Abstract
Lentil (Lens culinaris Medik.) is a nutritionally valuable pulse crop that enhances food security and soil fertility through biological nitrogen fixation. However, its productivity is often constrained by poor germination and uneven seedling establishment. This study investigated biochar and humic acid as sustainable seed-priming agents to improve lentil seed performance. After preliminary optimization, the most effective concentrations of both agents were evaluated under controlled laboratory conditions. Germination percentage, seedling length, vigour indices, chlorophyll content, and oxidative stress response (DAB assay) were assessed. Both treatments significantly enhanced seed and seedling traits compared with unprimed controls (p < 0.05). Biochar priming increased germination by 14.7%, seedling length by 26.3%, and vigour index by 32.5% over control, alongside higher chlorophyll content and reduced hydrogen peroxide accumulation, indicating improved physiological efficiency and stress mitigation. Humic acid also improved performance, though to a lesser degree. The results demonstrate that biochar and humic acid priming enhance seed vigour and metabolic activity by promoting stress regulation and efficient energy utilization. Overall, biochar emerged as the superior and eco-friendly option for improving germination and early growth in lentil, offering a practical approach for sustainable pulse production and climate-resilient agriculture.
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References
Dixit, G. P., Kumar, A. & Parihar, A. K. Variability for harvest index and biomass in lentil (Lens culinaris Medik) varieties. Legume Research-An Int. J. 40 (6), 1093–1096. https://doi.org/10.18805/LR-3705 (2017). doi: 10.18805/LR-3705.
Tripathi, K. et al. Agro-morphological characterization of lentil germplasm of Indian National genebank and development of a core set for efficient utilization in lentil improvement programs. Front. Plant. Sci. 12, 751429. https://doi.org/10.3389/fpls.2021.7514 (2022).
FAO (Food and Agriculture Organization). Crop production and trade data. http://www.fao.org/faostat/en/#home29 (2022).
Ali, M. & Gupta, S. Carrying capacity of Indian agriculture: pulse crops. Curr. Scie. 874–881. https://doi.org/10.18805/LR-4632 (2012).
Kumar, J. et al. Heat tolerance in lentil under field conditions. Legume Genomics Genet. 7(1). https://doi.org/10.5376/lgg.2016.07.0001 (2016).
Okçu, G., Kaya, M. D. & Atak, M. Effects of salt and drought stresses on germination and seedling growth of pea (Pisum sativum L). Turkish J. Agric. Forestry. 29 (4), 237–242. https://doi.org/10.4236/as.2015.67064 (2005).
Sinha, S. et al. Plant growth–promoting traits of culturable seed Microbiome of citrus species from purvanchal himalaya. Front. Plant. Sci. 14, 1104927. https://doi.org/10.3389/fpls.2023.1104927 (2023).
Sita, K. et al. Identification of high-temperature tolerant lentil (Lens culinaris Medik.) genotypes through leaf and pollen traits. Front. plant. sci. 8, 744. (2017).
Rao, D. et al. Unveiling the potential of silicic and humic acid priming in alleviating salinity stress on lentil (Lens culinaris) seed germination in a hydroponic system. J. Food Legumes. 37 (3), 291–296. https://doi.org/10.3390/plants12203539 (2024).
Rao, D. et al. Silicic and humic acid priming improves Micro-and macronutrient Uptake, salinity stress tolerance, seed quality, and physio-biochemical parameters in lentil (Lens culinaris spp. culinaris). Plants 12 (20). https://doi.org/10.3390/plants12203539 (2023).
Lehmann, J. et al. Biochar effects on soil biota–a review. Soil biol and biochemistry, 43(9), 1812–1836. (2011). https://doi.org/10.1016/j.soilbio.2011.04.022. Rao, D., Yadav, S., Choudhary, R., Sushkova, S., Sachan, C. P., & Yadav, S. K. Enhancing of Early Seedling Vigour (ESV) parameters in Lentils through integrated priming with silicic and humic acid. (Eurasian J. Soil Sci,) 14(2), 157–167. https://doi.org/10.3390/plants12203539. (2025).
