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Trevisan, Sara (2009) A genomic approach for studying the biological activity of humic substances. [Tesi di dottorato]

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Abstract (inglese)

Humic substances (HS) represent the organic material mainly widespread in nature. HS have positive effects on plant physiology by improving soil structure and fertility and by influencing nutrient uptake and root architecture. The biochemical and molecular mechanisms underlying these events are only partially known. HS have been shown to contain auxin and an “auxin-like” activity of humic substances has been proposed, but support to this hypothesis is fragmentary. In the first part of this thesis the “auxin activity” of HS has been investigated in detail by studying the induction of lateral root development in Arabidopsis thaliana with a combination of genetic and molecular evidence. The data obtained, also by using specific inhibitors of auxin transport or action, showed that HS induce lateral root formation mostly through their “auxin activity”. These findings were
further supported by the fact that the humic substances used in this study activated the auxin synthetic reporter DR5::GUS and enhanced the transcription of the early auxin responsive gene IAA19 with a time course not different from that of auxin. The presence of additional factors, different from auxin and responsible for part of the measured HS
biological activity, was also shown and discussed.
Furthermore in the second part a comprehensive transcriptomic study was used in the
model plant Arabidopsis thaliana to investigate the global molecular regulation that plant
acts in response to HS, providing a broad overview of differentially expressed transcripts
involved in this process.
The cDNA-AFLP (amplification fragment length polymorphism) technique was used to
identify more than 100 genes, which were sequenced and sorted into functional categories
according to their gene ontology annotation.
This study provides the first wide transcriptional profile for the comprehension of key
events of the HS-plant interaction and highlights the regulative role for auxin from the early
phase of this process.

Abstract (italiano)

Le Sostanze Umiche (SU) costituiscono la parte di materia organica maggiormente diffusa in natura. E’ stato ampiamente dimostrato come le sostanze umiche, agendo sulla struttura e fertilità del suolo, sono in grado di manifestare numerosi effeti postitivi sulla fisiologia della pianta, modificandone l'architettura dell'apparato radicale e conseguentemente influenzando l'assorbimento dei nutrienti. Nonostante numerosi studi ne abbiano evidenziato una forte attività biologica, i meccanismi biochimici e molecolari che regolano la risposta della pianta alle SU sono solo parzialmente conosciuti. La presenza di auxina, accertata in frazioni umiche diverse, supporta l’ipotesi avanzata da diversi autori che le SU possano agire sulla pianta attraverso una via auxino-simile. Tuttavia questo non basta a chiarire il complesso meccanismo dell’interazione SU-pianta. La prima parte di questa tesi ha avuto come obiettivo quello di verificare l’attività auxinica esercitata dalle SU nella pianta modello Arabidopsis thaliana attraverso l’osservazione dello sviluppo delle radici laterali. I risultati ottenuti grazie anche all’utilizzo di specifici inibitori del trasporto o dell’azione auxinica hanno dimostrato come le SU inducano la formazione di primordi radicali principalmente attraverso una attività auxinica. Inoltre le SU usate in questo studio hanno sia attivato l’espressione del costrutto DR5::GUS che stimolato la trascrizione di un early auxin responsive gene, IAA19, con una time-course non diversa rispetto a quella osservata in risposta all’auxina. I risultati ottenuti, comunque, non escludono la presenza di fattori addizionali diversi dall’auxina, i quali potrebbero contribuire all’attività biologica riscontrata in presenza di SU.
Nella seconda parte di questa tesi lo studio si è concentrato sui meccanismi molecolari che regolano la risposta della pianta alle SU. Per capire quali geni siano coinvolti in questo processo è stato scelto un approccio trascrittomico ad ampio spettro, in grado di individuare una vasta gamma di trascritti differenzialmente espressi in Arabidopsis thaliana in seguito a trattamenti con SU. La tecnica del cDNA-AFLP (cDNA-amplification fragment length polymorphism) ha permesso di individuare più di 100 geni, i quali sono stati in seguito suddivisi in categorie funzionali secondo la loro annotazione di gene ontology. Questo studio ha permesso di ottenere il primo ampio profilo trascrizionale in risposta alle sostanze umiche, iniziando a chiarire alcuni degli eventi chiave nell’interazione SU-pianta e evidenziando il ruolo regolativo dell’auxina sin dalle prime fasi di questo processo.

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Tipo di EPrint:Tesi di dottorato
Relatore:Quaggiotti, Silvia - Nardi, Serenella
Dottorato (corsi e scuole):Ciclo 21 > Scuole per il 21simo ciclo > SCIENZE DELLE PRODUZIONI VEGETALI > AGROBIOTECNOLOGIE
Data di deposito della tesi:21 Gennaio 2009
Anno di Pubblicazione:2009
Parole chiave (italiano / inglese):humic substances, auxin, lateral root, cDNA-AFLP
Settori scientifico-disciplinari MIUR:Area 07 - Scienze agrarie e veterinarie > AGR/13 Chimica agraria
Struttura di riferimento:Dipartimenti > pre 2012 - Dipartimento di Biotecnologie Agrarie
Codice ID:1354
Depositato il:21 Gen 2009
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