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Mikropropagasi planlet tebu menggunakan sistem perendaman sesaat (SPS) Micropropagation of sugarcane plantlets using temporary immersion system (TIS) Hayati MINARSIH; Imron RIYADI; . SUMARYONO; Asmini BUDIANI
Menara Perkebunan Vol. 81 No. 1: 81 (1), 2013
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v81i1.53

Abstract

bstractTo achieve Indonesian sugar self-sufficiency in2014, the national production needs to be escalatedthrough land extensification that requires a largenumbers of cane planting materials. This can be achievedby mass propagation of sugarcane through in vitroculture. Solid medium is commonly used for callusproliferation in sugarcane tissue culture. However, solidmedium is considered inefficient in terms of plantletproduction level, labour and space. The use of liquidmedium may solve the problem by allowing automationto increase plantlet production scale and uniformity.Temporary immersion system (TIS) is based on a shortperiodic immersion of explants in a liquid medium for aspecific frequency and duration. Research on in vitromass propagation of sugarcane using TIS was conductedat the Indonesian Biotechnology Research Institute forEstate Crops. Callus initiated from immature unfoldedleaves of PSJT 941 and PS 881 was cultured on liquidMS medium in TIS with different frequencies (12 and24 h) and durations (1 and 3 min) of immersion. Eachtreatment was replicated three times. The callus biomassof two elite cane varieties (PSJT 941 and PS 881)cultured in TIS for six weeks was higher (2 – 4 times fold)than that of on solid medium. The PSJT 941 varietyreached the highest calli biomass with immersion forthree min every 24 h. However, PS 881 variety reachedits highest biomass with immersion for one minute every24 h. The propagation of sugarcane using TIS culturewas proven to produce higher calli biomass up to fourfolds and to form more numbers and uniform shootscompared to the solid medium culture. The callus wassuccesfully regenerated to shoots and plantlets.AbstrakUntuk mencapai swasembada gula, perlu dilakukanpeningkatan produksi gula nasional melalui perluasanareal pertanaman tebu sehingga diperlukan bibit dalamjumlah besar. Hal tersebut dapat diatasi antara laindengan perbanyakan tebu melalui kultur in vitro. Peng-gunaan medium padat pada perbanyakan kalus tebumelalui kultur in vitro merupakan teknik yang umumdigunakan saat ini. Akan tetapi penggunaan mediumpadat dianggap kurang efisien dalam hal jumlah planletyang diproduksi, tenaga kerja dan ruang digunakan.Penggunaan medium cair dapat mengatasi kelemahantersebut dengan dimungkinkannya otomatisasi sehinggadapat meningkatkan skala produksi secara massal dankeseragaman planlet. Sistem perendaman sesaat (SPS)merupakan teknik kultur in vitro dalam medium cairmenggunakan bejana bersekat dimana kontak antaraeksplan dan medium terjadi hanya secara sesaat danperiodik. Penelitian perbanyakan massal bibit tebumelalui SPS dilakukan di Balai Penelitian BioteknologiPerkebunan Indonesia. Kalus diinisiasi dari daun meng-gulung varietas PSJT 941 dan PS 881 yang ditumbuhkanpada media MS cair dalam kultur SPS dengan frekuensiyang berbeda (12 dan 24 jam) dan lama perendaman (1dan 3 menit). Setiap perlakuan diulang tiga kali. Bobotbasah (biomassa) kalus dari dua varietas tebu (PSJT 941dan PS 881) yang ditumbuhkan dengan metode SPSsetelah enam minggu menunjukkan pening-katan yanglebih tinggi yaitu antara 2 - 4 kali lipat dibandingkandengan kontrol (media padat). Peningkatan biomassatertinggi pada varietas PSJT 941 diperoleh pada per-lakuan SPS dengan interval perendaman 24 jam dan lamaperendaman tiga menit. Sedangkan pada PS 881,peningkatan tertinggi biomassa diperoleh pada intervalperendaman 24 jam dan lama perendaman satu menit.Perbanyakan dengan metode SPS terbukti dapat mening-katkan biomassa kalus lebih dari empat kali lipat danpembentukan tunas yang lebih seragam dibandingkandengan pada media padat. Kalus yang dihasilkan dapatdiregenerasikan menjadi tunas dan planlet.
Pengaruh jenis penutup botol kultur terhadap pertumbuhan planlet kelapa sawit (Elaeis guineensis Jacq.) Effect of different culture vessel closures on the growth of oil palm (Elaeis guineensis Jacq.) plantlets Masna Maya SINTA; Imron RIYADI; . UMARYONO
Menara Perkebunan Vol. 79 No. 1: 79 (1), 2011
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v79i1.68

