Teknik Isolasi dan Kultur Protoplas Tanaman Padi

Abstract

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

Spesies Padi Liar (Oryza sp.) sebagai Sumber Gen Ketahanan Cekaman Abiotik dan Biotik pada Padi Budidaya

“…Sitch (1990) melaporkan, persilangan padi budi daya dengan padi liar genom BB hingga CCDD memperoleh seed set F1 030%, dan persilangan dengan genome EE dan FF memperoleh seed set 0,23,8%. Studi yang dilakukan Abdullah et al (1995) (Sukmadjaya et al 2007). …”

“…Keberhasilan kultur protoplas dan regenerasinya ditentukan oleh beberapa faktor, antara lain genotipe dan jaringan yang digunakan, fisiologi jaringan, jenis dan konsentrasi enzim, masa inkubasi, media kultur, zat pengatur tumbuh, dan kondisi inkubasi (Sukmadjaya et al 2007). Persilangan somatik melalui fusi protoplas merupakan metode alternatif yang sangat berguna dan mempunyai prospek yang baik bagi kegiatan pemuliaan pada masa mendatang.…”

“…Protoplas dapat diisolasi dari hampir semua bagian tanaman seperti akar, daun, nodul, koleoptil, kultur kalus, dan daun in vitro (Husni et al 2004). Dalam isolasi dan purifikasi protoplas belum ada standar atau metode baku karena setiap individu sel atau jaringan yang akan digunakan sebagai sumber protoplas memerlukan perlakuan khusus (Sukmadjaya et al 2007).…”