Changes in SeMSC, Glucosinolates and Sulforaphane Levels, and in Proteome Profile in Broccoli (Brassica oleracea var. Italica) Fertilized with Sodium Selenate
Abstract
:1. Introduction
2. Results and Discussion
2.1. Selenate Fertilization Increases SeMSC Level But does not Affect Significantly Glucosinolates and Sulforaphane Levels in Broccoli
2.2. Selenate Fertilization Changes the Proteome Profile in Broccoli Florets
Normalized Spot Volume | |||
---|---|---|---|
Spot Nr. | Control | Fortified | p-value |
1 | 6.3 ± 2.1 | 10.8 ± 5.4 | 0.172 |
2 | 5.5 ± 0.9 | 8.8 ± 2.9 | 0.073 |
3 * | 8.9 ± 3.1 | 32.8 ± 6.3 | 0.001 * |
4 | 6.1 ± 1.6 | 5.1 ± 1.6 | 0.449 |
5 | 9.0 ± 5.0 | 15.6 ± 2.3 | 0.055 |
6 | 34.3 ± 12.8 | 44.5 ± 9.8 | 0.252 |
7 | 13.9 ± 4.0 | 12.8 ± 4.8 | 0.727 |
8 | 8.2 ± 3.3 | 13.0 ± 2.5 | 0.059 |
9 | 10.5 ± 5.3 | 8.1 ± 2.1 | 0.437 |
10 * | 10.0 ± 1.1 | 14.5 ± 1.1 | 0.001 * |
11 | 12.0 ± 2.7 | 6.7 ± 3.6 | 0.077 |
12 | 10.4 ± 3.5 | 6.4 ± 2.2 | 0.105 |
13 | 9.7 ± 2.0 | 6.8 ± 1.8 | 0.079 |
14 | 3.6 ± 1.01 | 2.5 ± 1.2 | 0.296 |
15 * | 104.1 ± 11.0 | 132.3 ± 8.8 | 0.007 * |
16 | 5.8 ± 1.9 | 6.4 ± 1.7 | 0.633 |
17 | 4.9 ± 1.7 | 3.5 ± 0.8 | 0.206 |
18 * | 21.2 ± 2.1 | 12.3 ± 1.2 | 0.000 * |
19 | 21.8 ± 2.7 | 23.9 ± 2.3 | 0.332 |
20 * | 57.0 ± 8.1 | 76.1 ± 4.8 | 0.015 * |
21 | 18.5 ± 6.0 | 12.6 ± 2.7 | 0.123 |
22 | 9.8 ± 3.0 | 8.3 ± 1.7 | 0.403 |
23 * | 25.6 ± 0.9 | 11.4 ± 5.2 | 0.002 * |
24 | 5.9 ± 0.9 | 5.7 ± 0.6 | 0.397 |
25 | 7.9 ± 5.7 | - | - |
26 | 23.8 ± 11.2 | 1.8 ± 0.4 | 0.059 |
27 | 13.5 ± 3.5 | 16.1 ± 5.1 | 0.432 |
28 * | 9.5 ± 6.5 | 19.5 ± 2.3 | 0.028 * |
29 | 7.0 ± 1.7 | 5.6 ± 1.4 | 0.371 |
30 | 10.7 ± 2.4 | 9.1 ± 0.3 | 0.332 |
31 * | 34.9 ± 2.7 | 26.9 ± 3.9 | 0.014 * |
32 | 8.4 ± 3.7 | 5.2 ± 1.8 | 0.227 |
33 | 7.5 ± 4.6 | 5.1 ± 2.6 | 0.390 |
34 | 7.7 ± 4.4 | 3.9 ± 0.5 | 0.205 |
35 | 115.5 ± 17.1 | 93.8 ± 43.1 | 0.387 |
36 | 18.0 ± 1.1 | 19.2 ± 4.4 | 0.608 |
37 | 70.7 ± 7.6 | 60.8 ± 4.5 | 0.104 |
38 | 73.8 ± 8.3 | 84.4 ± 15.6 | 0.276 |
39 | 44.7 ± 8.6 | 38.3 ± 10.8 | 0.388 |
40 | 104.6 ± 22.7 | 125.8 ± 21.8 | 0.227 |
41 | 18.1 ± 7.6 | 15.0 ± 5.6 | 0.531 |
42 * | 11.2 ± 2.0 | 6.6 ± 0.1 | 0.012 * |
SN | RC | Protein identification | Peptide sequences | Accession number | Theoretical | PS | |
---|---|---|---|---|---|---|---|
MW (KDa) | pI | ||||||
3 | + | Heat shock protein 70KDa (A. thaliana) | FEELNMDLFR | gi|9294373 | 71.3 | 5 | 640 |
NALENYAYNMR | |||||||
TTPSYVAFTDSER | |||||||
ARFEELNMDLFR | |||||||
