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Gravitational and Space Research
Édition 4 (2016): Edition 2 (December 2016)
Accès libre
Methods for On-Orbit Germination of
Arabidopsis thaliana
for Proteomic Analysis
Sarahann Hutchinson
Sarahann Hutchinson
,
Proma Basu
Proma Basu
,
Sarah E. Wyatt
Sarah E. Wyatt
et
Darron R. Luesse
Darron R. Luesse
| 18 juil. 2020
Gravitational and Space Research
Édition 4 (2016): Edition 2 (December 2016)
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Article Category:
Research Article
Publié en ligne:
18 juil. 2020
Pages:
20 - 27
DOI:
https://doi.org/10.2478/gsr-2016-0009
Mots clés
Arabidopsis
,
Proteomics
,
Transcriptomics
,
BRIC
,
ISS
,
RNAlater®
© 2020 Sarahann Hutchinson et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.
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.
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