dc.description.abstract |
Plants are sessile organisms and have evolved with numerous mechanisms for
overcoming various environmental stresses. In response to various abiotic stresses, plants are
used to transcribe a class of proteins known as heat shock proteins (HSPs). The mysterious and
relatively understudied mitochondrial small heat shock proteins (msHSPs) evolved in the plant
lineage, which are the extensive variety of HSPs found in all three domains of life, are
intimately linked to protein homeostasis and survival under stress conditions. The present
investigation systematically characterized a novel mitochondrial sHSP (SmsHSP24.1) from
Eggplant (Solanum melongena L.) by transgenic overexpression and CRISPR/Cas9 mediated
genome editing method. Differential expression of SmsHSP24.1 suggested that it plays a
constructive role under stressful circumstances. The SmsHSP24.1 overexpressed Escherichia
coli-BL21 cell line demonstrated exceptional thermo-tolerance, withstanding temperatures of up
to 52ºC. The protein had a multimeric structure that was unveiled by spectrometry and electron
microscopy assay, acting as a molecular chaperone at high temperatures. In constitutively
overexpressed Eggplant lines, overexpression of SmsHSP24.1 dramatically improved resilience
to heat, drought, and salt stresses and demonstrated quick germination and seedling vigour in
Eggplant. According to the RNAseq data, the glutathione (GHS) pathway's reactive oxygen
species (ROS) scavenging enzymes appear to be upregulated. Genes essential to the
mitochondrial electron transport chain (ETC) were also subject to transcriptional modifications.
Auxin biosynthesis, cell wall transport, and repairs-related genes were also found to be
at elevated levels. Quantitative PCR (qPCR), biochemical, and physiological parameters of
transgenic Eggplants were found to play a crucial role in a range of stress responses and plant
growth and development. Apart from that, approximately 2.0 kb upstream region predicted as
SmsHSP24.1 promoter was cloned, followed by the presence of numerous stress-related cisregulatory
elements
has
been
confirmed
by
GUS
expression
assay,
indicating
that
the
protein
is
likely
to
be
expressed
as
inducible
way in transgenic Eggplant lines. Furthermore,
CRISPR/Cas9 mediated knocked out of SmsHSP24.1 protein also indicated the indispensable
role of this protein in stress response. Our results indicated an entirely novel source of the
mitochondrial small heat shock protein (SmsHSP24.1) and its positive physiological role in
transgenic Eggplant. Consequently, this gene has enormous potential in engineering stressresilient
crop
plants
to
boost
agricultural
production
and
ensure
our
food
security. |
en_US |