Auxin-Induced Rooting and Seedling Quality Enhancement in Seedless Lemon (Citrus limon L.) under Nursery Conditions

Marium Khaskheli; Noor-Un-Nissa Memon; Afifa Talpur; Iqra Baloch; Muhammad Nawaz Baloch

Auxin-Induced Rooting and Seedling Quality Enhancement in Seedless Lemon (Citrus limon L.) under Nursery Conditions

Authors

Marium Khaskheli Department of Horticulture, Sindh Agriculture University Tandojam, Pakistan
Noor-Un-Nissa Memon Professor, Department of Horticulture, Sindh Agriculture University Tandojam, Pakistan
Afifa Talpur Department of Horticulture, Sindh Agriculture University, Tandojam, Pakistan
Iqra Baloch Department of Horticulture, Sindh Agriculture University Tandojam, Pakistan
Muhammad Nawaz Baloch Department of Horticulture, Sindh Agriculture University Tandojam, Pakistan

Keywords:

Asexual propagation, Auxins, Seedless lemon

Abstract

Seedless lemons (Citrus limon L.) are valued at the global level for the character of seedless. However, these cultivars are often characterised by poor rooting capability and low survival rates. In order to resolve this issue, the present study was conducted during the spring season of 2024 at the Nursery of Sindh Agriculture University (SAU), Tandojam, to evaluate the influence of auxin treatments on the sprouting and seedling growth of seedless lemon cuttings. A factorial experiment in a Completely Randomised Design (CRD) was laid out with four replications, comprising two varieties (Persian lime and Malaysian lemon) and four treatments: IBA gel dip, NAA powder dip, IBA gel + NAA powder dip, and control. All the seedling-related parameters, such as sprouting percentage, number of sprouts, days to sprouting, rooting percentage, root depth, seedling quality index, sturdiness quotient, shoot biomass and root biomass, were significantly affected by the auxin treatments. However, varieties and their interactive effect with auxin treatments were only significant for a few sprouting and rooting-related parameters. The statistical results revealed that the maximum sprouting percentage, number of sprouts per cutting, rooting percentage and minimum days to sprouting were observed in Persian lemon with the NAA powder dip method. The Dickson Quality Index, Sturdiness quotient, and biomass of shoot and root were not significantly affected by the varieties or their interactive effect with auxin treatments. However, Auxin as an independent factor had significant effects on all these parameters. The highest DQI, SQ, and biomass of shoot and root were observed with NAA powder dip treatments. However, the Number of sprouts and root depth traits were significantly influenced by treatments alone, with IBA gel dip resulting in more sprouts and the deepest roots. The findings suggest that NAA powder dip is the most effective treatment for enhancing rooting and overall seedling vigour in seedless lemon cuttings, particularly in the Persian variety.

References

Ahmed, S., Jaffar, M. A., Ali, N., Ramzan, M., & Habib, Q. (2017). Effect of Naphthalene Acetic Acid on the sprouting and rooting of the stem cuttings of grapes. Science Letters, 5(3), 225-232.

Ali, S., Khan, A. S., Raza, S. A., & Rehman, R. N. U. (2013). Innovative breeding methods to develop seedless citrus cultivars. International Journal of Biosciences, 3(8), 191-201.

Alotaibi, M. M., El Nagy, M. M., Almuziny, M., Alsubeie, M. S., Abo-Zeid, A. A., Alzuaibr, F. M., & Awad-Allah, M. M. (2024). The effect of foliar application with Naphthalene Acetic Acid and potassium nitrate on the growth, sex ratio, and productivity of cucumbers (Cucumis sativas L.) under high temperatures in semi-arid areas. Agronomy, 14(6), 1202.

Ayaz, N., Aman, F., Saleem, S., Rehman, M., & Fahim, M. (2021). Olive cuttings as affected by different concentrations of indole butyric acid. [Journal name missing].

Bhatt, B. B., & Tomar, Y. K. (2011). Effect of IBA and growing conditions on vegetative performance of Citrus aurantifolia (Swingle) cuttings. Journal of Hill Agriculture, 2(1), 98-101.

