Study of differential protein expression in plasma membrane of chickpea (Cicer arietinum L.) under dehydration stress
Student name: Ms Natasha Navet
Guide: Dr Anandita Singh
Year of completion: 2014
Host Organisation: National Institute of Plant Genome Research (NIPGR), New Delhi
Supervisor (Host Organisation): Dr Niranjan Chakraborty
Abstract: It is crucial for a plant to amend the protein expression pattern in order to acclimatize and resist
environment fluctuations. The alteration pattern triggered due to stress can either induce the
production of new protein or show over-expression of existing proteins that might play a pivotal
role to combat stress or switch the plant metabolism on ‘emergency mode’. Dehydration is the
most unfavourable environmental constraint that limits the plant productivity by affecting the
cellular homeostasis. Chickpea is a widely grown legume crop and has an extensive property of
susceptibility to drought. Despite its importance chickpea has remained outside the realm of
large-scale proteomics screening and functional genomics. The plasma membrane (PM) exists as
a fence between the environment and cytosol in all living cells and offer as one of the most
composite and differentiated membrane. Therefore characterization of PM proteome can impart
new perception of plant membrane system. In the present study 21 day old chickpea plants were
subjected to dehydration for 24, 72 and 144hrs which caused loss of turgor and cellular integrity
with increase in senescence and persistent drooping and wilting. The severity of stress was
confirmed by histochemical staining by DAB and NBT assay as well as through physiological
assay by computing the activity of catalase and APX. Standardization of two-phase PM
extraction protocol was done whose purity was then confirmed by immunological
characterization. Further the protein profile was obtained by running 1-D SDS PAGE using
varied acrylamide concentration. 2-DE analysis was used to generate differential PM proteome
with respect to dehydration stress. The expression pattern for control was then studied
computationally using PDQuest software where 241 putative spots were detected in the first level
match set. In the later part of the study characterization of CaDREPP1 was performed by
conducting PCR amplification, plasmid isolation and restriction digestion reaction.
Key words: Cellular homeostasis/Dehydration/ Legume crop/plasma membrane/proteome/2-D