Abstract

AbstractMicroenvironment inside the culture vessel such astemperature, light intensity, relative humidity, and aerationaffect growth and development of plantlets. This experimentwas conducted to determine the effect of different culturevessel closures on microenvironmental conditions inside thevessel and on growth of plantlets of oil palm. Shoots of oilpalm derived from somatic embryos were cultured on DFmedium for eight weeks in transparent culture bottlescovered with five different vessel closures e.i. screw cap withplastic wrap, screw cap, plastic wrap, aluminum foil, andautoclavable plastic. The culture vessels were placed in theculture room with light intensity 20 µmol/m 2 /sec for 12 hoursphotoperiod, at room temperature 26°C. Parametersobserved on plantlet growth were shoot height, biomass freshweight, leaf number, and leaf color grade, while onmicroenvironment were temperature and light intensity. Atthe end of experiment, the volume and fresh weight of theremaining medium were measured to determine evaporationrate of each treatment. Results show that the use of differentculture vessel closures affected the microenvironment insidethe vessel, the volume of the remaining medium, and thegrowth of the plantlets. The closure increased thetemperature by 1.6 – 2.6°C and decreased the light intensityby 1.7 – 8.7 µmol/m 2 /sec inside the culture vessels dependson the culture vessel closures. Culture vessels with aluminumfoil closure had the lowest temperature (28.9°C) and thelowest light intensity (10.8 µmol/m 2 /sec) gave the best resultin the growth of the plantlets. Better plantlets growth wasalso observed in the culture vessel with autoclavable plasticclosure that less expensive, therefore it can be used as analternative vessel closure for the growth of oil palm plantlets.AbstrakLingkungan mikro di dalam botol kultur seperti suhu,intensitas cahaya, kelembaban nisbi dan aerasi mem-pengaruhi pertumbuhan dan perkembangan planlet.Penelitian ini dilakukan untuk mengetahui pengaruhpenggunaan penutup botol kultur yang berbeda terhadapkondisi lingkungan mikro di dalam botol kultur danpertumbuhan planlet kelapa sawit. Planlet kelapa sawit asalembrio somatik dikulturkan dalam botol kultur bening berisimedium DF selama delapan minggu dan ditutup mengguna-kan lima jenis penutup botol yang berbeda yaitu tutup ulirdengan plastik wrap, tutup ulir, plastik wrap, aluminium foildan plastik tahan diautoklaf. Kultur diletakkan dalam ruangkultur, di bawah lampu TL dengan intensitas cahaya20 µmol/m 2 /detik dan suhu ruang 26 o C. Parameterpertumbuhan planlet yang diamati adalah tinggi planlet,bobot basah, jumlah daun dan kelas warna daun, sedangkanlingkungan mikro adalah suhu dan intensitas cahaya. Padaakhir eksperimen, volume dan bobot basah medium yangtersisa diukur untuk mengetahui tingkat penguapan padasetiap perlakuan. Hasil penelitian menunjukkan bahwapenggunaan penutup botol yang berbeda berpengaruhterhadap lingkungan mikro, volume medium tersisa dalambotol kultur dan pertumbuhan planlet. Penutup botolmeningkatkan suhu 1,6 – 2,6 o C dan menurunkan intensitascahaya 1,7 – 8,7 µmol/m 2 /detik di dalam botol tergantungpada jenis penutup botol yang digunakan. Botol kulturdengan penutup berbahan aluminium foil mempunyaiintensitas cahaya terendah (10,8 µmol/m 2 /detik) dan suhuterendah (28,9 o C) memberikan hasil terbaik pada pembesaranplanlet kelapa sawit. Pertumbuhan planlet yang baik jugaterdapat pada botol kultur dengan penutup plastik tahandiautoklaf yang lebih murah, sehingga penutup ini dapatdigunakan sebagai pilihan untuk pembesaran planlet kelapasawit.
Pembentukan akar in vitro planlet kelapa sawit (Elaeis guineensis Jacq.) dalam medium cair dengan penambahan auksin In vitro rooting of oil palm (Elaeis guineensis Jacq.) plantlets in a liquid medium supplemented with auxins Imron RIYADI; . SUMARYONO
Menara Perkebunan Vol. 78 No. 1: 78 (1), 2010
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v78i1.76

