Abstract
BackgroundWild medicinal plants are suffering natural environmental stresses and habitat destruction. The genetic diversity evaluation of wild accessions and their in vitro raised genotypes using molecular markers, as well as the estimation of substances of pharmaceutical value in wild plants and their regenerated genotypes are convenient approaches to test the genetic fidelity of regenerated plants as a source of substances of pharmaceutical value. In this study, the genetic diversity of 12 accessions of the medicinal plant Achillea fragrantissima, representing five sites in the mountains of South Sinai, Egypt, were estimated by the inter simple sequence repeats (ISSR) fingerprinting and their volatile oil components were identified using gas chromatography-mass spectrometry (GC-MS) analysis. The same accessions were regenerated in vitro and the genetic diversity and volatile oil components of propagated genotypes were determined and compared to their wild parents. ResultsClustering and principal component analyses indicated that the wild accessions and their regenerated genotypes were genetically differentiated, but the regenerated plants are relatively more diverse compared to their wild parents. However, genetic variation between wild accessions is inherited to their in vitro propagated genotypes indicating genotypic differentiation of the examined accessions. The number of volatile oil compounds in the wild A. fragrantissima accessions was 31 compounds while in the in vitro propagated plants only 24 compounds were detected. Four major compounds are common to both wild and regenerated plants; these are artemisia ketone, alpha-thujone, dodecane, and piperitone. ConclusionsGenome profiling and essential oil components analysis showed variations in A. fragrantissima accessions from different populations. Genetic differences between wild and regenerated genotypes were analyzed and validated with the final conclusion that in vitro conditions elicited higher genetic variation that is associated with reduced amount and diversity in the essential oil components.
Highlights
Wild medicinal plants are suffering natural environmental stresses and habitat destruction
A total of 118 inter simple sequence repeats (ISSR) bands were amplified in all wild plants using the 20 primers and the same number was recorded in the regenerated plants using 18 primers
The number and percentage of monomorphic markers are lower in wild plants compared to regenerated genotypes, whereas the number and percentage of unique alleles are higher in the regenerated plants compared to wild plants (Table 3)
Summary
Wild medicinal plants are suffering natural environmental stresses and habitat destruction. Conservation of wild medicinal plants using in vitro propagation is a vital approach to preserve their medicinal and pharmaceutical value For this objective, it is important to evaluate the genetic variability in natural populations and preserve genetic variability in the in vitro raised genotypes. The genome profiling of plants with pharmaceutical value, using different molecular fingerprinting methods, was reviewed by Gantait et al [6] Several of these methods have been applied to address the genetic fidelity of in vitro propagated plants, examples include Hedychium coronarium by inter simple sequence repeats (ISSR) [7], the endangered medicinal plant Rauwolfia tetraphylla, by start codon targeted polymorphism (SCoT), ISSR, and random amplified polymorphic DNA (RAPD) [8], Helianthus verticillatus in vitro raised plants using SSRs [9] and the blackberry (Rubus fruticosus L.) using RAPD and sequence-related amplified polymorphism (SRAP) [10]
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