In vitro propagation of the endangered species Marsilea quadrifolia L. - morphological and biochemical analysis of the regenerates

Abstract

The present study is part of the current concerns with biodiversity conservation of endangered gymnosperm species from The Natural Park of Comana. Marsilea quadrifolia species is protected both at a national and European level (through the Habitats Directive of the EU and the Bern Convention). The aspects of in vitro morphogenesis through all the stages from inoculation, multiplication to rooting and acclimatization have been studied. In order to long term conserve and multiply or to exploit somaclonal variation induced by the in vitro technique identification of some biochemical or molecular markers for fast evaluation of regenerates is required. In this respect, genetic stability or variability of Marsilea quadrifolia regenerates obtained by the in vitro culture was assessed by electrophoretical methods to relieve izoenzymes activity as esterase, acid and alkaline phosphatase, glutamate-oxaloacetate transferase, malate dehydrogenase and peroxidase. In all enzymes systems studied a higher activity correlated with reactivity of this species to in vitro culture conditions was noticed. The expression of esterase, phosphatase, malate dehidrogenase display changes in correlation with growth condition, while the peroxidase pattern is more stable in natural population than in the in vitro regenerated plantlet.