Abstract

AbstractAuxin affects the growth and development of in vitro plantlets including root induction. An experiment was conducted to determine the combination and concentration of auxin for rooting of oil palm plantlets in liquid medium.Unrooted plantlets of oil palm MK 649 clone with height 6 – 7 cm and 2 – 3 leaves were used as material source. The plantlets were cultured in de Fossard liquid medium. The treatments used were combinations of NAA and IBA at 0, 5,10 and 20 μM. The results show that 10 μM NAA combined with 20 μM IBA gave the highest percentage of rooting of oil palm plantlets (73.3%) in 10 weeks. NAA and IBA concentration influenced significantly rooting percentageand root quality and there was a significant interaction between the two auxins. Root initiation response of oil palm plantlets to NAA was higher than to IBA. The best of oil palm root class which indicates root quality was obtained in a medium with 10 μM NAA + 20 μM IBA. The aerial parts of the plantlets grew well in term of shoot height, leaf number and shoot diameter especially in a medium with 10 μM NAA + 20 μM IBA. AbstrakAuksin berpengaruh terhadap pertumbuhan dan perkembangan planlet in vitro, termasuk terhadap induksi akar. Penelitian ini bertujuan untuk menentukan kombinasi dan konsentrasi auksin yang tepat dalam pembentukan akarplanlet kelapa sawit in vitro dalam medium cair. Bahan yang digunakan berupa planlet kelapa sawit klon MK 649 tanpa akar dengan tinggi 6 – 7 cm dan jumlah daun 2 – 3 helai. Planlet dikulturkan dalam medium de Fossard cair. Perlakuan yang digunakan adalah kombinasi NAA dan IBA dengan konsentrasi 0, 5, 10 dan 20 μM. Hasil penelitian menunjukkan bahwa perlakuan NAA 10 μM dikombinasikan dengan IBA 20 μM menghasilkan persentase pembentukan akar planlet kelapa sawit tertinggi yaitu 73,3% dalam waktu 10 minggu. Konsentrasi NAA dan IBA secara nyata mempengaruhi persentase pembentukan dan kualitas akar serta terdapat interaksi yang nyata antara kedua perlakuan auksin. Respons induksi akar kelapa sawit terhadap NAA lebih tinggi daripada IBA. Kelas akar planlet kelapa sawit terbaik yang menunjukkan kualitas perakaran, juga diperoleh pada NAA 10 μM dan IBA 20 μM. Pertumbuhan dan perkembangan organ bagian atas yang meliputi tinggi tunas, jumlah daun dan diameter tunas menunjukkan peningkatan yang cukup baik terutama pada perlakuan NAA 10 μM + IBA 20 μM.
Pengaruh interval dan lama perendaman terhadap pertumbuhan dan pendewasaan embrio somatik tanaman sagu (Metroxylon sagu Rottb.) Effect of immersion interval and duration on the growth and maturation of somatic embryos of sago palm ( Metroxylon sagu Rottb.) Imron RIYADI; . SUMARYON
Menara Perkebunan Vol. 77 No. 2: 77 (2), 2009
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v77i2.97

Abstract

AbstractLiquid culture via temporary immersionsystem (TIS) has a potency for enhancingmaturity and uniformity of plant somatic embryos(SE). An experiment was conducted to determinethe effect of medium immersion interval andduration on the growth and maturation of sagoSE in TIS. A clump of SE at globular stagederived from sucker’s tip meristem culture wasused as material source. The SE were cultured ona modified Murashige and Skoog medium addedwith 0.01 mg/L ABA, 1.0 mg/L kinetin and0.1 mg/L GA 3 . The treatments used were TISwith immersion interval of 3, 6 and 12 hourswith duration of 1 and 3 minutes. Solid mediumwas used as a control. The results show that TISwith immersion interval 12 hours for threeminutes produced the highest SE biomass(14.6 g/flask) which had increased by 9.8-foldwithin six weeks. The longer of immersioninterval (less frequent) and the longer ofimmersion duration (three minutes) increasedsignificantly biomass fresh weight of of sago SE.SE biomass of sago on solid medium wassignificantly lower than those of in liquid mediaof TIS. The highest number of advanced stageembryos (torpedo, cotyledonary and earlygerminant) of 643 or 48.2% from the totalnumber of SE was achieved in TIS with 12 hoursinterval for three minutes. During the maturationof sago SE, the color of embryos has changedfrom mostly yellowish to greenish and reddish.AbstrakKultur cair dengan sistem perendaman sesaat(SPS) berpotensi untuk meningkatkanpendewasaan dan keseragaman embrio somatik(ES) tanaman. Penelitian ini bertujuan untukmenentukan pengaruh interval dan lamaperendaman terhadap proses pendewasaan ESsagu dalam SPS. Bahan yang digunakan berupaES fase globuler asal kultur pucuk tunas anakansagu. ES sagu dikulturkan dalam mediumMurashige dan Skoog yang dimodifikasiditambah ABA 0,01 mg/L; kinetin 1 mg/L danGA 3 0,1 mg/L. Perlakuan yang digunakan adalahkultur SPS dengan interval perendaman 3, 6 dan12 jam dengan lama perendaman 1 dan 3 menitserta kultur padat sebagai pembanding. Hasilpenelitian menunjukkan bahwa perlakuan SPSdengan interval perendaman 12 jam selama tigamenit menghasilkan bobot biomassa ES tertinggiyaitu 14,6 g/bejana yang meningkat 9,8 kalidalam waktu enam minggu. Interval peren-daman lebih lama (lebih jarang) dan lama peren-daman lebih panjang (tiga menit) meningkatkansecara nyata bobot segar biomassa ES sagu.Biomassa ES sagu pada medium padat secaranyata lebih rendah dibandingkan dengan kulturkotiledon dan kecambah dini) tertinggi yaitu 643atau 48,2% dari jumlah total ES diperoleh pada perlakuan SPS interval 12 jam dengan lama tigamenit. Seiring dengan pendewasaan ES sagu, terjadi perubahan warna dari sebagian besar kuning menjadi hijau dan merah.
Perbanyakan in vitro tanaman kina (Cinchona ledgeriana Moens) melalui tunas aksiler dan apikal In vitro propagation of cinchona (Cinchona ledgeriana Moens) from axillary and apical buds Imron RIYADI; J. S. TAHARDI TAHARDI
Menara Perkebunan Vol. 77 No. 1: 77 (1), 2009
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v77i1.114

