Fitohemijska i molekularno-genetička karakterizacija populacija kičice (Centaurium erythraea Rafn) sa Balkanskog poluostrva
Phytochemical and molecular characterization of centaury (Centaurium erythraea Rafn) populations from the Balkan Peninsula
2012
Authors:
Šiler, BranislavContributors
Mišić, DanijelaSabovljević, Aneta
Avramov, Stevan
Simonović, Ana
Document Type:
Doctoral thesis (Published version)
Metadata
Show full item recordAbstract:
Centaury (Centaurium erythraea Rafn) is a medicinal and
pharmacologically attractive plant species which has a wide range of biological
activity. The main objective of presented study was to estimate genetic and
phytochemical diversity of centaury within the Balkan Peninsula. With that
purpose, seed collection of natural centaury populations was established. The
study included 43 populations of C. erythraea, among which 41 originated from
the Balkan Peninsula. Molecular marker techniques, such as RAPD (Random
Amplification Polymorphic DNA) and TRAP (Target Region Amplification
Polymorphism) were optimized to investigate the genetic diversity among these
populations. DNA used in these analyses originated from in vitro grown plants
to ensure the authenticity of centaury DNA by eliminating the possibility of its
contamination by extraneous DNA. To acquire sufficient informational
characters, 16 decameric primers were applied in RAPD assays, which produced
408 different PCR products, while TRAP assays included the combinations of 3
specific and 3 arbitrary primers, resulting in 767 fragments of DNA. Specific
primers used for TRAP analyses were constructed according to the RNA
sequences, showing high homology with DNA sequences published in electronic
data bases for following enzymes: geraniol 10-hydroxylase, cytochrome P450
reductase and secologanin synthase. Molecular markers provided a high
percentage of polymorphism. The informativeness of single primers (RAPD) of
primer combinations (TRAP) was confirmed by various parameters. Low
within-population genetic variability of natural populations of centaury could be
explained by the high levels of selfing or geitonogamy in the process of
fertilization and by strong genetic drift. Due to the fact that centaury grows in
small, fragmented populations and that gene flow between them probably does
not exist, a high genetic differentiation of analyzed populations was expected
and this was confirmed by molecular markers. Moreover, molecular markers
Резиме/Summary
efficiently differentiated diploid populations from tetraploids. Tetraploid
populations were clearly clustered into groups related to their geographic origin,
especially when RAPD markers were analyzed. Genetic similarity data between
the Balkan Peninsula populations, obtained with either RAPD or TRAP markers,
significantly correlated with the geographic distances between localities of their
origin. Identification and quantification of secondary metabolite compounds,
secoiridoid glycosides, highly contributed to the estimation of diversity of
centaury populations in the analyzed region. A dominant secoiridoid glycoside
in aerial parts of plants grown under greenhouse conditions was swertiamarin,
which pointed to the interpopulation variability of centaury. Furthermore, a
significant correlation between the secoiridoid glycosides content and genetic
similarity of populations has been obtained, though a higher correlation was
recorded for TRAP markers. The genetic and phytochemical diversity estimation
of centaury from the Balkan Peninsula provides a basis for future biodiversity
conservation efforts and also for high-productive genotype selection and field
production improvement.
Keywords:
Centaurium erythraea; Balkan Peninsula; Habitat fragmentation; Populations; Molecular markers; RAPD; TRAP; Genetic variability; Secoiridoid glycosidesSource:
University of Belgrade, Faculty of Biology, 2012, 1-149Funding / projects:
- Physiological, chemical and molecular analysis of the diversity of selected rare and endangered plant species and application of biotechnology for ex situ conservation and production of biologically active compounds (RS-MESTD-Basic Research (BR or ON)-173024)
URI
http://eteze.bg.ac.rs/application/showtheses?thesesId=1271https://fedorabg.bg.ac.rs/fedora/get/o:2077/bdef:Content/download
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http://nardus.mpn.gov.rs/123456789/2134
https://radar.ibiss.bg.ac.rs/handle/123456789/2417