Micropropagation of Date Palm (Phoenix dactylifera L.) Cultivar Gulistan Using Immature Inflorescence Explants
DOI:
https://doi.org/10.53560/PPASB(62-4)1089Keywords:
Micropropagation, Somatic Embryogenesis, Plantlet Regeneration, In vitro Hardening, AcclimatizationAbstract
Current study described micropropagation of commercial date palm cv. Gulistan through juvenile inflorescence explants. Immature spathes (20 cm in length) were excised before emergence from leaf axils in the crown at particular time i.e. early of February. For sterilization, the undissected spathes were dipped in NaOCl solution (40%) for 10 minutes on laminar air flow hood. Medium used for callus formation in the inflorescence explants was consisted of 0.1 mg/L 2,4-D, 0.1 mg/L IAA, 5.0 mg/L NAA induced significantly highest callus (87%) in primary explants. Highest somatic embryogenesis (84%) was obtained on the medium contained 0.1 mg/L NAA, 0.1 mg/L Kinetin. Shoot induction and multiplication (91%) was recorded on the medium consisted of 0.1 mg/L NAA, 0.05 mg/L BA. Leaves number (4.1), leaves length (17 cm), roots number (4.0), roots length (7 cm) were recorded as significantly highest on the medium consisted of 0.1 mg/L NAA. Trimming of embryonic root at 1-2 mm distance from plantlets’ base produced 3-4 adventitious roots. In vitro hardening supported to choose healthy plantlets which survived well in greenhouse. Plantlets’ survival percentage in greenhouse after 1M (88%) as significantly highest was recorded on the soil mixture contained peatmoss, river sand, perlite (1:1:¼ v/v). 46 cm long plantlets produced 4 small fronds/compound leaves, were cultivated in open field. Fruiting in trees started after three years of cultivation in open field. Introduction of elite date palm cultivars like Gulistan via micropropagation will be an addition to the existing cultivars in the area.
References
1. A.A. Mirani, M.A. Jatoi, L. Bux, C.H. Teo, A.I. Kabiita, J.A. Harikrishna, G.S. Markhand, T. Jatt, N. Solangi, and S. Abro. Genetic stability analysis of tissue culture derived date palm cv. Dedhi plants using IRAP markers. Acta Ecologica Sinica 42(1): 76-81 (2022).
2. N. Solangi, A.A. Abul-Soad, G.S. Markhand, M.A. Jatoi, T. Jatt, and A.A. Mirani. Comparison among different auxins and cytokinins to induce date palm (Phoenix dactylifera L.) somatic embryogenesis from floral buds. Pakistan Journal of Botany 52(4): 1243-1249 (2020).
3. N. Solangi, M.A. Jatoi, N. Tunio, A.A. Mirani, A.A. Abul-Soad, and G.S. Markhand. Fruit Morphological and Biochemical Characterization of Three Saudi Arabian Date Palm (Phoenix dactylifera L.) Cultivars Grown in District Khairpur, Pakistan. Proceedings of the Pakistan Academy of Sciences: B Life and Environmental Sciences 61(1): 11-20 (2024).
4. A.A. Abul-Soad, S.M. Jain, and M.A. Jatoi. Biodiversity and conservation of date palm. In: Biodiversity and conservation of woody plants. M.R. Ahuja and S.M. Jain (Eds.). Sustainable Development and Biodiversity. Springer Cham 17: pp. 313-353 (2017).
5. M.T. Rashid, B. Safdar, M.A. Jatoi, N. Solangi, A. Wali, N. Ali, and K. Liu. Structure, rheology, and tribology of date fruit paste procured from different date palm cultivars. Journal of Food Process Engineering 44: e13891 (2021).
6. C.T. Chao and R.R. Krueger. The date palm (Phoenix dactylifera L.): overview of biology, uses, and cultivation. Horticultural Science 42(5): 1077-1082 (2007).
7. P.K. Vayalil. Date fruits (Phoenix dactylifera Linn): an emerging medicinal food. Critical Reviews in Food Science and Nutrition 52(3): 249-271 (2012).
8. J. Ahmed, F.M. Al-Jasass, and M. Siddiq. Date fruit composition and nutrition. M. Siddiq, S.M. Aleid, and A.A. Kader (Eds.). In: Dates: postharvest science, processing technology and health benefits, (First Edition). Wiley, Chichester, United Kingdom pp. 261-283 (2013).
9. R. Omran, Z. Al-Taee, H. Hashim, and M. Al-Jassani. Preventive effects of Phoenix dactylifera polyphenols against 7,12-dimethylbenz (a) anthracene-induced mammary cancer. Asian Journal of Pharmaceutical and Clinical Research 10(7): 172-181 (2017).
