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Somatic hybridization in okra

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Title Somatic hybridization in okra
 
Creator Zaman, Mariya S.
Parihar, Akarsh
 
Subject Protoplast
isolation
viability test
fluorescence microimaging
microcalli
regeneration
 
Description 302-319
Protoplast isolation and culture are extremely difficult in recalcitrant crop like okra due to high mucilage content. Hence,
procedure was required to be standardized for protoplast isolation. Protoplasts isolation was carried out from both in vitro
and ex vitro leaf materials through an enzymatic method from cultivated okra (Abelmoschus esculentus L.) and its wild
species (A. moschatus subsps. tuberosus). Plasmolysed pre-treatment was utilized for easy isolation of protoplasts.
Protoplasts were purified using CPW 13M solution followed by centrifugation. Protoplast viability, yield, diameter and
residual cell wall were determined by fluorescent dyes such as fluorescein diacetate (FDA), rhodamine, propidium iodide,
4’, 6 - diamidino -2 phenylindole (DAPI) and calcoflour white under fluorescence and UV microimaging. The viable
protoplasts obtained were further utilized in protoplast fusion, culture and regeneration of inter-specific microcalli. Plating
density, growth regulator concentration and the use of antioxidants were all demonstrated to have a significant effect on the
protoplast plating efficiency. Protoplasts labeled with a fluorescent marker were subjected to chemical fusion. Using
micromanipulator, heterokaryons formed during chemical fusion, were recovered. Such heterokaryons, when cultured
underwent division and formed microcalli which subsequently developed into calli with green shoot bud point. The hybrid
nature of such calli was confirmed by morphological and molecular characterizations.
 
Date 2022-09-21T09:28:06Z
2022-09-21T09:28:06Z
2022-09
 
Type Article
 
Identifier 0972-5849 (print) 0975-0967 (online)
http://nopr.niscpr.res.in/handle/123456789/60518
 
Language en
 
Publisher NIScPR-CSIR,India
 
Source IJBT Vol.20(3) [July 2021]