Abstract

AbstractThe use of the appropriate source of nodalbud explants in culture can increase theeffectiveness and efficiency of shootmultiplication. An experiment was conducted todetermine and compare the rate of in vitro shootmultiplication from apical and axillary budsin cinchona (Cinchona ledgeriana Moens) andtheir subsequent growth and development. Theplant material used was Cinchona ledgerianaoriginating from the Indonesian Tea andCinchona Research Institute, Gambung, WestJava. Explants were taken from apical andaxillary nodes from in vitro germinated seedlings.The cultures were incubated at 26 0 C and 60%relative humidity under a 14-h photoperiod withlight intensity of 30 µmol photon/m 2 /sec.provided by cool-white fluorescent tubes (TL40 W) for 4 - 8 weeks. The parameters observedwere shoot multiplication rate, shoot growth anddevelopment such as shoot length, leaf numberand rooting frequency. Apical and axillary nodesproduced shoots at different multiplication rateson Murashige-Skoog (MS) standard mediumcontaining 30 g/L sucrose and supplemented with1 – 5 mg/L BA in combination with 0.1 mg/L IBA.Furthermore, shoots or plantlets of cinchonagrew and developed on the same mediacontaining 5 – 10 mg/L IAA combined with0.5 mg/L IBA. The results showed that shootmultiplication rate was higher in axillary than inapical nodes. The highest multiplication rate inaxillary nodes was 24.6 shootlets with 3 mg/LBA treatment, whereas in apical nodes it was17.2 shootlets with 5 mg/L BA treatment for eightweeks. The highest rooting frequency ofcinchona plantlet was 90%, achieved with 5 mg/LIAA in combination with 0.5 mg/L IBA. Theplantlets were successfully acclimatized andtransplanted to the fieldAbstrakSumber eksplan berupa nodus/tunas padakultur in vitro umum digunakan untuk multi-plikasi tunas. Penelitian ini bertujuan untukmembandingkan tingkat multiplikasi antara tunasapikal dengan tunas aksiler tanaman kina Ledgersecara in vitro. Bahan tanaman yang digunakanadalah kina Ledger (Cinchona ledgeriana Moens)yang berasal dari Pusat Penelitian Teh dan Kina,Gambung, Jawa Barat. Eksplan berupa nodus/tunas apikal dan aksiler asal biji yang dikecam-bahkan secara in vitro. Kultur tersebut diinku-basikan dalam ruang terang pada intensitascahaya 30 μmol foton/m 2 /detik dengan periodepenyinaran 14 jam pada suhu 260 C dankelembaban relatif + 60% selama 4 – 8 minggu.Parameter yang diamati adalah perbandinganmultiplikasi tunas dan pertumbuhan tunas yangmeliputi rata-rata tinggi tunas, jumlah daun danfrekuensi pengakaran. Nodus apikal maupunaksiler menghasilkan tunas dengan tingkatMurashige-Skoog (MS) standar yang me-ngandung sukrosa30 g/L dan ditambahkan BA1 – 5 mg/L dikombinasikan IBA 0,1 mg/L.Selanjutnya tunas/planlet kina tersebut berhasiltumbuhdan berkembang pada medium sama yangdiberi IAA 5 – 10 mg/L dikombinasikan denganIBA 0,5 mg/L. Hasil penelitian menunjukkanbahwa tingkat multiplikasi tunas aksiler lebihtinggi dari pada tunas apikal. Multiplikasi tunasaksiler menghasilkan jumlah tunas rata-ratatertinggi sebesar 24,6 tunas per eksplan padaperlakuan BA 3 mg/L sedangkan multiplikasitunas apikal tertinggi sebesar 17,2 tunas pereksplan pada perlakuan BA 5 mg/L pada umurdelapan minggu. Frekuensi pengakaran planletkina tertinggi mencapai 90% pada perlakuan IAA10 mg/L yang dikombinasikan dengan IBA 0,5mg/L. Planlet yang dihasilkan telah berhasildiaklimatisasi dan dipindahkan ke tempatpersemaian lapang.
Direct somatic embryogenesis and plant regeneration in tea by temporary liquid immersion Embriogenesis somatik langsung dan regenerasi tanaman teh melalui perendaman sesaat J S TAHARDI; Tatik RAISAWATI; Imron RIYADI; W A DODD
Menara Perkebunan Vol. 68 No. 1: 68(1), 2000
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v68i1.133

