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Evaluation and Selection of Mutative Artemisia (Artemisia annua L.) According to the Altitude Variants ENDANG GATI LESTARI, ENDANG GATI LESTARI ENDANG GATI LESTARI; SYUKUR, MUHAMAD; PURNAMANINGSIH, RAGAPADMI; YUNITA, ROSSA; FIRDAUS, ROHIM
HAYATI Journal of Biosciences Vol 18, No 1 (2011): March 2011
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (46.74 KB) | DOI: 10.4308/hjb.18.1.16

Abstract

Induction of genetic variant of Artemisia annua L. was conducted through the application of gamma ray irradiation in 2007-2008. The aim was to obtain a plant with high artemisine content > 0.5% and late flowering period of about > 7 month after planting. Tweleve selected genotypes were subsequently examined to gain genetic stability on altitude of 1500, 950, and 540 m asl. The results showed that the plants had shorter flowering age in Cicurug (540 m asl) than that of  in Pacet (950 m asl) and Gunung Putri (1540 m asl). Genotype 8 had the latest age of flowering in the three locations than the other genotypes, however, the growth and biomass were the lowest. Vegetative growth of Artemisia in Pacet and Gunung Putri was better than those in Cicurug. Genotype of 15 in Cicurug and 5A genotype in Gunung Putri and Pacet had higher wet and dry weight than that of two other associates. Based on plant biomass, 5 genotypes from Gunung Putri and Pacet i.e. 1D, 3, 5A, 14, and 15 genotypes were selected, as well as 5 genotypes i.e. 1D, 3, 4, 5A, and 15 genotypes from Cicurug. Analisys on artemisin content successfully obtained 5 selected somaclone lines i.e. 1B, 2, 4, 14, and 3 somaclones.
Induksi dan Multiplikasi Tunas Jarak Pagar (Jatropha curcas L.) Secara In Vitro Lizawati, ,; Novita, Trias; Purnamaningsih, Ragapadmi
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol 37, No 1 (2009): Jurnal Agronomi Indonesia
Publisher : Bogor Agricultural University

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Abstract

The conventional propagation of physic nut (Jatropha curcas) is difficult, because it requires a high number of mother plant, which is very limited. In vitro culture is an alternative technique to conventional one to solve the problem.  An experiment was done to obtain the best in vitro culture media for shoot induction and multiplication. This research was separated into two steps, (1) in vitro induction of explant growth, and (2) in vitro shoot multiplication.  Results showed that medium of WPM + 2.0 ppm BAP induced shoot and leaf better than the control.  The highest number of leaf axillary´s multiplication was obtained from the medium WPM + 2.0 ppm BAP + 0.1 ppm NAA. Various medium formulations for the induction and multiplication of shoots resulted in highly leaf fall.  The use of DKW + 2.0 ppm BAP + 0.4 ppm TDZ + 3.0 ppm AgNO3 medium has effectively induced shoot multiplication and reduction of dehydrated leaf. Meanwhile, the used of DKW medium supplemented with 5 ppm kinetin resulted in the best shoot elongation.   Key words :  Induction, in vitro, Jatropha curcas, shoot, multiplication
Pengaruh Iradiasi Gamma dan Ethyl Methan Sulfonate Terhadap Pembentukan Embriosomatik Kedelai (Glycine max L.) Purnamaningsih, Ragapadmi; Mariska, Ika; Lestari, Endang Gati; Hutami, Sri; Yunita, Rossa
Jurnal Aplikasi Isotop Radiasi Vol 10, No 1 (2014): Juni 2014
Publisher : BATAN

