Teknik Isolasi dan Kultur Protoplas Tanaman Padi

Deden Sukmadjaja, Novianti Sunarlim, Endang G. Lestari, Ika Roostika, Tintin Suharlini

Abstract


Protoplast
fusion or somatic hybridization technology is an alternative
technology for production hybrids of plants that are difficult
to be produced by conventional methods due to their sexual
incompatibility. An experiment was conducted to develop
techniques for isolation, purification, and culture of rice
protoplasts of cultivar IR64 and a wild rice species (Oryza
officinalis). Optimization of protoplast isolation and purification
methods from both rice genotypes were successfully
done. The highest protoplast density was obtained by
digesting embryonic callus or stems of young seedling in an
enzyme solution containing of 2% cellulose, 0.1% pectolyase,
0.5% macerozyme, 0.5% driselase, 5 mM ES, and 13% mannitol
in CPW solution. The protoplast digestion was done for
three hours by soaking in the enzyme solution followed by
shaking at 50 rpm under a room temperature. Purification of
the protoplasts were done by separating them from plant
debris using a 25% sucrose solution. Protoplast regeneration
was not successful using although different media compositions
and conditions. Growth process from cell division to
cell aggregate was only successful on IR64 protoplast culture
on a medium that contained AgNO3.


Keywords


Protoplasts; isolation and culture; cultivated and wild rices.

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DOI: http://dx.doi.org/10.21082/jbio.v3n2.2007.p60-65

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P-ISSN : 1907-1094
E-ISSN : 2549-1547


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