Haghaninia., M., Javanmard, A., Rasouli, F., Radicetti, E. & Mashhouri, S. M. Optimizing camelina performance under drought conditions through the combined action of Nanochitosan-Encapsulated Rosemary oil and arbuscular mycorrhizal Fungl. Biocat Agri Biotechnol. 103682. https://doi.org/10.1016/j.bcab.2025.103682 (2025).
Bhuker, A., Jakhar, S. S. & Singh, N. Assess the effect of natural ageing and storage containers on seed quality of tomato seeds. J. Agric. Ecol. Res. Int. 14, 131–134. https://doi.org/10.58628/JAE-2214-218 (2022).
Abdul-Baki, A. A. & Anderson, J. D. Vigor determination in soybean seed by multiple criteria. Crop Sci. 13 (6), 630–633. https://doi.org/10.2135/cropsci1973.0011183X001300060013x (1973).
Daudi, A. & O’brien, J. A. Detection of hydrogen peroxide by DAB staining in Arabidopsis leaves. Bio-protocol 2(18), e263–e263. https://doi.org/10.1007/s40502-023-00775-z (2012).
Ajouri, A., Asgedom, H. & Becker, M. Seed priming enhances germination and seedling growth of barley under conditions of P and Zn deficiency. J. Plant Nutr. Soil Sci. 167(5), 630–636. https://doi.org/10.1002/jpln.200420425 (2004).
Zhang, K., Wang, Y., Mao, J. & Chen, B. Effects of Biochar nanoparticles on seed germination and seedling growth. Enviorn. Pollut. 256, 113409. https://doi.org/10.1016/j.envpol.2019.113409 (2020).
Rao, D., Yadav, S., Choudhary, R. & Singh, D. Integrated application of silicic and humic acid seed priming for enhanced germination and yield of lentil (Lens culinarisL). Legume Research-An Int. J. 1(5). https://doi.org/10.18393/ejss.1646812 (2024).
Zou, Z. et al. Response of plant root growth to Biochar amendment: a meta-analysis. Agronomy 11 (12), 2442. https://doi.org/10.1016/j.cej.2025.160812 (2021).
Egamberdieva, D., Li, L., Ma, H., Wirth, S. & Bellingrath-Kimura, S. D. Soil amendment with different maize biochars improves Chickpea growth under different moisture levels by improving symbiotic performance with mesorhizobium ciceri and soil biochemical properties to varying degrees. Front. Microbiol. 10, 2423. https://doi.org/10.3389/fmicb.2019.02423 (2019).
Reyes-Cabrera, J. et al. Biochar changes shoot growth and root distribution of soybean during early vegetative stages. Crop Sci. 57 (1), 454–461. https://doi.org/10.2135/cropsci2016.01.0075 (2017).
Jahan, M. et al. Effects of mesorhizobium ciceri and Biochar on the growth, nodulation and antifungal activity against root pathogenic fungi in Chickpea (Cicer arietinum L). J. Plant. Pathol. Microbiol. 11, 520. https://doi.org/10.35248/2157- (2020).
Haghaninia, M., Mashhouri, S. M., Najafifar, A., Soleimani, F. & Mirzaei, A. Impact of silicon nanoparticle priming on metabolic responses and seed quality of Chia (Salvia Hispanica L.) under salt stress. Food Bioscience. 65, 106119. https://doi.org/10.1016/j.fbio.2025.106119 (2025).
Zhang, K. et al. Biochar coating as a cost-effective delivery approach to promoting seed quality, rice Germination, and seedling establishment. Plants 12 (22), 3896. https://doi.org/10.3390/plants12223896 (2023).
Abbas, A. et al. The efficacy of organic amendments on maize productivity, soil properties and active fractions of soil carbon in organic-matter deficient soil. Span. J. Soil. Sci. 14, 12814 (2024).
1080/15226514.2019.1644286.