10. C. Selmani, D. Chabane, and N. Bouguedoura. Ethnobotanical survey of Phoenix dactylifera L. pollen used for the treatment of infertility problems in Algerian oases. African Journal of Traditional, Complementary and Alternative Medicines 14(3): 175-186 (2017).
11. K. Rajmohan. Date palm tissue culture: A pathway to rural development. In: Date Palm Biotechnology. S.M. Jain, J.M. Al-Khayri, and D.V. Johnson (Eds.). Springer, Dordrecht pp. 29-45 (2011).
12. B. Tisserat. Factors involved in the production of plantlets from date palm callus cultures. Euphytica 31: 201-214 (1982).
13. N. Al-Khalifah and E. Askari. Growth abnormalities associated with micropropagation of date palm. In: Date palm biotechnology. S.M. Jain, J.M. Al-Khayri and D.V. Johnson (Eds.). Springer, Dordrecht, Heidelberg, London, New York pp. 205–219 (2011).
14. N. Solangi, M.A. Jatoi, G.S. Markhand, A.A. Abul-Soad, M.A. Solangi, T. Jatt, A.A. Mirbahar, and A.A. Mirani. Optimizing Tissue Culture Protocol for In Vitro Shoot and Root Development and Acclimatization of Date Palm (Phoenix dactylifera L.) Plantlets. Erwerbs-Obstbau 64: 97-106 (2022).
15. A.A. Abul-Soad and J.M. Al-Khayri. Date palm somatic embryogenesis from inflorescence explant. Step Wise Protocols for Somatic Embryogenesis of Important Woody Plants S.M. Jain and P. Gupta (Eds.). Springer Cham pp. 329-347 (2018).
16. M.M. Saker and H.A. Moursi. Molecular Characterization of Egyptian date palm cultivars: RAPD fingerprints. Arab Journal of Biotechnology 2(1): 71-78 (1999).
17. J. Janick (Ed.). Horticultural Science. W.H. Freeman and Company, San Francisco (1979).
18. A.A. Abul-Soad and S.M. Mahdi. Commercial production of tissue culture date palm (Phoenix dactylifera L.) by inflorescence technique. Journal of Genetic Engineering and Biotechnology 8(2): 39-44 (2010).
19. D.A. De Mason. The occurrence and structure of apparently bisexual flowers in the date palm, Phoenix dactylifera L. (Arecaceae). Botanical Journal of the Linnean Society 81(4): 283-292 (1980).
20. D.R. Sharma, R. Kumari, and J.B. Chowdhury. In vitro culture of female Date Palm (Phoenix dactylifera L.) tissues. Euphytica 29: 169-174 (1980).
21. S. Bhaskaran and R. Smith. Somatic embryogenesis from shoot tip and immature inflorescence of Phoenix dactylifera L cv. Barhee. Plant Cell Report 12: 22-25 (1982).
22. J.M. Al-Khayri. Micropropagation of Date Palm Phoenix dactylifera L. In: Protocols for Micropropagation of Woody Trees and Fruits. S.M. Jain and H. Haggman (Eds.). Springer, Berlin, Germany pp. 509-526 (2007).
23. L. Abahmane. Micropropagation of date palm (Phoenix dactylifera L.) selected genotypes from inflorescence tissues by using somatic embryogenesis technique. Acta Horticulturae 882: 827-832 (2010).
24. N. Nimavat and P. Parikh. Innovations in Date palm (Phoenix dactylifera L.) micropropagation: Detailed review of in vitro culture methods and plant growth regulator applications. Plant Cell Tissue and Organ Culture 159: 6 (2024). https://doi.org/10.1007/s11240-024-02866-7.
25. L. Abahmane. Recent achievements in date palm (Phoenix dactylifera L.) micropropagation from inflorescence tissue. Emirates Journal of Food and Agriculture 25(11): 863-874 (2013).
26. N. Solangi, M.A. Jatoi, A.A. Abul-Soad, A.A. Mirani, M.A. Solangi, and G.S. Markhand. Factors influencing somatic embryogenesis and plantlet regeneration of date palm using immature floral buds. Sarhad Journal of Agriculture 39(2): 323-331 (2023).
27. A. Zaid, A.A. Al-Kaabi, and B. El-Korchi. Large scale in vitro propagation of rare and unique male date palm (Phoenix dactylifera L.). 3rd International Date Palm Conference, Abu Dhabi, Acta Horticulturae 736: 27 (2007). https://doi.org/10.17660/ActaHortic.2007.736.22
28. A.A. Mirani, C.H. Teo, A.A. Abul-Soad, G.S. Markhand, T. Jatt, A.A. Mirbahar, and N. Solangi. Phenotypic reversion of somaclonal variants derived from inflorescence of date palm (Phoenix dactylifera L.) in the open field trials. Sarhad Journal of Agriculture 35(3): 719-726 (2019).
29. A.A. Mirani, C.H. Teo, G.S. Markhand, A.A. Abul-Soad, and J.A. Harikrishna. Detection of somaclonal variations in tissue cultured date palm (Phoenix dactylifera L.) using transposable element-based markers. Plant Cell, Tissue and Organ Culture 141(1): 119-130 (2020).
30. B. Tisserat. Propagation of date palm (Phoenix dactylifera L.) in vitro. Journal of Experimental Botany 30: 1275-1283 (1979).
31. A. Othmani, C. Bayoudh, N. Drira, and M. Trifi. In vitro Cloning of Date Palm Phoenix Dactylifera L., Cv. Deglet Bey by Using Embryogenic Suspension and Temporary Immersion Bioreactor (TIB). Biotechnology and Biotechnological Equipment 23(2): 1181-1188 (2009).
32. M.S. Omar, M.K. Hameed, and M.S. Al-Rawi. Micropropagation of date palm (Phoenix dactylifera L.). In: High-Tech and Micropropagation II. Biotechnology in agriculture and forestry. Volume 18. Y.P.S. Bajaj (Ed.). Springer-Verlag, Berlin, Heidelberg pp. 471-492 (1992). https://doi.org/10.1007/978-3-642-76422-6_25.
33. M.A. Abohatem, A. Ba‑Asher, and M. Al‑Duais. In vitro germination of date palm somatic embryos and conversion into plants under dark culture conditions: novel protocol to reducing the time and production costs. Discover Plants 1(1): 40 (2024).
34. O. Heinisch. Steel, R.G.D. and J.H. Torrie: Principles and Procedures of Statistics (With special Reference to the Biological Sciences). Biometrical Journal 4(3): 207-208 (1962). https://doi.org/10.1002/bimj.19620040313.
35. A.M.A. Ali, A.A. Qahtan, J.M. Al-Khayri, and H.S. Ghazzawy. Effect of low 2,4-D concentration on enhancing indirect embryogenesis and genetic stability in date palm (Phoenix dactylifera L.). Scientific Reports 15: 43865 (2025).
36. J.M. Al-Khayri. Somatic Embryogenesis of Date Palm (Phoenix dactylifera L.) from Shoot Tip Explants. In: Step Wise Protocols for Somatic Embryogenesis of Important Woody Plants, Forestry Sciences. S.M. Jain and P. Gupta (Eds.). Springer, Cham pp. 231-244 (2018).
37. E.M. Badawy, A.M.A. Habib, A. El Bana, and G.M. Yosry. Propagation of date palm (Phoenix dactylifera L.) plants by using tissue culture technique. Arab Journal of Biotechnology 8: 343-354 (2005).
38. C. Eeuwens and J. Blake. Culture of coconut and date palm tissue with a view to vegetative propagation. Acta Horticulturae 78: 277-286 (1977).
39. A.A. Abul-Soad. Influence of inflorescence explant age and 2, 4-D incubation period on somatic embryogenesis of date palm. Emirates Journal of Food and Agriculture 24(5): 434-443 (2012).
40. M. Laaguidi, R. Meziani, and K. Sellam. In vitro culture of date palm: a review of challenges and solutions for managing endophytic bacteria contamination. Vegetos (2025). https://doi.org/10.1007/s42535-025-01287-x.
41. C.J. Eeuwens. Mineral requirements for growth and callus initiation of tissue explants excised from mature coconut palms (Cocosnucifera) and cultured in vitro. Physiologia Plantarum 36(1): 23-28 (1978).
42. A.A. Abul-Soad. Micropropagation of date palm using inflorescence explants. In: Date Palm Biotechnology. S.M. Jain, J.M. Al-Khayri, and D.V. Johnson (Eds.). Springer Dordrecht pp. 91-117 (2011). https://doi.org/10.1007/978-94-007-1318-5.
43. N. Drira and A. Benbadis. Multiplication végétative du palmierdattier (Phoenix dactylifera L.) par réversion en culture in vitrod’ébauchesflorales de piedsfemelles. Journal of Plant Physiology 119: 227-235 (1985).
