In Vitro Regeneration of Castor (Ricinus Communis L.) Using Cotyledon Explants.

Source: OAI

ABSTRACT An efficient plant regeneration protocol using cotyledon explants was established for castor (Ricinus communis L.), an important oilseed crop. Mature seed-derived cotyledon explants produced adventitious shoots when placed on Murashige and Skoog (MS) medium containing thidiazuron (TDZ). The rate of shoot regeneration was maximal (25 shoots per explant) when explants were cultured on shoot induction medium supplemented with 5 μM TDZ and preincubated in the dark for the first 7 days before transferring to the day/night cycle (16/8 h). Only the proximal ends of cotyledon explants produced adventitious shoots, although green calli were observed in cotyledon veins. After 4 weeks in culture, explants with well-developed shoot buds were transferred to MS medium without plant growth regulators for the shoot elongation and development. At 4 months after culture initiation, shoots (2 cm in length) were transferred to root induction medium (MS medium supplemented with 5 μM indole-3-butyric acid) where they developed roots in 4 to 6 weeks. Plantlets were transferred to soil and acclimatized to greenhouse conditions. Histological analysis showed the adventitious induction of the shoots originated from the cortical and epidermal cell layers of the cotyledon explants.

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    ABSTRACT: Successful in vitro plant regeneration protocol has been described for fusarium wilt resistant castor (Ricinus communis L.) parental line SKP-84 through apical meristem. Shoot apex containing apical meristems, were excised from 5-7 days old in vitro grown seedlings and cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of cytokinins either alone or in combination. Kinetin had marked effect on shoot initiation and shoot quality. Kinetin (2.325µM) in combination with BAP (1.111µM) produced maximum number of shoot (10.33) and shoot length (5.20 cm). In vitro produced shoots were transferred to rooting media containing half strength MS basal media supplemented with NAA at various concentrations. Well developed roots appeared in media supplemented with 0.537µM NAA (69.00 %). Rooted plants were transferred to the pots containing vermicompost with 64.5% survival rate in hardeninng. No genetic variation was detected in plantlets as revealed by RAPD at different stages proved the genetically stability during in vitro plant regeneration through apical meristem culture.
    The Bioscan. 02/2013; 8(2):403-408.
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    ABSTRACT: Vigna radiata or mungbean belongs to the legume family of plants. Mature mungbean seeds are rich source of dietary proteins for human nutrition. The present study was aimed to analyze the comparative protein profiles of two cotyledon types, Cot and Cot E, prior to and during early time points of shoot morphogenic induction to understand the unique differential regeneration response in these two explant types which was reported earlier. These explants were grown separately in vitro on the shoot induction medium (SIM) containing Gamborg’s B5 basal nutrient composition supplemented with 15 μM N6-benzyladenine. Isolation and characterization of the proteomes from Cot and Cot E explants at different time points, during early events of shoot differentiation, were performed using two dimensional gel electrophoresis following matrix assisted laser desorption-ionization tandem mass spectrometry. A total of 112 differentially identified proteins were classified according to their putative biological function. The differential control of protein synthesis between these explants under control condition, i.e. before in vitro culture, was also noted. In Cot E explants SIM induced prompt acquisition of competence for direct shoot morphogenesis probably through fast phytohormone signaling. Over accumulated proteins in Cot E indicated stimulation of several metabolic and associated pathways earlier than Cot explants. Abundance of stress and defense related proteins in Cot E explants was presumably to cope up with stressful cultural condition. Enhanced accumulation of folding-assisted proteins involved in organogenesis mediated cellular reprogramming in Cot E explants contributed further in rapid and efficient regeneration responsiveness.
    Plant Cell Tissue and Organ Culture 10/2013; · 2.61 Impact Factor


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