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Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian, Bogor

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THE EFFECT OF PICLORAM AND LIGHT ON SOMATIC EMBRYOGENESIS REGENERATION OF PINEAPPLE Roostika, Ika; Khumaida, Nurul; Mariska, Ika; Wattimena, Gustaaf Adolf
Indonesian Journal of Agricultural Science Vol 13, No 2 (2012): October 2012
Publisher : Indonesian Agency for Agricultural Research and Development - MOA

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Abstract

Smooth Cayenne is the largest pineapple type cultivated in Indonesia, but its vegetative planting materials for mass propagation are limited. Somatic embryogenesis is a potential method to be applied. The aim of this study was to investigate the somatic embryogenesis regeneration under the effect of picloram and light. Callus formation was induced by picloram (21, 41 and 62 μM) added with 9 μM thidiazuron. The calli were transferred onto MS or Bac medium  enriched with N-organic compounds with or without addition of 21 μM picloram under dark or light condition. The compact calli were subcultured onto MS medium supplemented with 4.65 μM kinetin, while the friable calli were  transferred onto BIG medium (modified MS + 1.1 μM benzyl adenine + 0.9 μM indole butyric acid + 0.09 μM giberelic acid) or B medium (MS + 0.018 mM benzyl adenine). The results showed that the events of somatic embryogenesis were started from cell polarization, asymmetrical division, proembryo formation as  embryogenic tissues and friable embryogenic tissues, and embryo development. The best treatment for callus induction was 21 μM picloram. The addition of 21 μM picloram on N-organic enriched medium and the use of light condition  proliferated embryogenic calli. The N-organic enriched Bac medium and light condition yielded the highest number of mature somatic embryos (17 embryos perexplant in 2 months). The B medium was better than BIG medium to develop  somatic embryos from friable embryogenic tissues. The somatic embryogenesis method presented is potential for pineapple mass propagation and artificial seedproduction.Abstrak Bahasa IndonesiaSmooth Cayenne merupakan kultivar nenas yang banyak dibudidayakan di  Indonesia, namun ketersediaan benih untuk perbanyakan massal masih terbatas. Embriogenesis somatikadalah metode yang potensial untuk produksi bibit secara massal. Tujuan penelitian adalah untuk mempelajari pengaruh pikloram dan pencahayaan terhadap regenerasi embriogenesis somatik nenas. Kalus diinduksi menggunakan pikloram (21, 41, dan 62 μM) dan penambahan thidiazuron 9 μM. Selanjutnya, kalus dipindahkan ke media MS atau Bac yang diperkaya dengansenyawa N-organik dengan atau tanpa penambahan pikloram 21 μM dalam kondisi gelap atau dengan pencahayaan. Kalus kompak disubkultur pada media MS yang mengandung kinetin 4,65 μM, sedangkan kalus remah dipindahkan ke media BIG (MS modifikasi + bensil adenin 1.1 μM + indole butyric acid 0,9 μM + giberelic acid 0,09 μM) atau media B (MS + bensil adenin 0,018 μM). Hasil penelitian  menunjukkan bahwa tahapan embriogenesis somatik diawali dengan polarisasi sel, pembelahan asimetris, pembentukan proembrio sebagai jaringan embriogenik danjaringan embriogenik remah, serta perkembangan embrio. Perlakuan terbaik untuk induksi kalus adalah pikloram 21 μM. Penambahan pikloram 21 μM pada media yang diperkaya dengan senyawa N-organik mampu meningkatkan jumlah kalusembriogenik. Media Bac yang diperkaya senyawa N-organik dan kondisi pencahayaan menghasilkan jumlah embrio somatik dewasa terbanyak (17 embrio per eksplan dalam 2 bulan). Media B lebih baik daripada media BIG untuk regenerasi embrio somatik dari jaringan embriogenik remah. Metode embriogenesis somatik yang dihasilkan dari penelitian ini berpotensi diterapkan untukperbanyakan massal dan produksi benih nenas.
Pembentukan Benih Sintetik Tanaman Nenas Roostika, Ika; Purnamaningsih, R; Supriati, Y; Mariska, Ika; Khumaida, N; Wattimena, GA
Jurnal Hortikultura Vol 22, No 4 (2012): Desember
Publisher : Indonesian Center for Horticultural Research and Development

