Enhancing the Establishment of Black Cumin (Nigella sativa L.): The Role of Seed Improvement Techniques and Cultivation Substrate

Document Type : Original Article

Authors

1 Academic Center for Education, Culture and Research of Kashmar, Kashmar, Iran

2 Department of Crop Production and Plant Breeding, Faculty of Agriculture, Shahed University, Tehran, Iran

3 Department of Biotechnology and Plant Breeding, Faculty of Agriculture, Ferdowsi University, Mashahd, Iran

10.22126/atic.2026.12951.1243

Abstract

Seed coating is an economically viable method for seed enhancement, and biological seed treatments in the form of coating can substantially improve plant performance. This study, conducted in Bardaskan city, Razavi Khorasan province, Iran, aimed to evaluate the effects of seed enhancement and substrate on black cumin (Nigella sativa) and employed a two-factor factorial design based on a completely randomized design (CRD) and included three replications. The first factor included four treatments: control (field soil), field soil + farmyard manure (25% v/v), field soil + chemical fertilizers (ammonium sulfate as nitrogen source and triple superphosphate as phosphorus source), and field soil + biofertilizers. The second factor (seed pretreatment) consisted of four treatments: uncoated seeds (control), micronutrient-coated seeds, Azotobacter chroococcum -inoculated seeds, and combined micronutrient and A. chroococcum coating. The optimal treatment (A. chroococcum+ micronutrients under field soil + biofertilizer) resulted in 80% seed emergence, shoot length (5.08 cm), root length (3.27 cm), and a seedling vigor index of 6.68. This treatment also reduced oxidative stress markers, with hydrogen peroxide and malondialdehyde contents decreasing to 0.2 μmol g-1 FW and 15.24 nmol g-1 FW, respectively, while enhancing antioxidant capacity (86.6%) and phenolic content (87.4 mg GAE g-1 FW). Correlation and principal component analyses revealed a clear inverse relationship between oxidative stress markers and growth parameters, indicating that growth enhancement was achieved through increasing antioxidant capacity and phenolic content, and reducing oxidative stress. The substrate amended with biofertilizer consistently performed better than those treated with chemical fertilizers, emphasizing the significance of biological complexity rather than just focusing on nutrient supplementation. These findings show that integrated bio-fertilization strategies can fundamentally alter seedling physiology, offering a sustainable approach for medicinal plant cultivation that simultaneously enhances productivity, antioxidant properties, and environmental sustainability.

Graphical Abstract

Enhancing the Establishment of Black Cumin (Nigella sativa L.): The Role of Seed Improvement Techniques and Cultivation Substrate

Highlights

  • The integrated seed coating of Azotobacter and micronutrients within a biofertilizer-amended growth substrate was identified as the optimal treatment, significantly enhancing seedling emergence and seedling vigor index.
  • Seed coating with Azotobacter and micronutrients in cultivation substrate treatment fundamentally reprogrammed seedling physiology by significantly boosting antioxidant capacity and phenolic content, which in turn effectively reduced oxidative stress markers (H2O2 and MDA content).
  • Multivariate analysis revealed a clear inverse relationship between oxidative stress markers and plant growth parameters, establishing that the observed growth enhancement was mediated through the activation of robust antioxidant defense systems.
  • The study conclusively demonstrates that biofertilizer-amended substrates consistently surpass chemical fertilization, underscoring the critical role of biological complexity in sustainable cultivation over mere inorganic nutrient supplementation.

