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Plant regeneration through direct shoot formation from leaf cultures and from protocorm-like bodies derived from callus of Encyclia mariae (Orchidaceae), a threatened Mexican orchid

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Abstract

Two procedures for the in vitro propagation of Encyclia mariae, a threatened Mexican orchid, were developed. In the first procedure, leaves from in vitro germinated seedlings were cultured on Murashige and Skoog medium (MS) supplemented with the range of 2.21–4.4 μM 6-benzylaminopurine (BA) in combination with 2.69–10.74 μM naphthalene acetic (NAA), 2.07–8.29 μM indole-3-butyric (IBA), or 2.85–11.42 μM indole-3-acetic acid (IAA) to determine the best medium for the induction of shooting. Maximum direct shoot formation from leaves was observed on MS containing 22.21 μM BA and 10.74 μM NAA (25 shoots/explant). The second procedure began with the culture of protocorms on media containing NAA, IBA, or IAA, which induced callus formation with high regenerative potential in the form of protocorm-like-bodies (PLBs) that eventually differentiated into shoots. The optimal response was attained when these structures were cultured on medium with 4.14 μM IBA (30 shoots/PLB). To promote the elongation of shoots derived from PLBs, the material was subcultured onto MS medium containing 22.21 μM BA and 5.37 μM NAA. Through the exploration of the effects of auxins and matrix on the rooting of shoots, it was determined that the optimal rooting occurred on media supplemented either with 5.71 μM IAA or 4.14 μM IBA either on agar-gelled medium or in liquid media with coir as the matrix. Rooting was found to be 20% higher in liquid media than in agar-gelled medium.

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Acknowledgments

This work was carried out with financial support from FAI-UASLP (C06-FAI-03-25.28). The authors thank CONACYT for the scholarship to CCA (no. 173383).

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Authors and Affiliations

  1. Centro de Investigación y Estudios de Postgrado, Facultad de Ciencias Químicas de la Universidad Autónoma de San Luis Potosí, Manuel Nava 6, CP 78210, San Luis Potosí, Mexico

    María del Socorro Santos Díaz & Candy Carranza Álvarez

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  1. María del Socorro Santos Díaz
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  2. Candy Carranza Álvarez
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Correspondence to María del Socorro Santos Díaz.

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Editor: Edward C. Yeung

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Santos Díaz, M.d.S., Carranza Álvarez, C. Plant regeneration through direct shoot formation from leaf cultures and from protocorm-like bodies derived from callus of Encyclia mariae (Orchidaceae), a threatened Mexican orchid. In Vitro Cell.Dev.Biol.-Plant 45, 162–170 (2009). https://doi.org/10.1007/s11627-009-9201-2

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