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Nenas merupakan tanaman buah tropis dan subtropis yang komersial. Kultivar Smooth Cayenne memiliki tipe dan jumlah propagul yang terbatas, sehingga diperlukan dukungan teknologi lainnya untuk produksi benih secara masal. Teknologi benih sintetik dapat diterapkan untuk produksi benih secara masal dan konservasi. Tujuan  penelitian ialah untuk mengetahui pengaruh kombinasi auksin dan sitokinin terhadap morfogenesis eksplan nenas yang terenkapsulasi, mengetahui pengaruh interaksi antara suhu penyimpanan dengan konsentrasi paklobutrazol atau manitol terhadap pertumbuhan eksplan nenas yang terenkapsulasi dan masa simpan. Penelitian dilaksanakan dari Bulan April sampai dengan Desember 2011 di Laboratorium Kultur Jaringan, Kelompok Peneliti Biologi Sel dan Jaringan, Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian, Bogor. Percobaan disusun secara faktorial dalam rancangan acak lengkap terdiri atas enkapsulasi eksplan, pertumbuhan minimal menggunakan paklobutrazol, atau manitol yang dikombinasikan dengan suhu penyimpanan. Enkapsulasi dilakukan terhadap batang semu dan basal daun menggunakan Na-alginat 3% yang berisi media MS dengan penambahan BA (0, 1, 2, dan 3 mg/l) yang dikombinasikan dengan NAA (0, 1, 2, dan 3 mg/l). Untuk memacu proses diferensiasi, basal daun diberi praperlakuan menggunakan media MS yang mengandung BA 0,5 mg/l dan NAA 0,5 mg/l sebelum dienkapsulasi dengan perlakuan BA dan NAA pada konsentrasi 0; 0,5; dan 1 mg/l.  Pertumbuhan minimal dilakukan menggunakan paklobutrazol (0, 1, 2, dan 3 mg/l) atau manitol (0, 1, 2, 3, 4, dan 5%) pada suhu penyimpanan 15 dan 25 0C. Hasil penelitian menunjukkan bahwa basal daun nenas yang terenkapsulasi mampu berdiferensiasi setelah praperlakuan. Tidak terdapat interaksi yang nyata antara konsentrasi paklobutrazol dengan suhu penyimpanan terhadap daya hidup dan daya tembus kapsul tunas nenas. Biakan tersebut hanya dapat disimpan selama 1 bulan. Interaksi yang nyata juga tidak dijumpai antara konsentrasi manitol dengan suhu penyimpanan terhadap daya hidup dan daya tembus kapsul embrio somatik nenas. Manitol 4% mampu memperpanjang masa simpan hingga 4 bulan. Manitol dapat menggantikan aplikasi suhu rendah dalam penyimpanan kultur nenas yang terenkapsulasi. Pineapple is a commercial tropical and subtropical fruit crop. Smooth Cayenne cultivar has limited type and number of propagules so that it should be supported by the other technology to produce plenty seedlings. Artificial seed can be applied for seed production and conservation. The objectives of the study were to know the effect of combination treatments between auxin and cytokinin to the morphogenesis of encapsulated pineapple cultures, to know the effect of paclobutrazol, mannitol, and temperature of storage to the growth of encapsulated pineapple cultures. The experiment was conducted from April to December 2011 at Tissue Culture Laboratory, Researchers Group of Cell and Tissue of Biology, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Bogor. Factorial of a completey randomized design was used. The study consisted of encapsulation, minimal growth by using paclobutrazol or mannitol combined with storage temperature. Encapsulation was conducted by using 3% Na-alginat containing of MS medium with addition of BA (0, 1, 2, and 3 mg/l) combined with NAA (0, 1, 2, and 3 mg/l). To promote differentiation, leaf bases were pre-cultured on MS media containing BA and NAA at concentration of 0.5 mg/l respectively prior to encapsulated by BA and NAA (0; 0.5; and 1 mg/l). Minimal growth was conducted by using paclobutrazol (0, 1, 2, and 3 mg/l), or mannitol (0, 1, 2, 3, 4, and 5%), and combined with storage temperature (15 and 25 0C). The results showed that encapsulated leaf bases of pineapple could differentiate after pre-treatment. There was no interaction between paclobutrazol and temperature to the survival rate and emergence rate of the encapsulated cultures. The encapsulated shoots could be stored for 1 months. There was also no interaction between mannitol and temperature to the survival rate and emergence rate of the encapsulated cultures. By using somatic embryos and 4% mannitol, the storage period could be prolonged for 4 months. Mannitol could substitute the use of low temperature in the conservation of encapsulated pineapple cultures.
Regenerasi Tanaman Sedap Malam Melalui Organogenesis dan Embriogenesis Somatik Rostika, Ika; Mariska, Ika; Purnamaningsih, R
Jurnal Hortikultura Vol 15, No 4 (2005): Desember 2005
Publisher : Indonesian Center for Horticultural Research and Development