Binotto, A. F., Lúcio, A. D. C., & Lopes, S. J. (2010). Correlations between growth variables and the Dickson quality index in forest seedlings. Cerne, 16, 457-464. https://doi.org/10.1590/S0104-77602010000400005

Budiman, B., Sudrajat, D. J., Lee, D. K., & Kim, Y. S. (2015). Effect of initial morphology on field performance in white Jabon seedlings at Bogor, Indonesia. Forest Science and Technology, 11(4), 206-211.

Cohen, J. D., Slovin, J. P., & Hendrickson, A. M. (2024). Auxin: A central regulator of plant growth and development. The Plant Cell, 36(5), 1410–1425.

Dhand, A., Kaur, V., & Kaur, A. (2019). Effect of IBA and sucrose on the performance of cuttings in the pear cv. Patharnakh. International Journal of Current Microbiology and Applied Sciences, 8(7), 545-551.

Dickson, A., Leaf, A. L., & Hosner, J. F. (1960). Quality appraisal of white spruce and white pine seedling stock in nurseries. Forest Chronicle, 36, 10-13.

Edralin, D. I., & Mercado, A. Jr. (2010). Ensuring seedling quality through nursery accreditation. In Harrison, S. R., Bosch, A., Gregorio, N. O., & Herbohn, J. L. (Eds.), Improving the effectiveness and efficiency of the Philippines tree nursery sector. Proceedings from the Mid-term Workshop held on 13 Feb 2009 in Baybay, Leyte, Philippines (pp. 157–162).

Flasiński, M., & Hąc-Wydro, K. (2014). Natural vs synthetic auxin: Studies on the interactions between plant hormones and biological membrane lipids. Environmental Research, 133, 123-134.

Gautam, P., Tripathi, S. K., Kumar, A., Prakash, S., Sengar, R. S., Awasthi, M., ... & Kumar, A. (2022). Effect of different concentrations of PGRs on shooting and survival of stem cuttings in lemon (Citrus limon Burm.) cv. Pant lemon-1 under Western UP conditions. Biological Forum–An International Journal, 14(3), 1084-1088.

Ghosh, A., Dey, K., Mani, A., Bauri, F. K., & Mishra, D. K. (2017). Effects of IBA on the rooting of branch cuttings of Chinese jujube (Zizyphus jujuba Mill.) and changes to nutrients and endogenous hormones. International Journal of Chemical Studies, 5(6), 567-571.

Gnawali, P., Gurung, M., Kadel, S., Bhantana, S., Chand, N. B., Pathak, R., & Poudel, P. R. (2022). Vegetative propagation of Eureka seedless lemon (Citrus limon L. cv. Eureka Seedless) using different types of stem cutting and concentrations of Indole-3-butyric acid in winter. Nepalese Horticulture, 16(1), 81-90.

Guilfoyle, T. J. (2015). The PB1 domain in auxin response factor and Aux/IAA proteins: a versatile protein interaction module in the auxin response. Plant Cell 27, 33–43.

Hartmann, H. T., Kester, D. E., Davis, F. T., & Genev, R. L. (2002). Plant propagation: Principles and practices. Englewood Cliffs: Prentice Hall.

Harshita, G. T., & Singh, G. (2022). The use of auxin and media for rooting of citrus stem cuttings: A review. The Pharma Innovation Journal, 11(3S), 1268–1273.

Hawramee, O. A., Aziz, R. R., & Hassan, D. A. (2019, November). Propagation of white mulberry (Morus alba L.) fruitless cultivar using different cutting times and IBA. In IOP Conference Series: Earth and Environmental Science (Vol. 388, No. 1, p. 012069). IOP Publishing.

Jaiswal, E., Tripathi, V. K., Trivedi, H., & Kumar, R. (2023). Influence of IBA and NAA on regeneration of stem cuttings of sweet lime (Citrus limettioides Tanaka). Progressive Agriculture, 23(2), 191–197.

