Drought-induced molecular changes in crown of various barley phytohormone mutants

Figures & data

Table 1. Description of plant material.

	Genotype^a	Characteristics
1	Bowman	Two-rowed U.S. spring-type cultivar used for development of NILs: line BW827 (with sdw1.a allele), line BW828 (with sdw1.d allele) and line BW885 (carrying uzu1.a allele)
2	sdw1.a (GA)	NIL line BW827 developed in the genetic background of ‘Bowman’ cultivar, carrying mutation in HvGA20ox2 gene (allele derived from semi-dwarf mutant induced in cv. Jotun
3	sdw1.d (GA)	NIL line BW828 developed in the genetic background of ‘Bowman’ cultivar, carrying mutation in HvGA20ox2 gene (allele derived from cv. Diamant)
4	uzu1.a (BR)	NIL line BW885 line developed in the genetic background of ‘Bowman’ cultivar, characterized by defect in BR signaling pathway caused by mutation in the HvBRI1 gene (spontaneous mutation derived from cv. Baitori 11)
5	sdw1.d/uzu1.a (GA/BR)	A double mutant obtained through a cross between the mutants sdw1.d and uzu1.a, identified on the basis of HvGA20ox2 and HvBRI1 sequencing data
6	Sebastian	Two-rowed, spring cultivar, the parental cultivar for HorTILLUS TILLING population and thus of hvd14.d mutant
7	hvdwarf14.d (SL)	Mutant in HvD14 gene, impaired in strigolactone signaling

^aThe symbol of the hormone whose pathway is disturbed by the mutation is given in brackets.

Table 2. Number of SNPs between genotypes with effect on protein translation assigned by VEP tool.

Number of SNPs	Effects predicted by VEP (Ensembl Plants) for 45,456 SNP
	HIGH	LOW	MODERATE	MODIFIER
46170	89	17756	11601	16010

Figure 1. (a) Number of down- (green) and upregulated (red) genes for seven genotypes in drought vs. control comparison at two time points T1 and T2, (b) Venn diagrams visualizing the number of differentially expressed genes in crown tissue for seven genotypes specific and shared between time points T1 and T2.

Table 3. Number of DEGs related to GA, BR, SL, and drought response identified in seven genotypes under early (T1) and late (T2) drought.

Genotype	Time point	Number of DEGs annotated to:
		GA	BR	SL	Drought
Bowman	T1	1	3	0	2
	T2	27	11	6	24
sdw1.a	T1	0	0	0	2
	T2	20	12	10	23
sdw1.d	T1	2	0	0	1
	T2	25	21	11	27
uzu1.a	T1	1	2	0	7
	T2	5	2	2	16
sdw1.d/uzu1.a	T1	4	1	0	10
	T2	21	10	6	26
Sebastian	T1	2	0	0	0
	T2	24	17	10	28
hvdwarf14.d	T1	4	1	1	4
	T2	2	2	2	13

Figure 2. Heatmap of log2FC values with grouping of differentially expressed genes (DEGs) related to gibberellins (GAs), brassinosteroids (BRs), and strigolactones (SLs) in drought and control comparison (contrasts D vs C) for seven genotypes in two time points (T1, T2); DEGs annotated to transcription factor bHLH family are in bold.

Figure 3. Heatmap of log2FC values with grouping of differentially expressed genes (DEGs) related to drought in drought and control comparison (contrasts D vs C) for seven genotypes in two time points (T1, T2).

Figure 4. (a) Number of down- (green) and upregulated (red) proteins for seven genotypes in drought vs. control comparison at two time points T1 and T2, (b) Venn diagrams visualizing the number of differentially abundant proteins in crown tissue for seven genotypes specific and shared between time points T1 and T2.

Figure 5. Heatmap of log2FC values with grouping of selected differentially abundant proteins (DAPs) in drought and control comparison (contrasts D vs C) for seven genotypes in two time points (T1, T2).

Figure 6. Heatmap of log2FC values with grouping of phytohormones in drought and control comparison (contrasts D vs C) for seven genotypes in two time points (T1, T2).

Figure 7. The network of differentially expressed genes related to GAs, BRs, SLs (blue), differentially abundant proteins (green) and hormones (red) with substantially correlated expression profiles (adjacency coefficient > 0.3).

Figure 8. Grouping of post-harvest traits (T1–22; see table S1A) in drought and control comparison (contrasts D vs C) for seven genotypes.

Data availability statement

The authors confirm that the data supporting the findings of this study are available within the article and its supplementary materials are available in Figshare at https://figshare.com/s/0dfe5e7e8b95f4c87b19.

Additionally, RNA-seq data used in this paper are available in the ArrayExpress repository, accession number E-MTAB-13358 (https://www.ebi.ac.uk/biostudies/arrayexpress/studies/E-MTAB-13358).