Life Sciences & Biotechnology
Title : | Functional characterization of the Cicer arietinum Short Vegetative Phase (SVP) genes, their promoters, and potential microRNAs regulating them towards elucidating their role in floral transition. |
Area of research : | Life Sciences & Biotechnology |
Principal Investigator : | Dr. Konda Aravind Kumar, Indian Institute Of Pulses Research, Uttar Pradesh |
Timeline Start Year : | 2023 |
Timeline End Year : | 2026 |
Contact info : | aravindbio@gmail.com |
Details
Executive Summary : | MADS-box transcription factors (TF) are a well-studied family of TFs that regulate several processes directing the development of both vegetative and reproductive organs, particularly those of flowers and fruit. In our preliminary work, we identified 83 MADS-box genes in chickpea by genome-wide identification. Prediction of microRNA targets in MADS-box genes showed an intriguing fact: just a single novel microRNA (to be referred to as Car miRNA) targets three genes, all three of which are SVP genes: a single copy of the agl (Agamous-like) 24 and two copies of the agl22 genes. SVP-like genes play an important role in floral transition and floral meristem identity. agl22 and agl24 have opposite roles in the control of flowering time but later act redundantly to control floral meristem identity. Although SVP-like genes have been characterized from a variety of plant species, their role in flowering time and floral primordia development appear to be conserved. However, the SVP genes can have rather diverse functions and their role in chickpea floral development is yet to be identified. In addition, their regulation by the identified miRNA needs to be understood. Therefore the proposed project aims to characterize the function of the Cicer arietinum SVP (CaSVP) genes, promoters, and the novel microRNA Arabidopsis will be chosen as a model organism to address the problems since the SVP genes are well characterized, agl22 and agl24 mutants are available, and it lacks the Car miRNA gene. Initially, quantitative PCR was used to determine the expression patterns of the CaSVP genes and the Car miRNA in various vegetative and floral tissues of chickpea. The Car pri-miRNA expression pattern will be investigated further in Arabidopsis using heterologous expression of the gfp fused with the Car pri-miRNA promoter. Complementation experiments will be carried out to see if Car SVP genes fused with fluorescent tags can restore floral characteristics under native and constitutive promoters. Arabidopsis mutant lines complemented by heterologous CaSVP gene expression will be studied further for Car miRNA characterisation. The Car pri-miRNA will be overexpressed constitutively in Arabidopsis wild type and mutant lines complemented with CaSVP genes. Fluorescence microscopy and western blotting will be utilized to assess the Car miRNA's cleavage of the target CaSVP genes fused with fluorescent tags. Further degradome sequencing will offer a thorough understanding of the miRNA's other targets, which will be verified by quantitative PCR of endogenous Arabidopsis SVP and other floral pathway genes. The detailed characterization of the CaSVP genes and the Car miRNA will give us a comprehensive understanding of their roles in floral development, which may then be employed in breeding and genome editing programs. |
Total Budget (INR): | 35,79,724 |
Organizations involved