Raw data for Insight into GABA shunt-associated aldehyde dehydrogenases (ALDH) and stress responses of ALDH superfamily in moss and barley

Description of the data and file structure This supplementary dataset, integral to our research paper, contains data covering properties, interactions, and expression of studied aldehyde dehydrogenases (ALDHs). The dataset comprises gel permeation chromatography profiles of the studied plant ALDHs, microscale thermophoresis data used to describe the binding curves, as well as substrate screening profiles and saturation curves for the selected substrates, which were used to determine Km and kcat values. Thermal stability curves measured by nano-differential scanning fluorimetry are provided to show differences in stability. Quantitative PCR (qPCR) data are provided for the expression of barley ALDH gene superfamily and moss ALDH gene superfamily under various stress conditions and hormone treatments. Finally, for the aldh10A1, aldh21A1 and aldh5F2 mutants vs WT we provide RNA-seq data sets showing differentially expressed genes, phenotype and photosynthesis parameter data. Files and variables File: Gel_permeation_chromatography_dPpALDH10F1.csv Description Gel filtration of highly concentrated PpALDH10F1 (at 10 mg ml-1) on Superdex S200 column. The chromatogram shows the existence of a single peak corresponding to the dimer. Variables Elution time for dPpALDH10F1 (min): Elution time in minutes dPpALDH10F1 (Abs at 280 nm): Absorbance of protein at 280 nm Elution time for standards (min): Elution time in minutes Bio-Rad standards (Abs at 280 nm): Absorbance of protein at 280 nm Amy+Apo STD (Abs at 280 nm): Absorbance of protein at 280 nm File: Gel_permeation_chromatography_PpALDH5F1.csv Description Gel filtration of highly concentrated PpALDH5F1 (at 10 mg ml-1) on Superdex S200 column. The chromatogram shows the existence of a single peak corresponding to the tetramer. Variables Elution time (min): Elution time in minutes PpALDH5F1 (Abs at 280 nm): Absorbance of protein at 280 nm Bio-Rad standards (Abs at 280 nm): Absorbance of protein at 280 nm File: Moss_knockout_phenotype_mannitol.csv Description Growth after point inoculation on modified Knop medium and 1% plant agar. Cultivated for one month with mannitol (500 mOsm/kg) at 19 °C. Graph values indicate mean ± S.D. (n=15). Data were analyzed in a t-test (*, **, and *** correspond to P-values of 0.05 > p > 0.01, 0.01 > p > 0.001, and p < 0.001, respectively). Variables Day WT colony diameter (cm) WT colony diameter st deviation (cm) ppaldh5F2 colony diameter (cm) ppaldh5F2 colony diameter st deviation (cm) ppaldh5F2 vs WT colony diameter t-test ppaldh10A1 colony diameter (cm) ppaldh10A1 colony diameter st deviation (cm) ppaldh10A1 vs WT colony diameter t-test ppaldh21A1 colony diameter (cm) ppaldh21A1 colony diameter st deviation (cm) ppaldh21A1 vs WT colony diameter t-test File: Moss_knockout_phenotype_NaCl.csv Description Growth after point inoculation on modified Knop medium and 1% plant agar. Cultivated for one month with 300 mM NaCl at 19 °C. Graph values indicate mean ± S.D. (n=15). Data were analyzed in t-test (*, **, and *** correspond to P-values of 0.05 > p > 0.01, 0.01 > p > 0.001, and p < 0.001, respectively). Variables Day WT colony diameter (cm) WT colony diameter st deviation (cm) ppaldh5F2 colony diameter (cm) ppaldh5F2 colony diameter st deviation (cm) ppaldh5F2 vs WT colony diameter t-test ppaldh10A1 colony diameter (cm) ppaldh10A1 colony diameter st deviation (cm) ppaldh10A1 vs WT colony diameter t-test ppaldh21A1 colony diameter (cm) ppaldh21A1 colony diameter st deviation (cm) ppaldh21A1 vs WT colony diameter t-test File: Moss_knockout_phenotype_standard_conditions.csv Description Growth after point inoculation on modified Knop medium and 1% plant agar. Cultivated for one month under normal conditions at 19 °C. Graph values indicate mean ± S.D. (n=15). Data were analyzed in t-test (*, **, and *** correspond to P-values of 0.05 > p > 0.01, 0.01 > p > 0.001, and p < 0.001, respectively). Variables Day WT colony diameter (cm) WT colony diameter st deviation (cm) ppaldh5F2 colony diameter (cm) ppaldh5F2 colony diameter st deviation (cm) ppaldh5F2 vs WT colony diameter t-test ppaldh10A1 colony diameter (cm) ppaldh10A1 colony diameter st deviation (cm) ppaldh10A1 vs WT colony diameter t-test ppaldh21A1 colony diameter (cm) ppaldh21A1 colony diameter st deviation (cm) ppaldh21A1 vs WT colony diameter t-test File: Photosynthetic_parameters NPQ.csv Description Non-photochemical quenching (NPQ) in PSII in moss aldh mutants. Data were recorded in cultures dark-adapted for 30 min and then exposed to actinic light (278 µmol photons m–2 s–1). The parameters were calculated using saturating red-light pulses (300 ms, 10,000 µmol photons m–2 s–1) during actinic light exposure, followed by a dark relaxation phase. Values are means ± S.D. (n=7). Recorded on a Dual-PAM100 system. Variables