Osjetljivost mutanata i nadekspresora za auksinske amidohidrolaze uročnjaka (Arabidopsis thaliana) na solni, osmotski i temperaturni stres

master's thesis

• Title (english): The sensitivity of Arabidopsis thaliana auxin amidohydrolase mutants and overexpressors on salt, osmotic and temperature stress
• Author: Matko Tudor
• Mentor: Nataša Bauer (mentor)
• Committee member: Nataša Bauer (predsjednik povjerenstva)
• Committee member: Višnja Besendorfer (član povjerenstva)
• Committee member: Ivana Šola (član povjerenstva)
• Committee member: Dunja Leljak-Levanić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2022-09-29, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Za optimalni učinak, koncentracija auksina u stanicama je strogo regulirana transportom i
metabolizmom auksina. Konjugati auksina predstavljaju skladišnu ili razgradnu formu auksina.
Auksinske amidohidolaze su enzimi koji hidroliziraju konjugate te oslobađaju aktivni oblik
auksina. U ovom istraživanju, odredio sam relativnu ekspresiju gena za auksinske
amidohidrolaze ILR1, ILL2 i IAR3 u nadekspresorskim linijama ILR1 13.3, ILR1 3.1, ILL2
25.6, ILL2 27.2, ILL2 8.2, IAR3 11.5 te IAR3 13.4 vrste Arabidopsis thaliana, ekotipa
Wassilewskija. Istražio sam utjecaj solnog (150 mM NaCl), osmotskog (300 mM manitola) i
temperaturnog stresa (38 °C) na produžni rast i razvoj korijena kod nadekspresora i mutanata
(ilr1-2, ill2-1, iar3-2, ill2-1iar3-2 i ilr1-2ill2-1iar3-2) auksinskih amidohidrolaza. Uz to,
analizirao sam lokalizaciju rekombinantnog proteina ILL2-GFP u korijenu biljke
nadekspresora ILL2 8.2 uzgajanih u uvjetima solnog, osmotskog i temperaturnog stresa.
Nadekspresorske linije ILR1 3.1, ILR1 13.3 i ILL2 27.2 imale su 731.17, 182.5 odnosno 350.93
puta veću ekspresiju odgovarajućeg gena auksinske amidohidrolaze u odnosu na divlji tip.
Navedene linije su imale i značajnu promjenu u arhitekturi korijena u uvjetima solnog stresa u
odnosu na divlji tip. U uvjetima solnog i osmotskog stresa nakupljanje proteina ILL2 bilo je
povećano u zoni sazrijevanja korijena, dok se u vršku korijena nije mijenjalo u odnosu na
kontrolne uvjete.

Abstract (english)

Auxins act in concentration dependant manner, that is tightly controled by auxin metabolism
as well as its transportation. Auxin conjugates are storage or degradable forms of auxins. Auxin
amidohydrolases are enzymes that hydrolyse conjugates and release the active form of auxin.
In this research, I measured relative expression of auxin amidohydrolases ILR1, ILL2 i IAR3 in
Arabidopsis thaliana, ecotype Wassilewskija, overexpressing lines ILR1 13.3, ILR1 3.1, ILL2
25.6, ILL2 27.2, ILL2 8.2, IAR3 11.5 and IAR3 13.4. To reveal function of auxin
amidohydrolases in plant stress response I investigated the influence of salt (150 mM NaCl),
osmotic (300 mM mannitol) and temperature stress (38 °C) on growth and development of
roots in overexpressing lines and auxin amidohydrolase mutants (ilr1-2, ill2-1, iar3-2, ill2-
1iar3-2 i ilr1-2ill2-1iar3-2). Further, I analysed localisation of recombinant protein ILL2-GFP
in the overexpressing line ILL2 8.2, grown in conditions of salt, osmotic and temperature stress.
Overexpressing lines ILR1 3.1, ILR1 13.3 and ILL2 27.2 had 731.17, 182.5 and 350.93 times
higher expression of corresponding native gene compared to wild type, respectively. The root
architecture of those overexpressing lines was significantly changed in salt stress condition,
compared to wild type. In salt and osmotic stress, the accumulation of ILL2 was significanly
increased in the root maturation zone and did not change in the root tip compared to the control
conditions.

• Keywords: indol-3-octena kiselina (IAA); konjugati indol-3-octene kiseline; arhitektura korijena
• Keywords (english): indole-3-acetic acid (IAA); conjugates of indole-3-acetic acid; root architecture
• Language: croatian
• URN:NBN: urn:nbn:hr:217:252137
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-10-26 10:55:50