Analysis of Stored mRNA Degradation in Acceleratedly Aged Seeds of Wheat and Canola in Comparison to Arabidopsis
Abstract
:1. Introduction
2. Results
2.1. Germination Kinetics for Wheat, Canola, and Arabidopsis Seeds
2.2. Similar Quality of RNA Samples for Aged Wheat and Canola Seeds
2.3. Correlation of mRNA Degradation with Aging Time and Fragment Length
2.4. Similar Rate of mRNA Degradation in Aged Seeds of Wheat, Canola and Arabidopsis
3. Discussion
3.1. Stored mRNAs Were Gradually Degraded at a Constant Rate over Aging Time in Wheat and Canola Seeds
3.2. The β Value Can Be Used to Describe the Rate of mRNA Degradation Quantitatively in Different Plants
3.3. β Values for AA Seeds of Arabidopsis Increased Non-Linearly with High Temperatures
3.4. Practical Implications for Plant Germplasm Conservation
4. Materials and Methods
4.1. Comparing Seed Germination Kinetics in Wheat, Canola, and Arabidopsis Seeds
4.2. Total RNA Analysis for Differently Aged Seeds of Wheat, Canola, and Arabidopsis
4.3. qPCR Analysis of mRNA Degradation in Aged Seeds
4.4. Estimating the Rate of Stored mRNA Degradation in Aged Seeds
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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AA Day | Wheat | Canola | |||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
0 | 6 | 12 | 18 | 24 | 30 | 0 | 6 | 12 | 18 | 24 | 30 | ||
Germination Percentage | 100.0 | 92.7 | 76.0 | 34.7 | 0.0 | 0.0 | 100.0 | 94.0 | 89.0 | 58.0 | 15.5 | 0.0 | |
Yield (ng/μL) | a | 228.0 | 144.0 | 444.0 | 212.1 | 122.0 | 217.9 | 363.0 | 390.0 | 374.0 | 334.7 | 173.0 | 593.0 |
b | 347.0 | 551.0 | 338.0 | 296.3 | 335.2 | 276.3 | 354.0 | 415.0 | 478.6 | 347.6 | 238.0 | 601.0 | |
A280/260 | a | 2.02 | 2.02 | 2.03 | 2.07 | 2.05 | 2.10 | 2.10 | 2.11 | 2.04 | 2.07 | 1.95 | 2.10 |
b | 1.98 | 1.98 | 2.06 | 2.01 | 2.09 | 2.08 | 2.09 | 2.11 | 2.07 | 2.10 | 2.03 | 2.13 | |
A280/230 | a | 2.27 | 2.24 | 2.23 | 2.21 | 2.13 | 2.20 | 2.40 | 2.31 | 2.23 | 2.31 | 2.44 | 2.21 |
b | 2.30 | 2.32 | 2.24 | 2.23 | 2.25 | 2.10 | 2.38 | 2.32 | 2.32 | 2.37 | 2.39 | 2.34 |
Fragment | β Value (Break per Nucleotide per Day × 10−4) | |||||
---|---|---|---|---|---|---|
Day 6 | Day 12 | Day 18 | Day 24 | Average | Overall | |
W2950bp | 3.30 ± 0.10 | 2.68 ± 0.04 | 2.62 ± 0.06 | 3.14 ± 0.18 | 2.94 ± 0.15 | 2.26 ± 0.51 |
W3950bp | 1.86 ± 0.22 | 1.49 ± 0.11 | 1.71 ± 0.16 | 2.46 ± 0.36 | 1.88 ± 0.18 | |
W10955bp | 2.12 ± 0.15 | 1.94 ± 0.03 | 2.15 ± 0.02 | 2.42 ± 0.11 | 2.16 ± 0.09 | |
W12950bp | 1.80 ± 0.31 | 1.99 ± 0.25 | 2.41 ± 0.06 | 2.07 ± 0.02 | 2.07 ± 0.11 | |
Average | 2.27 ± 0.30 | 2.03 ± 0.21 | 2.22 ± 0.17 | 2.52 ± 0.19 |
Fragment | β Value (Break per Nucleotide per Day × 10−4) | ||||||
---|---|---|---|---|---|---|---|
Day 6 | Day 12 | Day 18 | Day 24 | Day 30 | Average | Overall | |
Bn111490bp | 1.73 ± 0.04 | 1.54 ± 0.06 | 1.28 ± 0.04 | 1.17 ± 0.04 | 1.33 ± 0.06 | 1.41 ± 0.05 | 1.37 ± 0.31 |
Bn121500bp | 2.46 ± 0.24 | 1.84 ± 0.04 | 1.54 ± 0.07 | 1.35 ± 0.03 | 1.40 ± 0.05 | 1.72 ± 0.09 | |
Bn131486bp | 1.51 ± 0.05 | 1.44 ± 0.02 | 1.15 ± 0.02 | 1.03 ± 0.00 | 1.29 ± 0.02 | 1.28 ± 0.02 | |
Bn141500bp | 1.44 ± 0.10 | 1.20 ± 0.10 | 1.02 ± 0.02 | 0.89 ± 0.02 | 1.20 ± 0.02 | 1.15 ± 0.05 | |
Bn171500bp | 1.35 ± 0.20 | 1.39 ± 0.01 | 1.17 ± 0.03 | 1.44 ± 0.01 | 1.12 ± 0.06 | 1.29 ± 0.06 | |
Average | 1.70 ± 0.13 | 1.48 ± 0.05 | 1.23 ± 0.04 | 1.18 ± 0.02 | 1.27 ± 0.04 |
Gene | AA Days | β Value (Break per Nucleotide per Day × 10−4) | ||||
---|---|---|---|---|---|---|
22 °C | 30 °C | 33 °C | 37 °C | 40 °C | ||
At1G743102000bp | 8 | 0.32 ± 0.06 | 0.61 ± 0.07 | 0.37 ± 0.05 | 1.33 ± 0.11 | 2.05 ± 0.15 |
16 | 0.46 ± 0.09 | 0.73 ± 0.06 | 0.73 ± 0.03 | 1.49 ± 0.08 | 2.86 ± 0.24 | |
Overall | 0.39 ± 0.10 | 0.67 ± 0.09 | 0.55 ± 0.18 | 1.41 ± 0.13 | 2.46 ± 0.45 |
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Zhao, L.; Wang, H.; Fu, Y.-B. Analysis of Stored mRNA Degradation in Acceleratedly Aged Seeds of Wheat and Canola in Comparison to Arabidopsis. Plants 2020, 9, 1707. https://doi.org/10.3390/plants9121707
Zhao L, Wang H, Fu Y-B. Analysis of Stored mRNA Degradation in Acceleratedly Aged Seeds of Wheat and Canola in Comparison to Arabidopsis. Plants. 2020; 9(12):1707. https://doi.org/10.3390/plants9121707
Chicago/Turabian StyleZhao, Liang, Hong Wang, and Yong-Bi Fu. 2020. "Analysis of Stored mRNA Degradation in Acceleratedly Aged Seeds of Wheat and Canola in Comparison to Arabidopsis" Plants 9, no. 12: 1707. https://doi.org/10.3390/plants9121707