Poomani, S. et al. Seed priming with humic acid modifies seedling Vigor and biochemical response of lentilunder heat stress conditions. Turk. J. Abric. 47 (6), 1043–1057. https://doi.org/10.55730/1300-011X.3147 (2023).
Murtaza, G. et al. Biochar enhances the growth and physiological characteristics of medicago sativa, Amaranthus caudatus and Zea Mays in saline soils. BMC Plant. Biol. 24 (1), 304. https://doi.org/10.3390/separations10030197 (2024).
Obadi, A. et al. Effect of Biochar application on morpho-physiological traits, yield, and water use efficiency of tomato crop under water quality and drought stress. Plants 12 (12), 2355. https://doi.org/10.3390/plants12122355 (2023).
Adetunji, A. E., Adetunji, T. L., Varghese, B., Pammenter, N. W. & Sershen, & Oxidative stress, ageing and methods of seed invigoration: An overview and perspectives. Agronomy 11 (12), 2369. https://doi.org/10.3390/agronomy11122369 (2021).
Tan, L. et al. Effect of three artificial aging techniques on physicochemical properties and Pb adsorption capacities of different biochars. Sci. Total Environ. 699, 134223. https://doi.org/10.1007/s42773-025-00460-4 (2020).
Hasanuzzaman, M. et al. Application of Biochar and humic acid improves the physiological and biochemical processes of rice (Oryza sativa L.) in conferring plant tolerance to arsenic-induced oxidative stress. Environ. Sci. Pollut. Res. 31(1), 1562–1575. https://doi.org/10.1007/s11356-023-31119-x (2024).
Mehdizadeh, L., Farsaraei, S. & Moghaddam, M. Biochar application modified growth and physiological parameters of ocimum ciliatum L. and reduced human risk assessment under cadmium stress. J. Hazard. Mater. 409, 124954. https://doi.org/10.1016/j.jhazmat.2020.124954 (2021).
Zhao, X., Chen, Y., Hu, J., Wang, H., Ye, Z., Zhang, J., Chen, Z. Efficacy of nitrate and biochar@birnessite composite microspheres for simultaneous suppression of As(III) mobilization and greenhouse gas emissions in flooded paddy soils. Environ. Res. 279, 121757. https://doi.org/10.1016/j.envres.2025.121757. (2025).
Zeng, Y., Wang, H., Hu, J., Zhang, J., Wang, F., Wang, T., Chen, Z. Illuminated fulvic acid stimulates denitrification and As(III) immobilization in flooded paddy soils via an enhanced biophotoelectrochemical pathway. Sci. Total Environ. 912, 169670. https://doi.org/10.1016/j.scitotenv.2023.169670. (2024).
Acknowledgements
The authors wish to thank the AIOA (AMITY Institute of Organic AGRICULTRE) Amity University Uttar Pradesh, Noida (India), Crop Nanobiology and Molecular Stress Physiology Lab for providing the required necessary facilities to conduct the studies and I express my heartfelt gratitude to my esteemed faculty guide, Dr. Deepak Rao, for his unwavering support, valuable guidance, and constant encouragement throughout my research work.
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Conceptualization, DR; methodology, DR, software, DR; validation, DR; formal analysis and KB; investigation, DR and KB; data curation, DR, KB and SC; writing—original draft preparation, KB; writing—review and editing, KB, DR, NP, RK, AAL, and MR; visualization, D.R.; supervision, DR. All authors have read and agreed to the published version of the manuscript.
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This study adhered to all relevant institutional, national, and international guidelines and legislation in India. Plant materials were collected was taken from ICAR – IARI, New Delhi. Permission for the collection of these plant materials was obtained from Amity Institute of Organic Agriculture, Amity University, which does not require special permits for academic research without commercial use. All plant materials used in the study were in compliance with Indian regulations and legislation.
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Chauhan, S., Bhardwaj, K., Rao, D. et al. Enhanced seed quality and physio-biochemical parameters in lentil through biochar and humic acid- based seed priming. Sci Rep (2026). https://doi.org/10.1038/s41598-026-39380-z
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DOI: https://doi.org/10.1038/s41598-026-39380-z
