Optimizing Germination Protocols for Citrullus colocynthis: A Study on Seed Dormancy Breakthrough through Chemical and Physical Treatments
DOI:
https://doi.org/10.14720/aas.2025.121.3.22753Keywords:
breaking seed dormancy, physiological dormancy, diurnal temperature alternation, gibberellic acidAbstract
Bitter apple (Citrullus colocynthis), a desert plant with medicinal properties, typically exhibits low seed germination due to dormancy. This study investigated physical and chemical treatments to improve C. colocynthis seed germination. A completely randomized design with factorial arrangement and three replications was used to test the effects of gibberellic acid (GA3) at concentrations of 0, 10, 50, 100, and 150 ppm, under four diurnal temperature and light (TL) regimes: (1) 20°C (12h light/dark), (2) 25°C (12h light/dark), (3) 35°C (12h light) / 15°C (12h dark), and (4) 30°C (12h light) / 20°C (12h dark). Results showed that TL regime, GA3, and their interaction significantly affected all measured germination parameters. The highest germination percentage, relative germination percentage, mean daily germination, germination index, germination value, and seedling dry mass were achieved with 100 ppm GA3 under the 35-15°C regime. The study concluded that average germination speed, germination time variation coefficient, germination speed coefficient, and synchronization index were unreliable for assessing seed quality under low germination conditions, suggesting that physiological dormancy in Bitter Apple seeds may stem from embryo immaturity, inhibitory factors, or both.
Metrics
Downloads
References
Afzal, O., Hassan, F., Ahmed, M., Shabbir, G., & Ahmed, S. (2022). Temperature affects germination indices of safflower (Carthamus tinctorius L.). Journal of Animal & Plant Sciences, 32(6), 1691-1702. https://doi.org/:10.36899/JAPS.2022.6.0577
Al-Turki, T.A., Davy, A.J., Al-Ammari, B.S., & Basahi, M.A. (2022). Seed germination characteristics of some medicinally important desert plants from the Arabian Peninsula. Journal of Arid Environments, 198, 104689. https://doi.org/:10.1016/j.jaridenv.2021.104689
Ali-Rachedi, S., Bouinot, D., Wagner, M.-H., Bonnet, M., Sotta, B., Grappin, P., & Jullien, M. (2004). Changes in endogenous abscisic acid levels during dormancy release and maintenance of mature seeds: studies with the Cape Verde Islands ecotype, the dormant model of Arabidopsis thaliana. Planta, 219(3), 479-488. https://doi.org/:10.1007/s00425-004-1251-4
Amini, E., Mehrabi, A.A., & Alizadeh, Y. (2019). Effect of gibberellic acid on germination and seedling growth in different sorghum genotypes under salt stress condition. Seed Research, 32(9), 35-44.
Asaadi, A.m., & Heshmati, g. (2015). The effect of different treatments on seeds dormancy and germination of Thymus transcaucasicus Ronn. and Zataria moltiflora Boiss. Journal of Plant Research (Iranian Journal of Biology), 28(1), 12-22.
Aziz, A., et al. (2023). Novel copper oxide phyto-nanocatalyst utilized for the synthesis of sustainable biodiesel from Citrullus colocynthis seed oil. Processes, 11(6), 1857. https://doi.org/:10.3390/pr11061857
Banerjee, A., & Roychoudhury, A. (2020). Gibberellic acid-priming promotes fluoride tolerance in a susceptible Indica rice cultivar by regulating the antioxidant and phytohormone homeostasis. Journal of Plant Growth Regulation, 39(4), 1476-1487. https://doi.org/:10.1007/s00344-020-10110-7
Baskin, C.C., & Baskin, J.M. (2014). Chapter 12 - Biogeographical and Evolutionary Aspects of Seed Dormancy. Pages 1005-1073 in Baskin CC, Baskin JM, eds. Seeds (Second Edition). San Diego: Academic Press.
