Arabidopsis thaliana KO1 and KO3 deletion lines of the selective autophagy receptor AtNBR1

We used the CRISPR/Cas9 technique to construct nbr1-KO lines (KO1 and KO3) in order to test the effects of AtNBR1 depletion. Reduced expression of several ABA-up regulated genes were observed in shoots of the two KO lines. Grant number: 2014/15/B/NZ3/04854 Grant funding source: National Science Centre (Poland) Grantee Name: Agnieszka Sirko Grant title: Upstream regulators and cellular targets of the selective autophagy cargo receptor AtNBR1 in different phases of sulfur deficiency in plants. Plant material for transcript profiling was grown in Araponic boxes (16 plants/box) kept in short day conditions (22 °C 8h day/18 °C 16h night) for about 4 weeks, with weekly changes of the 0.5 x Hoagland medium. Individual plants were divided into shoot and root tissue and frozen in liquid nitrogen. Portions of plant material, drawn from at least 3 plants, were homogenised in TRI Reagent (Molecular Research Centre), and genomic DNA was removed with a RapidOut DNA removal kit (ThermoScientific) and total RNAs were separately isolated from shoots and roots. We looked for effects of the NBR1 gene deletion in the KO1 and KO3 lines compared to the the wild type plants (Col-0). These effects were investigated in roots and shoots separately. Totally, 18 samples of RNA were prepared.