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Abstract

Kedelai merupakan salah satu sumber protein dan lemak nabati yang penting. Perubahan iklim global berpengaruh terhadap produktivitas kedelai, sehingga diperlukan kultivar-kultivar baru yang mempunyai sifatunggul tertentu agar produktivitas kedelai dapat ditingkatkan. Teknik in vitro dengan mutasi dan keragaman somaklonal merupakan meoda alternatif untuk memperoleh varietas baru apabila material genetik sebagai bahan seleksi tidak tersedia. Induksi mutasi dapat dilakukanpada populasi sel embriogenik dengan menggunakan iradiasi sinar gamma atau senyawa kimia, antara lain Ethyl Methan Sulfonate (EMS). Kedua metoda tersebut telah banyak digunakan untuk meningkatkan keragaman genetik tanaman dan telah dihasilkan galur-galurbaru dengan sifat unggul. Salah satu masalah penting yang harus dikuasai dalam penerapan teknologi tersebut adalah meregenerasikan sel somatik hasil mutasi dan keragaman somaklonal agar dapat ditumbuhkan menjadi planlet (tunas in vitro). Beberapa faktor yangmempengaruhi regenerasi tanaman adalah jenis bahan tanaman, genotipe, komposisi media, dll. Perlakuan keragaman somaklonal dan mutasi yang diberikan dapat menyebabkan kerusakan pada sel sehingga diperlukan modifikasi pada metoda regenerasi yang sudah diketahui agar populasi sel yang hidup setelah perlakuan mutasi dapat tumbuh menjadi tunastunas mutan. Tujuan penelitian adalah untuk mendapatkan planlet mutan hasil perlakuan mutasi dengan iradiasi gamma dan EMS. Varietas kedelai yang digunakan adalah Wilis, Burangrang, Baluran dan aksesi No. B 3592. Eksplan yang digunakan adalah embriozigotik muda berasal dari polong yang berumur 12-20 hari setelah penyerbukan. Induksi kalusembriogenik dilakukan dengan menggunakan media MS + vitamin Gamborg (B5) dengan penambahan 2,4-D 20 mg/l dan sukrosa 3%. Kalus yang didapatkan diberi perlakuan mutasi menggunakan sinar gamma pada dosis 400 rad atau direndam dalam larutan EMS (0.1 %, 0.3%, dan 0.5 %) selama 1, 2 dan 3 jam. Selanjutnya kalus dipindahkan pada media untuk menginduksi pembentukan benih somatik. Hasil penelitian menunjukkan bahwa pembentukan kalus dipengaruhi oleh genotipe tanaman. Pembentukan kalus tertinggi dihasilkan dari Baluran (93.40%) dan terendah Burangrang (75.90%). Perlakuan iradiasigamma menurunkan pembentukan struktur torpedo, dimana struktur torpedo tertinggi diperoleh dari Burangrang (25.4-26.3/eksplan). Aksesi B 3592 mempunyai kemampuan membentuk struktur torpedo paling tinggi pada semua perlakuan EMS yang digunakan. Perendaman kalus dalam larutan EMS 0.5% selama 1, 2, dan 3 jam menurunkan regenerasinya membentuk struktur torpedo pada semua genotipe. Perlakuan EMSmenyebabkan kerusakan sel yang lebih besar dibandingkan dengan iradiasi sinar gamma, ditunjukkan dengan persentase pembentukan struktur torpedo setelah perlakuan EMS (0-15/eksplan) lebih kecil dibanding dengan iradiasi sinar gamma (10.3-26.3/eksplan).Kata kunci : Glycine max, iradiasi sinar gamma, Ethyl Methan Sulfonate, embriogenesis somatik  ABSTRACTThe Effect of Gamma Irradiation and Ethyl Methan Sulfonate on Somatic Embryo Formation of Soybean (Glycine max L.). Soybean is a source of protein and vegetable oil. Global climate change affect the productivity of soybean, so that new cultivars that have superior characteristic can be produced. In vitro techniques through somaclonal variation andmutation is one alternative for obtaining new varieties when genetic material as the material selection is not available. Mutation induction can be performed on embryogenic cell populations using gamma irradiation or chemical compounds, such as Ethyl Methane Sulfonate (EMS). Both of these methods have been widely used to increase the geneticdiversity of plants and have produced new clones with superior characteristic. The main component that must be controlled in the implementation of these technologies is somatic cells regeneration after mutation treatment in order to get in vitro shoots. Regenerationmethods which are successfully applied to certain varieties, often is not successfully for other varieties of the same species. Some factors that influence it, are such as explants source, genotype, medium composition, genotype, medium composition, etc. Somaclonal variation and mutation treatment can cause cell damage that is sometimes necessary need modifications of the regeneration method that has been produced before. The aim of the experiment was to get cell population and planlet mutation with gamma iradiadion and Ethyl Methan Sulfonate (EMS). Young embryozygotic was used as explant came from young pod that was harvested at 12-20 days after fertilization of Wilis, Burangrang and Baluran varieties and accession No B 3592. Embryogenic callus induction was done by using MS media with vitamin B5 added with 20 mg/l of 2,4-D and 3% sucrose. The callus were irradiated by gamma rays 400 rad or dilutein EMS solution with 0.1%, 0.3% and 0.5% concentration for 1, 2, and 3 hours. After mutation treatment, the callus were sub culture for seed somatic induction. The results showed that callus formation was influenced by plant genotype. All genotipe were able produced callus, where the highest percentage callus production was Baluran (93.40 %) and the lowest of that was Burangrang (75.90 %). Gamma irradiation reduces formation of torpedo structure. The highest torpedo structure after gamma irradiation was obtained from Burangrang (25.4-26.3/eksplan). Accession B 3592 had the ability to form torpedo structure highest among all treatments EMS used. Callus immersion in a solution of 0.5% EMS for 1, 2, and 3 hoursdecreased callus regeneration to formed torpedo structure in all  genotypes. EMS treatment causes greater cell damage than the gamma-ray, indicated by the percentage of the torpedo structure formation after EMS treatment (0-15/eksplan) which was smaller than the gammaray irradiation (10.3-26.3/explant). Key words : Glycine max, irradiation gamma rays, Ethyl Methan Sulfonate, somatic embryogenesis
Genetic Variation of the First Generation of Rodent Tuber (Typhonium flagelliforme Lodd.) Mutants Based on RAPD Molecular Markers SIANIPAR, NESTI FRONIKA; LAURENT, DANNY; PURNAMANINGSIH, RAGAPADMI; DARWATI, IRENG
HAYATI Journal of Biosciences Vol 22, No 2 (2015): April 2015
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1930.619 KB) | DOI: 10.4308/hjb.22.2.98

