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Journal : Jurnal AgroBiogen

Multiplikasi Tunas dan Induksi Perakaran pada Ubi Kelapa (Dioscorea alata L.) dan Gembili (Dioscorea esculenta L.) Secara In Vitro Hutami, Sri; Purnamaningsih, Ragapadmi; Mariska, Ika; Diantina, Surya
Jurnal AgroBiogen Vol 10, No 2 (2014): 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.v10n2.2014.p53-60

Abstract

Dioscorea sp. (yam) is one of the minor tuber crops whichgrows wildly in the forest and only a few of its species arecultivated and used as main or secondary food.Conservation is needed to preserve plant genetic material.The objective of this research was to obtain methods ofplantlets propagation of D. alata L. and D. esculenta L.through in vitro culture. The research was conducted atTissue Culture Laboratory of ICABIOGRAD in 2012. Theresearch consisted of three stages. First, shoot emergence.In this experiment, young shoots were planted in MS basicmedium combined with benzyl adenine (BA) (0, 1, 3, and 5mg/l) and gibberelic acid (GA) (0 and 5 mg/l). Second, shootmultiplication. Shoots of Dioscorea which were planted inthe best medium of the first experiment were subcultured inMS medium combined with thidiazuron (0, 0.1, 0.5, 1, 2, and3 mg/l). Third, root initiation. Shoots of Dioscorea whichwere planted in the best medium of the second experimentwere subcultured in MS medium (½ MS and 1 MS)combined with indole-3-butyric acid (IBA) (0, 1, 3, and 5mg/l). Result of these experiments showed that shootemergence of D. alata L. and D. esculenta L. began at 2weeks after planting in MS medium. More plantlets of D.alata L. and D. esculenta L. were obtained by shootmultiplication in MS media. Root initiation of the Dioscoreabegan at 4 weeks after planting in MS media. The addition ofIBA (3–5 mg/l) on D. esculenta L. could not stimulate rootingbut led to the formation of callus at the base of the stembuds.
Pengaruh Sumber Karbon dan Kondisi Inkubasi terhadap Pertumbuhan Kultur In Vitro Purwoceng (Pimpinella pruatjan Molk.) Roostika, Ika; Purnamaningsih, Ragapadmi; Noviati, Arief V.
Jurnal AgroBiogen Vol 4, No 2 (2008): Oktober
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.v4n2.2008.p65-69

Abstract

Pruatjan (Pimpinella pruatjan Molk.) is an Indonesianmedicinal plant which is categorized as endangeredplant and included in Appendix I based on CITES. The invitro conservation techniques have been studied. However,the storage period was very short (4 months) when plantgrowth retardant and media dilution were applied. Besidethat, the residual effect of growth retardant was strongenough so that it needed more than 4 months for recovery.Thus, the use of certain carbon source may prolong thepreservation period with shorter time for recovery. Theobjective of the study was to know the effects of carbonsources (sucrose and mannitol) and culture conditions (cultureroom and growth chamber) to the growth of pruatjancultures. This application was hoped to prolong preservationperiod of pruatjan longer than 4 months and to cut therecovery period after presservation. The study was conductedat Tissue Culture Laboratory in Indonesian Center forAgricultural Biotechnology and Genetic Resources Researchand Development from August 2006 to July 2007. Theactivities included propagation of in vitro shoot grown invitro as explants source, preservation of in vitro shoots ofpruatjan, and regeneration of the cultures after preservation.The experiment was designed as factorial in RandomizedCompletely Block Design with 6 replications. The DKW basalmedia containing 1 ppm BA, 0.2 ppm thidiazuron, and 100ppm arginine were supplemented with mannitol or sucroseat the level of 1, 2, 3, 4, and 5%. The observed variables weretotal number of leaves, number of shoot, and number of wiltleaves. The result revealed that pruatjan cultures could bestored longer than 4 months. Generally, the effect ofmannitol or sucrose was more dominant than that of culturescondition. The mannitol (1-5%) strongly inhibited thegrowth of pruatjan cultures so that only a few culturessurvived at 7 months preservation period and needed about1 month for recovery. On the contrary, the effect of sucrose(at the same level) was better than mannitol. The 2.5%sucrose optimally inhibited pruatjan cultures. At that condition,the cultures could be stored for 10 months withoutmorphological changes so that they could recover spontaneously.
Induksi Kalus dan Optimasi Regenerasi Empat Varietas Padi melalui Kultur In Vitro Purnamaningsih, Ragapadmi
Jurnal AgroBiogen Vol 2, No 2 (2006): Oktober
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.v2n2.2006.p74-80