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Secara konvensional perbanyakan tanaman sedap malam dilakukan melalui umbi. Semakin kecil ukuran umbi semakin lama tanaman berbunga. Penerapan teknik kultur in vitro diharapkan dapat membantu perbanyakan tanaman secara masal. Hingga saat ini, teknik kultur in vitro tanaman sedap malam belum pernah dilaporkan di Indonesia. Penelitian ini bertujuan memperoleh formulasi media yang efektif menginduksi organogenesis dan embriogenesis kultur in vitro tanaman sedap malam serta memacu regenerasinya. Percobaan dibagi menjadi 4 tahap, yaitu (1) induksi tunas, (2) multiplikasi tunas, (3) induksi kalus embriogenik, dan (4) regenerasi kalus embriogenik. Media induksi tunas yang diuji adalah MS+BA 0 ppm, MS+BA 3 ppm, MS+BA 5 ppm, dan MS+BA 7 ppm. Pemacuan multiplikasi tunas lanjut dilakukan pada media subkultur MS+BA 7 ppm+glutamin 100 ppm, MS+BA 7 ppm, DKW+TDZ 7 ppm, dan DKW+TDZ 7 ppm+glutamin 100 ppm. Untuk induksi kalus embriogenik, media induksi kalus yang diujikan adalah MS+2,4-D 2,5 ppm, MS +2,4-D 5 ppm, dan MS+2,4-D 10 ppm. Untuk meregenerasikan kalus embriogenik, media yang diujikan MS+BA 2 ppm+TDZ 0,2 ppm, MS+BA 3 ppm+TDZ 0,4 ppm, MS+zeatin 1ppm+kinetin 1ppm, dan MS+zeatin 0,5 ppm+kinetin 2 ppm. Hasil percobaan menunjukkan bahwa pembentukan tunas terbanyak diperoleh dari media BA 3 ppm (80%) namun inisiasi tunas tercepat dihasilkan pada media BA 0 ppm. Formula media MS+BA 7 ppm+glutamin 100 ppm menghasilkan jumlah tunas dan akar terbanyak. Penggunaan MS+2,4-D 5 ppm dapat menginduksi kalus embriogenik dengan persentase pembentukan nodul sebesar 18,75% dan jumlah nodul yang terbentuk sebanyak 3,6 dengan visual kalus yang paling baik. Setelah disubkultur, calon tunas terbanyak (17) dihasilkan dari perlakuan MS+BA 2 ppm+TDZ 0,4 ppm. Kalus embriogenik pada media MS+zeatin 0,5 ppm+kinetin 2 ppm dapat berkembang membentuk benih somatik.Regeneration of tuberose through organogenesis and embryogenesis. Tuberose is normally propagated by the tuber. The smaller size of tuber the longer time plant to flower. The application of in vitro culture technique might be used for mass propagation. Up to know, the research of in vitro culture of tuberose in Indonesia has not been reported. The objective of the study was to find out media formulation for organogenesis and embryogenesis. The experiments consisted of 4 steps of (1) shoot induction, (2) shoot multiplication, (3) induction of embryogenic callus, and (4) regeneration of embryogenic callus. The treatments for shoot induction were MS+BA 0 ppm, MS+BA 3 ppm, MS+BA 5 ppm, and MS+BA 7 ppm. The shoots were multiplied on media MS+BA 7ppm+glutamine 100ppm, MS+BA 7 ppm, DKW+TDZ 7 ppm, and DKW+TDZ 7 ppm+glutamin 100 ppm. For induction of embryogenic callus, the treatments were MS+2.4-D 2.5 ppm, MS+2,4-D 5 ppm, and MS+2.4-D 10 ppm. For regeneration of embryogenic callus, the treatments were MS+BA 2 ppm+TDZ 0.2 ppm, MS+BA 3 ppm +TDZ 0.4 ppm, MS+zeatin 1ppm+kinetin 1ppm, and MS+zeatin 0.5 ppm+kinetin 2 ppm. The results showed that the highest shoot formation was obtained from media MS+BA 3 ppm but the earliest shoot initiation was obtained from media MS+BA 0 ppm. The media formulation of MS+BA 7 ppm+glutamine 100 ppm gave the highest number of shoot and root. The application of media MS+2.4-D 5 ppm could induce embryogenic callus with high percentage of nodul formation (18.75%) and high number of nodul (3.6) with the best visual calli. After subculturing, the highest number of nodul (17) was obtained from media MS+BA 2 ppm+TDZ 0.4 ppm. The embryogenic callus from media MS+zeatin 0.5 ppm+kinetin 2 ppm could develop to form somatic seed.
Indirect Organogenesis and Somatic Embryogenesis of Pineapple Induced by Dichlorophenoxy Acetic Acid Roostika, Ika; Mariska, Ika; Khumaida, Nurul; Wattimena, Gustaaf A
Jurnal AgroBiogen Vol 8, No 1 (2012): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

