Dataset Information

An optimised clearing protocol for the quantitative assessment of sub-epidermal ovule tissues within whole cereal pistils.

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.

SUBMITTER: Wilkinson LG

PROVIDER: S-EPMC5558753 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

An optimised clearing protocol for the quantitative assessment of sub-epidermal ovule tissues within whole cereal pistils.

Wilkinson Laura G LG Tucker Matthew R MR

Plant methods 20170815

<h4>Background</h4>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 ...[more]

PMID: 28824704

Similar Datasets

Project description:Abstract Objectives Health outcomes in previous fiber intervention studies have been variable, potentially due to differences in gut microbiome composition. This study aimed to determine if the effect of fiber intervention on the microbiome differs by initial microbiome or the quantity of fiber provided. Methods This study was designed as a randomized, un-blinded, cross-over trial of fiber cereal dosage. The cross-over design tested the effect of two 2-week long interventions with a High (28g) and Low (14g) level of daily supplemental fiber from whole wheat and bran cereal. Analysis was also completed on the overall study as a single arm, non-randomized, intervention of fiber cereal. The study enrolled 31 healthy adults. The microbiome was assessed at baseline and after intervention for changes in diversity, composition, and stability. Results Across all individuals, fiber intervention increased microbiome alpha-diversity (paired t-test, P = 0.047), but the microbiome was otherwise resistant to the effects of the intervention. Increasing fiber dose (High v. Low) was not associated with consistent changes in beta-diversity (linear mixed models). Approximately 20% of subjects were identified as responders based on beta diversity effect size. At baseline, responders had higher Prevotella copri and lower Bacteroides abundance than non-responders (Wilcoxon rank sum, qval < 0.05). In responders, fiber intake caused increased abundance of Bacteroides and Alistipes and reduced Prevotella (paired Wilcoxon, q < 0.2). In all subjects, fiber intervention decreased microbiome stability (paired Wilcoxon signed rank test, P = 0.006). In responders, there was a significant effect of the fiber level on stability, with higher fiber further lowering stability (linear mixed model, P = 0.05). Conclusions Our data suggest a responder/non-responder microbiome signature for this whole wheat and bran fiber cereal. We find that many effects were not additive by dosage level. Overall, microbiome diversity was increased and stability was decreased during the fiber cereal intervention and in responders this was dose dependent; the clinical implications of the impact of changes in stability remain unknown, and it is possible that the microbiome would stabilize in a longer intervention study. ClinicalTrials.gov identifier: NCT03623308. Funding Sources General Mills, Inc.

Dataset Information

An optimised clearing protocol for the quantitative assessment of sub-epidermal ovule tissues within whole cereal pistils.

Background

Results

Conclusion

Publications

An optimised clearing protocol for the quantitative assessment of sub-epidermal ovule tissues within whole cereal pistils.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets