Abstract
Date palm (Phoenix dactylifera L.) production is severely hampered due to several pests and diseases. Biotechnological tools such as protoplast fusion appear as an alternative to ensure rapid genetic improvement and multiplication of this species. However, establishment of an effective system of plant regeneration from protoplasts culture is a prerequisite for date palm somatic hybridization. In this chapter, we describe an effective protocol to induce microcalli in protoplasts isolated from nodular callus of important Algerian date palm cultivars. In this protocol, the main factors influencing the isolation (i.e., enzymatic solution, mannitol concentration, duration, and mode of maceration) of protoplasts from the calli of Algerian date palm cultivars were optimized. Purified protoplasts were cultured on a semisolid medium supplemented with a hormonal balance of auxin and cytokinin to obtain microcalli formation.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Assani A, Chabane D, Shittu H, Bouguedoura N (2011) Date palm cell and protoplast culture. In: Jain SM, Al-Khayri JM, Johnson DV (eds) Date palm biotechnology. Springer, Netherland, pp 605–629
Bouguedoura N, Michaux-Ferrière N, Bompar J-L (1990) Comportement in vitro de bourgeons axillaires de type indéterminé du palmier dattier (Phoenix dactylifera). Can J Bot 68(9):2004–2009
Chabane D (2007) Amélioration du palmier dattier Phoenix dactylifera L. par fusion de protoplastes de deux cultivars Deglet Noor sensible et Takerboucht résistant au Bayoud. Dissertation, University of Science and Technology Houari Boumediene, Algiers, Algeria
El Hadrami A, El Idrissi-Tourane A, El Hassni M, Daayf F, El Hadrami I (2005) Toxin-based in-vitro selection and its potential application to date palm for resistance to the bayoud Fusarium wilt. C R Biol 328(8):732–744
Chabane D, Assani A, Bouguedoura N, Haicour R, Ducreux G (2007) Induction of callus formation from difficile date palm protoplasts by means of nurse culture. C R Biol 330(5):392–401
Boufis N, Khelifi-Slaoui M, Djillali Z, Zaoui D, Morsli A, Bernards MA, Khelifi L (2014) Effects of growth regulators and types of culture media on somatic embryogenesis in date palm (Phoenix dactylifera L. cv. Degla Beida). Sci Hortic 172(9):135–142
Titouh K, Khelifi L, Slaoui M, Boufis N, Morsli A, Titouh-Hadj Moussa K et al (2015) A simplified protocol to induce callogenesis in protoplasts of date palm (Phoenix dactylifera L.) cultivars. Iran J Biotechnol 13(1):26–35
Davey MR, Anthony P, Power JB, Lowe KC (2005) Plant protoplasts: status and biotechnological perspectives. Biotechnol Adv 23(2):131–171
Sun Y, Zhang X, Huang C, Nie Y, Guo X (2005) Plant regeneration via somatic embryogenesis from protoplasts of six explants in Coker (Gossypium hirsutum). Plant Cell Tissue Organ Cult 201(82):309–315
Rizkalla AA, Badr-Elden AM, Nower AA (2007) Protoplast isolation, salt stress and callus formation of two date palm genotypes. J Appl Sci Res 3(10):1186–1194
Assani A, Haïcour R, Wenzel G, Foroughi-Wehr B, Bakry F, Côte F-X et al (2002) Influence of donor material and genotype on protoplast regeneration in banana and plantain cultivars (Musa spp.) Plant Sci 162(3):355–362
Borgato L, Pisani F, Furini A (2007) Plant regeneration from leaf protoplasts of Solanum virginianum L. (Solanaceae). Plant Cell Tissue Organ Cult 88(3):247–252
Gabr MF, Tisserat B (1984) Parameters involved in the isolation, culture, cell wall regeneration and callus formation from palm and carrot protoplasts. Date Palm J 3(2):359–365
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Zhang J, Shen W, Yan P, Li X, Zhou P (2011) Factors that influence the yield and viability of protoplasts from Carica papaya L. Afr J Biotechnol 10(26):5137–5142
Monteiro M, Appezzato-Da-Glória B, Valarini MJ, De Oliveira CA, Vieira MLC (2003) Plant regeneration from protoplasts of alfalfa (Medicago sativa) via somatic embryogenesis. Sci Agric 60:683–689
Sané D, Aberlenc-Bertossi F, Gassama-Dia YK, Sagna M, Trousslot MF, Duval Y et al (2006) Histological analysis of callogenesis and somatic embryogenesis from cell suspensions of date palm (Phoenix dactylifera). Ann Bot 98:301–308
Sinha A, Wetten AC, Caligari PDS (2003) Effect of biotic factors on the isolation of Lupinus albus protoplasts. Aust J Bot 51(1):103–109
Horine RK, Ruesink AW (1972) Cell wall regeneration around protoplasts isolated from Convolvulus tissue culture. Plant Physiol 50(4):438–445
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Titouh, K., Boufis, N., Khelifi, L. (2017). Microcalli Induction in Protoplasts Isolated from Embryogenic Callus of Date Palm. In: Al-Khayri, J., Jain, S., Johnson, D. (eds) Date Palm Biotechnology Protocols Volume I. Methods in Molecular Biology, vol 1637. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7156-5_19
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7156-5_19
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7155-8
Online ISBN: 978-1-4939-7156-5
eBook Packages: Springer Protocols