Keywords

Main Subjects

References

Abdel Latef A.A., Omer A.M., Badawy A.A., Osman M.S., Ragaey M.M. 2021. Strategy of salt tolerance and interactive impact of Azotobacter chroococcum and/or Alcaligenes faecalis inoculation on canola (Brassica napus L.) plants grown in saline soil. Plants 10(1): 110. https://doi.org/10.3390/plants10010110
Ali A.S., Elozeiri A.A. 2017. Metabolic processes during seed germination. In: Advances in Seed Biology. InTech. https://doi.org/10.5772/intechopen.70653
Ammar E.E., Rady H.A., Khattab A.M., Amer M.H., Mohamed S.A., Elodamy N.I., Al-Farga A., Aioub A.A. 2023. A comprehensive overview of eco-friendly bio-fertilizers extracted from living organisms. Environmental Science and Pollution Research 30(53): 113119-113137. https://doi.org/10.1007/s11356-023-30260-x
Asoegwu C.R., Awuchi C.G., Nelson K.C., Orji C.G., Nwosu O.U., Egbufor U.C., Awuchi C.G. 2020. A review on the role of biofertilizers in reducing soil pollution and increasing soil nutrients. Himalayan Journal of Agriculture 1(1): 34-38.
Atapattu A.J., Nuwarapaksha T.D., Dissanayaka D.M.N.S., Udumann S.S. 2025. Unraveling the role of plant-microbe interactions in sustainable agriculture. In: Babalola O.O., Amund O.O., Akanmu A.O. (eds) Ecofriendly Frontiers. Springer, Cham. https://doi.org/10.1007/978-3-031-98700-7_1
Bao J., Cai Y., Sun M., Wang G., Corke H. 2005. Anthocyanins, flavonols, and free radical scavenging activity of Chinese bayberry (Myrica rubra) extracts and their color properties and stability. Journal of Agricultural and Food Chemistry 53(6): 2327-2332. https://doi.org/10.1021/jf048312z
Colombo F., Paolo D., Cominelli E., Sparvoli F., Nielsen E., Pilu R. 2020. MRP transporters and low phytic acid mutants in major crops: main pleiotropic effects and future perspectives. Frontiers in Plant Science 11: 1301. https://doi.org/10.3389/fpls.2020.01301
Davar Zareii F., Roozbahani A., Hosnamidi A. 2014. Evaluation the effect of water stress and foliar application of Fe nanoparticles on yield, yield components and oil percentage of safflower (Carthamus tinctorius L.). International Journal of Advanced Biological and Biomedical Research 2(4): 11-20. https://www.ijabbr.com/article_7295.html 
Ekren S., Paylan I.C., Gokcol A. 2023. Seed quality improvement applications in black cumin seeds (Nigella sativa L.). Frontiers in Sustainable Food Systems 7: 1212958. https://doi.org/10.3389/fsufs.2023.1212958
Fazeli-Nasab B., Piri R., Rezaei A., Shakoory-Moghadam V., Parray J.A. 2025. The effect of elements and fertilizers on health and quality of soil, maternal plant nutrition, stress tolerance and increasing seed yield and quality. In: Parray J.A., Shameem N., Haghi A.K. (eds) Producing Healthy Food with Healthy Soils. Sustainable Landscape Planning and Natural Resources Management. Springer, Cham. https://doi.org/10.1007/978-3-031-82536-1_17
Forti C., Shankar A., Singh A., Balestrazzi A., Prasad V., Macovei A. 2020. Hydropriming and biopriming improve Medicago truncatula seed germination and upregulate DNA repair and antioxidant genes. Genes 11: 242. https://doi.org/10.3390/genes11030242
Gao C., El-Sawah A.M., Ismail Ali D.F., Alhaj Hamoud Y., Shaghaleh H., Sheteiwy M.S. 2020. The integration of bio and organic fertilizers improve plant growth, grain yield, quality and metabolism of hybrid maize (Zea mays L.). Agronomy 10(3): 319. https://doi.org/10.3390/agronomy10030319
Guo X., He Z., Ren H., Ren B., Zhang J., Liu P., Zhao B. 2025. Long-term effects of coated fertilizers: enhancing soil properties, optimizing microbial structures, and boosting soil fertility and crop productivity. Agriculture, Ecosystems & Environment 385: 109564. https://doi.org/10.1016/j.agee.2025.109564