Begum, N., Wang, L., & Zhao, T. (2022). Effects of temperature regimes on seed germination and early growth of different soybean cultivars. International Journal of Applied and Experimental Biology, 1(2),75-85. https://doi.org/:10.56612/ijaeb.v1i2.15
Bhatla, S.C., & Lal, M.A. (2023). Seed Dormancy and Germination. Pages 625-640. Plant Physiology, Development and Metabolism. Singapore: Springer Nature Singapore.
Chauhan, A., AbuAmarah, B.A., Kumar, A., Verma, J.S., Ghramh, H.A., Khan, K.A., & Ansari, M.J. (2019). Influence of gibberellic acid and different salt concentrations on germination percentage and physiological parameters of oat cultivars. Saudi Journal of Biological Sciences, 26(6), 1298-1304. https://doi.org/:10.1016/j.sjbs.2019.04.014
Czabator, F.J. (1962). Germination value: An index combining speed and completeness of pine seed germination. Forest Science, 8(4), 386-396. https://doi.org/:10.1093/forestscience/8.4.386
Edelstein, M., & Kigel, J. (1990). Seed germination of melon (Cucumis melo) at sub- and supra-optimal temperatures. Scientia Horticulturae, 45(1), 55-63. https://doi.org/:10.1016/0304-4238(90)90068-P
El-Keblawy, A., Shabana, H.A., Navarro, T., & Soliman, S. (2017). Effect of maturation time on dormancy and germination of Citrullus colocynthis (Cucurbitaceae) seeds from the Arabian hyper-arid deserts. BMC Plant Biology, 17(1), 263. https://doi.org/:10.1186/s12870-017-1209-x
Esanejad, N.S., Omidi, H., & Paraver, A. (2015). Effect of safflower seeds priming with abscisic and gibberellic acid on germination indices in salinity stress condition. Agroecology Journal, 11(4), 1-11. https://doi.org/:10.22034/aej.2016.521052
Gairola, K., Nautiyal, A., & Dwivedi, A. (2011). Effect of temperatures and germination media on seed germination of Jatropha curcas Linn. Advances in Bioresearch, 2(2), 66-71.
Gharehmatrossian, S., Popov, Y., & Ghorbanli, M. (2014). Seed germination, dormancy breaking techniques of Citrullus colocynthis (L.) Schrab plant. Iranian Journal of Plant Physiology, 4(4), 1167-1171. https://doi.org/:10.30495/ijpp.2014.540662
Guragain, R.P., Baniya, H.B., Pradhan, S.P., Pandey, B.P., Shrestha, B., Fronczak, M., Kierzkowska-Pawlak, H., & Subedi, D.P. (2023). Growth enhancement of radish sdeed induced by low-temperature argon plasma. Plasma Chemistry and Plasma Processing, 43(1), 111-137. https://doi.org/:10.1007/s11090-022-10291-x
Huarte, H.R., & Benech-Arnold, R.L. (2010). Hormonal nature of seed responses to fluctuating temperatures in Cynara cardunculus (L.). Seed Science Research, 20(1), 39-45. https://doi.org/:10.1017/S0960258509990249
Hung, L.Q., Hong, T.D., & Ellis, R.H. (2004). Factors influencing the germination of myrtle (Lagerstroemia speciosa (L.) Pers. and L. floribunda Jack) seeds. Seed Science and Technology, 32(1), 35-41. https://doi.org/:10.15258/sst.2004.32.1.05
Jafarizadeh, A., Raeisi, S.A., Ghassab-Abdollahi, N., Yarani, R., Araj-Khodaei, M., & Mirghafourvand, M. (2022). Effect of Citrullus colocynthis on glycemic factors and lipid profile in type II diabetic patients: a systematic review and meta-analysis. Journal of Diabetes & Metabolic Disorders, 21(2), 1785-1795. https://doi.org/:10.1007/s40200-022-01045-9
