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Methods for On-Orbit Germination of Arabidopsis thaliana for Proteomic Analysis Cover

Methods for On-Orbit Germination of Arabidopsis thaliana for Proteomic Analysis

Gravitational and Space Research
Volume 4 (2016): Issue 2 (December 2016)
By: Sarahann Hutchinson,  Proma Basu,  Sarah E. Wyatt and  Darron R. Luesse  
Open Access
|Jul 2020
Figures & tables

Figures & Tables

Figure 1

Protocol flow chart for proteomic analysis of BRIC-grown material. Blue ovals indicate aspects of the approach that are addressed in this work.

Figure 2

Comparison of mean protein yield by starting seed weight. Seed sets weighing between 0.026 g (~1300 seeds) to 0.006 g (~300 seeds) were grown on plates for 96 h prior to protein extraction. * indicates p<0.03 using a Student's T-test. Bars represent an average of three separate extractions, each using approximately 1300 seeds. Error bars represent SEM.

Figure 3

Cold storage of planted seed delays germination for up to 120 h. Approximately 100 seeds were planted on ½ MS agar +1% sucrose plates. Plates were cold treated for 18 h, followed by 2 h white light. Plates were then returned to cold, with a subset of plates removed and imaged every 24 h. Representative plates are shown. Arrows indicate seedlings emerging from the seed.

Figure 4

Germination efficiency after return to room temperature. Plates were cold treated for 18 h, exposed to 2 h white light, followed by an additional 6 days at 4°C. After return to room temperature, germination percentage was recorded every 6 h for 44 h. * (p<0.03) and # (p<0.01) indicate significant result from a Student's T-test. n=3 replicates of approximately 1000 seeds each. Bars represent standard error.

Figure 5

Impact of growth time on mean protein yield. Plates containing 0.016 g of seed were grown for 1, 2, or 3 d prior to protein extraction and quantification of total protein. * indicates p<0.03 using a Student's T-test. n=6 plates of approximately 1,300 seedlings. Error bars represent SEM.

Figure 6

RNAlater™ stabilizes protein yield. Plates containing 0.016 g of seed were grown in darkness for 96 h. Seedlings were either soaked in RNAlater™ for 12 h prior to freezing at −80°C, or subject to immediate protein extraction. Results indicate no signifiant difference in total protein yield between the two approaches. n=3 plates of approximately 1300 seedlings. Bars represent SEM.
DOI: https://doi.org/10.2478/gsr-2016-0009 | Journal eISSN: 2332-7774
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Language: English
Page range: 20 - 27
Published on: Jul 18, 2020
Published by: American Society for Gravitational and Space Research
In partnership with: Paradigm Publishing Services
Publication frequency: Volume open
Keywords:
Arabidopsis,
Proteomics,
Transcriptomics,
BRIC,
ISS,
RNAlater®
Related subjects:
Life sciences,
Life sciences, other,
Materials sciences,
Materials sciences, other,
Physics,
Physics, other

© 2020 Sarahann Hutchinson, Proma Basu, Sarah E. Wyatt, Darron R. Luesse, published by American Society for Gravitational and Space Research
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Volume 4 (2016): Issue 2 (December 2016)
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