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Pengaruh periode pra-kondisi dan penutupan sungkup terhadap daya hidup planlet karet (Hevea brasiliensis Muell. Arg) Effect of pre-condition period and vessel closure on the survival rate of rubber (Hevea brasiliensis Muell. Arg) plantlets Masna Maya SINTA; . NURHAIMI-HARIS; . SUMARYONO
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.v81i1.47

Abstract

Acclimatization of plantlets is a critical stage in the micropropagation of many plants. An experiment was conducted to determine the effect of pre-condition period and vessel closure on the growth and survival rate of rubber (Hevea brasiliensis Muell.Arg.) plantlets derived from in vitro microcutting during acclimatization. Plantlets were planted in plastic pots containing mixed growing media after being conditioned in ex vitro environment for 0, 3 and 6 days. Five closure vessel treatments were closed pots placed in opened container, opened pots in closed glass container, closed pots in closed glass container, opened pots in closed plastic container, and closed pots in closed plastic container. Observation on leaf conditions, rooting frequency, and plant height were conducted at 1.5 months and on the percentage of survive plantlets at 1.5 and 3 months after acclimatization. The results showed that pre-condition was required to increase survival rate and growth of the plantlets. Pre-condition period of six days gave a higher survival rate than 0 and 3 days which reached 100% and 93% on opened pot in closed plastic container and closed pot in opened container, respectively after 1.5 months and was reduced to 80% after three months of acclimatization. The highest formation of new leaves and roots were also obtained on six days pre-condition period. Plantlets with pre-condition for six days and were planted on closed pots in an opened container had the best rooting frequency which was 90%. The result showed that the highest survival rate (80%) of rubber plantlets after three months was obtained when the plantlets were pre-conditioned in ex vitro conditions for six days before acclimatization and planted on opened pots in a closed plastic container or closed pots in an opened container.
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.
Peningkatan laju multiplikasi tunas dan keragaan planlet Stevia rebaudiana pada kultur in vitro Increasing shoot multiplication rate and plantlet vigor of Stevia rebaudiana in vitro culture . SUMARYONO; Masna Maya SINTA
Menara Perkebunan Vol. 79 No. 2: 79 (2), 2011
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v79i2.59

Abstract

AbstractStevia (Stevia rebaudiana Bertoni) is a natural zero-calorie sweetener plant grown in a high population density.Tissue culture technique is useful for rapid mass propagationof plants to provide superior planting materials. Experimentswere conducted to increase growth and multiplication ofshoots and vigor of plantlets of stevia. Explants used wereapical and axillary buds from plantlets grown on MS mediumwithout plant growth regulators. Combinations of BA andIAA at different concentrations were used for shoot growthand multiplication, whereas plant growth retardants(ancymidol and paclobutrazol) and light intensity were usedfor plantlet vigor. The results showed that stevia explantscultured on MS medium without plant growth regulatorsproduced the highest shoots (4.5 cm) with two shoots perexplant. The best multiplication rate of shoots were found onMS medium added with 1.13 mg/L BA combined with0.35 mg/L IAA which produced on average 4.5 shoots and11.9 nodes per initial explant. Ancymidol and paclobutrazolconcentrations affected significantly growth and vigor ofstevia plantlets. Increasing the concentration of ancymidoland paclobutrazol decreased plantlet height and biomassfresh weight, but increased stem diameter. Paclobutrazol at0.1 mg/L was the best treatment to increase the vigor ofstevia plantlets. Light intensity at 20 µmol/m 2 /s gave betterplantlet vigor than other light intensities. It can be concludedthat multiplication of stevia shoots should be grown on MSmedium supplemented with 1.13 mg/L BA + 0.35 mg/L IAAand the vigor of the shoots can be increased by culturing onMS medium containing 0.1 mg/L paclobutrazol underfluorescence lamps with 20 µmol/m 2 /s light intensity.AbstrakStevia (Stevia rebaudiana Bertoni) adalah tanamanpemanis alami nir-kalori yang ditanam dengan kerapatanpopulasi yang sangat tinggi. Teknik kultur jaringan dapatdigunakan untuk perbanyakan tanaman secara massal dancepat untuk menyediakan bahan tanam unggul. Penelitiantelah dilakukan untuk meningkatkan pertumbuhan danmultiplikasi tunas dan keragaan planlet stevia. Eksplan yangdigunakan adalah tunas pucuk dan tunas samping dari planletyang ditumbuhkan pada medium MS tanpa zat pengaturtumbuh. Kombinasi BA dan IAA dengan konsentrasi yangberbeda digunakan untuk pertumbuhan dan multiplikasitunas, sedangkan zat penghambat tumbuh (ansimidol danpaklobutrazol) serta intensitas cahaya digunakan untukkeragaan planlet. Hasil penelitian menunjukkan bahwaeksplan stevia yang ditumbuhkan pada medium MS tanpa zatpengatur tumbuh menghasilkan tunas paling tinggi (4,5 cm)dengan dua tunas per eksplan. Multiplikasi tunas terbaikdiperoleh pada medium dengan BA 1,13 mg/L yangdikombinasikan dengan IAA 0,35 mg/L yang menghasilkan4,5 tunas dan 11,9 ruas per eksplan awal. Konsentrasiansimidol dan paklobutrazol berpengaruh nyata terhadappertumbuhan dan keragaan planlet stevia. Meningkatnyakonsentrasi ansimidol dan paklobutrazol menurunkan tinggiplanlet dan bobot basah biomassa, tetapi meningkatkandiameter batang. Paklobutrazol pada konsentrasi 0,1 mg/Lmerupakan perlakuan terbaik untuk meningkatkan keragaanplanlet stevia. Intensitas cahaya pada 20 µmol/m 2 /detikmemberikan keragaan planlet yang lebih baik dibandingkanintensitas cahaya yang lain. Dapat disimpulkan bahwamultiplikasi tunas stevia sebaiknya dilakukan pada mediumMS ditambah BA 1,13 mg/L + IAA 0,35 mg/L dan keragaanplanlet dapat ditingkatkan dengan menanam planlet padamedium MS ditambah paklobutrazol 0,1 mg/L di bawahlampu fluoresen dengan intensitas cahaya 20 µmol/m 2 /detik.
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.
Perkembangan kalus embriogenik sagu (Metroxylon sagu Rottb.) pada tiga sistem kultur in vitro Development of embryogenic callus of sago (Metroxylon sagu Rottb.) on three systems of in vitro culture Pauline D KASI; . SUMARYONO
Menara Perkebunan Vol. 76 No. 1: 76 (1), 2008
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v76i1.88