Kaur, S. (2017). Evaluation of different doses of indole-3-butyric acid (IBA) on the rooting, survival, and vegetative growth performance of hardwood cuttings of Flordaguard peach (Prunus persica L. Batch). Journal of Applied and Natural Science, 9, 173-180.

Kondle, R., Singh, G., Mirza, A. A., & Kotiyal, A. (2022). IBA effect on hardwood stem cuttings of Morus alba in open conditions in the Punjab region.

Kour, H., Beniwal, B., Chhabra, A., & Pathlan, N. (2022). Response of IBA on the rooting behaviour of rough lemon (Citrus × jambhiri Lush) under various growing conditions.

Kurd, A. A., Khan, S. A., Shah, B. H., & Khetran, M. A. (2010). Effect of indole butyric acid (IBA) on rooting of olive stem cuttings. Pakistan Journal of Agricultural Research, 23, 3-4.

Lalramhluna, P., & Prasad, V. M. (2016). Effect of different levels of Indole-3-butyric acid on growth, development, survival and establishment of air-layered lemon (Citrus limon L. Burm.) cv. Assam Lemon under the Allahabad agro-climatic condition. International Journal of Life Science and Scientific Research, 2(5), 599-603.

Loushambam, R. S., Thakur, C., Bharadwaj, D. R., & Singh, N. R. (2014). Evaluation of mulberry (Morus indica L.) provenances of Himachal Pradesh under nursery conditions. Ecology, Environment and Conservation, 20(4), 1945-1948.

Liu, Y., Zhang, H., Wang, X., & Chen, L. (2025). Auxin regulates stem growth and vascular development in citrus (Poncirus trifoliata). Plant Science, 345, 111234.

Luna, R. K., & Chamouli, N. (2006). Effect of root trainer size on the quality of seedling production in Albizia procera (Roxb.) Benth., Eucalyptus tereticornis Sm., and Acacia catechu Wild. Annals of Forestry, 14(2), 184-193.

Melgarejo, P., Martinez, J., Martinez, J. J., & Sanchez, M. (2008). Preliminary survival experiments in transplanting pomegranate. In Production, Processing and Marketing of Pomegranate in the Mediterranean Region: Advances in Research and Technology. Zaragoza, CIHEAM Publication, Europe, 163-167.

Mishra, S., Pandey, S. K., Pandey, C. S., Shakya, H. K., & Dwivedi, K. (2025a). Response of different types of cuttings of seedless lemon to indole butyric acid levels on propagation, germination and growth. Journal of Experimental Agriculture International, 47(4), 151–158. https://doi.org/10.9734/jeai/2025/v47i43365

Mishra, S., Pandey, S. K., Pandey, C. S., & Shakya, H. K. (2025b). Influence of IBA levels on rooting, growth, physiological traits and survival of different seedless lemon (Citrus limon (L.) Burm. f.) cuttings. International Journal of Agriculture, Biological and Research (IJABR), 9(4), 531–537. https://www.doi.org/10.33545/26174693.2025.v9.i4g.4146

Patel, H. R., & Patel, M. J. (2018). Role of auxins on the rooting of different types of cuttings in fig. International Journal of Current Microbiology and Applied Sciences, 7(3), 1317-1322.

Patel, S. K., Singh, J. P., Yadav, A. K., & Vivekanand. (2021). Effect of IBA and NAA levels on growth traits of Kagzi lime (Citrus aurantifolia Swingle). Journal of Pharmacognosy and Phytochemistry, 10(2), 892–897.

Polat, A. A., & Caliskan, O. (2009). Effect of Indole Butyric Acid (IBA) on rooting of cutting in various pomegranate genotypes. Acta Horticulturae (ISHS), 818, 187-192.

Rathour, S. S., Tomar, K. S., Poonam, S. B., & Katoriya, R. S. (2021). Impact of auxins on rooting and establishment of seedless lemon (Citrus limon L. Burm.) air-layers for successful propagation. International Journal of Plant & Soil Science, 33(8), 10-19.

Reddy, K. V. R., Reddy, P., & Goud, P. V. (2008b). Role of auxins synergists in the rooting of hardwood and semi-hardwood cuttings of fig (Ficus carica L.). Indian Journal of Agricultural Research, 42(1), 47-51.

