Tesi etd-10212024-103503
Link copiato negli appunti
Tipo di tesi
Dottorato
Autore
FIORITI, FRANCESCO
URN
etd-10212024-103503
Titolo
Identification of bioactive compounds through a molecular high throughput screening approach
Settore scientifico disciplinare
BIO/04
Corso di studi
Istituto di Scienze della Vita - PHD IN AGROBIOSCIENZE
Commissione
relatore Prof. PERATA, PIERDOMENICO
Parole chiave
- abiotic/biotic stress
- bioactive molecules
- Biologicals
- Chemicals
- Genomics
- Hormones
- Reporter lines
- Screening HTS
Data inizio appello
13/12/2024;
Disponibilità
parziale
Riassunto analitico
The present work a screening platform aimed at scouting active molecules in plants has been developed, which allows the rapid identification of a high number of molecules and the selection of those involved in a specific biological process. This platform is based on the the use of transgenic plants expressing a reporter system capable of producing a visible signal when subjected to a specific treatment condition. To achieve this goal, a large collection of reporter lines has been developed, preceded by an in-depth transcriptomic analysis to correlate certain physiological response conditions with the expression of specific genes. The resulting reporter lines will serve as the fundamental element for a molecular bioassay based on growing plants in 96-well plates, employed in a high-throughput screening context.
A chemical library purchased from MedChem Express, consisting of 2,235 molecules, was screened using two reporter lines, pIAA2::3xGFP and pARR5::GFP, which specifically respond to auxins and cytokinins. This screening allowed for the identification of a total of 42 biologically active molecules in plants, approximately 1.8% of the total. Of these, 64% are represented by molecules that have not yet been characterized in plant or published.
There is significant commercial, publishing, or patenting interest in molecules with auxin-like activity. For this reason, an in-depth study was performed on compound 20. This study revealed that Compound 20 can be considered a bioactive molecule with auxinic activity, capable of significantly influencing the distribution and active transport of auxins through carriers that mediate efflux and influx in root tissues.
In conclusion, this work enabled the development of a high-throughput molecular screening bioassay for molecules, from which phenotypic and molecular information can be obtained using transgenic reporter plants activated by specific conditions. The scientific approach adopted in this work represents a proof of concept that can be used in the future for scouting new active molecules, as well as for studying complex raw materials that are widely used in agriculture today in the form of biological formulations.
A chemical library purchased from MedChem Express, consisting of 2,235 molecules, was screened using two reporter lines, pIAA2::3xGFP and pARR5::GFP, which specifically respond to auxins and cytokinins. This screening allowed for the identification of a total of 42 biologically active molecules in plants, approximately 1.8% of the total. Of these, 64% are represented by molecules that have not yet been characterized in plant or published.
There is significant commercial, publishing, or patenting interest in molecules with auxin-like activity. For this reason, an in-depth study was performed on compound 20. This study revealed that Compound 20 can be considered a bioactive molecule with auxinic activity, capable of significantly influencing the distribution and active transport of auxins through carriers that mediate efflux and influx in root tissues.
In conclusion, this work enabled the development of a high-throughput molecular screening bioassay for molecules, from which phenotypic and molecular information can be obtained using transgenic reporter plants activated by specific conditions. The scientific approach adopted in this work represents a proof of concept that can be used in the future for scouting new active molecules, as well as for studying complex raw materials that are widely used in agriculture today in the form of biological formulations.
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