Abstract

Ringkasan Perbanyakan tanaman teh [Camellia sinen­sis (L.) O. Kuntze] melalui stek tunas berdaun tunggal hanya dapat menghasilkan klon unggul dalam jumlah terbatas. Oleh sebab itu diperlukan metode alternatif dengan teknik kultur sel dan jaringan untuk perbanyakan klonal secara cepat. Dalam penelitian ini dikembangkan metode yang lebih efektif untuk regenerasi tanaman teh melalui embriogenesis somatik langsung. Massa pro­embriogenik dari eksplan kotiledon dihasilkan dengan frekuensi 56,7% dalam media MS padat setengah konsentrasi yang mengandung BAP 2 mg1L. Proliferasi, perkembangan, pendewasaan dan perkecambahan embrio somatik diperoleh dengan sistem perendaman sesaat (SPS) yang menggunakan media MS cair setengah konsen­trasi, yang diperkaya dengan zat pengatur tumbuh dengan berbagai konsentrasi. Proliferasi embrio meningkat 4,3 kali dalam media yang diberi BAP 2 mglL; perkembangan dan pendewasaannya meningkat dengan penambahan kinetin dan ABA masing-masing pada konsentrasi 0,1 mg1L yang 30% diantaranya berkecambah dan membentuk planlet tanpa penambahan zat pengatur tumbuh. Protokol SPS tersebut merupakan sistem in vitro yang berpotensi bagi proliferasi dan perkembang­an embrio somatik tanaman teh yang cepat dan sinkron dari kultur kotiledon, serta regenerasinya menjadi planlet tanpa melalui fase kalus.Summary Tea propagation by single-leaf bud cuttings has limited applications for rapid dissemination of planting materials from new elite clones. An alternative method for rapid cloning by cell and tissue culture technique is necessary. In this study we have established an improved method for tea [Camellia sinensis (L.) O. Kuntze] plant regenera­tion via direct somatic embryogenesis. Clumps of proembryogenic masses were initiated at a fre­quency of 56.7% from cotyledonary slices cul­tured on a half-strength MS agar-gelled medium supplemented with 2 mg/L BAP. Proliferation, development, maturation and germination of so­matic embryos were achieved using the temporary immersion system (TIS) provided with half­strength MS liquid media supplemented with varying concentrations of growth regulators. Em­bryo proliferation increased by 4.3-fold in me­dium provided with 2 mg/L BAP; their develop­ment and maturation were enhanced by the presence of both kinetin and ABA at 0.1 mg/L each. Germination and plant recovery were achieved at a frequency of about 30% without the use of growth regulators. The TIS protocol des­cribed above represents an in vitro system poten­tial for rapid proliferation and synchronized development of tea somatic embryos from cotyledon cultures, and their regeneration into plantlets without an intervening callus phase.
Pertumbuhan dan perkembangan kalus embriogenik dan embrio somatik kelapa sawit (Elaeis guineensis Jacq.) pada sistem perendaman sesaat Growth and differentiation of embryogenic callus and somatic embryos of oil palm (Elaeis guineensis Jacq.) in a temporary immersion system . SUMARYONO; Imron RIYADI; Pauline D. KAS; Gale GINTING
Menara Perkebunan Vol. 75 No. 1: 75 (1), 2007
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v75i1.152