Abstract

A study was conducted at the Tissue Culture Laboratory of ICABIOGRAD, Bogor, to obtain an optimum medium formulation for calli regenerations of for rice varities (Ciherang, Cisadane, IR64, and T-309). The research activities were done in five steps, i.e., callus induction, callus regeneration, shoot multiplication, root formation, and plant acclimatization. The type of explants used in the study was embriozygotic explants. Five media formulations were used for the callus induction, while four media formulations were used for the callus regeneration. The results showed that the best medium formulation for induction of callus formation was MS + 2,4-D 2 mg/l + casein hidrolisat 3 mg/l, while the best medium formulation for callus regeneration was MS + BA 3 mg/l + thidiazuron 0,1 mg/l.
In Vitro Culture Manipulation on Pruatjan for Secondary Metabolite Production Ika Roostika; Ragapadmi Purnamaningsih; Ireng Darwati; Ika Mariska
Jurnal AgroBiogen Vol 3, No 2 (2007): Oktober
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.v3n2.2007.p55-59

Abstract

Purwoceng (Pimpinella pruatjan Molk. atau Pimpinellaalpina KDS.) adalah tanaman obat langka yang dapat dimanfaatkansebagai bahan obat afrodisik, diuretik, dan tonik.Kultur in vitro tidak hanya dapat digunakan untuk konservasidan perbanyakan tanaman, melainkan dapat juga diterapkanuntuk produksi metabolit sekunder. Melalui teknik ini,produksi metabolit sekunder tidak bergantung kepada sumbertanaman di lapang. Penelitian ini dilakukan dengan tujuanuntuk meningkatkan kadar stigmasterol melalui kultur invitro dengan menggunakan prekursor asam mevalonat. Penelitiandibagi menjadi dua tahap, yaitu induksi kalus danmanipulasi kultur in vitro untuk meningkatkan kadar stigmasterol.Pada tahap induksi kalus, terdapat 16 perlakuan yangmerupakan kombinasi perlakuan 2,4-D dan piklorammasing-masing pada taraf 0,5; 1,0; 1,5; dan 2,0 ppm. Untukmeningkatkan kadar stigmasterol, digunakan asam mevalonatpada taraf 0, 250, 500, dan 750 ppm dengan masa inkubasiselama 4 dan 6 minggu. Kandungan stigmasterol dianalisismenggunakan GC-MS. Hasil penelitian menunjukkanbahwa media P2 (DKW + 2,4-D 0,5 ppm + pikloram 1,0ppm) adalah media terbaik untuk induksi kalus. Eksplan daunlebih baik daripada eksplan petiol. Hasil analisis GC-MSmenunjukkan bahwa kandungan stigmasterol tertinggi(0,0356 ppm) diperoleh dari kalus dengan masa inkubasi 4minggu pada media dengan penambahan asam mevalonat250 ppm. Peningkatan taraf asam mevalonat tidak mampumeningkatkan kandungan stigmasterol. Kadar tersebut miripdengan kandungan stigmasterol pada planlet dari GunungPutri (0,0365 ppm) dan Dieng (0,0414 ppm). Dibandingkandengan kadarnya dalam akar tanaman dari lapang, kandungantersebut sekitar 10-100 kali lipat lebih tinggi.
Keragaman Somaklonal untuk Perbaikan Tanaman Artemisia (Artemisia annua L.) melalui Kultur In Vitro Endang G Lestari; Ragapadmi Purnamaningsih; Muhammad Syukur; Rosa Yunita
Jurnal AgroBiogen Vol 6, No 1 (2010): 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.v6n1.2010.p26-32

