Utjecaj ekstrakta grmaste glavulje (Globularia alypum L.) na klijavost sjemenki i razinu neenzimskih antioksidansa u klijancima rotkvice (Raphanus sativus L.) uzgojenima u prisutnosti suviška iona bakra(II)

master's thesis

• Title (english): The effects of Globularia alypum L. on radish (Raphanus sativus L.) seed germination and on the level of non-enzymatic antioxidants in radish seedlings grown in the presence of excess copper(II) ions
• Author: Andrea Babić
• Mentor: Maja Friščić (mentor)
• Committee member: Maja Friščić (predsjednik povjerenstva)
• Committee member: Ana-Marija Domijan (član povjerenstva)
• Committee member: Miranda Sertić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of pharmaceutical botany); Zagreb
• Defense date and country: 2022-07-22, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Cilj ovog rada bio je ispitati učinak različitih koncentracija metanolnog ekstrakta grmaste glavulje (Globularia alypum L., Plantaginaceae) na klijavost sjemenki i razinu neenzimskih antioksidansa u klijancima rotkvice (Raphanus sativus L., Brassicaceae) koji su bili izloženi oksidacijskom stresu izazvanom suviškom iona bakra(II). Sjemenke su bile predtretirane dva sata destiliranom vodom i odabranim koncentracijama ekstrakta grmaste glavulje otopljenog u destiliranoj vodi (50, 500 i 5 000 μg/mL), a nakon toga tretirane su s 3,5 mL 0,005 M CuSO4 ili destilirane vode (negativna kontrola). Nakon 5 dana klijanja na dnevnoj svjetlosti pri temperaturi 20 ± 2 °C, izračunati su parametri klijavosti (ukupna klijavost, prosječno vrijeme klijanja i prosječna brzina klijanja), izmjerena duljina korjenčića i hipokotila te izvagana masa suhih klijanaca. Nakon toga su pripremljeni homogenati klijanaca rotkvice u 5%-tnoj (w/v) trikloroctenoj kiselini. Nakon centrifugiranja i odvajanja supernatanta, određen je sadržaj ukupnih fenola i koncentracija reduciranog glutationa. Tretman 0,005 M CuSO4 statistički je značajno smanjio klijavost sjemenki, duljinu korjenčića i hipokotila te suhu masu klijanaca rotkvice u odnosu na negativnu kontrolu. Također, smanjio je i razinu neenzimskih antioksidansa u klijancima rotkvice, sadržaj ukupnih fenola i koncentraciju reduciranog glutationa. Dvije primijenjene koncentracije ekstrakta grmaste glavulje (50 i 500 μg/mL) statistički su značajno povećale klijavost sjemenki rotkvice u tretmanu s bakrom (p < 0,05). U istom tretmanu, sve su koncentracije ekstrakta statistički značajno povećale duljinu hipokotila klijanaca, dok su dvije koncentracije (500 i 5 000 μg/mL) povećale duljinu hipokotila i u usporedbi s negativnom kontrolom. Nadalje, sve su koncentracije ekstrakta povećale razinu neenzimskih antioksidansa, ali nisu imale utjecaj na duljinu korjenčića i suhu masu klijanaca. Dobiveni rezultati ukazuju na to da vrsta G. alypum može smanjiti negativne učinke oksidacijskog stresa izazvanog suviškom iona bakra(II) te potaknuti rast rotkvice.

Abstract (english)

The aim of this study was to explore the effects of different concentrations of Globularia alypum L. (Plantaginaceae) methanolic extract on radish (Raphanus sativus L.) seed germination and on the level of non-enzymatic antioxidants in radish seedlings which were exposed to oxidative stress in the presence of excess copper(II) ions. Seeds were pre-treated for two hours with distilled water and chosen concentration of G. alypum extract dissolved in distilled water (50, 500 and 5000 μg/mL) and afterwards treated with 3.5 mL of 0.005 M CuSO4 or distilled water (negative control). After five days of germination in daylight at 20 ± 2 °C, germination parameters (total germination, mean germination time and mean germination rate) were assessed, the root and hypocotyl lengths measured, and the dry masses of radish seedlings weighed. Afterwards, seedling homogenates were prepared using 5% (w/v) trichloroacetic acid. After centrifugation and separation of supernatant, total phenolic content and reduced glutathione concentration were determined. Treatment with 0.005 M CuSO4 significantly decreased seed germination, the root and hypocotyl lengths, and dry mass of radish seedlings compared to negative control. It also decreased the level of non-enzymatic antioxidants in radish seedlings, total phenolic content and reduced glutathione concentration. Two applied concentrations of G. alypum extract (50 and 500 μg/mL) significantly increased germination percentages of radish seeds in the copper treatment (p < 0.05). In the same treatment, all extract concentrations significantly increased hypocotyl lengths, while two concentrations (500 and 5000 μg/mL) increased hypocotyl lengths also compared to negative control. Furthermore, all extract concentrations significantly increased the level of non-enzymatic antioxidants but had no effect on root length and seedling dry mass. Obtained results indicate that G. alypum may reduce the negative effects of oxidative stress induced by excess of copper(II) ions and promote radish growth.

• Keywords: Globularia alypum; bakar; oksidacijski stres; antioksidativni potencijal; rotkvica
• Keywords (english): Globularia alypum; copper; oxidative stress; antioxidant potential; radish
• Language: croatian
• URN:NBN: urn:nbn:hr:163:982227
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-07-26 10:52:26