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Tri P. Priyatno
Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian, Jl. Tentara Pelajar 3A, Bogor 16111 Telp. (0251) 8337975; Faks. (0251) 8338820
Agriculture, Biological Sciences & Forestry

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Journal : Jurnal AgroBiogen

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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).
Pemurnian Parsial dan Karakterisasi Kitinase Asal Jamur Entomopatogen Beauveria bassiana Isolat BB200109 Yadi Suryadi; Tri P. Priyatno; I Made Samudra; Dwi N. Susilowati; Nuni Lawati; Eman Kustaman
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.p77-84

Abstract

Beauveria bassiana is one of theentomopathogenic fungus that produces chitinase wheninfecting its host. This study was aimed to purify, isolate andcharacterize chitinase of B. bassiana isolate BB200109.Pathogen identity was determined both morphologically andmolecularly using ITS primer, whilst characterization wasdone at various conditions i.e. temperature, pH, metal ionand incubation time. Results showed that the BB200109isolate belonged to B. bassiana. The isolate producedextracellular chitinase with chitinolytic index of 1.035. Partialpurification of three saturated ammonium sulphateprecipitation (10, 30, and 70%) showed maximum purity of1.2 times, while dialysis could increase the purity of 1.9times compared to that of crude enzyme extract.Characterization results showed that the chitinase isolatedfrom B. bassiana isolate BB200109 had an optimum activityat pH 4, temperature 50oC, and optimum incubation time of90 minutes. The effect of metal ions (60 mM) Mn2+ served asactivator, while EDTA, K+, Mg2+, Cu2+, Fe2+, Zn2+, and Na+acted as inhibitors. The chitinase demonstrated loweraffinity to chitin substrate as indicated by high Km value of0.266 mg/l and a Vmax of 0.067 mg/l sec. Based on SDS-PAGE,chitinase from B. bassiana isolate BB200109 had molecularweight of 60.25 kDa. The study implied the potency ofB. bassiana isolate BB200109 as extracellular chitinaseproducer with its enzyme charateristics seems to bedeveloped as an insect biocontrol agent.
Co-Authors Deden Sukmadjaja Dwi N. Susilowati Eman Kustaman I Made Samudra Nuni Lawati RAGAPADMI PURNAMANINGSIH Yadi Suryadi