Document Type : Research Paper
Authors
1 PHD.Department of Biology, Faculty of Sciences, Golestan University, Gorgan, Iran
2 Professor.Department of Agronomy, Faculty of Plant Production, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran
3 Professor.Department of Human Genetics, Gorgan University of Medical Sciences
4 Associate Professor, Department of Biology, Golestan University, Gorgan, Iran
Abstract
Introduction: Long-term storage of seeds under unfavorable conditions leads to their deterioration associated with decreased germination. Several studies have shown increased lipolysis in deteriorating seeds, but little is known about the role of protease activity during deterioration. In the present study, the effect of the controlled deterioration (CD) on the mobilization of storage proteins and the effects of pretreatment with protease inhibitor (PMSF) on the germination percentage of walnut kernels after CD were investigated. Materials and methods: Kernels were adjusted to 15% and 20% MC with water (control) or PMSF solution and then incubated for 3 and 6 days at 45°C for CD. Results: CD increased soluble proteins, solubility of 19-24 kDa glutelins, increased activity of a 80 kDa protease, accumulation of total amino acid, proline and increased protein carbonylation. Although aging-dependent decrease of germination was similar in both control and PMSF pretreatment, the aged kernels in PMSF pretreatment had lower proline, amino acid and carbonyl groups contents compared to the control. Discussion: These results suggest that CD causes increased solubility of protein reserves, but the inhibition of serine protease(s), unlike lipase, has no effect on improving the adverse physiological effects of CD including germination loss, and only at the biochemical level, it partially improved kernel response to stress. Identification of different metabolic pathways operating during CD of kernels can draw a picture of deterioration mechanism and also leads to introducing protocols for the quality and viability maintenance of kernels during storage.
Keywords
Main Subjects
[2] Xiang, F., Liu, W. C., Liu, X., Song, Y., Zhang, Y., Zhu, X., Wang, P., Guo, S. & Song, C-P. (2023). Direct balancing of lipid mobilization and reactive oxygen species production by the epoxidation of fatty acid catalyzed by a cytochrome P450 protein during seed germination. New Phytologist, 237 (6), 2104-2117.
[3] He, D., Cai, M., Liu, M., & Yang, P. (2023). TMT-based quantitative proteomic and physiological analyses on lotus plumule of artificially aged seed in long-living sacred lotus Nelumbo nucifera. Journal of Proteomics, 270, 104736.
[4] Bewley, J. D., Bradford, K. J., Hilhorst, H. W. & Nonogaki, H. (2013). Longevity, storage, and deterioration. In Seeds (pp. 341-376). Springer, New York, NY.
[5] Gerna, D., Ballesteros, D., Arc, E., Stöggl, W., Seal, C.E., Marami-Zonouz, N., Na, C.S., Kranner, I. & Roach, T. (2022). Does oxygen affect ageing mechanisms of Pinus densiflora seeds? A matter of cytoplasmic physical state. Journal of Experimental Botany, 73 (8), 2631–2649.
[6] Kibinza, S., Bazin, J., Bailly, C., Farrant, J. M., Corbineau, F. & El-Maarouf-Bouteau, H. (2011). Catalase is a key enzyme in seed recovery from ageing during priming. Plant science, 181 (3), 309-315.
[8] Galleschi, L., Capocchi, A., Ghiringhelli, S., Saviozzi, F., Calucci, L., Pinzino, C. & Zandomeneghi, M. (2002). Antioxidants, free radicals, storage proteins, and proteolytic activities in wheat (Triticum durum) seeds during accelerated aging. Journal of Agricultural and Food Chemistry, 50 (19), 5450-5457.
[9] Rajjou, L., Lovigny, Y., Groot, S. P., Belghazi, M., Job, C. & Job, D. (2008). Proteome-wide characterization of seed aging in Arabidopsis: a comparison between artificial and natural aging protocols. Plant Physiology, 148 (1), 620-641.
[10] Parkhey, S., Naithani, S. C. & Keshavkant, S. (2014). Protein metabolism during natural ageing in desiccating recalcitrant seeds of Shorea robusta. Acta Physiologiae Plantarum, 36 (7), 1649-1659.
[11] Nguyen, T. P., Cueff, G., Hegedus, D. D., Rajjou, L. & Bentsink, L. (2015). A role for seed storage proteins in Arabidopsis seed longevity. Journal of Experimental Botany, 66 (20), 6399-6413.
[12] Dell'Aquila, A. (1994). Wheat seed ageing and embryo protein degradation. Seed Science Research, 4 (3), 293-298.
[13] Viñegra de la Torre, N., Kaschani, F., Kaiser, M., van der Hoorn, R. A., Soppe, W. J. & Misas Villamil, J. C. (2019). Dynamic hydrolase labelling as a marker for seed quality in Arabidopsis seeds. Biochemical Journal, 476 (5), 843-857.
[14] Sze‐Tao, K. W. C. & Sathe, S. K. (2000). Walnuts (Juglans regia L.): proximate composition, protein solubility, protein amino acid composition and protein in vitro digestibility. Journal of the Science of Food and Agriculture, 80 (9), 1393-1401.
[15] Raoufi, A., Vahdati, K., Karimi, S. & Roozban, M. R. (2020). Optimizing seed germination and growth of seedlings in persian walnut. Journal of Nuts, 11 (3), 185-193.
[16] Vahdati, K., Aslamarz, A. A., Rahemi, M., Hassani, D. & Leslie, C. (2012). Mechanism of seed dormancy and its relationship to bud dormancy in persian walnut. Environmental and experimental botany, 75, 74-82.
[17] Lotfi, N., Soleimani, A., Vahdati, K. & Çakmakçı, R. (2019). Comprehensive biochemical insights into the seed germination of walnut under drought stress. Scientia Horticulturae, 250, 329-343.
