characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-01-31T00:19:42Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-01-31T00:19:42Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA110401644
characteristics.characteristic: External Id
characteristics.text: PhD student
characteristics.characteristic: INSDC center name
characteristics.text: 2023-01-31T00:19:42Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-01-31T00:19:42Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS12499836
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11986:LN2-6
characteristics.characteristic: Submitter Id
characteristics.text: 18
characteristics.unit: day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: bread wheat
characteristics.characteristic: common name
characteristics.text: Cadenza
characteristics.characteristic: cultivar
characteristics.text: Protocols: In this study, the basal node is called the 0.5 cm long section from the base of the main shoot, which includes the shoot apical meristem, lateral buds and leaf meristems. After removing all the roots from the shoot, the outgrown primary tillers were also carefully removed, and 0.5 cm of the main shoot base was dissected. Samples were quickly frozen in liquid nitrogen and stored at -80 oC until processing. Tissue samples from 6 plants were pooled per biological replicated. Plants were grown hydroponically in a controlled environment chamber using a custom hydroponic system. Individual plants were grown held by foam buds on top of 1 L black pots containing the nutrient solution. The nutrient solution was aerated throughout the experiment by an aeration pump tubing system. The growing conditions were 20/10 oC day and night temperature, respectively, and 14 h day length. Lighting was provided by fluorescent bulbs at an intensity of 550 µmol m-2 sec-1. The humidity was stable at 65% during the day and 75% during the night. Total RNA was extracted from approximately 100 mg of ground frozen tissue using TRIzol reagent (Invitrogen) following the manufacturer's protocol. To ensure no DNA carryover, DNase treatment was performed using RQ1 RNase-Free DNase (Promega). After DNase treatment RNA was further purified by phenol/chloroform/isoamyl-alcohol purification. Extracted total RNA from basal node samples was further purified with Plant RNeasy kit (Qiagen) following the provided RNA clean-up protocol. Quality control, poly-A selection for rRNA removal, library preparation, multiplexing were performed by Azenta/Genewiz UK according to their standard workflow. Briefly, RNA library preparation was prepared using NEBNext Ultra II Library Prep Kit for Illumina following the manufacturer's protocol (NEB). Firstly, oligo-dt beads were used for mRNA enrichment, and the mRNAs were fragmented for 15 min at 94 oC. First and second strand cDNA were subsequently synthesized. Indexed adapters were ligated to cDNA fragments after adenylation of the 3' end of the cDNA fragments. Limited cycle PCR was used for library amplification. Sequencing libraries were validated using NGS Kit on the Agilent 5300 Fragment Analyzer (Agilent Technologies) and quantified by using Qubit 4.0 Fluorometer (Invitrogen) or equivalent.
characteristics.characteristic: description
characteristics.text: 2 formation of axillary shoot stage
characteristics.characteristic: developmental stage
characteristics.text: low nitrogen - nitrogen 0.1 millimolar; phosphorous 1 millimolar
characteristics.characteristic: environmental stress
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: Triticum aestivum
characteristics.characteristic: organism
characteristics.text: Triticum aestivum
characteristics.characteristic: organism
characteristics.text: stem node
characteristics.characteristic: organism part
characteristics.text: basal node
characteristics.characteristic: sampling site
characteristics.text: Triticum aestivum
characteristics.characteristic: scientific_name
characteristics.text: LN2-6
characteristics.characteristic: title