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Hermawaty, D.

Publications and source records attributed to Hermawaty, D..

4 recordsLinked to original sources

Seasonal changes in the physiology and metabolism of grapevine perennating buds

Grapevine (Vitis vinifera L.) buds undergo seasonal dormancy to survive unfavourable conditions and synchronize growth with environmental cues. While dormancy transitions have been widely studied in temperate woody perennials, the physiological and metabolic dynamics underlying these transitions in grapevine remain poorly understood. Our study investigates seasonal changes in the physiology and metabolism of perennating buds in V. vinifera cv. Cabernet Sauvignon, focusing on dormancy depth, respiration, sugar metabolism, and cell cycle activity. We identified three distinct phases of bud quiescence: (i) para-dormancy (early summer), characterized by active metabolism and high levels of tricarboxylic acid cycle intermediates, shikimate, and myo-inositol; (ii) endo-dormancy (late summer to autumn), where dormancy depth peaked in late summer and was marked by reduced respiration and bud water content, and a sharp decline in hexose levels with a concomitant increase in raffinose; and (iii) eco-dormancy (winter to spring), featuring increased respiration, sugar mobilization (notably sucrose, glucose, and trehalose), and reactivation of cell division, with a shift of cells into the G2 phase. Just prior to bud burst, we observed a significant accumulation of sugar-phosphates, providing evidence that supports their role in promoting bud burst also in grapevine. Our findings provide new insights into the biochemical and physiological regulation of bud dormancy and bud burst, contributing to a deeper understanding of dormancy transitions in perennial crops.

plant biology↗

The physiology of bud dormancy and cell cycle status in grapevine

Evaluating the cell cycle status during dormancy of multicellular organisms is problematic. This is particularly so for woody perennial buds, where dormant and quiescent states are diffuse, and the organ may remain visibly unchanged for six to nine months of the year. In this study, we investigate cell cycle status of dormant grapevine buds by measuring mitotic index using an optimised method developed for grapevine bud tissue. The experimental material showed a dynamic range in the depth of dormancy, declining from 200 days in March to less than 60 days in May and 30 days in August, measured as the time to reach 50% bud burst in forcing conditions. Despite these differences, flow cytometry analysis showed that most nuclei isolated from these buds were arrested at the G1 phase. Ultrastructure analysis of the cells in the region of the shoot apical meristem confirmed that the mitotic activities of buds remained low at all time points, together with the development of starch grains and the relative absence of organelle development. HIGHLIGHTThe cell cycle and ultrastructure data suggest interesting evidence correspond to the growth resumption capacity of grapevine cv. Cabernet Sauvignon buds, i.e., absence of mitosis activities regardless of dormancy depth and starch accumulation irrespective of chilling accumulation.

plant biology↗

Systematic evaluation of chromatin immunoprecipitation sequencing to study histone occupancy in dormancy transitions of grapevine buds

1.The regulation of DNA accessibility by histone modification has emerged as a paradigm of developmental and environmental programming. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a versatile tool widely used to investigate in vivo protein-DNA interaction. The technique has been successfully demonstrated in several plant species and tissues; however, it has remained challenging in woody tissues. Here we developed a ChIP method specifically for mature dormant grapevine buds (Vitis vinifera cv. Cabernet Sauvignon). Each step of the protocol was systematically optimised, including crosslinking, chromatin extraction, sonication, and antibody validation. Analysis of histone H3-enriched DNA was performed to evaluate the success of the protocol and identify occupancy of histone H3 along grapevine bud chromatin. To our best knowledge, this is the first ChIP experiment protocol optimised for grapevine bud system.

plant biology↗

Physiological regulation of bud burst in grapevine

The physiological constraints on bud burst in woody perennials, including the prerequisite for vascular development remain unresolved. Both light and tissue oxygen status have emerged as important cues for vascular development in other systems, however, light requirement appears to be facultative in grapevine, and the information related to the spatial variability of oxygen in buds is unclear. Here, we analysed apoplastic development at early stages of grapevine bud burst and combined molecular modelling with histochemical techniques to determine the pore size of cell walls in grapevine buds. The data demonstrate that quiescent grapevine buds were impermeable to apoplastic dyes (acid fuchsin and eosin Y) until after bud burst was established. The molecular exclusion size was calculated to be 2.1 nm, which would exclude most macromolecules except simple sugars and phytohormones. In vivo experiments show that grapevine buds were able to resume growth even following excision from the cane, and that the outer scales of grapevine buds may participate in the biochemical repression of bud burst. Furthermore, we demonstrate that the tissue oxygen partial pressure data correlated well with structural heterogeneity within the bud and differences in tissue density. These data consolidate evidence that the meristematic core becomes rapidly oxygenated during bud burst. Taken together, and when put in the context of earlier studies, these data provide solid evidence that the physiological and biochemical events that initiate bud burst reside within the bud, and question the role of long distance signalling in this developmental transition.\n\nHighlightsO_LIThe apoplastic pore size between the grapevine bud and the mother vine is dynamically regulated in the transition to bud burst.\nC_LIO_LIThe molecular exclusion size of the apoplastic connection between the bud and cane is calculated 2.1 nm prior to the initiation of bud burst.\nC_LIO_LIThe structural heterogeneity of the bud explains the spatial variance in tissue oxygen status, and the meristematic core is oxygenated during the initiation of bud burst.\nC_LIO_LILong distance maternal signals are not a requirement for bud burst.\nC_LI

plant biology↗