44. L. Fki, R. Masmoudi, N. Drira, and A. Rival. An optimized protocol for plant regeneration from embryogenic suspension cultures of date palm, Phoenix dactylifera L., cv. Deglet Nour. Plant Cell Report 21: 517-524 (2003).
45. M. Mazri and R. Meziani. Micropropagation of date palm: a review. Cell and Developmental Biology 4(3): 160 (2015).
46. P. Eshraghi, R. Zarghami, and M. Mirabdulbaghi. Somatic embryogenesis in two Iranian date palm cultivars. African Journal of Biotechnology 4(11): 1309-1312 (2005).
47. J.M. Al-Khayri. Date Palm (Phoenix dactylifera L.). In: Protocols of somatic embryogenesis in woody plants. S.M. Jain and P.K. Gupta (Eds.). Springer, Dordrecht pp. 309-319 (2005). https://doi.org/10.1007/1-4020-2985-3_25.
48. J.M. Al-Khayri. Somatic embryogenesis of date palm (Phoenix dactylifera L.) improved by coconut water. Biotechnology 9(4): 477-484 (2010).
49. I. El-Hadrami and M. Baaziz. Somatic embryogenesis and analysis of peroxidases in Phoenix dactylifera L. Biologia Plantarum 37(2): 197-203 (1995).
50. J. Aslam, S.A. Khan, A.J. Cheruth, A. Mujib, M.P. Sharma, and P.S. Srivastava. Somatic embryogenesis, scanning electron microscopy, histology and biochemical analysis at different developing stages of embryogenesis in six date palm (Phoenix dactylifera L.) cultivars. Saudi Journal of Biological Sciences 18: 369-380 (2011).
51. A. Othmani, C. Bayoudh, N. Drira, and M. Trifi. Somatic embryogenesis and plant regeneration in date palm Phoenix dactylifera L., cv. Boufeggous is significantly improved by fine chopping and partial desiccation of embryogenic callus. Plant Cell, Tissue and Organ Culture 97: 71-79 (2009).
52. A. Othmani, C. Bayoudh, N. Drira, and M. Trifi. In vitro cloning of date palm Phoenix dactylifera L. cv. Deglet Bey using embryogenesis suspension and temporary immersion bioreactor (TIB). Biotechnology & Biotechnological Equipment 23(2): 1181-1188 (2009).
53. M.H. Hassan and R.A. Taha. Callogenesis, somatic embryogenesis and regeneration of date palm (Phoenix dactylifera L.) cultivars affected by carbohydrate sources. International Journal of Agricultural Research 7: 231-242 (2012).
54. F.C. Steward, A.E. Kent, and M.O. Mapes. Growth and organization in cultured cells: sequential and synergestic effects of growth regulating substances. Annals of the New York Academy of Sciences 144: 326-334 (1967).
55. T. Fujimura and A. Komamine. Effects of various growth regulators on the embryogenesis in a carrot cell suspension culture. Plant Science Letters 5(6): 359-364 (1975).
56. P.V. Ammirato, and F.V. Steward. Some effects of the environment on the development of embryos from cultured free cells. Botanical Gazette 132(2): 149-158 (1971). https://doi.org/10.1086/336573
57. S.F. El-Sharabasy, H.A. Bosila, and I.A. Ibrahim. Micropropagation studies on Zaghlool and Sewi cvs. of date palm (Phoenix dactylifera L.): III. Plantlet acclimatization. Proceedings of the Second International Conference on Date Palm, Al Ain, UAE (2001).
58. A. Zaid and B. Tisserat. In vitro shoot tip differentiation in Phoenix dactylifera L. Date Palm Journal 2(2): 163-182 (1983).
59. B. Tisserat. Propagation of date palm by shoot tip cultures. Horticultural Science 19: 230-231 (1984).
60. M.S. Omar. In vitro response of various date palm explants. Date Palm Journal 2: 371-389 (1988).
61. K.W. Al-Maari and A.S. Al-Ghamdi. Micropropagation of five date palm cultivars through in vitro axillary buds’ proliferation. Journal of Agricultural Science 13: 55-71 (1997).
62. A.A. Abul-Soad and M.A. Jatoi. Factors affecting in vitro rooting of date palm (Phoenix dactylifera L.). Pakistan Journal of Agricultural Science 51: 467-474 (2014).
63. M.M. Hassan, I.A. Ibrahim, N.M. Fathy, M.K.H. Ebrahim, and E. Komor. Protocol for Micropropagated Date Palm Acclimatization: Effect of Micropropagated Plantlet Type, Soil Composition, and Acclimatization Season. International Journal of Fruit Science 14(2): 225-233 (2014).