Abstract

Rodent tuber (Typhonium flagelliforme Lodd.) is a herbal plant from the Araceae family. The plant has high medical potential and is effective to cure cancer. However, the low level of its genetic variation limits its exploration for desirable traits. The low level of genetic variation in Rodent tuber is mainly due to its asexual reproduction system. It usually reproduces vegetatively via tuber separation. Therefore, gamma irradiation had been applied to rodent tuber in vitro to increase its genetic diversity. The objective of this study was to analyze the genetic diversity of the first generation (MV1) of gamma irradiated rodent tuber mutant using random amplified polymorphic DNA (RAPD) markers. A total of 14 mutant DNA samples were analyzed with 14 RAPD primers. The result showed that 67 out of 123 DNA bands were polymorphic among mutant lines. Based on cluster analysis these mutants showed 0.78-0.97 genetic similarity. Cutting of dendogram at genetic distance of 0.89 produced four main clusters. Mutants with high genetic variation are now available. This increases the opportunity of obtaining mutant lines with high anti-cancer activity.
PENGEMBANGAN TANAMAN KELADI TIKUS (TYPHONIUM FLAGELLIFORME LODD.) ASAL INDONESIA SEBAGAI OBAT ANTIKANKER Sianipar, Nesti Fronika; Purnamaningsih, Ragapadmi; Rosiana, Rosaria
ETHOS (Jurnal Penelitian dan Pengabdian) Vol 4 No.1 (Januari 2016) Ethos: Jurnal Penelitian dan Pengabdian (Sains & Teknologi)
Publisher : Universitas Islam Bandung