Javaid, M.M., Florentine, S., Ali, H.H., & Weller, S. (2018). Effect of environmental factors on the germination and emergence of Salvia verbenaca L. cultivars (verbenaca and vernalis): An invasive species in semi-arid and arid rangeland regions. PLoS One, 13(3), e0194319. https://doi.org/:10.1371/journal.pone.0194319
Javaid, M.M., et al. (2022). Influence of environmental factors on seed germination and seedling characteristics of perennial ryegrass (Lolium perenne L.). Scientific Reports, 12(1), 9522. https://doi.org/:10.1038/s41598-022-13416-6
Joshi, K., Kotadiya, P., Singh, A., Shrimali, G., & Rami, E. (2023). Effect of different pre-treatments on seed germination and seedling growth of Adenanthera pavonina. Research on Crops, 23(1), 229-234. https://doi.org/:10.31830/2348-7542.2022.032
Kahrom, N., Farzam, M., & Mesdaghi, M. (2022). Autecology of colocynth (Citrullus colocynthis L. Schrad) in Gonabad Desert, Iran. Journal of Rangeland Science, 12(2), 129-140. https://doi.org/:10.30495/rs.2022.682499
Kharshiing, E., Sreelakshmi, Y., & Sharma, R. (2019). The Light Awakens! Sensing Light and Darkness. Pages 21-57 in Sopory S, ed. Sensory Biology of Plants. Singapore: Springer Singapore.
Klupczyńska, E.A., & Pawłowski, T.A. (2021). Regulation of seed dormancy and germination mechanisms in a changing environment. International Journal of Molecular Sciences, 22(3), 1357. https://doi.org/:10.3390/ijms22031357
Kumar, B., Yadav, R., Singh, S.C., & Singh, H.P. (2016). Seed germination behavior of Withania spp. under different temperature regimes. Journal of Crop Improvement, 30(3), 287-292. https://doi.org/:10.1080/15427528.2016.1151849
Kumari, N., Rai, P.K., Bara, B.M., & Singh, I. (2017). Effect of halo priming and hormonal priming on seed germination and seedling vigour in maize (Zea mays L) seeds. Journal of Pharmacognosy and Phytochemistry, 6(4), 27-30.
Li, Q.-Y., Munawar, M., Saeed, M., Shen, J.-Q., Khan, M.S., Noreen, S., Alagawany, M., Naveed, M., Madni, A., & Li, C.-X. (2022). Citrullus colocynthis (L.) Schrad (Bitter Apple Fruit): Promising traditional uses, pharmacological effects, aspects, and potential applications. Frontiers in Pharmacology, 12. https://doi.org/:10.3389/fphar.2021.791049
Nakamura, S., Okasako, Y., & Yamada, E. (1955). Effect of light on the germination of vegetable seeds. Journal of the Japanese Society for Horticultural Science, 24(1), 17-28. https://doi.org/:10.2503/jjshs.24.17
Nedunchezhiyan, V., Palanivel, M., Akhila Jabeen, P.A., Thangavel, P., Ramakrishnan, B., Velusamy, M., Muthusamy, S., & Abasianyanga Edm, I. (2023). Effects of gibberellic acid on seed dormancy of black gram (Vigna mungo L.). 3.
Nejad, S.R., Bistgani, Z.E., & Barker, A.V. (2022). Enhancement of seed germination of yarrow with gibberellic acid, potassium nitrate, scarification, or hydropriming. Journal of Crop Improvement, 36(3), 335-349. https://doi.org/:10.1080/15427528.2021.1968553
Nogueira, F., Bezerra, M., Pinheiro, C., Brito, S., & Filho, S. (2014). Piptadenia stipulacea (Benth.) Ducke Seed germination in response to temperature, light and water stress. American Journal of Plant Sciences, 5(26), 3796-3904. https://doi.org/:10.4236/ajps.2014.526397
Omidi, H., Jafarzadeh, L., & Shirpoor, S. (2012). Adjustment effects of drought stress on the herbal plantago (Plantago psyllium L.) with salicylic acid. Paper presented at First national congress in medicinal plants, Kish Island, Iran.
