Sample 6
Hordeum vulgare subsp. vulgare
Protocols: The sample is a fragment of a barley leaf, collected from plants during the experiment. Leaves were excised using sterile scissors to avoid contamination and immediately frozen in liquid nitrogen to preserve RNA integrity. Samples were stored at -80°C until further processing.  To evaluate the impact of specific mutations in the HvSNAC1 gene on the stress response, both mutants and the parent variety 'Sebastian', which is wild type (WT) for mutants, underwent drought treatment using a protocol extensively detailed by Daszkowska-Golec et al. (2017). The drought treatment involved planting barley seeds in pots filled with a mixture of sandy loam, with sand (w/w; 7:2), and supplemented with a nutrient medium. Initially, barley seeds were germinated in Petri dishes containing water-soaked vermiculite and incubated in darkness at 4°C for two days. Subsequently, the germinated seedlings were transferred to a greenhouse for an additional two days before being transplanted into the soil, with 15 seedlings per pot. One pot was one biological replicate, and three biological replicates were tested for each genotype and condition. Soil moisture levels were monitored daily throughout the experiment using Time-domain reflectometer (TDR) EasyTest equipment provided by the Institute of Agrophysics, Polish Academy of Sciences, Poland.  The drought assay comprised of two main phases:  1. Control Growth (CG): Seedlings were grown under optimal conditions with soil moisture at 12% vwc for ten days after transplantation. 2. Drought Stress (DS): Seedlings were subjected to severe drought stress, first soil moisture was gradually reduced from 12 to 3% vwc over four days for adaptation to water deficit (AWD), then soil moisture was maintained at 3-1.5% vwc for ten days.  The entire drought test spanned 24 days. During the CG and AWD, seedlings were cultivated in a greenhouse at 20˚C/18˚C with a 16/8 hour photoperiod and 400 μEm-2s-1 light intensity from high pressure sodium lamps. After AWD, seedlings were transferred to a growth chamber with a temperature regime of 25˚C/20˚C, maintaining the same photoperiod and light intensity as in the greenhouse.   The RNAqueous™ Total RNA Isolation Kit (ThermoFisher Scientific) was used per the manufacturer protocol, but instead of a lysis buffer, a TriPure Isolation Reagent (Roche Life Science) was applied. Stranded cDNA library preparation (poly-A selection) was used.
ENA first public
2025-02-12
INSDC center name
Institute of Biology, Biotechnology and Environmental Protection, Faculty of Natural Sciences, University of Silesia in Katowice, Katowice, Poland
INSDC status
public
Submitter Id
E-MTAB-14768:Sample 6
broker name
ArrayExpress
collection date
not collected
common name
domesticated barley
cultivar
Sebastian
environmental stress
drought
genotype
hvsnac1.d
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
leaf
sample name
E-MTAB-14768:Sample 6
scientific_name
Hordeum vulgare subsp. vulgare