ABSTRACT
Background: Cyclamen persicum is an economically important ornamental crop that is propagated exclusively through seeds as vegetative propagation using cuttings is not possible. Therefore a micropropagation method through somatic embryogenesis is of high interest; however the method suffers from low reliability concerning quality and quantity of the produced plantlets. A crucial step of the protocol is the removal of plant growth regulators (PGRs) that triggers embryo development. In order to get a better insight in this crucial step of the propagation process, a gene expression analysis has been set up using five different genes of glutathione S-transferases (GST) as these are known to be auxin responsive as well as stress reactive. Results: One out of the five genes of glutathione S-transferases (CpGST1) displayed a clear down-regulation 72 hrs after removal of PGRs compared to 4 hrs after, implying auxin responsiveness. However, a more detailed analysis including the time points 0, 4 and 72 hrs revealed an initial strong up-regulation after 4 hrs before it was down-regulated after 72 hrs. In comparison fold-changes of the additional four GST-genes were marginal. Comparing cultures on semisolid medium to that in suspension, transcript abundances of CpGST1 were clearly decreased in suspension culture. Conclusions: Against the initial hypothesis CpGST was not auxin responsive but stress reactive, probably especially indicating drought stress imposed on the cells upon transfer from submerged suspension culture to semisolid medium. Mechanical stress caused by shaking of suspensions cultures seemed to be less important.
Subject(s)
Cyclamen/enzymology , Cyclamen/genetics , Gene Expression , Glutathione Transferase/genetics , Cell Culture Techniques , Cyclamen/embryology , Polymerase Chain Reaction , Stress, Physiological , Time FactorsABSTRACT
In the present study we examined the possibility of propagating different Cyclamen species (C. africanum Boiss. and Reut., C. cilicium Boiss. and Heldr., C. coum Mill., C. hederifolium Ait., C. persicum Mill., C. purpurascens Mill.) including some of their subspecies and cultivars in vitro using explants of adult plants. For this purpose two protocols have been applied to eleven genotypes combined with mostly four explant types (placentas with ovules, leaves, petioles and peduncles). The use of these protocols has given rise to either somatic embryo-like structures and/or adventitious shoots in all genotypes. This way it was possible to propagate each of the examined genotypes in vitro using explants of adult plants in a time less than one year. These results may be used in breeding and propagation of Cyclamen as an ornamental plant and as a medicinal plant.