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Genomic DNA was extracted from T-DNA insertion mutant leaves using the Shorty method.
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These mutants were inoculated on detached barley leaves using droplets of conidial suspensions or on barley plants by spraying.
To compare the expression of PRX genes between the dxr mutant and WT plants, we first balanced the transcripts between their leaves using OsUbi5 as we have previously described (Chandran et al. 2016b).
To screen mutants, AA levels were analyzed qualitatively in small pieces of two-week-old rosette leaves using the nitroblue tetrazolium assay previously described.
Reverse transcription RT-PCR analysis of the total RNA from mutant leaves or flowers was performed using gene-specific primers.
As a case study of metabolic pathways or functional modules assessment using MapMan tool, we carried out transcriptome profiling of rice mutant leaves in 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) gene compared to its wild type control.
Genomic DNA was extracted from leaves of mutant 3790 using the DNeasy Plant Mini kit (Qiagen), digested with restriction enzyme EcoRI (Thermo, product # ER0275) or BamHI (Thermo, product # ER0051) and subsequently ligated with T4 DNA ligase (Fermentas, product # EL0011).
Rice mutant leaves were homogenised in liquid nitrogen.
FW, fresh weight; WT, wild type; z3 (dg), dark-green sectors of the z3 mutant leaves; z3 (g), green sectors of the z3 mutant leaves.
Analysis of reactive oxygen species (ROS) in the z3 mutant leaves.
However, the z3 mutant leaves did not show any of these defects.
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