Univerza na Primorskem Fakulteta za matematiko, naravoslovje in informacijske tehnologije
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Vsebina projekta / Project content
Smilj Helichrysum italicum (Roth) G. Don izvira iz Sredozemlja in vsebuje številne fito-spojine, pomembne za zdravje ljudi. Odkritje različnih kemotipov smilja je prispevalo k njegovi priljubljenosti. Poleg uporabe v ljudski tradicionalni medicini je rastlina zanimiva tudi za farmacevtsko, dišavno, kozmetično in živilsko industrijo. Karakteristike kserofita in njegova odlična prilagoditev stresnim okoljskim razmeram v Sredozemlju so prispevali k širjenju pridelave smilja v Jadranski regiji, vključno s Slovensko Istro. Kljub naraščajočemu pomenu v sredozemskem kmetijstvu, pa celostne raziskave smilja in njegovih potencialov bistveno zaostajajo za drugimi aromatičnimi rastlinami.
Visoko ekonomsko vrednost H. italicum gre pripisati raznovrstnim rastlinskim izvlečkom in eteričnemu olju, pripravljenih iz različnih delov te rastline, ki se kemično razlikujejo glede na ekstrakcijske parametre, izvor rastlinskega materiala, razvojno stopnjo, taksonomijo rastline in okoljske dejavnike. Različne fito-spojine ali njihove kombinacije sprožajo dokumentirane blagodejne zdravstvene učinke.
Kljub izjemnemu pomenu sekundarnih metabolitov v življenju človeka in visokemu gospodarskemu pomenu H. italicum, je za H. italicum na voljo zelo malo genomskih in transkriptomskih podatkov. Študije genetske diferenciacije, s poudarkom na delitvi podvrst in ločevanju med naravno rastočimi in gojenimi ekotipi, so izjemno skromne, čeprav je fenotip, skupaj s kemotipom in njegovimi terapevtskimi karakteristikami, tesno povezan z genotipom. In dejstvo, da za vrste Helichrysum ni na voljo kodominantnih, visoko informativnih molekulskih markerjev in le 105 nukleotidnih zaporedij v podatkovni bazi NCBI, potrjuje, da so raziskave smilja v izjemno začetni fazi. Posledično ni na razpolago informacij o genih, ki se vključujejo v biosintezo sekundarnih metabolitov in v odgovore na okoljske dejavnike. Zaradi tega sta nujna identifikacija genov, ki so odgovorni za sintezo najdragocenejših, biološko pomembnih molekul, ter razvoj z njimi povezanih molekulskih markerjev, in sicer s poglobljenimi raziskavami transkriptomskih in genomskih podatkov.
Glavni cilj projekta je določiti in karakterizirati najobetavnejše genotipe H. italicum na območju severnega Jadrana. Ta namen bo dosežen preko naslednjih specifičnih ciljev: 1) morfološko vrednotenje in agronomsko vrednotenje različnih populacij jadranskega H. italicum, ki so zbrane v kolekcijskem nasadu Purissima (vzpostavljenem maja 2018 v Slovenski Istri, Ankaran, SVN); 2) sekvenciranje celotnega transkriptoma H. italicum s tehnologijo naslednje generacije sekvenciranja (NGS), analiza transkriptoma (mRNA), anotacija in identifikacija glavnih genov, vključenih v biosintezo sekundarnih metabolitov; 3) razvoj genomskih (iz DNA-seq) in genskih (iz mRNA-seq) mikrosatelitskih markerjev za genotipizacijo, za zanesljivo identifikacijo taksonov, za razlikovanje različnih vrst in podvrst Helichrysum in za druge študije raznolikosti; 4) vrednotenje genetske raznolikosti rastline v Jadranski regiji za določitev strategije ohranjanja.