[18] Vahdati, K., Lotfi, N., Kholdebarin, B., Hassani, D., Amiri, R., Mozaffari, M. R. & Leslie, C. (2009). Screening for drought-tolerant genotypes of persian walnuts (Juglans regia L.) during seed germination. HortScience, 44 (7):, 1815-1819.
[19] Ortiz, C.M., Vicente, A.R., Fields, R.P., Grilo, F., Labavitch, J.M., Donis-Gonzalez, I. & Crisosto, C.H. (2019) Walnut (Juglans regia L.) postharvest deterioration as affected by pellicle integrity, cultivar and oxygen concentration. Postharvest Biology and Technology, 156, 110948.
[20] Ziaolhagh, S., Fatemian, H. & Goodarzi, F. (2020). The effect of thermal pretreatment and packaging conditions on the shelf-life of walnut kernels. Journal of Nuts, 11 (2), 159-168.
[21] Pournik, S., Abbasi-Rostami, M., Sadeghipour, H. R. & Ghaderi-Far, F. (2019). True lipases beside phospholipases contribute to walnut kernel viability loss during controlled deterioration and natural aging. Environmental and Experimental Botany, 164, 71-83.
[22] Einali, A. R. & Sadeghipour, H. R. (2007). Alleviation of dormancy in walnut kernels by moist chilling is independent from storage protein mobilization. Tree physiology, 27(4), 519-525.
[23] Shahmoradi, Z., Tamaskani, F., Sadeghipour, H. R. & Abdolzadeh, A. (2013). Redox changes accompanying storage protein mobilization in moist chilled and warm incubated walnut kernels prior to germination. Journal of plant physiology, 170 (1), 6-17.
[24] Heussen, C. & Dowdle, E. B. (1980). Electrophoretic analysis of plasminogen activators in polyacrylamide gels containing sodium dodecyl sulfate and copolymerized substrates. Analytical biochemistry, 102 (1), 196-202.
[25] Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical biochemistry, 72 (1-2), 248-254.
[26] Fling, S. P. & Gregerson, D. S. (1986). Peptide and protein molecular weight determination by electrophoresis using a high-molarity tris buffer system without urea. Analytical biochemistry, 155 (1), 83-88.
[27] Kobrehel, K., Wong, J. H., Balogh, A., Kiss, F., Yee, B. C. & Buchanan, B. B. (1992). Specific reduction of wheat storage proteins by thioredoxin h. Plant Physiology, 99 (3):, 919-924.
[28] Stone, S. L. & Gifford, D. J. (1997). Structural and biochemical changes in loblolly pine (Pinus taeda L.) seeds during germination and early-seedling growth. I. Storage protein reserves. International Journal of Plant Sciences, 158 (6), 727-737.
[29] Markwell, M. A. K., Haas, S. M., Tolbert, N. E. & Bieber, L. L. (1981). [16] Protein determination in membrane and lipoprotein samples: Manual and automated procedures. In Methods in enzymology, 72, 296-303.
[30] Sun, W. Q. & Leopold, A. C. (1995). The Maillard reaction and oxidative stress during aging of soybean seeds. Physiologia Plantarum, 94 (1), 94-104.
[31] Xia, Q., El-Maarouf-Bouteau, H., Bailly, C. & Meimoun, P. (2016). Determination of protein carbonylation and proteasome activity in seeds. Methods in Molecular Biology, 1450, 205-12.
[32] Bates, L.S., Waldren, R.P. & Teare, I.D. (1973). Rapid determination of free proline for water-stress studies. Plant Soil, 39, 205–207.
[33] Omokolo, N. D., Nankeu, D. J., Niemenak, N. & Djocgoue, P. F. (2002). Analysis of amino acids and carbohydrates in the cortex of nine clones of Theobroma cacao L. in relation to their susceptibility to Phytophthora megakarya Bra. and Grif. Crop Protection, 21 (5), 395-402.
[34] Yemm, E. W., Cocking, E. C. & Ricketts, R. E. (1955). The determination of amino-acids with ninhydrin. Analyst, 80 (948), 209-214.
[36] Calucci, L., Capocchi, A., Galleschi, L., Ghiringhelli, S., Pinzino, C., Saviozzi, F. & Zandomeneghi, M. (2004). Antioxidants, free radicals, storage proteins, puroindolines, and proteolytic activities in bread wheat (Triticum aestivum) seeds during accelerated aging. Journal of Agricultural and Food Chemistry, 52 (13), 4274-4281.
[37] Palma, J. M., Sandalio, L. M., Corpas, F. J., Romero-Puertas, M. C., McCarthy, I. & del Río, L. A. (2002). Plant proteases, protein degradation, and oxidative stress: role of peroxisomes. Plant Physiology and Biochemistry, 40 (6-8), 521-530.
[38] Chen, C., Wang, R., Dong, S., Wang, J., Ren, C. X., Chen, C. P., et al. (2022). Integrated proteome and lipidome analysis of naturally aged safflower seeds varying in vitality. Plant Biol. 24 (2), 266-277.
[39] Zhang, L. & Becker, D. F. (2015). Connecting proline metabolism and signaling pathways in plant senescence. Frontiers in Plant Science, 6, 552.
[40] Ciacka, K., Tymiński, M., Gniazdowska, A. & Krasuska, U. (2020). Carbonylation of proteins—an element of plant ageing. Planta, 252 (1), 1-13.
[41] Dębska, K., Krasuska, U., Budnicka, K., Bogatek, R. & Gniazdowska, A. (2013). Dormancy removal of apple seeds by cold stratification is associated with fluctuation in H2O2, NO production and protein carbonylation level. Journal of plant physiology, 170 (5), 480-488.
)