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ABSTRACTIndirect Organogenesis and Somatic Embryogenesis ofPineapple Induced by Dichlorophenoxy Acetic Acid. IkaRoostika, Ika Mariska, Nurul Khumaida, and Gustaaf A.Wattimena. This research aimed to study the effect of 2,4-D,AdS, and basal media to the regeneration of pineapplethrough indirect organogenesis and somatic embryogenesis,and to study the complete event of somatic embryogenesis.Callus formation was induced by 21, 41, and 62 μM 2,4-Dwith addition of 9 μM TDZ. The non embryogenic calli weretransferred onto 4.65 μM Kn containing medium.Embryogenic callus formation was induced on MS or Bacbasal media consisted of N-organic compounds withaddition of AdS (0, 0.05 and 0.1 μM). The embryogenic calliwere regenerated on modified MS medium with addition of0.9 μM IBA, 1.1 μM BA, 0.09 μM GA3 or MS mediumsupplemented with 0.018 mM BA. The result proved that thesingle auxin of 2,4-D was not enough to induce embryogeniccells. Therefore the non embryogenic calli were regeneratedthrough organogenesis. The 21 μM 2,4-D yielded high level ofcallus formation (80%), higher fresh weight (0.2 g/explant)and higher number of shoot (25 shoots/explant in twomonths). Embryogenic calli were produced on N-organiccompounds enriched media. The regeneration mediumsignificantly affected the level of browning, where the MSmedium with addition of 0.018 mM BA yielded lower level ofbrowning. There was an interaction of embryogenic callusinduction medium and regeneration medium to the numberof mature somatic embryos. The embryogenic callusinduction on MS medium enriched with N-organiccompounds and 0.05 μM AdS followed by the regenerationof somatic embryos on MS medium with addition of 0.018mM BA was the best treatment which yielded 17 maturesomatic embryos/explant
Pengaruh Radiasi Sinar Gamma dan Asam Fusarat untuk Meningkatkan Ketahanan Abaka (Musa textilis Nee) terhadap Fasariurn oxysporum Damayanti, Fitri; Suharsono, Suharsono; Mariska, Ika
JURNAL BIOLOGI INDONESIA Vol 3, No 4 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (707.082 KB) | DOI: 10.14203/jbi.v3i4.3285

