Cloning and characterization of dehydration stress responsive proteins in chickpea (Cicer arietinum L.).
Student name: Ms Divya Jain
Guide: Dr Sonika Gupta
Year of completion: 2017
Host Organisation: National Institute of Plant Genome Research, New Delhi
Supervisor (Host Organisation): Dr Niranjan Chakraborty
Abstract: Dehydration is not only the most common environmental stress imposed on plants, but also one of the most prominent factor causing losses in crop production. Chickpea, a legume with multitude of nutrient benefits due to its high protein content is susceptible to dehydration. With increasing necessity for studier varieties that can withstand a barrage of onslaught from the environment as well as pathogens has led to compound interest in stress dedicated studies with proteins as their focal point. Importance of plasma membrane in maintenance of cell homeostasis and ion concentration has been known for years and hence, exploration of plasma membrane for better understanding of changes in cellular dynamics during stress are of ultimate prominence. Plasma membrane proteome analysis of chickpea subjected to144h of dehydration revealed many differentially regulated proteins with probable functions in dehydration resistance. Amongst these, two novel proteins NCL and ANK were cloned into YFP containing destination vector and studied for protein localization. Also, deletion variants of a well characterized dehydration responsive protein CaDREPP were examined to understand the feature each amino acid has in the process of localization. Cell wall lacking plant protoplast as means of confirmation for protein localization to the plasma membrane after transformation with naked DNA was also carried out. Physiochemical studies to visualize stress as ROS accumulation with NBT and DAB assay highlighted the severity of the plant state after 144hrs of dehydration. Corroboration of the proteomics data with transcript fold change through RT-PCR provided a new outlook at the early or late nature of the dehydration response put forth by the plant through these proteins. Protein characterization with In-Silico recognition of conserved domains brought insights about possible functions of NCL as sodium calcium exchanger and ANK as ankyrin repeat containing mediator of protein-protein interactions respectively. Thus, the pathway for further characterization of these proteins amounts a fascinating future prospect.
Key words: Dehydration, Chickpea, Proteome, Plasma membrane, Protein localization.