Abstract

SummaryIn temporary immersion system (TIS),plant materials are exposed to the medium fora short time, therefore they are more exposedto the air and a lack of oxygen frequentlyexperienced by a liquid culture can be avoided.This experiment was conducted to determinethe procedure for callus proliferation up tosomatic embryo germination of oil palm(Elaeis guineensis Jacq.) in TIS culture.Embryogenic calli of oil palm clone MK 638from Marihat Research Institute were culturedon solid medium in the dark culture room andthen used as materials for TIS. Immersion timefor all cultures was three minutes every sixhours. Callus proliferation was conducted inDF liquid culture with 5 mg/L 2,4-D and0.1 mg/L kinetin with transfer interval of 4, 6and 8 weeks. The treatments for somaticembryo maturation were kinetin and ABA,whereas for somatic embryo germination wasIBA, kinetin and GA 3 . The results show thatthe best transfer interval for callus proli-feration was four weeks. In this treatment therelative growth rate of callus was0.38 g/g/week. Somatic embryo initiation fromthe callus was done in DF mediumsupplemented with 1 mg/L 2,4-D and 0.1 mg/Lkinetin. The percentage of somatic embryowas 80% based on biomass fresh weight afterthe fourth subculture. The addition of 0.5 mg/Lkinetin and 0.05 mg/L ABA improved somaticembryo maturation of oil palm; the averagenumber of somatic embryos at advanced stages(torpedo and cotyledonary) was 16.3 embryosper flask. The addition of 2 mg/L IBA and0.5 mg/L kinetin in DF medium with half-strength macro-salt enhanced significantly thegermi-nation of somatic embryos. GA 3 at0.1 mg/L increased the total number ofgerminants.RingkasanPada sistem perendaman sesaat (SPS),bahan tanam hanya terpapar sebentar dalammedium sehingga paparan dengan udara lebihlama dan kekurangan oksigen yang seringterjadi pada kultur cair dapat diatasi. Penelitianini bertujuan menetapkan prosedur untukperbanyakan kalus embriogenik sampai denganperkecambahan embrio somatik kelapa sawit(Elaeis guineensis Jacq.) dalam kultur SPS.Kalus embriogenik kelapa sawit klon MK 638yang diperoleh dari Balai Penelitian Marihatdiperbanyak pada medium padat di ruang gelapyang kemudian digunakan sebagai bahan untukkultur cair SPS. Lama perendaman semuakultur di SPS diatur tiga menit denganfrekuensi setiap enam jam. Perbanyakan kalusdalam medium cair DF dengan 2,4-D 5 mg/Ldan kinetin 0,1 mg/L dilaksanakan denganinterval subkultur 4, 6 dan 8 minggu.Perlakuan pematangan embrio somatik adalahkinetin dan ABA sedangkan perlakuan untukperkecambahan embrio somatik adalah IBA,kinetin dan GA 3 . Hasil penelitian menunjuk-kan bahwa untuk proliferasi kalus embriogenikkelapa sawit, interval subkultur terbaik adalahempat minggu. Pada perlakuan ini laju tumbuhrelatif kalus mencapai 0,38 g/g/minggu.Inisiasi embrio somatik dari kalus dilakukanpada medium DF ditambah 2,4-D 1 mg/L dankinetin 0,1 mg/L. Persentase embrio somatikmencapai 80% dari total bobot basah biomassasetelah subkultur keempat. Penambahan kinetin0,5 mg/L dan ABA 0,05 mg/L meningkatkanpematangan embrio somatik kelapa sawit; rata-rata jumlah embrio somatik fase lanjut (torpedodan kotiledon) adalah 16,3 embrio per bejana.Penambahan IBA 2 mg/L dan kinetin 0,5 mg/Lpada medium DF dengan setengah garammakro meningkatkan perkecambahan embriosomatik secara nyata. GA 3 0,1 mg/L mening-katkan jumlah kecambah yang terbentuk.
The development of somatic embryos of sago palm (Metroxylon sagu Rottb.) on solid media *) Perkembangan embrio somatik tanaman sagu (Metroxylon sagu Rottb.) pada medium padat Imron RIYADI; J.S. TAHARDI TAHARDI; . SUMARYONO
Menara Perkebunan Vol. 73 No. 2: 73 (2), 2005
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v73i2.155

