Effect of nurse culture on including division of isolated pollen protoplasts of Hevea brasiliensis
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TextPublication details: Journal of Plantation Crops 2013Description: 123-129Subject(s): Summary: Haploids are of grate relevance in crop improvement of Hevea, a highly heterozygous tree species with a long breeding cycle. The isolation and culture of pollen protoplasts may be a viable proposition for raising haploid plants/homozygous lines in Hevea. The present work envidages the developement of a method for the isolation and culture of pollen protoplasts of Hevea. Effecct of different nurse cultures on the developement of cultured protoplasts has been studied. Intact pollen grains were isolated from mature male flowere of Heavea prior to opening. Viable protopasts in high yield could be isolated from these pollen grains when exposed to a mixture of 0.5 per cent cellulase and 0.05 per cent pectolyase in the presence of the osmotic stabilizers 0.6 M mannitol and 0.3 M sorbitol. These protoplasts were partially purified and cultured in the nutrient medium with three different nurse cultures nmely embryogenic calli from Hevea, tobacco and carrot. Divisiion of the cultured protoplasts leading to the formation of a few micro-colonies was obseved in the medium containing 0.8 mg 1-12, 4-D and 0.5 mg1-1 BA and enriched with Hevea nurse culture. Cultures with micro-colonies are dark incubated for further developement. This is the first report of division of pollen protoplasts and micro-colony formation in Hevea brasiliensis.
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Journals
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RRII Library | Volume 41, Issue 2 | Journals |
Haploids are of grate relevance in crop improvement of Hevea, a highly heterozygous tree species with a long breeding cycle. The isolation and culture of pollen protoplasts may be a viable proposition for raising haploid plants/homozygous lines in Hevea. The present work envidages the developement of a method for the isolation and culture of pollen protoplasts of Hevea. Effecct of different nurse cultures on the developement of cultured protoplasts has been studied. Intact pollen grains were isolated from mature male flowere of Heavea prior to opening. Viable protopasts in high yield could be isolated from these pollen grains when exposed to a mixture of 0.5 per cent cellulase and 0.05 per cent pectolyase in the presence of the osmotic stabilizers 0.6 M mannitol and 0.3 M sorbitol. These protoplasts were partially purified and cultured in the nutrient medium with three different nurse cultures nmely embryogenic calli from Hevea, tobacco and carrot. Divisiion of the cultured protoplasts leading to the formation of a few micro-colonies was obseved in the medium containing 0.8 mg 1-12, 4-D and 0.5 mg1-1 BA and enriched with Hevea nurse culture. Cultures with micro-colonies are dark incubated for further developement. This is the first report of division of pollen protoplasts and micro-colony formation in Hevea brasiliensis.
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