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Abstract

Keladi tikus merupakan tanaman obat yang bermanfaat sebagai obat kanker. Keladi tikus memiliki kandungan kimia diantaranya adalah alkaloid, flavonoid, saponin, steroid dan glikosida. Keladi tikus memiliki keragaman genetik yang rendah. Untuk meningkatkan keragaman telah dilakukan mutasi secara fisik melalui iradiasi sinar gamma pada kultur in vitro keladi tikus. Media MS dengan penambahan 0.5 mg/l 2.4D dan 0.1 mg/l kinetin menghasilkan jumlah tunas terbanyak yaitu 14.38 plantlet. Kalus embriogenik mutan dihasilkan melalui induksi dengan sinar gamma 6 gray. Kalus embriogenik diregenerasikan menjadi 59 tunas mutan. Plantlet mutan yang dihasilkan telah dideteksi dengan marka molekuler RAPD dan menunjukan perubahan genetik. Beberapa galur mutan yang telah diperoleh memiliki keragaman tinggi secara morfologi dan genetik. Keragaman genetik klon mutan generasi pertama (MV1) sampai generasi keempat (MV4) yang diperoleh telah dideteksi secara genetik molekuler dengan RAPD. Beberapa klon MV5 (mutan stabil) memiliki kandungan senyawa bioaktif antikanker tinggi berdasarkan GC-MS. Analisis GC-MS menunjukkan bahwa tanaman mutan mengalami peningkatan kandungan senyawa antikanker dibanding kontrol serta senyawa baru yang tidak ditemukan di kontrol. Beberapa klon unggul yang dihasilkan perlu dikembangkan sebagai bahan baku obat antikanker. Keladi tikus unggul bermanfaat untuk mencegah dan mengobati penyakit kanker pada masyarakat Indonesia.
Pengaruh Iradiasi Gamma dan Ethyl Methan Sulfonate Terhadap Pembentukan Embriosomatik Kedelai (Glycine max L.) Purnamaningsih, Ragapadmi; Mariska, Ika; Lestari, E.G.; Hutami, Sri; Yunita, Rossa
Jurnal Ilmiah Aplikasi Isotop dan Radiasi Vol 10, No 1 (2014): Juni 2014
Publisher : BATAN

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (170.167 KB) | DOI: 10.17146/jair.2014.10.1.2746

