Optimasi Sistem Regenerasi dan Transformasi Padi Varietas Elit Indonesia

Aniversari Apriana, Toto Hadiarto, Ahmad Dadang
2016 Jurnal AgroBiogen  
<p>New rice<br />variety can be generated by means of transgenic approach.<br />Transgenic rice researches have been conducted in many<br />institutions worldwide using Japonica, Indica, and Javanica<br />varieties. The most cultivated rice in Indonesia is Indica.<br />Indica type is known to have low responsive tissues in<br />culture and transformation media when compared to<br />Japonica rice. This research activity is aimed to optimize<br />regeneration and transformation systems of the
more » ... systems of the Indonesian<br />elite rice varieties, so that good method can be achieved to<br />be used in the generation of transgenic elite rice varieties of<br />Indica type. The research consisted of two activities:<br />regeneration and transformation optimizations in varieties of<br />Dodokan (upland rice) and Inpari 6 (irrigated rice).<br />Immature embryo was used as the explant in this research.<br />The optimization studies used 2 types of media, NBH (N6<br />salts and vitamins, cassamino acid 0.5 g/l, L-proline 0.5 g/l,<br />sucrose 20 g/l, D-glucose 10 g/l, 2.4-D 2 mg/l, NAA 1 mg/l, BA<br />1 mg/l, agarose Type I 5.5 g/l) and NBH-M (N6 macro salts,<br />B5 micro salts, and vitamins, 0.3 g/l cassamino acid, 3 g/l Lproline,<br />20 g/l sucrose, 3 mg/l 2.4-D, 1 mg/l NAA, 1 mg/l BAP,<br />5.5 agarose Type I), 2 types of regeneration media, R1 (MS<br />base media and vitamis, 0.3 g/l glutamine, 30 g/l sucrose, 2<br />mg/l kinetin, 1 mg/l NAA, 3 g/l phytagel) and R2 (MS base<br />media and vitamins, 2 g/l cassamino acid, 20 g/l sucrose, 30<br />mg/l sorbitol, 2.5 mg/l kinetin, 0.25 mg/l NAA, 3 g/l phytagel).<br />Optimization transformation of Indonesian elite rice varieties<br />used developed an empty plasmid pCAMBIA 1301 containing<br />hpt gene. The transformation was conducted using two<br />types of co-cultivation media, K1 (N6 macro salts, B5 micro<br />salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l L-proline,<br />20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l NAA, 1<br />mg/l BAP, 0.1 mM acetosyringone) and K2 (N6 macro salts,<br />B5 micro salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l Lproline,<br />20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l<br />NAA, 1 mg/l BAP, 0.2 mM acetosyringone). The results<br />showed that Inpari 6 could form embryonic calli in NBH<br />media and further regenerated well in R1 media (13.8%).<br />The co-cultivation media K1 generated more selected calli<br />which then generated green plant of young embryo<br />compared to K2. Inpari 6 showed higher regeneration rates<br />after transformation (3.6%) compared to Dodokan (0%).<br />Molecular analysis showed that all 11 transformants (Inpari</p><p>6) tested contained the hpt gene. These results are expected<br />to support the development of transgenic Indica rice<br />generation in Indonesia.</p>
doi:10.21082/jbio.v9n1.2013.p1-10 fatcat:js6gxmccv5g63f4qwbdqhff3ne