Ozden, E., Light, M.E., & Demir, I. (2021). Alternating temperatures increase germination and emergence in relation to endogenous hormones and enzyme activities in aubergine seeds. South African Journal of Botany, 139, 130-139. https://doi.org/:10.1016/j.sajb.2021.02.015
Payal, K., Maikhuri, R.K., Rao, K.S., & Kandari, L.S. (2014). Effect of gibberellic acid- and water-based pre-soaking treatments under different temperatures and photoperiods on the seed germination of Allium stracheyi Baker: An endangered alpine species of Central Himalaya, India. Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology, 148(6), 1075-1084. https://doi.org/:10.1080/11263504.2013.823131
Pellizzaro, V., Omura, M.S., Furlan, F.F., da Costa, D.S., Ponce, R.M., Freiria, G.H., Neto, H.F.I., & Takahashi, L.S.A. (2019). Physiological potential of Physalis peruviana L. seeds under different temperatures and light wavelengths. Semina: Ciências Agrárias, 40(5), 1737-1746. https://doi.org/:10.5433/1679-0359.2019v40n5p1737
Perveen, S., Ashfaq, H., Ambreen, S., Ashfaq, I., Kanwal, Z., & Tayyeb, A. (2021). Methanolic extract of Citrullus colocynthis suppresses growth and proliferation of breast cancer cells through regulation of cell cycle. Saudi Journal of Biological Sciences, 28(1), 879-886. https://doi.org/:10.1016/j.sjbs.2020.11.029
Ramirez, A.H.M., Jhala, A.J., & Singh, M. (2014). Factors affecting germination of citronmelon (Citrullus lanatus var. citroides). Weed Science, 62(1), 45-50. https://doi.org/:10.1614/WS-D-13-00041.1
Rath, D., Dash, A.K., & Padhi, S. (2023). Effect of low frequency (50 Hz) electromagnetic field on germination process of wheat (Triticum aestivum L.). Scholars Academic Journal of Biosciences, 11(4), 147-158. https://doi.org/:10.36347/sajb.2023.v11i04.003
Rhie, Y.H., Lee, S.Y., & Kim, K.S. (2015). Seed dormancy and germination in Effersonia dubia (Berberidaceae) as affected by temperature and gibberellic acid. Plant Biology, 17(2), 327-334. https://doi.org/:10.1111/plb.12235
Saberi, M., Niknahad, H., Heshmati, G., Barani, H., & Shahriyaei, A. (2017). Investigating morphological characteristics and effect of different treatments on improving seed germination of two Citrullus colocynthis stands in Sisitan and Balouchestan. Rangeland, 11(3), 353-364.
Saberi, M., shahriari, A., & bozorgmehr, M. (2020). Effect of priming and salinity on the germination and seedling growth of Stipagrostis plumosa. Journal of Range and Watershed Managment, 73(3), 513-524. https://doi.org/:10.22059/jrwm.2020.277900.1364
Saberi, M., Shahriari, A., Tarnian, F., & Noori, S. (2011). Comparison the effect of different treatments for breaking seed dormancy of Citrullus colocynthis. Journal of Agricultural Science, 3(4), 62–67. https://doi.org/:10.5539/jas.v3n4p62
Sarwar, G., Anwar, T., Qureshi, H., Younus, M., Hassan, M.W., Sajid-ur-Rehman, M., Khalid, F., Faiza, Zaman, W., & Soufan, W. (2024). Optimizing germination: comparative assessment of various growth media on dragon fruit germination and early growth. BMC Plant Biology, 24(1), 533. https://doi.org/:10.1186/s12870-024-05247-6
Shahmoradi, S., Chaichi, M., Mozafari, J., Mazaheri, D., & Sharif Zadeh, F. (2015). Breaking seed dormancy in Hordeum spontaneum L. Journal of Plant Research (Iranian Journal of Biology), 27(5), 872-884. https://doi.org/:27511