Riaz, A., Rahman, K. U., Ilyas, M., Ibrahim, M., & Rauf, M. A. (2007). Effect of indole butyric acid concentrations on the rooting of kiwi cuttings. Sarhad Journal of Agriculture, 23, 293-296.

Roller, K. J. (1977). Suggested minimum standards for containerised seedlings in Nova Scotia. Canadian Forestry Service, Department of Environment, Information Report, M-X(69).

Roychoudhry, S., & Kepinski, S. (2022). Auxin in root development. Cold Spring Harbour Perspectives in Biology, 14(4), a039933.

Rymbai, H., & Sathyanarayana, R. G. (2010). Effect of IBA concentrations on guava stooling and plantlet survival under open and polyhouse conditions.

Saroj, P. L., Awasthi, O. P., Bhargava, R., & Singh, U. V. (2008). Standardisation of pomegranate propagation by cutting under a mist system in a hot arid region. Indian Journal of Horticulture, 65, 25–30.

Seiar, Y. A. (2016). Effect of growth regulators on rooting of cuttings in pomegranate (Punica granatum L.) cv. Bhagwa. Journal of Horticultural Sciences, 11(2), 156–160.

Singh, K. K., Choudhary, T., & Kumar, A. (2014). Effect of various concentrations of IBA and NAA on the rooting of stem cuttings of mulberry (Morus alba L.) under mist house conditions in the Garhwal hill region. Indian Journal of Hill Farming, 27(1), 74–77.

Singh, K. K., Choudhary, T., & Kumar, P. (2013). Effect of IBA concentrations on growth and rooting of Citrus limon cv. Pant Lemon cuttings. HortFlora Research Spectrum, 2(3), 268–270.

Singh, K. K., & Tomar, Y. K. (2015). Effect of planting time and indole butyric acid levels on rooting of woody cuttings of phalsa (Grewia asiatica L.). Horti Flora Research Spectrum, 4, 39–43.

Singh, S., & Singh, K. K. (2016). Effect of various concentrations of IBA and types of stem cuttings on the performance of rooting in sweet orange (Citrus sinensis L. Osbeck) cv. Malta under mist. The Bioscan, 11(2), 903–906.

Sinha, N. K., Kumar, S., Santra, P., Raja, P., & Mertia, D. (2014). Temporal growth performance of Indian myrrh (Commiphora wightii) raised by seedlings and cuttings from the same genetic stocks in the extremely arid Thar desert of India. The Ecoscan, 8, 241–244.

Statistix. (2006). Statistics 8.1 user guide, version 1.0. Analytical Software, P.O. Box 12185, Tallahassee, FL 32317 USA.

Thota, S., Madhavi, K., & Vani, V. S. (2012). Effect of the type of cuttings and IBA concentrations on the propagation of fig. International Journal of Tropical Agriculture, 32, 89–94.

Wu, G. A., Terol, J., Ibáñez, V., López-García, A., Pérez-Román, E., Borredá, C., Domingo, C., Tadeo, F. R., Carbonell-Caballero, J., Alonso, R., et al. (2018). Genomics of the origin and evolution of Citrus. Nature, 554, 311–316.

Yadav, P. B., & Mate, V. N. (2023). Physical and textural properties of lemon (Citrus limon). Pharma Innovation, 12(6), 2634–2637.

Zenginbal, H., Haznedar, A., & Dolgun, O. (2014). Effects of Indole-3-Butyric Acid (IBA) and cutting type on rooting of Camellia sinensis (L.) O. Kuntze. American Journal of Experimental Agriculture, 4(12), 1935–1943.

Zhong, G., & Nicolosi, E. (2023). Citrus origin, diffusion, and economic importance. In Gentile, A., La Malfa, S., & Deng, Z. (Eds.), The Citrus Genome (pp. 5–21). Springer, Cham, Switzerland.

Auxin-Induced Rooting and Seedling Quality Enhancement in Seedless Lemon (Citrus limon L.) under Nursery Conditions