Isolation and characterization of HSP family gene from cardamom (Elettaria cardamomum Maton)
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Date
2020
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Department of Plant Biotechnology, College of Agriculture, Vellayani
Abstract
The study entitled “Isolation and characterization of Hsp family gene from cardamom
(Elettaria cardamomum Maton)” was conducted at the Biotechnology and
Bioinformatics Division, KSCSTE - Jawaharlal Nehru Tropical Botanic Garden and
Research Institute (JNTBGRI), Palode, Thiruvananthapuram, during 2019 – 2020. The
primary objective of this study was to analyze differentially expressed genes belonging
to small heat shock (sHsp) family under heat stress from cardamom.
During the study, six month old cardamom plantlets were exposed to thermal stress by
gradually increasing the temperature from 22 °C to 40 °C to understand the mode of
expression of small heat shock protein (sHsp) transcripts namely sHsp 17.8, sHsp 17.9,
sHsp 18.1 and sHsp 18.6. After attaining 40 °C, the heat stress treatment was continued
for a period of 3 hrs and leaf tissues were collected at instants viz. 30 min, 1 hr, 2 hr
and 3 hr intervals. Total RNA was isolated from control as well as treated samples and
converted to cDNA. Real-Time PCR analysis was performed using gene specific
primers designed from cardamom transcriptome data.
The highest level of transcript accumulation was evident after the exposure of plants to
thermal stress at 40 °C for 30 min. Among the four sHsps studied, sHsp 18.1 and sHsp
17.8 showed the highest expression at 30 min with a fold change of 5571 and 5000
respectively compared to the control. The two remaining sHsps also showed
considerable upregulation during the initial period of heat stress. After 30 min, a sharp
reduction in the expression of all sHsp transcripts has been observed. In addition to
this, the overall transcript kinetics were different among the sHsp genes studied. These
differentially expressed genes were isolated, purified and sequenced. Sequence
analysis using various bioinformatics tools confirmed their identity as small heat shock
proteins. Physico-chemical parameters as well as secondary structure were predicted
using Protparam and PSIPRED respectively.
In line with our results, it can be speculated that 17.8 and 18.1 sHsps could be used as
potential biomarker candidates for screening tolerance to heat stress in cardamom. The
findings obtained from this study can be used as a foundation for future lines of research
related to rational improvement of cardamom against thermal stress.
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