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Abstract

Centaurium erythraea plants obtained by indirect organogenesis are described in the paper. The plants were initiated from a single adventitious shoot regenerated from callus derived from the cotyledon of a 30-day-old seedling. The shoot was multiplied on MS medium supplemented with IAA (0.1 mg·L-1) and BAP (1.0 mg·L-1). The multiplication rate (28 shoots per culture within 4 weeks) was highest at the first subculture and decreased in further subcultures. The shoots were rooted on MS medium. The effect of IBA (0.1 mg·L-1) on the number of shoots forming roots differed depending on the composition of the basal medium (MS). The rooted shoots were transplanted to soil and grown in a greenhouse with 90% effectiveness. RAPD analysis was done with adventitious shoots of C. erythraea from in vitro culture. In shoots and whole plants regenerated from the callus tissue, secoiridoid content was determined by the HPLC method. We showed significant differences in morphology (leaf size, fresh and dry weight and height of plants) and changes in the DNA profiles as compared to earlier reports for shoot tip-derived shoots and plants of C. erythraea, but the two groups of plants biosynthesized the same qualitative pattern and similar levels of secoiridoids, up to 150 mg·g-1 dry weight; the increased biomass of plants regenerated from callus tissue makes them a better source of secondary metabolites.

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Authors and Affiliations

Ewelina Piątczak
Aleksandra Królicka
Halina Wysokińska
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Abstract

Hladnikia pastinacifolia RCHB., a narrow endemic, has an extremely restricted distribution in Trnovski gozd (Slovenia), despite the presence of many sites with suitable habitats. We compared the morphological traits of plants from different populations and habitats. The overall pattern showed that the smallest plants, with low fruit number, are found on Èaven (locus classicus or type locality); the largest individuals, with high fruit number, grow in the Golobnica gorge. As judged by plant size and seed set, the optimal habitats are screes. We used RAPD markers to estimate genetic variation between and within populations, as well as between and within the northern and the southern parts of the distribution area. Hladnikia showed only a low level of RAPD variability. AMOVA partitioned the majority of genetic diversity within selected populations. The low genetic differentiation between populations and their genetic depauperation indicates survival in situ, since the Trnovski gozd plateau most likely was a nunatak region in the southern Prealps during Pleistocene glaciations. Later range expansion of extant populations was limited by poor seed dispersal. We also analyzed the cpDNA trnL-F intergenic spacer to check whether the sequence is useful for studying the phylogenetic relationships of Hladnikia within the family Apiaceae (Umbelliferae). Our results support the assertion that H. pastinacifolia is an old taxon.

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Authors and Affiliations

Nina Šajna
Tatjana Kavar
Jelka Šuštar-Vozliè
Mitja Kaligariè

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