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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
 oraz 
Darron R. Luesse
Darron R. Luesse
   | 18 lip 2020
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Article Category: Research Article
Data publikacji: 18 lip 2020
Zakres stron: 20 - 27
DOI: https://doi.org/10.2478/gsr-2016-0009
Słowa kluczowe
© 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.
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.
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.
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.
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.
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.
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.
eISSN:
2332-7774
Język:
Angielski
Częstotliwość wydawania:
2 razy w roku
Dziedziny czasopisma:
Life Sciences, other, Materials Sciences, Physics
Kanał RSS czasopisma