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Jasmin Šutković, Ajdina Karic, A. Yildirim
2 6. 10. 2020.

In silico identification and expression analysis of Metal-nicotianamine transporter (YSL3) and Oligopeptide transporter 3 (OPT3) under Cd stress in Brassica oleracea var. acephala

Background: Metal-nicotianamine transporter (YSL) family protein belongs to the oligopeptide heavy metal transporter group, as characterized in Arabidopsis thaliana. Oligopeptide transporters (OPTs) are a group of membrane-localized proteins, involved in different transport mechanisms, contributing to nitrogen mobilization, glutathione transport and long-distance metal distribution. Metal-nicotianamine transporter gene 3 (YSL3) incorporates the oligopeptide transporter domain, found to transfer several heavy metals in diverse plant species, and among them cadmium transport in Brassica oleracea. Objective: To evaluate and confirm the expression of Metal-nicotianamine transporter (YSL3) under cadmium stress. Studied species: Brassica oleracea var. acephala Study site and dates: Brassica oleracea var. acephala samples were collected from Blagaj region, Bosnia and Herzegovina. Methods: Through a simple bioinformatic approach the interactome partner of Metal-nicotianamine transporter (YSL3) was discovered and annotated. Oligopeptide transporter 3 (OPT3) and Metal-nicotianamine transporter (YSL3) genes were checked for expression levels under cadmium stress. Results: We have identified a strong interacting partner of YSL3, later confirmed as Oligopeptide transporter 3 (OPT3) protein in Brassica oleracea. The in vitro expression analysis by using a qRT-PCR revealed a significant upregulation of YSL3 and OPT3, during Cd stress. Conclusions: These findings indicate that the represented in-silico approach, followed by in vitro gene expression study, successfully confirmed YSL3 and identified OPT3 as a new gene, in correlation to cadmium stress.

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