Abstract
Cattleya tigrina is endemic to the Atlantic forest biome and classified as vulnerable in the Red Book of Brazilian Flora. In vitro techniques comprise valuable tools for the conservation of endangered plant species. The aim of the present study was to evaluate the morphological features, global DNA methylation levels and free polyamines during protocorm-like bodies (PLBs) induction of C. tigrina . Along with that, an efficient protocol for in vitro propagation of this species is proposed. The first evidences of PLBs induction in C. tigrina occurred at seven days in culture, starting from the basal portion of the leaf abaxial surface. A hypomethylation marked the beginning of cell differentiation, followed by an increased global DNA methylation at 35 days in culture, coinciding with a subtle change in the structures morphogenetic development. During PLBs induction, putrescine exhibited higher levels as compared to spermidine and spermine, and apparently presents a major role during the PLBs induction in C. tigrina . Due to the apparent secondary PLBs formation, this protocol can represent a highly efficient method for in vitro propagation of this species.
Highlights
The Atlantic Forest biome is worldwide recognized by its high diversity and endemism of species
Hypomethylation coincided with the beginning of cell differentiation (Figure 1B), and was maintained during the developmental stages, that showed an increase in the number and volume of protocorm-like bodies (PLBs) (Figure 1C, D and E)
The increased global DNA methylation at 35 days in culture coincided with a subtle change in the PLBs morphogenetic development (Figure 1F)
Summary
The Atlantic Forest biome is worldwide recognized by its high diversity and endemism of species. Considered a biodiversity hotspot, it holds about 20,000 different plant species, of which 8,000 are endemic (Myers, Mittermeier, Mittermeier, Da Fonseca, & Kent, 2000). The family Orchidaceae includes 27,801 species distributed in 899 genera. Biological Sciences (The Plant List, 2013), being one of the largest and specialized families in the plant kingdom, spread throughout almost all continents, especially in tropical and subtropical regions (Hossain et al, 2013). Richard is an endemic orchid species to Atlantic forest biome, and classified as vulnerable in the Red Book of Brazilian Flora (Martinelli & Moraes, 2013)
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