Abstract

Kedelai merupakan salah satu sumber protein dan lemak nabati yang penting. Perubahan iklim global berpengaruh terhadap produktivitas kedelai, sehingga diperlukan kultivar-kultivar baru yang mempunyai sifat unggul tertentu agar produktivitas kedelai dapat ditingkatkan. Teknik in vitro dengan mutasi dan keragaman somaklonal merupakan meoda alternatif untuk memperoleh varietas baru apabila material genetik sebagai bahan seleksi tidak tersedia. Induksi mutasi dapat dilakukan pada populasi sel embriogenik dengan menggunakan iradiasi sinar gamma atau senyawa kimia, antara lain Ethyl Methan Sulfonate (EMS). Kedua metoda tersebut telah banyak digunakan untuk meningkatkan keragaman genetik tanaman dan telah dihasilkan galur-galur baru dengan sifat unggul. Salah satu masalah penting yang harus dikuasai dalam penerapan teknologi tersebut adalah meregenerasikan sel somatik hasil mutasi dan keragaman somaklonal agar dapat ditumbuhkan menjadi planlet (tunas in vitro). Beberapa faktor yang mempengaruhi regenerasi tanaman adalah jenis bahan tanaman, genotipe, komposisi media, dll. Perlakuan keragaman somaklonal dan mutasi yang diberikan dapat menyebabkan kerusakan pada sel sehingga diperlukan modifikasi pada metoda regenerasi yang sudah diketahui agar populasi sel yang hidup setelah perlakuan mutasi dapat tumbuh menjadi tunas-tunas mutan. Tujuan penelitian adalah untuk mendapatkan planlet mutan hasil perlakuan mutasi dengan iradiasi gamma dan EMS. Varietas kedelai yang digunakan adalah Wilis, Burangrang, Baluran dan aksesi No. B 3592. Eksplan yang digunakan adalah embriozigotik muda berasal dari polong yang berumur 12-20 hari setelah penyerbukan. Induksi kalus embriogenik dilakukan dengan menggunakan media MS + vitamin Gamborg (B5) dengan penambahan 2,4-D 20 mg/l dan sukrosa 3%. Kalus yang didapatkan diberi perlakuan mutasi menggunakan sinar gamma pada dosis 400 rad atau direndam dalam larutan EMS (0.1 %, 0.3 %, dan 0.5 %) selama 1, 2 dan 3 jam. Selanjutnya kalus dipindahkan pada media untuk menginduksi pembentukan benih somatik. Hasil penelitian menunjukkan bahwa pembentukan kalus dipengaruhi oleh genotipe tanaman. Pembentukan kalus tertinggi dihasilkan dari Baluran (93.40%) dan terendah Burangrang (75.90%). Perlakuan iradiasi gamma menurunkan pembentukan struktur torpedo, dimana struktur torpedo tertinggi diperoleh dari Burangrang (25.4-26.3/eksplan). Aksesi B 3592 mempunyai kemampuan membentuk struktur torpedo paling tinggi pada semua perlakuan EMS yang digunakan. Perendaman kalus dalam larutan EMS 0.5% selama 1, 2, dan 3 jam menurunkan regenerasinya membentuk struktur torpedo pada semua genotipe. Perlakuan EMS menyebabkan kerusakan sel yang lebih besar dibandingkan dengan iradiasi sinar gamma, ditunjukkan dengan persentase pembentukan struktur torpedo setelah perlakuan EMS (0-15/eksplan) lebih kecil dibanding dengan iradiasi sinar gamma (10.3-26.3/eksplan). Kata kunci :   Glycine max, iradiasi sinar gamma, Ethyl Methan Sulfonate, embriogenesis somatik
In Vitro Culture Manipulation on Pruatjan for Secondary Metabolite Production Roostika, Ika; Purnamaningsih, Ragapadmi; Darwati, Ireng; Mariska, Ika
Jurnal AgroBiogen Vol 3, No 2 (2007): Oktober
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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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 Lestari, Endang G; Purnamaningsih, Ragapadmi; Syukur, Muhammad; Yunita, Rosa
Jurnal AgroBiogen Vol 6, No 1 (2010): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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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%.
Penyimpanan In Vitro Tanaman Obat Daun Dewa melalui Pertumbuhan Minimal Lestari, Endang G.; Purnamaningsih, Ragapadmi
Jurnal AgroBiogen Vol 1, No 2 (2005): Oktober
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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Abstract

Daun dewa (Gynura Procumbens) is a medicinal crop commonly used to remedy cancer, diabetes, and dermatitis. It has a bright prospect for future used. Plant preservations through tissue culture is done to anticipate an urgent need. An experiment was carried out to in vitro preservation of Daun Dewa by the minimum growth and regeneration to examine viability of the culture after the preservation. Terminal shoots (±1 cm) were cultured on a 1/2 MS basic medium + paclobutrazol (0, 1, 2, 3, and 4 mg/l) or ABA (1, 2, and 5 mg/l). The trial was arranged in a, completely randomized with 10 replications. The results showed a three-month preservation of the culture on a medium containing ABA inhibited proliferation and expansion of the plant shoots. Increasing ABA concentrations up to 5 mg/l, according to the shoot-growth inhibition, resulted in the height of 0.6 cm. After three month preservation, the shoots were able to produce roots. After 12 month preservation, the optimum capacity of growth inhibition was shown on 1/2 MS medium + ABA (1, 3, and 5 mg/l). The application of paclobutrazol (1, 2, 3, and 4 mg/l) in the medium produced low multiplication level of shoots, the length of the shoots remains higher than those on 1/2 MS medium without paclobutrazol. Seven months after preservation, viability of the plants was still high when cultured on MS medium + 2 mg/l BA combined with paclobutrazol and ABA as previously given. In addition, the rooted culture could be directly acclimatized in the glasshouse. The lowest number of shoot and shortest shoot after 12 month preservation period was found on the medium containing 5 mg/l ABA and 4 mg/l paclobutrazol, this treatment produced two shoots of 4 cm long. The best medium for the explant regeneration after 7 month preservation was MS + 2 mg/l BA. The plant shoots produced roots directly after they were acclimatized in the glasshouse.
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

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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.