Silva, D.Y.B.d.O., Silva, A.M.P.d., Farias, S.G.G.d., Silva, R.B.e., Matos, V.P., & Silva, L.S.d. (2018). Substrate and temperature on germination and performance of Albizia niopoides Benth. Seedlings. Ciência Rural, 48(3), e20161043. https://doi.org/:10.1590/0103-8478cr20161043
Silva, E.C.D., Villa, F., Silva, D.F.D., Possenti, J.C., Silva, L.S.D., & Ritter, G. (2021). Araticum accessions: Effect of gibberellic acid concentrations and soaking times on seed dormancy overcoming. Revista Caatinga, 34(3), 614-620. https://doi.org/:10.1590/1983-21252021v34n313rc
Thanos, C.A., & Mitrakos, K. (1992). Watermelon seed germination. 1. Effects of light, temperature and osmotica. Seed Science Research, 2(3), 155-162. https://doi.org/:10.1017/S0960258500001288
Tsegay, B.A., & Andargie, M. (2018). Seed priming with gibberellic acid (GA3) alleviates salinity induced inhibition of germination and seedling growth of Zea mays L., Pisum sativum var. abyssinicum A. Braun and Lathyrus sativus L. Journal of Crop Science and Biotechnology, 21(3), 261-267. https://doi.org/:10.1007/s12892-018-0043-0
Vicente, M.J., Martínez-Díaz, E., Martínez-Sánchez, J.J., Franco, J.A., Bañón, S., & Conesa, E. (2020). Effect of light, temperature, and salinity and drought stresses on seed germination of Hypericum ericoides, a wild plant with ornamental potential. Scientia Horticulturae, 270, 109433. https://doi.org/:10.1016/j.scienta.2020.109433
Yan, A., & Chen, Z. (2020). The control of seed dormancy and germination bytemperature, light and nitrate. The Botanical Review, 86(1), 39-75. https://doi.org/:10.1007/s12229-020-09220-4
Yari, L., Hashemi, S., & Hasani, F. (2014). Effects of different storage temperatures on sunflower seeds vigor. Iranian Journal of Seed Research, 1(1), 11-19. https://doi.org/:10.29252/yujs.1.1.11
Yousefi, F., Sihampoosh, A., Bakhshandeh, A., & Mousavi, S.A. (2021). The effect of hormone seed priming using gibberellic acid on seed germination characteristics and seedling growth of coneflower (Echinacea purpurea). Iranian Journal of Seed Research, 8(1), 173-188. https://doi.org/:10.52547/yujs.8.1.173
Zhang, T., Yan, Y., Li, C., Liu, J., Yin, D., Xiong, X., Liu, W., & Yang, Y. (2021). Influence of illumination time and soil moisture on seed germination and seedling establishment of Magnolia sprengeri Pamp. HortScience, 56(11), 1381-1386. https://doi.org/:10.21273/hortsci16144-21
Zheng, Y., Xie, Z., Gao, Y., Jiang, L., Xing, X., Shimizu, H., & Rimmington, G.M. (2005). Effects of light, temperature and water stress on germination of Artemisia sphaerocephala. Annals of Applied Biology, 146(3), 327-335. https://doi.org/:10.1111/j.1744-7348.2005.040039.x
Zhu , G., An, L., Jiao, X., Chen, X., Zhou, G., & McLaughlin, N. (2019). Effects of gibberellic acid on water uptake and germination of sweet sorghum seeds under salinity stress. Chilean Journal of Agricultural Research, 79(3), 415-424. https://doi.org/:10.4067/S0718-58392019000300415
Downloads
Published
Issue
Section
License
Copyright (c) 2025 Nasir ABADDAR, Hassanali NAGHDI BADI, Majid AMINI DEHAGHI, Elias SOLTANI, Alireza REZAZADEH

This work is licensed under a Creative Commons Attribution 4.0 International License.