Life Sciences & Biotechnology
Title : | Molecular and genetic analysis of phosphorus (P) deficiency related phospholipases for enhanced P deficiency tolerance in rice |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Amarjeet Singh, National Institute Of Plant Genome Research (NIPGR), New Delhi |
Timeline Start Year : | 2023 |
Timeline End Year : | 2026 |
Contact info : | amarjeet.singh@nipgr.ac.in |
Equipments : | 4°C Refrigerator
4°C Refrigerator |
Details
Executive Summary : | Phosphorus (P) is an essential macronutrient for plant growth and development, and it accounts for upto 0.5% of plant dry weight. It is an integral constituent of Important macromolecules, including nucleic acids, proteins, and phospholipids. Low availability of P adversely affects photosynthesis, respiration and root growth which could be detrimental for plant development and yield. Globally, most agricultural soils, especially rice fields are P deficient thus, rice cultivation demands for huge amount of expensive P fertilizer application. Extensive use of chemical fertilizers enforces a financial burden, and it also poses an environmental hazard. To minimize the use of chemical fertilizers and cost of farming, generation of rice plants with better nutrient uptake and use efficiency is required. Membrane lipid remodelling (MLR) mediated changes in root system architecture (RSA) is a crucial adaptive mechanism in response to P deficiency in plants. Phospholipase-mediated lipid hydrolysis is one of the important ways of phospholipid disintegration during MLR. Members of phospholipase C (PLC) and PLD family are mainly implicated in regulation of RSA via MLR, under P deficiency in Arabidopsis. In our preliminary analysis, promoters of some of the rice PLDs and NPCs were found to have nutrient stress related cis-regulatory elements. In-silico expression analysis showed that some of these genes were significantly expressed under P deficiency in rice roots. This suggests a crucial role of phospholipases in P deficiency tolerance in rice. We hypothesize that these phospholipases could be the key regulators of RSA changes, and they might play an important role in improving P uptake and utilization efficiency in rice. Thus, we aim at molecular and genetic analysis of key phospholipases to achieve desired RSA for P deficiency tolerance. For this aim, following objectives are designed: (1) Identification of P-deficiency related phospholipase genes in rice through expression analysis (2) Sub-cellular localization of P-deficiency responsive rice phospholipase(s) (3) Generation of CRISPR/Cas9 edited mutant rice lines for selected phospholipase(s) (4) Phenotype analysis of mutant lines to assess overall plant growth and root system architecture under P-deficient and P-sufficient conditions (5) Molecular analysis of mutant lines to understand the expression pattern of lipid remodelling and P transport related genes under P-deficient and P-sufficient conditions. To achieve these objectives various methods will be employed, including expression analysis by Q-PCR, subcellular localization in Nicotiana benthamiana by confocal microscopy. CRISPR-Cas9 construct preparation and rice transformation will be done to get phospholipase edited mutants. Molecular and functional characterization of rice mutant plants will be done in P sufficient and deficient conditions to assess the role of phospholipases in RSA changes and in regulating P deficiency response. |
Total Budget (INR): | 39,61,828 |
Organizations involved