Abstract

Summary Embryogenic callus of sago (Metroxylon sagu Rottb.) has been grown on three systems of in vitro culture i.e. agar-solidified medium, liquid medium, and temporary immersion system (TIS) medium to observe and compare the development of embryogenic callus over one passage of six weeks.  A-half gram of embryogenic callus was cultured on a modified MS medium containing 10 mg/L   2,4-D and 0.1 mg/L kinetin. For histological studies, embryogenic callus was fixed in FAA and embedded in paraplast wax. Serial sections were stained with safranin 1% and observed microscopically. By the end of culture period, the development of embryogenic callus in TIS medium was relatively better than those of the other two media.  Fresh weight of callus in liquid medium and TIS increased by 6.5-fold, while on agar-solidified medium increased by 5.4-fold in six weeks.  About 40% of callus in liquid medium and TIS and 20% of callus on agar solidified medium have changed into somatic embryos at globular stage. Histology structure of embryogenic callus of the three systems of in vitro culture shows different pattern. On agar-solidified medium, secondary callus and friable embryogenic callus that consist of meristematic cells were formed. In contrast, more embryogenic cells were formed in liquid medium and TIS to support maturation process to somatic embryos. Therefore, temporary immersion system and liquid medium are recommended for maturation of embryogenic callus, whereas agar-solidified medium is for proliferation of embryogenic callus of sago.  Ringkasan Kalus embriogenik sagu (Metroxylon sagu Rottb.) telah ditumbuhkan pada tiga sistem kultur in vitro yaitu medium padat, medium cair, dan medium dengan sistem perendaman sesaat (SPS) untuk mempelajari dan mem-bandingkan perkembangan dari kalus embrio-genik selama periode enam minggu. Setengah gram kalus embriogenik dikulturkan pada medium MS modifikasi yang mengandung  2,4-D 10 mg/L dan kinetin 0,1 mg/L.  Untuk studi histologi, kalus embriogenik difiksasi dengan FAA dan embedding menggunakan lilin paraplast. Irisan diwarnai dengan safranin 1% dan diamati menggunakan mikroskop. Pada akhir periode kultur, pertumbuhan kalus pada medium dengan SPS lebih baik dibandingkan dengan medium cair dan padat. Bobot basah kalus pada  medium cair dan SPS meningkat 6,5 kali sedangkan pada medium padat meningkat 5,4 kali dalam waktu enam minggu. Sebanyak 40% kalus pada medium cair dan SPS serta 20% kalus pada medium padat berubah menjadi embrio somatik fase globuler. Struktur histologi kalus embriogenik pada ketiga jenis sistem kultur in vitro menunjukkan pola yang berbeda. Pada medium padat terjadi pembentukan kalus sekunder dan kalus embriogenik remah yang terdiri atas sel-sel meristematik. Sebaliknya pada medium cair dan SPS pembentukan sel embriogenik lebih banyak yang menunjang proses pendewasaan menjadi embrio somatik. Oleh karena itu, medium cair dan SPS direkomendasikan untuk pendewasaan kalus embriogenik, sedangkan medium padat untuk proliferasi kalus embriogenik sagu. 
Pengaruh bahan pra-sterilan, tutup tabung kultur, dan musim terhadap tingkat kontaminasi eksplan pada kultur microcutting karet Effect of pre-sterilization agent, culture tube closure, and season on the contamination level of rubber microcutting culture . NURHAIMI-HARIS; . SUMARYONO; M.P. CARRON CARRON
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.v77i2.96