Podoktorski projekt se bo izvajal v štirih delovnih sklopih (DS), naslovljenih: Vrednotenje genskih virov smilja v kolekcijskem nasadu (DS1), Sekvenciranje celotnega transkriptoma H. italicum in funkcijska anotacija (DS2), Razvoj mikrosatelitov in validacija markerjev (DS3) ter Aplikacija SSR za genotipizacijo in populacijske študije (DS4). Projekt bo zagotovil ključne, edinstvene in široko uporabne informacije o H. italicum in bo združil pristope raziskovalnih področij kmetijstva, botanike, taksonomije rastlin in genetike (genomike in transkriptomike). Uvod v podrobno analizo transkriptoma ter razvoj novih genskih in genomskih molekulskih markerjev predstavljajo nove pristope pri H. italicum, ki bodo prispevali k razvoju znanstvenih ved žlahtnjenja rastlin, funkcijske genomike in biokemije.
ANG
The native Mediterranean plant of immortelle Helichrysum italicum (Roth) G. Don contains numerous phytochemicals beneficial for human health. Discovery of distinct chemotypes has contributed to its popularity. In addition to the traditional use in folk medicine, the plant is also interesting for pharmaceutical, fragrance, cosmetic and food industry. Xerophyte characteristics and its excellent adaptation to stressed environmental conditions have contributed to its cultivation in the Adriatic region, including Slovenian Istria. Despite its growing importance in the Mediterranean agriculture, the integrated research of immortelle and its potential is significantly lagging behind of other aromatic plants.
The high economic value of H. italicum is attributed to various extracts and essential oils prepared from different parts of the plant, which chemically differ depending on the extraction process parameters, the origin of plant material, developmental stage, plant taxonomy, and environmental conditions. Various phytochemicals or their combinations may exert the documented beneficial health effects of these extracts.
Regardless of huge importance of secondary metabolites in human life and high economic relevance of H. italicum, the availability of genomic and transcriptomic data for H. italicum is still limited. Studies on genetic differentiation with the attempted division into subspecies and separation of wild and cultivated ecotypes are very scarce, although the phenotype together with chemotype and its overall therapeutic characteristics is always intrinsically linked to the genotype. Furthermore, no codominant, high informative molecular markers and only 105 nucleotide sequences at NCBI database confirming very initial phase of genetic research of Helichrysum species. Consequently, no information on genes responsible for biosynthesis of secondary metabolites and on their response to the environment is available. Therefore, we urgently need to identify genes dictating the synthesis of most valuable, bioactive molecules, and to develop the associated molecular markers through indepth investigation of transcriptomic and genomic data.
The main goal of the postdoctoral project is to identify and characterize the most promising genotypes of H. italicum from the North Adriatic growing region. This aim will be reached through specific objectives: 1) Morphological and agronomical evaluation of different populations of Adriatic H. italicum, grown in the field collection Purissima (established in May 2018 in the Slovenian Istria, Ankaran, SVN); 2) Deep transcriptome sequencing of H. italicum with next generation sequencing (NGS) technology, detailed transcriptome (mRNA) analysis, annotation and identification of major genes involved in biosynthesis of secondary metabolites; 3) Development of genomic (from DNA-seq) and genic (from mRNA-seq) microsatellite markers for genotyping, reliable taxon identification, differentiation of different Helichrysum species/subspecies and other diversity studies; 4) Assessment of genetic diversity of the plant in the Adriatic region for conservation strategy determination.
The postdoctoral project will be implemented in four work packages (WPs), namely: Field collection of immortelle germplasm evaluation (WP1), Deep transcriptome sequencing of H. italicum and functional annotation (WP2), Microsatellite discovery and markers validation (WP3) and SSR application in genotyping and population studies (WP4). The project is going to deliver crucial, unique and widely applicable information on H. italicum as well as to link the approaches from research fields of agriculture, botany, plant taxonomy and genetics (genomics and transcriptomics). The introduction of in-depth transcriptome analysis and development of new genome-wide and gene-based molecular markers represent new approaches for H. italicum, enabling development of scientific fields such as plant breeding, functional genomics and biochemistry.