time [s] WT average WT SD ppaldh21A1 average ppland21 SD ppaldh5F2 average ppaldh5F2 SD ppaldh10A1 average ppaldh10A1 SD File: Photosynthetic_parameters YI.csv Description The effective quantum yield of photosystem I photochemistry Y(I) in moss aldh mutants. Data were recorded in cultures dark-adapted for 30 min and then exposed to actinic light (278 µmol photons m–2 s–1). The parameters were calculated using saturating red-light pulses (300 ms, 10,000 µmol photons m–2 s–1) during actinic light exposure, followed by a dark relaxation phase. Values are means ± S.D. (n=7). Recorded on a Dual-PAM100 system. Variables time [s] WT average WT SD ppaldh21A1 average ppland21 SD ppaldh5F2 average ppaldh5F2 SD ppaldh10A1 average ppaldh10A1 SD File: Photosynthetic_parameters YNA.csv Description The effective quantum yield of non-photochemical energy dissipation in PSI due to donor (Y(ND)) side limitation in moss aldh mutants. Data were recorded in cultures dark-adapted for 30 min and then exposed to actinic light (278 µmol photons m–2 s–1). The parameters were calculated using saturating red-light pulses (300 ms, 10,000 µmol photons m–2 s–1) during actinic light exposure, followed by a dark relaxation phase. Values are means ± S.D. (n=7). Recorded on a Dual-PAM100 system. Variables time [s] WT average WT SD ppaldh21A1 average ppland21 SD ppaldh5F2 average ppaldh5F2 SD ppaldh10A1 average ppaldh10A1 SD File: Photosynthetic_parameters YND.csv Description The effective quantum yield of non-photochemical energy dissipation in PSI due to acceptor (Y(ND)) side limitation in moss aldh mutants. Data were recorded in cultures dark-adapted for 30 min and then exposed to actinic light (278 µmol photons m–2 s–1). The parameters were calculated using saturating red-light pulses (300 ms, 10,000 µmol photons m–2 s–1) during actinic light exposure, followed by a dark relaxation phase. Values are means ± S.D. (n=7). Recorded on a Dual-PAM100 system. Variables time [s] WT average WT SD ppaldh21A1 average ppland21 SD ppaldh5F2 average ppaldh5F2 SD ppaldh10A1 average ppaldh10A1 SD File: qPCR_barley.csv Description Gene expression screening in barley upon hormone or stress treatments. Barley was grown in Hoagland's solution and expression was determined by qPCR using dual-labeled FAM-TAM probes and RT Luna Universal Probe qPCR Master Mix on a QuantStudio 5 Real-Time PCR System. Fold change in relative expression vs control in biological triplicates was statistically analyzed with a t-test; *, **, and *** correspond to p-values of 0.05 > p > 0.01, 0.01 > p > 0.001, and p < 0.001, respectively. Variables Gene name PEG (Fold change in relative expression vs control) PEG SD (Fold change) PEG t-test NaCl (Fold change in relative expression vs control) NaCl SD (Fold change) NaCl t-test ABA (Fold change in relative expression vs control) ABA SD (Fold change) ABA t-test MeJA (Fold change in relative expression vs control) MeJA SD (Fold change) MeJA t-test BA (Fold change in relative expression vs control) BA SD (Fold change) BA t-test 2,4-D (Fold change in relative expression vs control) 2,4-D SD (Fold change) 2,4-D t-test Excess of nitrogen (Fold change in relative expression vs control) Excess of nitrogen SD (Fold change) Excess of nitrogen t-test Lack of nitrogen (Fold change in relative expression vs control) Lack of nitrogen SD (Fold change) Lack of nitrogen t-test Heat (Fold change in relative expression vs control) Heat SD (Fold change) Heat t-test Heat Recovery for 1h (Fold change in relative expression vs control) Heat Recovery for 1h SD (Fold change) Heat Recovery for 1h t-test Heat Recovery for 4h (Fold change in relative expression vs control) Heat Recovery for 4h SD (Fold change) Heat Recovery for 4h t-test Cold for 5h (Fold change in relative expression vs control) Cold for 5h SD (Fold change) Cold for 5h t-test File: qPCR_moss.csv Description Gene expression screening in moss upon hormone or stress treatments. Moss was grown in Knop's medium and expression was determined by qPCR using dual-labeled FAM-TAM probes and RT Luna Universal Probe qPCR Master Mix on a QuantStudio 5 Real-Time PCR System. Fold change in relative expression vs control in biological triplicates was statistically analyzed with a t-test; *, **, and *** correspond to p-values of 0.05 > p > 0.01, 0.01 > p > 0.001, and p < 0.001, respectively. Variables Gene name Mannitol (Fold change in relative expression vs control) Mannitol SD (Fold change) Mannitol t-test NaCl (Fold change in relative expression vs control) NaCl SD (Fold change) NaCl t-test ABA (Fold change in relative expression vs control) ABA SD (Fold change) ABA t-test MeJA (Fold change in relative expression vs control) MeJA SD (Fold change) MeJA t-test BA (Fold change in relative expression vs control) BA SD (Fold change) BA t-test 2,4-D (Fold change in relative expression vs control) 2,4-D SD (Fold change) 2,4-D t-test Excess of nitrogen (Fold change in rela