Abstract

SummarySago palm (Metroxylon sagu Rottb.) isusually propagated vegetatively by suckers.However, the limited availability of uniformsuckers is a major obstacle in the establishmentof cultivated sago plantations. Tissue culture hasthe potential for large-scale mass clonalpropagation of superior genotypes of sago palm.In vitro culture of sago palm has been establishedthrough somatic embryogenesis. Embryogeniccallus derived from shoot apical tissue of youngsuckers was cultured on a modified Murashigeand Skoog (MMS) medium containing 30 g/Lsucrose, 2 g/L Gelrite, 1 g/L activated charcoal,5.0 mg/L 2,4-D, and 0.1 mg/L kinetin to inducesomatic embryos. Callus clumps formed somaticembryos within four weeks. In the subsequentculture, approximately 0.3 g initial globularcallus grown on MMS medium containing 1.0mg/L kinetin, 0.01 mg/L ABA and 0.1 mg/L GA 3produced 140 to 200 somatic embryos at differentdevelopmental stages four weeks later. All stagesof developing embryos with different sizesand colors were present at any one time ofculture. Secondary (repetitive) somatic embryo-genesis was also found in the culture.Transferring of the mature stage of somaticembryos to solid media with half-strength macro salts and with sucrose at concentration of 20 or 30 g/L without growth regulators led to the development of normal plantlets.RingkasanTanaman sagu (Metroxylon sagu Rottb.)biasanya diperbanyak secara vegetatif dengantunas anakan. Namun, terbatasnya ketersediaantunas anakan yang seragam merupakanhambatan utama dalam pembukaan perkebunansagu. Teknologi kultur jaringan mempunyaipotensi untuk perbanyakan klonal tanaman saguunggul dalam skala besar. Kultur in vitrotanaman sagu telah dikembangkan melaluiembriogenesis somatik. Kalus embriogenik yangberasal dari eksplan pucuk tunas anakandikulturkan pada medium modifikasi Murashigedan Skoog (MMS) dengan sukrosa 30 g/L,Gelrite 2 g/L, arang aktif 1 g/L, 2,4-D 5 mg/Ldan kinetin 0,1 mg/L untuk menginduksi embriosomatik. Kalus membentuk embrio somatikdalam waktu empat minggu. Dalam kulturberikutnya, dari kurang-lebih 0,3 g embrio faseglobuler yang dikulturkan pada medium MMSdengan kinetin 1,0 mg/L, ABA 0,01 mg/L danGA 3 0,1 mg/L menghasilkan 140 sampai 200embrio somatik dengan fase perkembangan yangberbeda-beda. Embrio somatik dalam semuafase perkembangan dengan ukuran dan warnayang berbeda-beda ditemukan setiap saat dalamkultur. Di samping itu, embriogenesis somatiksekunder (berulang) juga terjadi dalam kultursagu. Embrio somatik fase dewasa biladipindah ke medium padat dengan garam makrosetengah konsentrasi dan sukrosa padakonsentrasi 20 atau 30 g/L tanpa zat pengaturtumbuh akan menjadi planlet normal.
Pertumbuhan biak kalus dan suspensi sel tanaman kina (Cinchona ledgeriana Moens) Growth of callus and cell suspension cultures of cinchona (Cinchona ledgeriana Moens) . SUMARYONO; Imron RIYADI
Menara Perkebunan Vol. 73 No. 1: 73 (1), 2005
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v73i1.158

Abstract

SummaryIn vitro technology of plants can be used topropagate plants and to produce secondarymetabolites with a short and continuousproduction cycle. Callus cultures of cinchona(Cinchona ledgeriana Moens) on solid media andcell cultures in liquid media have beenestablished. Callus could be easily initiated fromvarious explants of cinchona clone CB5, GA22and QRC312. The best callus initiation andproliferation were obtained on a Woody Plant(WP) solid medium supplemented with 15 µMpicloram,0.5 µM BAP and 1 µM phloroglucinol.In this medium the fresh weight of callusincreased by 12 to 14-fold within 5 to 6 weeks.Callus that constantly grew fast was selected as amaterial source for cell suspension cultures. InWP liquid medium with the same composition,the cells remained to grow fast where cell volumeafter sedimentation (CVS) increased by almost4-fold in two weeks. However, repeated sub-cultures decreased cell growth rate. The cellsuspension culture was then scaled-up in a 5-Lbioreactor. The culture medium was the same asin Erlenmeyer flasks. Cells in a bioreactor grewvery slowly, the cell biomass fresh weight andpacked cell volume (PCV) increased by 34% and50% respectively after 21 days of culture,although most of the cells remained viable.RingkasanTeknologi in vitro tanaman dapat digunakanuntuk memperbanyak tanaman dan memproduksisenyawa sekunder dengan siklus sangat singkatdan berkelanjutan. Biak kalus tanaman kina(Cinchona ledgeriana Moens) pada mediumpadat dan biak sel di medium cair telahdikembangkan. Kalus dengan mudah dapatdiinduksi dari berbagai jenis eksplan tanamankina klon CB5, GA22 dan QRC312. Inisiasi danproliferasi kalus terbaik diperoleh pada mediaWoody Plant (WP) padat dengan pikloram 15µM, BAP 0,5 µM dan floroglusinol 1 µM. Padamedium ini bobot basah kalus meningkat 12-14kali lipat dalam waktu 5-6 minggu. Kalus yangtetap tumbuh cepat dipilih sebagai sumber bahanuntuk biak suspensi sel. Dalam medium cair WPdengan komposisi yang sama, sel tetap tumbuhdengan pesat, volume sel setelah pengendapan(CVS) meningkat hampir empat kali lipat dalamwaktu dua minggu. Namun subkultur berulangmenurunkan laju pertumbuhan sel. Skala biaksuspensi sel kemudian diperbesar dalam bio-reaktor kapasitas 5 L. Medium kultur yangdigunakan sama dengan medium pada labuErlenmeyer. Pertumbuhan sel dalam bioreaktorsangat lambat, bobot basah sel dan packed cellvolume (PCV) hanya bertambah berturut-turutsebesar 34% dan 50% setelah 21 hari dalamkultur, walaupun sebagian besar sel tetap viabel.
Morphological variations during the development of somatic embryos of tea (Camellia sinensis L.) in vitro Keragaman morfologi selama perkembangan embrio somatik teh (Camellia sinensis L.) in vitro . SUMARYONO; Imron RIYADI; J.S. TAHARDI
Menara Perkebunan Vol. 69 No. 2: 69 (2), 2001
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v69i2.167

