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Histological Analysis of the Developmental Stages of Direct Somatic Embryogenesis Induced from In Vitro Leaf Explants of Date Palm

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Date Palm Biotechnology Protocols Volume I

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1637))

Abstract

Somatic embryogenesis is an ideal technique for the micropropagation of date palm using different explant tissue; however, histological studies describing the ontogenesis of plant regeneration are limited. This chapter provides a simple protocol for the histological analysis of the successive developmental stages of direct somatic embryogenesis induced from in vitro leaf explants. Direct somatic embryos are obtained from Murashige and Skoog (MS) medium containing 2 mg/L 6-benzylaminopurine. In order to observe the different developmental stages, histological analysis is carried out on samples at 15-day intervals for 60 days. Samples are fixed in formalin acetic alcohol and embedded in paraffin wax. Stain serial transverse and longitudinal sections, 8 μm thick, are stained with safranin-Fast Green. After 15 days on the induction medium, somatic embryos exhibit multicellular origin directly from the procambium cells, whereas the mesophyll and the epidermal cells are not involved in this process. After 2 months, several developmental stages (pre-globular, globular, early bipolar, bipolar, and cotyledonary-shaped) are observed. These embryos germinate after transferring to MS medium without plant growth regulators and rooting on 2 mg/L NAA-containing medium resulting in complete plantlets.

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References

  1. Al Kaabi H, Rhiss A, Hassan M (2001) Effect of auxins and cytokinins on the in vitro production of date palm bud generative tissues and on the number of differentiated buds. In: Proceedings second international conference on date palm Al Ain, UAE, pp 47–86

    Google Scholar 

  2. Al-Khateeb A (2006) Role of cytokinin and auxin on the multiplication stage of date palm (Phoenix dactylifera L.) cv. Sukry. Biotech 5:349–352

    Article  CAS  Google Scholar 

  3. Quiroz-Figueroa FR, Rojas-Herrera R, Galaz-Avalos RM, Loyola-Vargas VM (2006) Embryo production through somatic embryogenesis can be used to study cell differentiation in plants. Plant Cell Tissue Organ Cult 86(3):285–301

    Article  Google Scholar 

  4. Al-Khayri JM (2003) In vitro germination of somatic embryos in date palm: effect of auxin concentration and strength of MS salts. Curr Sci 84:680–683

    CAS  Google Scholar 

  5. Othmani A, Bayoudh C, Drira N, Marrakchi M, Trifi M (2009) 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 Organ Cult 97(1):71–79

    Article  Google Scholar 

  6. Tisserat B, DeMason D (1980) A histological study of development of adventive embryos in organ cultures of Phoenix dactylifera L. Ann Bot 46(4):465–472

    Article  Google Scholar 

  7. Zayed ME, Abd Elbar OH (2015) Morphogenesis of immature female inflorescences of date palm in vitro. Ann Agric Sci 60(1):113–120

    Google Scholar 

  8. Al-Khayri JM (2005) Date palm Phoenix dactylifera L. In: Jain SM, Gupta PK (eds) Protocol for somatic embryogenesis in woody plants. Springer, Dordrecht, pp 309–319

    Chapter  Google Scholar 

  9. Sudhersan C, Abo El-Nil M, Al-Baiz A (1993) Occurrence of direct somatic embryogenesis on the sword leaf of in vitro plantlets of Phoenix dactylifera L. cultivar Barhee. Curr Sci 65(11):887–889

    Google Scholar 

  10. Fki L, Masmoudi R, Kriaa W, Mahjoub A, Sghaier B, Mzid R et al (2011) Date palm micropropagation via somatic embryogenesis. In: Jain SM, Al-Khayri JM, Johnson DV (eds) Date palm biotechnology. Springer, Dordrecht, pp 47–68

    Chapter  Google Scholar 

  11. Abd El Bar OH, El Dawayati MM (2014) Histological changes on regeneration in vitro culture of date palm (Phoenix dactylifera) leaf explants. Aust J Crop Sci 8(6):848–855

    CAS  Google Scholar 

  12. Alkhateeb A (2008) Comparison effects of sucrose and date palm syrup on somatic embryogenesis of date palm (Phoenix dactylifera L.) Am J Biochem Biotech 4(1):19–23

    Article  CAS  Google Scholar 

  13. Kunert K, Baaziz M, Cullis C (2003) Techniques for determination of true-to-type date palm (Phoenix dactylifera L.) plants: a literature review. Emir J Food Agric 15:1–16

    Article  Google Scholar 

  14. Sané D, Aberlenc-Bertossi F, Gassama-Dia YK, Sagna M, Trouslot MF, Duval Y, Borgel A (2006) Histocytological analysis of callogenesis and somatic embryogenesis from cell suspensions of date palm (Phoenix dactylifera). Ann Bot 98:301–308

    Article  PubMed  PubMed Central  Google Scholar 

  15. El Dawayati MM, Abd El Bar OH, Zaid ZE, El Din AFZ (2012) In vitro morpho-histological studies of newly developed embryos from abnormal malformed embryos of date palm cv. Gundila under desiccation effect of polyethelyne glycol treatments. Ann Agric Sci 57(2):117–128

    Google Scholar 

  16. Kwaaitaal MA, de Vries SC (2007) The SERK1 gene is expressed in procambium and immature vascular cells. J Exp Bot 58(11):2887–2896

    Article  CAS  PubMed  Google Scholar 

  17. Nolan KE, Irwanto RR, Rose RJ (2003) Auxin up-regulates MtSERK1 expression in both Medicago truncatula root-forming and embryogenic cultures. Plant Physiol 133(1):218–230

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497

    Article  CAS  Google Scholar 

  19. Berlyn GP, Miksche JP, Sass JE (1976) Botanical microtechnique and cytochemistry. Iowa State University Press, Ames, IA, pp 24–100

    Google Scholar 

  20. Sass JE (1951) Botanical microtechnique, 2nd edn. The Iowa State College Press, Ames, IA, pp 197–198

    Google Scholar 

  21. Caboni E, Tonelli M, Falasca G, Damiano C (1996) Factors affecting adventitious shoot regeneration in vitro in the apple rootstock 'Jork 9′. Adv Hortic Sci 10(3):146–150

    Google Scholar 

  22. Welander M (1988) Plant regeneration from leaf and stem segments of shoots raised in vitro from mature apple trees. J Plant Physiol 132(6):738–744

    Article  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Agricultural Botany, Faculty of Agriculture, Ain Shams University, 68 Hadayek Shoubra, 11241, Cairo, Egypt

    Ola H. Abdelbar

Authors
  1. Ola H. Abdelbar
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Correspondence to Ola H. Abdelbar .

Editor information

Editors and Affiliations

  1. Department of Agricultural Biotechnology, King Faisal University, Al-Hassa, Saudi Arabia

    Jameel M. Al-Khayri

  2. Agricultural Sciences, University of Helsinki, Helsinki, Finland

    S. Mohan Jain

  3. Cincinnati, Ohio, USA

    Dennis V. Johnson

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Abdelbar, O.H. (2017). Histological Analysis of the Developmental Stages of Direct Somatic Embryogenesis Induced from In Vitro Leaf Explants 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_13

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  • DOI: https://doi.org/10.1007/978-1-4939-7156-5_13

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  • 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

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