Abstract

ABSTRACTEffect of gamma radiation and fusaric acid for resistance to wild Fusarium disease onabaca plant (Mum textillis Nee). The problem in abaca production is wilt disease infectioncaused by Fusarium oxysporzim. The resistant variety against the pathogen has not beenavailable yet. The disease resistance character of the species might be improved throughsomaclonal variation and in vitro selection. Different pure toxin of fusaric acid concentration(0, 15,30,45,60, and 75 mg/l) was used as component selection to get new hope numbers ofresistant abaca to wilt Fusarium disease through in vitro selection. Concentration of 45 mgllfusaric acid is lethal for abaca, so we used this concentration as dose of selection to selectshoots from irradiated calli. Gamma irradiation was used as mutagent to increase somaclonalvariation on abaca. Six levels of gamma-ray radiation (0, 0.5, 1, 2, and 3 Krad) were appliedto embriogenic calli. Increasing dose of radiation decreased the viability of calli. LDso wasfound between 1-1.5 Krad of radiation dose. In vitro selection was carried out in two stages.The concentration of selection of hsaric acid in stage I1 was increased one level to theconcentration in stage 1. Stage I selection of shoots from irradiated calli on mediumcontaining pure toxin 45 mg/l fusaric acid, showed that the survival capacity decreasing withthe increasing doses of gamma irradiation. In stage 11, shoots from irradiated calli (at 0.5 and1 Krad) could survive on medium containing 60 mg/l fusaric acid. In medium selectioncontaining 50% filtrate F. oxysporum, fusaric acid resistant shoots were also filtrate resistant.There was a correlation between in vitro fusaric acid and filtrate of F. oxysporum resistantplant and conidia suspension of F. oxysporum resistkt plant in the greenhouse.Keywords: Gamma radiation, in vitro selection, fusaric acid, Musa textilis, Fusariumoxysporum
REGENERASI TANAMAN PEPAYA HASIL TRANSFORMASI DENGAN GEN ACC OKSIDASE ANTISENSE [Regeneration of Transforman Papaya Plant with ACC Oxidase Antisense Gene] Purnamaningsih, Ragapadmi; Mariska, Ika; Hutami, Sri
BERITA BIOLOGI Vol 7, No 5 (2005)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (497.521 KB) | DOI: 10.14203/beritabiologi.v7i5.873

Abstract

Papaya is climacteric fruit. As the other climacteric fruit, papaya has hight speed ripening, so papaya fruit can not stored in long period. Genetic enginering is one alternative technology to solve the problem by introducing antisense oxidase ACC gen to the papaya plant genome to get delay ripening characteristic. Success of genetic enginering technology depend on plant regeneration system.There were two ways of plant regeneration: organogenesis and somatic embryogenesis. The aim of this experiment was to induce root formation of papaya planlet which trasformated by ACC oxidase antisense gene.The former experiment showed that explant which transformated by ACC oxidase antisense gene can regenerated to be shoot/planlet with P6 medium.But when the shoot transferred to root induction medium the root was difficult to formed, callus was formed at the base of shoot, the leaves turn to yellow and fall down.Many media formulations were tried in this experiment with different basic medium for root induction and development.MS (1, Vi) DKW (1, A) and WPM (1, Vi) were used as basic media combined with sucrose (2 % and 3 %) and plant growth regulators (kinetin, IAA, and paclobutrazol) adding with some organic compound. Result of the experiment showed that MS Vi + paclobutrazol 0.5 mg/1 induced root formation 80 %, inhibited callus formation and decreased yellowing and falling of the leaves.
MIKROPROPAGASI SUKUN (Artocarpus communis Forst), TANAMAN SUMBER KARBOHIDRAT ALTERNATIF Supriati, Yati; Mariska, Ika; Hutami, Sri
BERITA BIOLOGI Vol 7, No 4 (2005)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (467.716 KB) | DOI: 10.14203/beritabiologi.v7i4.1049

