@article{504c4277b6e644a38629650b60c94212,
title = "An Evaluation of Nuclei Preparation of the Dormant Axillary Bud of Grapevine for Cell Cycle Analysis by Flow Cytometry",
abstract = "Whether the division of cells of a dormant meristem may be arrested, e.g., in the G1 phase, has proven to be an extremely difficult hypothesis to test. This is particularly so for woody perennial buds, where dormant and quiescent states are diffuse, and the organ may remain visibly unchanged for 6–9 months of the year. Flow cytometry (FCM) has been widely applied in plant studies to determine the genome size and endopolyploidy. In this study, we present the application of FCM to measure the cell cycle status in mature dormant buds of grapevine (Vitis vinifera cv. Cabernet Sauvignon), which represent a technically recalcitrant structure. This protocol illustrates the optimisation and validation of FCM data analysis to calculate the cell cycle status, or mitotic index, of dormant grapevine buds. We have shown how contamination with debris can be experimentally managed and give reference to the more malleable tomato leaves. We have also given a clear illustration of the primary pitfalls of data analysis to avoid artefacts or false results. Data acquisition and analysis strategies are detailed and can be readily applied to analyse FCM data from other recalcitrant plant samples.",
keywords = "cell cycle, dormancy, flow cytometry, grapevine buds, mitotic index",
author = "Dina Hermawaty and Considine, {John A.} and Considine, {Michael J.}",
note = "Funding Information: DH was a recipient of The Indonesia Endowment Fund for Education Scholarships. The authors acknowledge the funding support of the Australian Research Council (DP150103211 and FT180100409). The authors acknowledge the facilities and scientific and technical assistance of the National Imaging Facility, a National Collaborative Research Infrastructure Strategy (NCRIS) capability, at the Centre for Microscopy, Characterisation and Analysis, The University of Western Australia. Funding Information: DH was a recipient of The Indonesia Endowment Fund for Education Scholarships. The authors acknowledge the funding support of the Australian Research Council (DP150103211 and FT180100409). The authors acknowledge the facilities and scientific and technical assistance of the National Imaging Facility, a National Collaborative Research Infrastructure Strategy (NCRIS) capability, at the Centre for Microscopy, Characterisation and Analysis, The University of Western Australia. Publisher Copyright: Copyright {\textcopyright} 2022 Hermawaty, Considine and Considine.",
year = "2022",
month = feb,
day = "24",
doi = "10.3389/fpls.2022.834977",
language = "English",
volume = "13",
journal = "Frontiers in Plant Science",
issn = "1664-462X",
publisher = "Frontiers Media SA",
}