In vitro Propagation of Citrus aurantifolia and its Conservation in Pakistan


  • Abdul Momin Department of Botany, Kohat University of Science & Technology, Kohat-26000 Author
  • Rafia Naheed Kohat University of Science & Technology, Kohat-26000, Pakistan Author
  • Wajahat Khan Department of Botany, Kohat University of Science and Technology, Kohat, Pakistan Author
  • Hira Shujaat Kohat University of Science & Technology, Kohat-26000, Pakistan Author
  • Muhammad Ghazanfar Javed Khan Faculty of Bioinformatics and Biotechnology. International Islamic University, Islamabad, Pakistan Author



Citrus aurantifolia, in vitro propagation, tissue culture, conservation, growth regulators, Pakistan


Citrus varieties are propagated by both asexual and sexual methods. Generally, rootstocks are propagated sexually through seeds, while most of the commercial varieties are propagated by various asexual methods such as grafting and budding. The technology called plant tissue culture is used for commercial production of plants that are microbe and virus free and to save germplasm of infrequent or threatened species of plants. Fast reproduction techniques have the potential to produce plants free of viruses. Likewise, traditional propagation methods are old, time consuming and are difficult. Tissue culture techniques are assumed for reliable commercial production of economically important plants. In present study shoot tips of Citrus aurantifolia were collected from the field of BCI, NARC. Different disinfectant (Ethanol and NaOCl) with the concentration ranging from 10 to 20 percent were used for culture establishment. Maximum survival percentage was observed in 10% NaOCl. After 5 weeks, established cultures were shifted to hormonal growth media (BAP, GA3 and NAA) at the concentration ranging from 0.1 to 0.8 mg/L. Our finding results that maximum phenotypic characters (plant height, number of leaves and number of shoots were observed on BAP at the concentration of 0.1 to 0.3 mg/L, while maximum number of roots were observed on NAA at the concentration of 0.5mg/L. In present study, for in vitro conservation different conservants like sorbitol and mannitol (10, 20 and 30 g/L) were used. Incubation of cultures was maintained at 21°C under the light intensity of 1000 lux. Old cultures of Citrus aurantifolia were shifted in conservant media. Overall, remarkable difference was observed. In present study mannitol at the concentration of (10, 20 and 30 g/L) showed slow growth after 35 days. In future the plants in conservation media will be used for further propagation and for growing them in fields.


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Author Biographies

  • Rafia Naheed, Kohat University of Science & Technology, Kohat-26000, Pakistan

    Department of Botany

  • Hira Shujaat, Kohat University of Science & Technology, Kohat-26000, Pakistan

    Department of Botany


Sagone, J. A., Greenwald, J., Kraut, E. H., Bianchine, J., & Singh, D. (1983). Glucose: a role as a free radical scavenger in biological systems. The Journal of laboratory and clinical medicine, 101(1), 97-104.

Anwar, F., Naseer, R., Bhanger, M. I., Ashraf, S., Talpur, F. N., & Aladedunye, F. A. (2008). Physico-chemical characteristics of citrus seeds and seed oils from Pakistan. Journal of the American Oil Chemists' Society, 85(4), 321-330.

Patil, J. R. (2009). Studies on isolation and characterization of bioactive compounds in lime [Citrus aurantifolia (Christm) Swingle], their antioxidant and anticancer properties (Doctoral dissertation, UAS Dharwad).

Sethpakdee, R. (1992). Citrus aurantifolia (Christm. & Panzer) Swingle. Plant Resources of South-East Asia. Edible fruits and nuts. Prosea Foundation, Bogor, Indonesia, 2, 126-128.

Golob, P. (2013). Alphabetical List of Plant Families with Insecticidal and Fungicidal Properties. The use of spices and medicinals as bioactive protectants for grains. Food and Agriculture Organization of the United Nations, 13.

Ali, S., & Mirza, B. (2006). Micropropagation of rough lemon (Citrus jambhiri Lush.): Effect of explant type and hormone concentration. Acta Botanica Croatica, 65(2), 137-146.

Duke J.A., duCellier J.L. (1993): CRC handbook of alternative cash crops (page 139- 145)

Rajasekharan, P. E., & Sahijram, L. (2015). In vitro conservation of plant germplasm. In Plant biology and biotechnology (pp. 417-443). Springer, New Delhi.

Micheli, M., & Standardi, A. (2016). From somatic embryo to synthetic seed in Citrus spp. through the encapsulation technology. In In Vitro Embryogenesis in Higher Plants (pp. 515-522). Humana Press, New York, NY

Siddique, M. I., & Garnevska, E. (2018). Citrus value chain (s): A survey of Pakistan citrus industry. Agric. Value Chain, 37.

Paunescu, A. (2009). Biotechnology for endangered plant conservation: a critical overview. Romanian biotechnological letters, 14(1), 4095-4103.

Sarker, I., Islam, J., Shaekh, M. P. E., Rahman, M. M., Khan, H., Chowdhury ASMHK, M. M., ... & Haque, R. (2015). Establishment of a standard protocol for in vitro meristem culture and plant regeneration of Citrus aurantifollia. IOSR J. Pharm. Biol. Sci, 10(2), 61-69.

Siddiqui, M. W., Bhattacharjya, A., Chakraborty, I., & Dhua, R. S. (2011). 6- Benzylaminopurine improves shelf life, organoleptic quality and healthpromoting compounds of fresh-cut broccoli florets.

de Saint Germain, A., Ligerot, Y., Dun, E. A., Pillot, J. P., Ross, J. J., Beveridge, C. A., & Rameau, C. (2013). Strigolactones stimulate internode elongation independently of gibberellins. Plant physiology, 163(2), 1012-1025.

Riley, J. M. (2012). Gibberellic acid for fruit set and seed germination. CRFG Journal, 19, 10-12.

Morikawa, H., & Takahashi, M. (2004). U.S. Patent No. 6,800,482. Washington, DC: U.S. Patent and Trademark Office

Shibli, R. A., Shatnawi, M. A., Subaih, W. S., & Ajlouni, M. M. (2006). In vitro conservation and cryopreservation of plant genetic resources: a review. World Journal of Agricultural Sciences, 2(4), 372-382.




How to Cite

Momin A, Naheed R, Khan W, Shujaat H, Khan MGJ. In vitro Propagation of Citrus aurantifolia and its Conservation in Pakistan. IRABCS [Internet]. 2024 Jul. 2 [cited 2024 Jul. 20];2(1):40-6. Available from:

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