Abstract

Bread fruit (Artocarpus communis Forst) is one of tropical fruit, which has a high contain of carbohydrate.In certain area, it becomes an alternative staple food when the main staple foods are scarce.The amount of carbohydrate in breadfruit is almost the same with the one in sweet potato, but it is higher than in potato. The main constraint of the development of breadfruit is the limited of seedling availability.Tissue culture technique has been known for its excellent result for plant propagation, because this technique has ability in producing seedling in a large quantity, in uniform growth rate and in a relative short time.The experiment was conducted at Cell Tissue Culture Division, Indonesian Center Agricultural Biotechnology and Genetic Resource Research and Development (1CAB1OGRAD) from February 2003 until December 2004.There were some steps experiments with series of combination medium as treatments. The first steps was shoot multiplication at Sk-2 medium with WPM + BA (0; 0,5; 1,0; 1,5 and 2,0 mg/1) + Thidiazuron (0; 0,4 mg/1);The second step was elongation shoot at Sk-3 with WPM + kinetin (1,2 and 3 mg/1) + GAa(0 and 5 mg/1), and the third was root initiation and proliferation, by comparing WPM + IBA (0, 2, 4 and 6 mg/1) + charcoal (0;0,5 %) and WPM (1; 0,5) + BA (0; 1,5 and 5 mg.l) or NAA (1,2 and 3 mg/1). For the step of acclimatization, soil and compost were used in comparison of (1;1 and 1:2).The result showed that the best media for shoot multiplication of breadfruit was WPM + BA 2 mg/1 + TDZ 0 4 with shoot number of 15,5., while the best media for shoot elongation was WPM + Kinetin 1 mg/1 + GA, 5 mg/1.WPM + IBA 3 mg/1 was the best formula for root proliferation with the highest root number about 6.5 and percentage of shoot producing root about 60%. For acclimatization, soil and compost in combination of 1:1 was the best media for planlet of breadfruit with the success rate about 70%. Charcoal is not necessary in root initiation and proliferation.
PENGGUNAAN PACLOBUTRAZOLDAN ABA DAL AM PERBANYAKAN X NENAS SIMADU MELALUI KULTUR IN VITRO Purnamaningsih, Ragapadmi; Mariska, Ika; Supriati, Yati
BERITA BIOLOGI Vol 9, No 6 (2009)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (510.62 KB) | DOI: 10.14203/beritabiologi.v9i6.852

Abstract

Pineapple (Ananas comosus L. Merr.), represents an important crop in Subang. Somaclonal variation is one of the problem to develop pineapple, especially Simadu variety. Probability to conduct Simadu progeny from the mother plant is very low (5%).Its caused by chimeric of the somatic cells that form meristem.In vitro culture is the alternative method to solve the problem by using the meristem cells from Simadu fruit as explant. Unfortunately, genetic diversity has been observed in many spesies during tissue culture.This phenomenon is usually termed somaclonal variation. Many studies on pineapple demonstrsted that some in vitro propagated materials differ from the source materials from which they are derived.To minimize genetic variability, the use of growth inhibitor such as paclobutazol and absisic acid hopefully would gave the important role in genetic stability. The aim of the research is to multiply Simadu pineapple by using tissue culture technic. In vitro shoot induce from crown of the Simadu fruit until get the sterile shoots. Combination of kinetin (0-5 ppm) with paclobutrazol ( 0-0.1 ppm) or ABA (0-1 ppm) was used in the multiplication stage. Result showed that there are no interaction between kinetin and paclobutrazol or ABA, but there is influence of the single factor. Kinetin increase leave number but decrease plant height and root number. Paclobutrazol increase shoot and leave number, but decrease plant height and root number. There is no influence of ABA to plant height, shoot and root number but decreased leaves number.
INDUKSI POLIPLOIDI DENGAN KOLKISIN PADA HIBRID F1 HASIL PERSILANGAN ANTAR SPECIES PADA TANAMAN PANILI ASAL CIAMIS Damayanti, Fitri; Mariska, Ika
BERITA BIOLOGI Vol 6, No 4 (2003)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v6i4.3455

