Differential DNA Endoreduplication and Protein Profile during Cotyledon Ontogeny of Vigna radiata

Differential regeneration response of the two cotyledon types. ‘Cot E’ (attached to the embryo) and ‘Cot’, of Vigna radiata have been reported earlier. The present preliminary study addresses V. radiata cotyledon development with respect to patterns of endoreduplication and protein accumulation. In this communication two distinct types of cotyledon (in relation to their attachment with the embryonal axis), differing in regeneration responses, were characterized in terms of polyploidy levels and profiles, and extent of protein synthesis/accumulation. The embryo development was studied histologically from the first day after fertilization till seed coat formation and divided into 8 different stages to determine stages of cotyledon development. Early cotyledonary stage of embryo was recorded on the 6 DAF, at this stage ‘Cot’ and ‘Cot E’ were inseparable and referred to as stage VI. ‘Cot’ and ‘Cot E’ could be distinguished from 9 DAF onwards. Two major events, endoreduplication of DNA and protein synthesis/accumulation that occur during the cotyledon development of grain-legumes, were analysed to probe the differential status, if any, in these two cotyledon types. The cell division phase of cotyledon development continues upto stage VII, while cell expansion phase starts at the stage VIII. The cotyledonary cells began to undergo the endoreduplicating cell cycle (ECC) from stage VII and continue until seed maturity. During the cell division phase the mitotic cycle and ECC occur simultaneously; whereas, only ECC continues in the cell expansion phase. Analysis of protein content indicated that ‘Cot E’ always contained comparatively higher amount during in vivo development than that of ‘Cot’. Similarity indices between ‘Cot’/’Cot E’ were 46.15%,82.35% and 90.9% at stage VII, stage VIII and at maturity, respectively, as computed from the presence oftotal polypeptides. The differential temporal pattern of DNA-endoreduplication and storage protein accumulation clearly dictates the influence of differential gene expression and regulation control in the developmental- type determination of the two cotyledons.