Hamoud Y.A., Shaghaleh H., Sheteiwy M., Guo X., Elshaikh N.A., Khan N.U., Oumarou A., Rahim S.F. 2019. Impact of alternative wetting and soil drying and soil clay content on the morphological and physiological traits of rice roots and their relationships to yield and nutrient use-efficiency. Agricultural Water Management 223: 105706. https://doi.org/10.1016/j.agwat.2019.105706
Hichem H., Mounir D. 2009. Differential responses of two maize (Zea mays L.) varieties to salt stress: changes on polyphenols composition of foliage and oxidative damages. Industrial Crops and Products 30(1): 144-151. https://doi.org/10.1016/j.indcrop.2009.03.003
Hoseini A., Salehi A., Sayyed R.Z., Balouchi H., Moradi A., Piri R., Fazeli-Nasab B., Poczai P., Ansari M.J., Al Obaid S., Datta R. 2022. Efficacy of biological agents and fillers seed coating in improving drought stress in anise. Frontiers in Plant Science 13: 955512. https://doi.org/10.3389/fpls.2022.955512
Hosseini S.S., Nadjafi F., Asareh M.H., Rezadoost H. 2018. Morphological and yield related traits, essential oil and oil production of different landraces of black cumin (Nigella sativa) in Iran. Scientia Horticulturae 233: 1-8. https://doi.org/10.1016/j.scienta.2018.01.038
Hosseini-Moghaddam M., Moradi A., Piri R., Glick B.R., Fazeli-Nasab B., Sayyed R.Z. 2024. Seed coating with minerals and plant growth-promoting bacteria enhances drought tolerance in fennel (Foeniculum vulgare L.). Biocatalysis and Agricultural Biotechnology 58: 103202. https://doi.org/10.1016/j.bcab.2024.103202
Huang Y.C., Fan R., Grusak M.A., Sherrier J.D., Huang C.P. 2014. Effects of nano-ZnO on the agronomically relevant Rhizobium-legume symbiosis. Science of the Total Environment 497-498: 78-90. https://doi.org/10.1016/j.scitotenv.2014.07.100
Javed T., Afzal I., Shabbir R., Ikram K., Zaheer M.S., Faheem M., Ali H.H., Iqbal J. 2022. Seed coating technology: An innovative and sustainable approach for improving seed quality and crop performance. Journal of the Saudi Society of Agricultural Sciences 21(8): 536-545. https://doi.org/10.1016/j.jssas.2022.03.003
Kara N., Katar D., Baydar H. 2015. Yield and quality of black cumin (Nigella sativa L.) populations: the effect of ecological conditions. Turkish Journal of Field Crops 20(1): 9-14. https://doi.org/10.17557/.23190 
Kundan R., Pant G., Jadon N., Agrawal P.K. 2015. Plant growth promoting rhizobacteria: mechanism and current prospective. Journal of Fertilizers and Pesticides 6: 155. https://doi.org/10.4172/2471-2728.1000155
Maximize Market Research (MMR). 2025. Black cumin seed oil market: global industry analysis and forecast (2024–2030). https://www.maximizemarketresearch.com/market-report/global-black-cumin-seed-oil-market/83552/
Miljakovic D., Marinkovic J., Balesevic-Tubic S. 2020. The significance of Bacillus spp. in disease suppression and growth promotion of field and vegetable crops. Microorganisms 8(7): 1037. https://doi.org/10.3390/microorganisms8071037
Mirshekari B., Asadi Rahmani H., Mirmozafari Rodsari A. 2010. The effect of seed inoculation with Azospirillum strains and coating with microelements on seed yield and essence of cumin (Cuminum cyminum L.). Iranian Journal of Medicinal and Aromatic Plants Research 25(4): 470-481. (In Farsi). https://doi.org/10.22092/ijmapr.2010.7021
Motamednezhad M., Eslami S.V., Sayyari Zahan M.H., Mahmoodi S. 2016. The effect of enrichment with bio-fertilizers and three nutrients of iron, zinc and manganese on germination characteristics of ajowan (Carum copticum L.). Journal of Horticultural Science 29(4): 564-571. (In Farsi). https://doi.org/10.22067/jhorts4.v29i4.29951