Abstract

Steem root disease caused by Fusarium oxysporum Schl. f. sp. Vanillae (Tucker) Gordon represent one internal issues of vanilla development (Vanilla planifolia Andrews). To obtain resistance clone to the disease, it can exploit resource of wild vanilla (V. albida B. L Syn) through crosses. Hybrids which were interspecific crossed generally were sterile. To overcome sterility problem of the hybrids, chromosome doubling was made by colchicine application. Explant used globular structure of proembryo from F1 seed result from a cross between wild vanilla of Ciamis as female parents and cultivated vanilla clone of Ciamis as male parent. Concentration level colchicine used were 0.00%, 0.05%, 0.10%, 0.20% and 0.25% with period of treatment of 3 and 6 days. After colchicine treatment embryo cultures were subcultured into new mwdium that was basal media Murashige-Skoog enriched with 2.5 mg/l BAP. Result of the experiment showed that colchicine treatment,globular structure were F1 embryo tending to inhibit early regeneration. The cultures showed variabilities from treatment of colchicin 0.20% during of 6 day and 0.25% for 3 days. Phenotypic performance of the chromosome doubled hybrids showing great variation in color and vigor of the culture. Tetraploid plant(2n=4x=64) was obtained from the colchicine treatment of 0.25% for 6 days. Chromosome addtion was followed by improvement of cell dimensions and organ magnification.
PENGARUH RADIASI SINAR GAMMA DAN ASAM FUSARAT UNTUK MENINGKATKAN KETAHANAN ABAKA (MUSA TEXTILIS NEE) TERHADAP FASARIURN OXYSPORUM Damayanti, Fitri; Suharsono, Suharsono; Mariska, Ika
JURNAL BIOLOGI INDONESIA Vol 3, No 4 (2002): JURNAL BIOLOGI INDONESIA
Publisher : Perhimpunan Biologi Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/jbi.v3i4.3285

Abstract

ABSTRACTEffect of gamma radiation and fusaric acid for resistance to wild Fusarium disease onabaca plant (Mum textillis Nee). The problem in abaca production is wilt disease infectioncaused by Fusarium oxysporzim. The resistant variety against the pathogen has not beenavailable yet. The disease resistance character of the species might be improved throughsomaclonal variation and in vitro selection. Different pure toxin of fusaric acid concentration(0, 15,30,45,60, and 75 mg/l) was used as component selection to get new hope numbers ofresistant abaca to wilt Fusarium disease through in vitro selection. Concentration of 45 mgllfusaric acid is lethal for abaca, so we used this concentration as dose of selection to selectshoots from irradiated calli. Gamma irradiation was used as mutagent to increase somaclonalvariation on abaca. Six levels of gamma-ray radiation (0, 0.5, 1, 2, and 3 Krad) were appliedto embriogenic calli. Increasing dose of radiation decreased the viability of calli. LDso wasfound between 1-1.5 Krad of radiation dose. In vitro selection was carried out in two stages.The concentration of selection of hsaric acid in stage I1 was increased one level to theconcentration in stage 1. Stage I selection of shoots from irradiated calli on mediumcontaining pure toxin 45 mg/l fusaric acid, showed that the survival capacity decreasing withthe increasing doses of gamma irradiation. In stage 11, shoots from irradiated calli (at 0.5 and1 Krad) could survive on medium containing 60 mg/l fusaric acid. In medium selectioncontaining 50% filtrate F. oxysporum, fusaric acid resistant shoots were also filtrate resistant.There was a correlation between in vitro fusaric acid and filtrate of F. oxysporum resistantplant and conidia suspension of F. oxysporum resistkt plant in the greenhouse.Keywords: Gamma radiation, in vitro selection, fusaric acid, Musa textilis, Fusariumoxysporum