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Type: Journal article
Title: An optimized clearing protocol for the quantitative assessment of sub-epidermal ovule tissues within whole cereal pistils
Author: Wilkinson, L.
Tucker, M.
Citation: Plant Methods, 2017; 13(1):67-1-67-10
Publisher: BioMed Central
Issue Date: 2017
ISSN: 1746-4811
Statement of
Laura G. Wilkinson and Matthew R. Tucker
Abstract: Background: Seed development in the angiosperms requires the production of a female gametophyte (embryo sac) within the ovule. Many aspects of female reproductive development in cereal crops are yet to be described, largely due to the technical difficulty in obtaining phenotypic information at the cellular or sub-cellular level. Hoyer’s solution is currently well established as a solution for clearing thin tissues samples, such as sections or whole tissues of bryophytes, mycorrhizal fungi, and small model organisms (e.g. Arabidopsis thaliana). Results: Here we report a Hoyer’s solution-based clearing method to facilitate clearing of the whole barley pistil, with high reproducibility. The clearing process takes 10 days from fixation to visualisation, whereupon tissue is sufficiently clear to obtain multiple phenotypic measurements from sub-epidermal tissues and cells within the ovule. Conclusion: Visualisation of cereal ovules that have not been dissected from the pistil allows an unprecedented capability to collect quantitative morphological information from the developing ovule, integument, nucellus and embryo sac. This will enable comparisons with genetic data to reveal the contribution of pre-fertilisation ovule tissues towards downstream seed development.
Keywords: Microscopy; Hoyer’s solution; barley, ovule; monocot; development; cereal; clearing
Rights: © The Author(s) 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( applies to the data made available in this article, unless otherwise stated.
DOI: 10.1186/s13007-017-0217-z
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Appears in Collections:Agriculture, Food and Wine publications
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