Paravar A., Piri R., Balouchi H., Ma Y. 2023. Microbial seed coating: an attractive tool for sustainable agriculture. Biotechnology Reports 37: e00781. https://doi.org/10.1016/j.btre.2023.e00781
Piri R., Moradi A., Balouchi H., Salehi A. 2019. Improvement of cumin (Cuminum cyminum) seed performance under drought stress by seed coating and biopriming. Scientia Horticulturae 257: 108667. https://doi.org/10.1016/j.scienta.2019.108667
Piri R., Moradi A., Salehi A., Balouchi H.R. 2021. Effect of seed biological pretreatments on germination and seedling growth of cumin (Cuminum cyminum L.) under drought stress. Iranian Journal of Seed Science and Technology 9(4): 11-26. https://doi.org/10.22092/ijsst.2019.109182.1054
Rocha I., Ma Y., Souza-Alonso P., Vosátka M., Freitas H., Oliveira R.S. 2019. Seed coating: a tool for delivering beneficial microbes to agricultural crops. Frontiers in Plant Science 10: 1357. https://doi.org/10.3389/fpls.2019.01357
Samal K., Kar S., Trivedi S. 2019. Ecological floating bed (EFB) for decontamination of polluted water bodies: design, mechanism and performance. Journal of Environmental Management 251: 109550. https://doi.org/10.1016/j.jenvman.2019.109550
Seleiman M.F., Almutairi K.F., Alotaibi M., Shami A., Leonardo Battaglia M. 2021. Nano-fertilization as an emerging fertilization technique: why can modern agriculture benefit from its use? Plants 10(1): 2. https://doi.org/10.3390/plants10010002
Seyedi S.M., Rezvani Moghaddam P., Khajeh Hosseini M., Shahandeh H. 2016. Improving the physiological aspects of black seed (Nigella sativa L.) in calcareous soil: the role of seed phosphorus under the influence of mother plant nutrition. Iranian Journal of Seed Science and Technology 4(1): 27-38. (In Farsi). https://ijsst.areeo.ac.ir/article_106706.html?lang=en
Singh P., Vaishnav A., Liu H., Xiong C., Singh H.B., Singh B.K. 2023. Seed biopriming for sustainable agriculture and ecosystem restoration. Microbial Biotechnology 16(12): 2212-2222. https://doi.org/10.1111/1751-7915.14322
Srivastav A.L., Patel N., Rani L., Kumar P., Dutt I., Maddodi B.S., Chaudhary V.K. 2024. Sustainable options for fertilizer management in agriculture to prevent water contamination: A review. Environment, Development and Sustainability 26(4): 8303-8327. https://doi.org/10.1007/s10668-023-03117-z
Szerement J., Szatanik-Kloc A., Mokrzycki J., Mierzwa-Hersztek M. 2022. Agronomic biofortification with Se, Zn, and Fe: an effective strategy to enhance crop nutritional quality and stress defense—a review. Journal of Soil Science and Plant Nutrition 22(1): 1129-1159. https://doi.org/10.1007/s42729-021-00719-2
Velikova V., Yordanov I., Edreva A. 2000. Oxidative stress and some antioxidant systems in acid rain-treated bean plants: protective role of exogenous polyamines. Plant Science 151(1): 59-66. https://doi.org/10.1016/S0168-9452(99)00197-1
Weiss O., Minixhofer P., Werner N., Riedenbauer S., Olesko E., Lubke K., Scharf B., Pitha U. 2021. Alternative engineered soils and seed mixes used for seepage troughs. Land 10(11): 1152. https://doi.org/10.3390/land10111152
Wilson S.A., Roberts S.C. 2014. Metabolic engineering approaches for production of biochemicals in food and medicinal plants. Current Opinion in Biotechnology 26: 174-182. https://doi.org/10.1016/j.copbio.2014.01.006
Yimer E.M., Tuem K.B., Karim A., Ur-Rehman N., Farooq A. 2019. Nigella sativa L. (black cumin): a promising natural remedy for wide range of illnesses. Evidence-Based Complementary and Alternative Medicine 2019: 1528635. https://doi.org/10.1155/2019/1528635
Agrotechniques in Industrial Crops

Articles in Press, Accepted Manuscript
Available Online from 27 August 2026

Files

Share

How to cite

Statistics