NAVVTVPAYFNDSQR | |||||||
GVWEGQPHADIGRIDLGTTYSCVG | |||||||
EYQEGIFESRTYSDNQPGVLIQV | |||||||
10 | + | Mitochondrial F1 ATP synthase beta subunit (A. thaliana) | VVDLLAPYQR | gi|6686269 | 54.2 | 5.4 | 1060 |
AHGGFSVFAGVGER | |||||||
VGLTGLTVAEYFR | |||||||
FTQANSEVSALLGR | |||||||
LVLEVSHHLGQNVVR | |||||||
DAEGQDVLLFIDNIFR | |||||||
QISELGIYPAVDPLDSTSR | |||||||
IQPESRAVGYQPTLASDLGAL | |||||||
FHEPDTQREGLPPIMTSLEVQD | |||||||
FHEPDTQREGLPPIMTSLEVQD | |||||||
15 | + | Myrosinase- binding protein (A. thaliana) | VYVGQGDSGVVYVK | gi|9279646 | 32.3 | 4.8 | 97 |
18 | − | β-1,3-glucanase (O. europaea) | AIETYIFAMFDENQK | gi|51507325 | 23.4 | 8.8 | 68 |
20 | + | Chlorophyll protein (B. oleracea) | KEPFYGGIAYK | gi|27530934 | 22.2 | 6 | 322 |
TTAQYLILPLSPR | |||||||
LQPLCPLGISQSSVK | |||||||
YGWGKEVDESSSASEEPAILV | |||||||
23 | − | Photosystem II protein (B. oleracea) | FLVPSYR | gi|49359169 | 36.8 | 6.8 | 469 |
EREDGIDYAAVTVQLPGGE | |||||||
SDKTPDELGTGAEKVIGVFQSLQPS | |||||||
28 | + | Glutathione S-transferase (A. thaliana) | VYGPHFASPK | gi|3201613 | 24.9 | 5.6 | 121 |
YDLYALGVGDFPVIGSKLADLAHLPFT |
3. Experimental
3.1. Broccoli Culture
3.2. Quantification of SeMSC
3.3. Quantification of Total Glucosinolates
3.4. Quantification of Sulforaphane
3.5. Protein Extraction and Quantification of Myrosinase Activity
3.6. Protein Extraction for 2-D Gel Electrophoresis
3.7. Two-dimensional Gel Electrophoresis
3.8. Image Acquisition and Analysis
3.9. Protein Identification
3.10. Statistical Analyses
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Sepúlveda, I.; Barrientos, H.; Mahn, A.; Moenne, A. Changes in SeMSC, Glucosinolates and Sulforaphane Levels, and in Proteome Profile in Broccoli (Brassica oleracea var. Italica) Fertilized with Sodium Selenate. Molecules 2013, 18, 5221-5234. https://doi.org/10.3390/molecules18055221
Sepúlveda I, Barrientos H, Mahn A, Moenne A. Changes in SeMSC, Glucosinolates and Sulforaphane Levels, and in Proteome Profile in Broccoli (Brassica oleracea var. Italica) Fertilized with Sodium Selenate. Molecules. 2013; 18(5):5221-5234. https://doi.org/10.3390/molecules18055221
Chicago/Turabian StyleSepúlveda, Ignacio, Herna Barrientos, Andrea Mahn, and Alejandra Moenne. 2013. "Changes in SeMSC, Glucosinolates and Sulforaphane Levels, and in Proteome Profile in Broccoli (Brassica oleracea var. Italica) Fertilized with Sodium Selenate" Molecules 18, no. 5: 5221-5234. https://doi.org/10.3390/molecules18055221