Deden Sukmadjaja
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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Seleksi In Vitro dan Pengujian Mutan Tanaman Pisang Ambon Kuning untuk Ketahanan terhadap Penyakit Layu Fusarium Deden Sukmadjaja; Ragapadmi Purnamaningsih; Tri P. Priyatno
Jurnal AgroBiogen Vol 9, No 2 (2013): Agustus
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.v9n2.2013.p66-76

Abstract

Fusarium wilt of banana (Musa spp.) caused byFusarium oxysporum f. sp. cubense (Foc) is the most seriousproblem faced in banana cultivation in terms of plantproductivity and fruit quality. Mutation breeding is one of thealternative method that can be applied in producing newbanana cultivar. Mutants can be induced by chemicalmutagen such as ethyl methane sulfonate (EMS) followed byin vitro selection and then evaluation of the mutants tofusarium wilt disease in glasshouse and Foc infected field.The aim of this research was obtained EMS induced and invitro selected mutants of banana var. Ambon Kuning andevaluated Foc disease resistant clones in glasshouse andFoc infected field. The first step to obtain the explants forthis research was initiation and formation of multiple budclumps (MBC) using MS basal media supplemented with 5,10, and 20 mg/l of benzyladenin. Plant regeneration of MBCwas also studied by using MS media containing 0, 0.2, and 1mg/l of benzyladenin. To induce mutagenesis, MBC wassoaked in 0.1, 0.3, and 0.5% (v/v) EMS for 1, 2, and 3 hours.The assesment of resistant MBC mutants to Fusariumphytotoxin was conducted by using fusaric acid (FA) asselection agent in concentration of 30, 45, and 60 ppm.Putative mutant plants produced by in vitro selection werefurther tested using spore solution of Foc race 4 inglasshouse. Meanwhile, Foc resistance assesment in theinfected field was conducted in Pasirkuda ExperimentalStation, Bogor Agricultural University. The results showedthat MBC can be formed in MS basal media supplementedwith 10 or 20 mg/l benzyladenin. The EMS played a role inobtaining mutants by producing 68 MBC putative mutantstolerant to Foc based on FA selection. Further evaluation inthe glasshouse was obtained 64 Foc resistant plants from391 putative mutants produced by in vitro selection.Evaluation in the Foc infected field showed six clonessurvived until generative phase (12 month of age).
Transformasi Genetik Pisang Ambon dengan Gen Kitinase dari Padi Ragapadmi Purnamaningsih; Deden Sukmadjaja
Jurnal AgroBiogen Vol 8, No 3 (2012): Desember
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.v8n3.2012.p97-104

Abstract

One of the main constrains on theproductivity and quality enhancement of banana is wiltdiseases caused by Fusarium oxysporum (Foc). Productiondecrease by wilt disease was 63.33%. Therefore, an effort toobtain the banana new variety which is tolerant to fusariumwas absolutely necessary to be done. Genetic engineeringcan be used in new variety improvement, especially forproduction of pest and disease tolerant varieties.Transformation of banana with chi gene which expressedchitinase enzyme have been used in obtaining the plantresistant to Foc. The goals of the research were to obtain:determine lowest higromisin consentration inhibited nodulgrowth by tested four consentration of higromisin,determine optimum cocultivation time by tested three timescocultivation, tested asetosiringone added on two timescocultivation, and gen chi introduction at bananatransforman shoots with PCR. The explants used werenodule induced from pseudostem of banana cv. Ambonkuning. Genetic transformation done by sowing the explantsin bacterial suspension 0, 15, 30, and 45 minutes.The effectof asetosiringone (0 and 100 mg/l) on cocultivation mediumwas observed. The research results showed that the lowesthigromisin concentration inhibited nodule growth was 25mg/l for 5 weeks and the best time for inoculation of nodulewere 30 minute. Asetosiringone added on bacterialsuspension did not increase transformation efficiency.Chitinase gene transformation using Agrobacteriumtumefaciens on banana nodules produced 25 noduly ines ofputative transformant on selection media and 34 plantstransforman identification by PCR.