Agrobacterium-mediated transformation of the wild orchid Cattleya maxima Lindl

Protocorms are unique anatomical structures; they are akin to rhizoids and are formed by young orchid seedlings under physiological conditions. Explanted orchid tissues produce similar structures called protocorm-like bodies (PLBs) when exposed to appropriate in vitro growing conditions. Both the pr...

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Bibliographic Details
Published in:Universitas scientiarum 2018-03, Vol.23 (1), p.89-107
Main Authors: Cueva-Agila, Augusta Yadira, Cella, Rino
Format: Article
Language:English
Subjects:
Agrobacterium tumefaciens
Cattleya
Deoxyribonucleic acid
DNA
Embryonic growth stage
Embryos
Fluorescence
Gene expression
Genetic transformation
Hygromycin
Orchids
pCAMBIA
Plant tissues
Plantlets
Protocorm-Like-Bodies
Rhizoids
Seedlings
SERK
Somatic embryogenesis
T-DNA
Transformation
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Summary:Protocorms are unique anatomical structures; they are akin to rhizoids and are formed by young orchid seedlings under physiological conditions. Explanted orchid tissues produce similar structures called protocorm-like bodies (PLBs) when exposed to appropriate in vitro growing conditions. Both the propagative nature of PLBs and the easiness by which they can be generated, make these structures an attractive alternative to seed-mediated production for growing large numbers of plants. To increase somatic embryogenesis and optimize the procedure, PLBs of Cattleya maxima were transformed using the Agrobacterium tumefaciens method. The T-DNA carried a Hygromycin-resistance gene, a visible marker (GFP5-GUSA) and a rice gene encoding the Somatic Embryogenesis Receptor Kinase, deemed to be important for somatic embryogenesis. Treated PLBs generated somatic embryos developing Hygromycin-resistant plantlets. The insertion of T-DNA was confirmed by PCR, and GFP expression was observed using a fluorescent stereomicroscope. Transformed Cattleya maxima PLBs were more efficient in forming somatic embryos (60-80%) than untransformed controls (45-57%), and this contrast was maximized in hormone-free, Murashige and Skoog (MS) medium (80% of the transformed plants compared to 57% of the untransformed ones). This finding supports the notion that SERK plays an important role in Orchid embryogenesis.
ISSN:0122-7483
2027-1352
DOI:10.11144/Javeriana.SC23-1.amto