Figure legends:
Figure 1. Subcellular localization of TaA6b Hsf. The ORF of TaA6bwas cloned in CaMV35S::TaA6b-GFP vector in which GFP was fused at
the C-terminus. The TaA6b-GFP fusion protein and GFP control were
transiently expressed in N. benthamiana leaves and observed by
fluorescence microscopy. The left column is the green fluorescence
channel, the center column is the DIC and the right column is the merge
of the two images.
Figure 2. Effect of constant heat stress on transgenic lines (Line 2-1,
Line 2-2 and Line 6-2) and wild type control plants (WT) in greenhouse
with constant temperature at 35 ºC. Seeds were germinated in 2 inch
nursery pots and kept in temperature controlled greenhouse from sowing
and pictures were taken at specified time.
Figure 3.A. Detection of reactive oxygen species by NBT staining method
in the leaves of barley plants from wild type (WT) and transgenic (AP2-2
and AP6-2) plants over-expressing wheat HSF (TaHsfA6b ) under
control and heat stress conditions.
B. The activity of three different ROS scavenging enzymes viz ascorbate
peroxidase (APX), catalase and super oxide dismutase (SOD) in wild type
(WT) and transgenic (AP2-2 and AP6-2) plants under heat stress
conditions. Error bars SD (n=5), P values **= <0.05 and ***=
<0.001
Figure 4.A. Principle component analysis of RNA sequencing in WT (RCWT)
and transgenic plants of two lines AP 2-1 and AP 2-2, showing grouping
of biological replicates of WT and transgenic lines.
B. Volcano plot showing comparison of differentially expressed genes
(DEGs) in wild type and transgenic barley plants. NS- Not significant ,
Log2 fold change- transcripts with fold change of >2 but
non significant p-value, P-Transcript with p value<0.05,
log2FC& P differentially expressed transcripts with log2FC
>2 and p-value < 0.05.
Figure 5. qRT-PCR expression analysis of some upregulated genes in
RNA-seq data (Supplementary table S2). Y-axis represent relative fold
change calculated by 2^-ddct method.
Figure 6. Analysis of transgenic and WT plants under simulated drought
stress treatment.A. 4 weeks-old wild type and transgenic plants of
overexpression line AP2-2, under limited water condition.
B. Relative water content of leaves of wild type and transgenic plants
of line AP2-2 kept under water limitation condition.