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Organogenesis and encapsulation of in vitro-derived propagules of Carrizo citrange [Citrus sinensis (L.) Osb. × Poncirius trifoliata (L.) Raf]

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Abstract

Due to widespread polyembryony, Citrus rootstocks are usually propagated by open-pollinated seed germination, although micropropagation offers many advantages. Encapsulation technology has recently attracted the interest of researchers in the field of plant propagation because it combines the advantages of zygotic or gamic seeds with those of micropropagation. In this study, we examined the encapsulation of Carrizo citrange uninodal microcuttings (3–4 mm long) and evaluated the influence of the calcium alginate coating, a short time storage at cold temperature, and different sowing substrates on the viability and regrowth of the explants. A secondary aim was to develop an efficient protocol to induce root formation in the microcuttings. The results showed that encapsulation did not negatively affect the viability, providing a satisfactory regrowth, and storage potential for 30 days at low temperature. No differences in viability and regrowth were detected between the two different sowing substrates tested (agar-solidified medium and paper filter). To optimize the production of microcuttings required to perform the encapsulation experiments, in a preliminary experiment we assessed different factors affecting the in vitro shoot regeneration from epicotyl segments (obtained from seeds of Carrizo citrange germinated in vitro), including the influence of in vitro organogenesis, explant orientation, cut surface contact with the medium, treatments with different growth regulators, and distance of the organogenic explants from the cotyledonary node. The highest organogenic response was obtained from segments horizontally cultured (particularly from the basal portion), from segments with the cut surface in contact with the medium and from explants cultured on medium supplemented with 6-benzyladenine. No significant difference in regeneration efficiency was found in response to the distance of the epicotyl portions from the cotyledonary node.

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Abbreviations

BA:

6-benzyladenine

CSD:

Cut surface down, in contact with the medium

CSU:

Cut surface facing up

HP:

Horizontal position

IBA:

Indole-3-butyric acid

NAA:

α-naphthaleneacetic acid

TDZ:

Thidiazuron

VP:

Vertical position

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Acknowledgments

This work was supported by the project “CITRUS SYNSEED”, financed by the Assessorato Agricoltura e Foreste Regione Sicilia, XI Servizio Regionale allo Sviluppo.

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Authors and Affiliations

  1. Dipartimento DEMETRA, Università degli Studi di Palermo, Viale delle Scienze, 11, 90128, Palermo, Italy

    Maria Antonietta Germanà, Benedetta Chiancone & Laura Macaluso

  2. Dipartimento di Scienze Agrarie e Ambientali, Università degli Studi di Perugia, Borgo XX Giugno 74, 06121, Perugia, Italy

    Maurizio Micheli & Alvaro Standardi

Authors
  1. Maria Antonietta Germanà
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  2. Maurizio Micheli
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  3. Benedetta Chiancone
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  4. Laura Macaluso
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  5. Alvaro Standardi
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Correspondence to Maria Antonietta Germanà.

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Germanà, M.A., Micheli, M., Chiancone, B. et al. Organogenesis and encapsulation of in vitro-derived propagules of Carrizo citrange [Citrus sinensis (L.) Osb. × Poncirius trifoliata (L.) Raf]. Plant Cell Tiss Organ Cult 106, 299–307 (2011). https://doi.org/10.1007/s11240-011-9921-y

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