64. M.F. Gabr and M.M. Abd-Alla. Micropropagation of Phoenix dactylifera L. var. karama. New York Science Journal 3(12): 64-69 (2010).
65. T. Kozai, M. Hayashii, Y. Hirosawa, T. Kodama, and I. Watanabe. Environmental control for acclimatization of in vitro cultured plantlets (1) Development of acclimatization unit for accelerating the plantlet growth and test cultivation. Journal of Agricultural Meteorology 42(4): 349-358 (1987).
66. M. Mohammed, M. Munir, and H.S. Ghazzawy. Design and Evaluation of a Smart Ex Vitro Acclimatization System for Tissue Culture Plantlets. Agronomy 13(1): 78 (2023).
67. S.T. Hussein and E.B. Mohamed. Micropropagation of some Egyptian date palm dry cultivars 1-maturation of somatic embryos. Arab Journal of Biotechnology 10(2): 333-340 (2007).
68. S.S. Kurup, M.A. Aly, G. Lekshmi, and N.H. Tawfik. Rapid in vitro regeneration of date palm (Phoenix dactylifera L.) cv. Kheneizi using tender leaf explant. Emirates Journal of Food and Agriculture 26: 539-544 (2014).
69. B. Tisserat (Ed.). Date palm tissue culture. Volume 17. Advances in Agricultural Technology. Western Series. U.S. Department of Agriculture, Science and Education Administration, USA (1981).
70. A.M. Abdelghaffar, S.S. Soliman, T.A. Ismail, A.M. Alzohairy, A.A.H.A. Latef, K. Alharbi, J.M. Al-Khayri, N.I.M. Aljuwayzi, D.A. El-Moneim, and A.A. Hassanin. In Vitro Propagation of Three Date Palm (Phoenix dactylifera L.) Varieties Using Immature Female Inflorescences. Plants 12(3): 644 (2023).
71. A.A. Abul-Soad, S.M. Mahdi, and G.S. Markhand. Date Palm Status and Perspective in Pakistan. In: Date Palm Genetic Resources and Utilization. J.M. Al-Khayri, S.M. Jain, and D.V. Johnson (Eds.). Springer Dordrecht pp. 153-205 (2015).
72. V. Gurevich, U. Lavi, and Y. Cohen. Genetic variation in date palms propagated from offshoots and tissue culture. Journal of the American Society of Horticultural Science 130: 46-53 (2005).
73. I. Pinker, Y.M. Ibraheem, and M.H. Böhme. Propagation of some date palm cultivars by using tissue culture methods. Acta Horticulturae 839: 71-77 (2009).
74. S. Alavipour, E. Khaleghi, N. Moallemi, K. Mehdikhanlou, and A. Trohi. Heteromorphism of normal and abnormal flowers of date palm cv. Barhee and comparison of green and red Ghanami pollens as dominant pollens. South African Journal of Botany 163: 541-551 (2023).
75. Y. Cohen, R. Kochinsky, and E. Tripler. Flower abnormalities cause abnormal fruit setting in tissue culture-propagated date palm (Phoenix dactylifera L.). Journal of Horticultural Science and Biotechnology 79(6): 1007-1013 (2004).
76. Y. Cohen. Molecular detection of somaclonal variation in date palm. In: Date Palm Biotechnology. S.M. Jain, J.M. Al-Khayri, and D.V. Johnson (Eds.). Springer, Dordrecht pp. 221-235 (2011). https://doi.org/10.1007/978-94-007-1318-5_11.
77. N. Solangi, A.A. Mirani, M.A. Jatoi, A.A. Abul-Soad, L.B. Bhanbhro, G.S. Markhand, M. Hedayat, and G. Abdi. Field evaluation of tissue culture-derived and offshoot-grown date palm cultivars: a comparative analysis of vegetative and fruit attributes. Frontiers in Plant Science 16: 1516983 (2025).
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Upon acceptance of an article, its copyright will be assigned to the Pakistan Academy of Sciences.
This work is licensed under a Creative Commons Attribution 4.0 International License.
Creative Commons Attribution (CC BY). Allows users to: copy the article and distribute; abstracts, create extracts, and other revised versions, adaptations or derivative works of or from an article (such as a translation); include in a collective work (such as an anthology); and text or data mine the article. These uses are permitted even for commercial purposes, provided the user: includes a link to the license; indicates if changes were made; gives appropriate credit to the author(s) (with a link to the formal publication through the relevant DOI); and does not represent the author(s) as endorsing the adaptation of the article or modify the article in such a way as to damage the authors' honor or reputation.