Abstract

SummarySomatic embryo culture of tea (Camelliasinensis L.) on an agar-solidified medium consistsof embryos of different sizes, colors anddevelopmental stages. One gram of mostly globularsomatic embryos were cultured on a solidproliferation medium of WP containing 57.1 µMIAA and 4.4 µM BAP to observe theirmorphological variations with respect to embryosize, color, and developmental stage over oneculture passage of 6 weeks. Fresh weight ofsomatic embryos increased slowly during the first4 weeks and then sharply thereafter. At the fourthweek, the number of embryos increasedconsiderably although their weight did not increase,indicating the formation of secondary embryos.The average size of tea somatic embryos did notchange significantly over the culture period,however, the embryo size was already highly variedat the start and increased as the embryo developed.About one half of the embryos were yellowish and the rest were divided equally between the greenishand reddish embryos. At the initial culture, 60% ofthe embryos were at the globular, 30% at heart and10% at torpedo stage. Generally, globular embryosdeveloped into later-stage embryos as the cultureprogressed, however, on this proliferation mediumalmost 80% of the embryos remained at the globularand heart-shaped stages even after the sixth week.If single globular somatic embryos with a particularcolor were cultured on a solid regeneration mediumof WP with 0.47 µM kinetin, 0.69 µM ABA and0.29 µM GA 3 , some of them especially theyellowish embryos underwent color change. Mostof these single globular embryos developedgradually into the later stages. While the initialcolors of embryos affected the rate ofdevelopmental stage changes, yellowish globularembryos tended to develop more rapidly intocotyledonary or germinant stages than the greenishand reddish embryos.RingkasanBiak embrio somatik tanaman teh (Camelliasinensis L.) pada medium padat terdiri dari embriodalam berbagai ukuran, warna dan stadiaperkembangan. Satu gram embrio somatik yangsebagian besar dalam stadia globuler telahdibiakkan pada medium padat proliferasi (mediumWP dengan IAA 57,1 µM dan BAP 4,4 µM) untukmengamati keragaman morfologi embrio dalam halukuran, warna dan stadia perkembangan dalamsatu periode kultur 6 minggu. Berat basah embriosomatik meningkat perlahan pada 4 minggupertama kemudian meningkat dengan tajam. Padaminggu keempat, jumlah embrio melonjakwalaupun beratnya tidak meningkat, hal inimenunjukkan adanya pembentukan embriosekunder. Ukuran rata-rata embrio somatik tidakberubah secara nyata selama periode kultur, tetapiukuran embrio sudah sangat beragam sejak awalkultur dan terus meningkat sejalan denganberkembangnya embrio. Sekitar setengah dariembrio berwarna kuning dan sisanya terdiri dariembrio berwarna hijau dan merah. Pada awalkultur, 60% embrio berada pada stadia globuler,30% stadia bentuk-hati dan 10% stadia bentuk-torpedo. Pada umumnya embrio globulerberkembang ke stadia lebih lanjut sejalan denganwaktu, tetapi pada medium proliferasi ini hampir80% embrio masih dalam stadia globuler danbentuk-hati pada minggu keenam. Apabila embriosomatik globuler tunggal dengan warna tertentudibiakkan pada medium padat regenerasi(WP dengan kinetin 0,47 µM, ABA 0,69 µM danGA 3 0,29 µM, sebagian embrio terutama embriokuning akan mengalami perubahan warna.Sebagian besar embrio globuler tunggal iniberkembang secara bertahap kestadia per-kembangan lebih lanjut. Warna awal embrioberpengaruh terhadap kecepatan perubahan stadiaperkembangan embrio, dengan embrio globulerawal warna kuning cenderung lebih cepatberkembang kestadia kotiledon dan kecambahdibandingkan dengan embrio hijau dan merah. 
Co-Authors . PRIYONO . SISWANTO . Suharyo . SUMARYON . SUMARYONO . UMARYONO Asmini BUDIANI Asmini Budiani Asmini BUDIANI1 Barahima Abbas Daniel Happy Putra Deden Dewantara ERIS Diah Ratnadewi Dwi SUBIYARTI Gale Ginting HAPPY WIDIASTUTI Hayati MINARSIH Hayati Minarsih J S TAHARDI J. S. TAHARDI TAHARDI J.S. TAHARDI J.S. TAHARDI TAHARDI LAKSMI AMBARSARI Masna Maya SINTA Pauline D. KAS Riza Arief PUTRANTO Soekarno Mismana Putra Sri Wahyuni Tatik RAISAWATI Turhadi Turhadi W A DODD