Abstract

Somaclonal Variability for the Improvement of PlantsArtemisia (Artemisia annua L.) by In Vitro Culture.Endang G. Lestari, Rosa Yunita, and Ali Husni. Artemisiaannua L., a family member of Asteraceae, is medicinalplants originated from China. The plant has been widelyused by the local people for malaria remedy. Its active substance,artemisine, has been proved to hamper the malariabacteria incubation, Plasmodium sp. In accordance with theWHO recomendation, the Department of Health of Indonesiais now in the attempt of developing this plant as thesubtitute of chloroquin because of the malaria bacteriaresistance to this antidote. In Indonesia, the artemisinecontent of the plant less than 0,5% is the crucial problemleading no investors are interested in its economic value.Therefore, Indonesian Medicinal and Spice Crops ResearchInstitute; BPTO Tawangmangu, Indonesian Institute ofSciences; and PT Kimia Farma cooperate for obtaining theprime clone by breeding, selection, as well as environmentaladaptation. In coping with the problem, ICABIOGRAD in thecollaboration with Bogor Agricultural University haveconducted the research for genetic improvement throughmutative induction and field selection. This research onsomaclonal variation. was conducted from Januari 2006 toJuni 2008. Eksplan used for experiment were shoots radiatedwith 10-100 Gy gamma ray. The result showed that the shootradiated with the dosage of 70-100 Gy was unable to grow.On the other hand, the high level of multiplication wasacquired in the one radiated with 10-30 Gy. The optimumradiation for somaclonal radiation was eventually gainedwith 40-60 Gy. The somaclone lines with 10-60 Gy radiationhave been aclimatized and planted in Gunung Putri plot inthe elevation of 1545 asl. Artemisinin content at the highbiomases genotype is 0,49-0,52%.
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.
Co-Authors A G Wattimena Agus Purwito Ashrina, Misky Azrai, Muh. DANNY LAURENT Darliah Darliah Deden Sukmadjaja Diantina, Surya Didy Soepandi Didy Soepandi, Didy Didy Sopandie E.G. Lestari E.G. Lestari E.G. Lestari Endang G Lestari Endang G Lestari Endang Gati Lestari Endang Gati Lestari ENDANG GATI LESTARI ENDANG GATI LESTARI ENDANG GATI LESTARI Enny Sudarmonowati Enny Sudarmonowati GA Wattimena Hutami, Sri I Mariska I Roostika I. Darwati I. Darwati I. DARWATI I. Mariska I. Roostika I. Roostika I. ROOSTIKA Ika Mariska Ika Mariska Ika Mariska Ika Mariska Ika Mariska Ika Roostika Ika Roostika Ika Rostika Ika Rostika IRENG DARWATI Ireng Darwati Laela Sari laela Sari, laela Lestari, Endang Gati LESTARI, ENDANG GATI Lizawati . Mariska, I Mariska, Ika MARISKA, IKA Muhamad Syukur Muhammad Syukur N Khumaida N Khumaida NESTI FRONIKA SIANIPAR Nesti Fronika Sianipar NESTI FRONIKA SIANIPAR Noviati, Arief V. Nur, Amin Nurhayani, Siti Pardal, S.J. Pardal, Saptowo Jumali Rita Megia RITA MEGIA ROHIM FIRDAUS ROHIM FIRDAUS Rohim Firdaus Roostika, Ika Rosa Yunita Rosa Yunita Rosa Yunita Rosaria Rosiana Rosaria Rosiana, Rosaria ROSSA YUNITA Rossa Yunita S.J. Pardal Sari, Laela Siti Nurhayani Slamet . Slamet Slamet Slamet, nFN Soeranto, Soeranto Sri Hutami Sri Hutami Sri Hutami Subagio, Herman Sudarmonowati, Enny Sustiprajitno, Sustiprajitno Syarifah Iis Aisyah Tri P. Priyatno Trias Novita Trikoesoemaningtyas UTAMI, SRI Wahyu Handayati Wahyu Handayati Y Supriati Y Supriati Yati Supriati, Yati Yunita, Rossa