeks after chilling fulfillment, until the milder temperatures reactivated development. After dormancy, starch was accumulated in the connective tissue until tracheary element differentiation. Quantification of starch in the connective tissue of anthers revealed its importance in supporting pollen meiosis and subsequent anther growth.