Staiti, Annalisa (2024) Behind the colourful surface: a study of anthocyanin pathway regulation is Solanaceae. [Tesi di dottorato]

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Abstract

Anthocyanins are a group of polyphenolic water-soluble pigments responsible for the colouration of many flowers, fruits and vegetables in several plant families. Anthocyanins draw an outstanding interest not only for their role in pigmentation but also for their protective functions in plants against environmental and biotic stresses. Moreover, due to their role in human health, research is becoming more addressed in identifying health-promoting anthocyanins and in validating their functional activity against several non-communicable human diseases. For this reason, obtaining a rife resource of anthocyanins in vitro and in planta is an enticing achievement for food, pharmaceutical and cosmetic industries. The levels of anthocyanins in plants are controlled by both synthesis and degradation processes, which are genetically regulated in a time-specific and tissue-specific manner across different stages of development. In this thesis, we focused on anthocyanin metabolism, with a particular interest in the transcriptional factors implicated in the regulation of this pathway in Solanaceae. The first aim of this research was to knock-out the gene Inducer Silencing of Anthocyanins in Cell culture (StISAC), a novel R3-MYB repressor, using CRISPR-Cas9 editing, obtaining a stable platform for anthocyanin production. The second aim was to use the edited potato cell cultures as a unique platform for investigating the possible impact of editing on all the anthocyanin metabolism. In particular, we paid attention to anthocyanin catabolism, mainly focusing on enzymes involved in an active degradation of these pigments, and to post-biosynthetic modifications mediated by decorative enzymes. The third and last aim of this thesis was to use a traditional breeding method to identify transcriptional factors involved in anthocyanin retaining in pepper. This was included as model crop due to its simpler (diploid) genetic inheritance of traits. Our findings contribute to a better understanding of the counterbalance between anthocyanin biosynthesis and degradation in Solanaceae. Altogether, results obtained in this thesis provide different strategies for obtaining in vitro and in planta stable resources of these pigments.

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