Somatic hybridization in okra
NOPR - NISCAIR Online Periodicals Repository
View Archive InfoField | Value | |
Title |
Somatic hybridization in okra
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Creator |
Zaman, Mariya S.
Parihar, Akarsh |
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Subject |
Protoplast
isolation viability test fluorescence microimaging microcalli regeneration |
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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. |
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Date |
2022-09-21T09:28:06Z
2022-09-21T09:28:06Z 2022-09 |
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Type |
Article
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Identifier |
0972-5849 (print) 0975-0967 (online)
http://nopr.niscpr.res.in/handle/123456789/60518 |
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Language |
en
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Publisher |
NIScPR-CSIR,India
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Source |
IJBT Vol.20(3) [July 2021]
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