Določanje kemijske sestave terpenov in kanabinoidov ter genetske raznolikosti odbranih fenotipov navadne konoplje (Cannabis sativa L.).

Eržen, Marjeta (avtor), Čerenak, Andreja (mentor), Kreft, Samo (komentor)

V okviru doktorske disertacije smo na podlagi fenotipov znotraj treh sort konoplje (Carmagola selected (CS), Tiborszallasi in selekcija Finole) želeli ugotoviti ali med njimi obstajajo razlike tudi na kemijski in genetski ravni. Fenotipe smo med seboj najprej primerjali glede na vsebnosti komponent eteričnega olja (EO) in kanabinoidov. Najvišja vsebnost EO je bila pri selekciji Finole (2,81 mL/100 g suhe mase). Na podlagi povprečnih vsebnosti komponent EO smo lahko sorte ločili med seboj, fenotipov pa ne. Znotraj posameznih sort smo lahko ločili le fenotipa sorte CS. Fenotipi selekcije Finola so imeli najvišje vsebnosti kanabidiolne kisline (CBD-A) (6,36-6,56 %). Najvišja vsebnost tetrahidrokanabinolne kisline (THC-A) je bila pri fenotipu TV (1,39 %). Glede na povprečno vsebnost kanabinoidov smo lahko ločili selekcijo Finole od ostalih dveh sort, znotraj posameznih sort pa sta se tudi tukaj ločila le fenotipa sorte CS. Najvišja antioksidativna aktivnost je bila pri etanolnih ekstraktih sorte Tiborszallasi (73,5 %). Na podlagi antioksidativne aktivnosti tako pri vodnih kot pri etanolnih ekstraktih smo med seboj lahko ločili le fenotipa selekcije Finola. Najvišja vsebnost celokupnih polifenolov je bila pri etanolnih ekstraktih selekcije Finola (27,7 mg GAE/g). Glede na vsebnost celokupnih polifenolov so se pri etanolnih ekstraktih znotraj posameznih sort vsi fenotipi razlikovali med seboj, medtem ko sta se pri vodnih ekstraktih razlikovala le fenotipa sorte Tiborszallasi. Na podlagi 4631 odbranih SNP je bilo mogoče izdelati 4537 sond za lovljenje. Glede na sekvenčne podatke smo izvedli analizo populacijske strukture, kjer smo na genetski ravni genotipe razdelili v pet skupin (ena skupina pri CS ter dve skupini pri Tiborszallasi in selekciji Finole). Z analizo GWAS smo 45 fenotipskih lastnosti združili s končnimi 3670 pozicijami SNP ter izvedli MLM analizo. Na podlagi analize smo izrisali grafe tipa Manhattan s katerimi smo prikazali signifikantne pozicije SNP za posamezne lastnosti in določili gene, ki se na teh pozicijah nahajajo. Večino genov ni bilo neposredno povezanih s sintezo kanabinoidov ali komponent EO.

• Jezik: Slovenski jezik
• Ključne besede: Konoplja, Cannabis sativa, fenotipi, terpeni, kanabinoidi, polifenoli, antioksidativna aktivnost, genetska raznolikost, GWAS
• Vrsta gradiva: Doktorsko delo/naloga
• Tipologija: 2.08 - Doktorska disertacija
• Organizacija: BF - Biotehniška fakulteta
• Leto izida: 2023
• PID: 20.500.12556/RUL-152371
• COBISS.SI-ID: 173957123
• Datum objave v RUL: 23.11.2023

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Determination of chemical composition of terpenes and cannabinoids and genetic variability in selected hemp (Cannabis sativa L.) phenotypes.

Izvleček

Within the scope of this doctoral dissertation study, phenotypes of three different varieties of hemp (Carmagnola Selected (CS), Tiborszallasi, and Finola Selection) were examined to investigate potential differences at the chemical and genetic levels. Individual phenotypes were initially compared based on their terpene and cannabinoid content. The highest content of essential oil was found in Finola Selection (2.81 mL/100 g dry weight). Calculating the average terpene content enabled the differentiation between varieties, but not between phenotypes. The exception were phenotypes within the CS variety. Phenotypes of Finola Selection had the highest content of cannabidiolic acid (CBD-A) (6.36-6.59 %), the highest content of tetrahydrocannabinolic acid (THC-A) was in the TV phenotype (1.39 %). The average cannabinoid content distinguished the Finola Selection from the Tiborszallasi and CS varieties. Phenotypes of the CS variety were also distinguishable from one another. The highest antioxidant activity was in ethanol extract of Tiborszallasi (73.5 %). Antioxidant activity enabled the observation of differences between the phenotypes of the Finola Selection in both water and ethanol extracts. The highest total polyphenol content was in ethanol extracts of Finola Selection (27.7 mg GAE/g). Concerning the total polyphenol content, all phenotypes within individual varieties exhibited different characteristics in ethanol extracts, while only the phenotypes of the Tiborszallasi differentiated themselves in aqueous extracts. Based on the 4,631 single-nucleotide polymorphisms (SNP) that were selected, 4,537 capture probes were designed. Sequencing data were utilized to analyze the population structure allowing genotypes to be categorized into five distinct groups at the genetic level (one group at CS and two groups at Tiborszallasi and Finola Selection). Through a genome-wide association study (GWAS), 45 phenotypic traits were combined with 3,670 filtered SNP positions and a mixed linear model (MLM) analysis was done. Based on that analysis, Manhattan plots were made that illustrated the significant positions of SNPs for individual traits and identified the genes located at these positions. Most genes were not responsible for the synthesis of cannabinoids or components of essential oils.

• Ključne besede: Hemp, Cannabis sativa, phenotypes, terpenes, cannabinoids, polyphenols, antioxidative activity, genetic diversity, GWAS