Ahmad Dadang
Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian, Jl. Tentara Pelajar 3A, Bogor 16111 Telp. (0251) 8337975; Faks. (0251) 8338820

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Journal : Jurnal AgroBiogen

Seleksi dan Konfirmasi Alel Gen-gen Hd pada Padi Berumur Genjah dan Produktivitas Tinggi Persilangan Code x Nipponbare Ahmad Dadang; Tasliah Tasliah; Joko Prasetiyono
Jurnal AgroBiogen Vol 9, No 1 (2013): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v9n1.2013.p11-18

Abstract

To support the IP 300 program rice varietieswith both early maturity and high productivity are needed.The objective of the research was to improve those traits inCode variety using RIL (recombinant inbreed line) methods.The research was conducted in the year 2009-2011 at thegreenhouse and laboratory of Molecular Biology, IndonesianCenter for Agricultural Biotechnology and GeneticResources Research and Development. Materials consistedof 600 individuals of F2 derived from one of F1 Code xNipponbare crossed plants and they were planted up to F4with seed-to-seed, then confirmed by 8 microsatelitemarkers linked to loci of Hd genes. The molecular analysisshowed that 84 of 600 F2 plants produced. The F4 plants have50% of flowering time shorter than F2 and F3 plants. Twolines (CdNb_388 and CdNb_270) have higher productivitycompared to Code by the number of productive tillers (20)and the number of tiller grains (2240 and 1740) and haveclosely 50% of 60 days flowering time of Nipponbare. Threelines of F4 plants (CdNb_270, 364, and 388) were predictedto have allele of Hd7 gene, and CdNb_472 was predicted tohave alelle of Hd14 gene.
Optimasi Sistem Regenerasi dan Transformasi Padi Varietas Elit Indonesia Aniversari Apriana; Toto Hadiarto; Ahmad Dadang
Jurnal AgroBiogen Vol 9, No 1 (2013): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v9n1.2013.p1-10

Abstract

New ricevariety can be generated by means of transgenic approach.Transgenic rice researches have been conducted in manyinstitutions worldwide using Japonica, Indica, and Javanicavarieties. The most cultivated rice in Indonesia is Indica.Indica type is known to have low responsive tissues inculture and transformation media when compared toJaponica rice. This research activity is aimed to optimizeregeneration and transformation systems of the Indonesianelite rice varieties, so that good method can be achieved tobe used in the generation of transgenic elite rice varieties ofIndica type. The research consisted of two activities:regeneration and transformation optimizations in varieties ofDodokan (upland rice) and Inpari 6 (irrigated rice).Immature embryo was used as the explant in this research.The optimization studies used 2 types of media, NBH (N6salts and vitamins, cassamino acid 0.5 g/l, L-proline 0.5 g/l,sucrose 20 g/l, D-glucose 10 g/l, 2.4-D 2 mg/l, NAA 1 mg/l, BA1 mg/l, agarose Type I 5.5 g/l) and NBH-M (N6 macro salts,B5 micro salts, and vitamins, 0.3 g/l cassamino acid, 3 g/l Lproline,20 g/l sucrose, 3 mg/l 2.4-D, 1 mg/l NAA, 1 mg/l BAP,5.5 agarose Type I), 2 types of regeneration media, R1 (MSbase media and vitamis, 0.3 g/l glutamine, 30 g/l sucrose, 2mg/l kinetin, 1 mg/l NAA, 3 g/l phytagel) and R2 (MS basemedia and vitamins, 2 g/l cassamino acid, 20 g/l sucrose, 30mg/l sorbitol, 2.5 mg/l kinetin, 0.25 mg/l NAA, 3 g/l phytagel).Optimization transformation of Indonesian elite rice varietiesused developed an empty plasmid pCAMBIA 1301 containinghpt gene. The transformation was conducted using twotypes of co-cultivation media, K1 (N6 macro salts, B5 microsalts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l L-proline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l NAA, 1mg/l BAP, 0.1 mM acetosyringone) and K2 (N6 macro salts,B5 micro salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l Lproline,20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/lNAA, 1 mg/l BAP, 0.2 mM acetosyringone). The resultsshowed that Inpari 6 could form embryonic calli in NBHmedia and further regenerated well in R1 media (13.8%).The co-cultivation media K1 generated more selected calliwhich then generated green plant of young embryocompared to K2. Inpari 6 showed higher regeneration ratesafter transformation (3.6%) compared to Dodokan (0%).Molecular analysis showed that all 11 transformants (Inpari6) tested contained the hpt gene. These results are expectedto support the development of transgenic Indica ricegeneration in Indonesia.