Abstract

AbstractMicrobial contamination is a major obstaclein clonal propagation of hevea (Heveabrasiliensis) through microcutting technology;therefore the ability to reduce contamination willdetermine the success of the application of thistechnology. The aim of experiments was toincrease healthy and survived plantlets by testingpre-sterilization agents for cleaning explantsduring pre-sterilization step, culture tubeclosures suitable for explants growth and anappropriate time for introducing explants at theprimary culture phase. The pre-sterilizationagents tested were aganol, alcohol anddesogerme, the culture tube closures used wereparafilm and cotton, and the time for culturingexplants were determined by using rubbergenotypes introduced during the year of 2006 and2007. The results show that desogermedecreased significantly the level of explantcontamination compared to aganol and alcohol,meanwhile the type of culture tube closure didnot affect the level of explant contamination. Thetype of culture tube closure influencedsignificantly the survival of explants where thenumber of survived explants in culture tubescovered with cotton was higher than that of withparafilm. Season also affected the contaminationfrequency of the explants. Higher number ofhealthy plantlets were obtained whenintroduction of the explants were conducted fromJune to October considered as dry season inBogor compared to introduction of the explantsduring rainy season from January to May.Different genotypes of rubber introduced at theprimary culture phase did not affect thepercentage of explant contamination.AbstrakKontaminasi oleh mikroba merupakanmasalah utama pada perbanyakan klonal tanamankaret (Hevea brasiliensis) melalui teknologimicrocutting sehingga kemampuan mengurangikontaminasi menentukan keberhasilan aplikasiteknologi tersebut. Penelitian ini bertujuanmempelajari pengaruh jenis bahan pra-sterilanyang efektif untuk pencucian eksplan tahap pra-sterilisasi, mempelajari pengaruh tutup tabungterhadap perkembangan eksplan serta meng-identifikasi waktu yang tepat untuk melaksanakanintroduksi eksplan pada tahap kultur primer(kultur awal) sehingga jumlah eksplan sehat dantumbuh dapat ditingkatkan. Bahan pra- sterilanyang diuji adalah aganol, alkohol dan desogerme,tutup tabung yang digunakan adalah parafilm dankapas, sedangkan identifikasi waktu kulturdilakukan melalui introduksi eksplan sepanjang tahun 2006 dan 2007 terhadap berbagai genotipetanaman karet yang tersedia. Hasil penelitianmenunjukkan bahwa desogerme menurunkansecara nyata tingkat kontaminasi eksplandibandingkan dengan aganol dan alkohol,sedangkan jenis tutup tabung tidak berpengaruhterhadap persentase kontaminasi. Jenis tutuptabung berpengaruh sangat nyata terhadappersentase eksplan yang hidup dan membentuktunas, di mana persentase eksplan membentuktunas pada tabung dengan tutup kapas lebih tinggidibandingkan dengan tutup parafilm. Musim jugasangat mempengaruhi tingkat kontaminasieksplan. Eksplan sehat jauh lebih banyakdiperoleh apabila penanaman eksplan dilakukanpada bulan Juni sampai Oktober, yang merupakanmusim kemarau di Bogor dibandingkan denganintroduksi eksplan pada bulan Januari sampaiMei, yang merupakan musim hujan. Jenisgenotipe yang ditanam pada tahap kultur primertidak berpengaruh terhadap persentasekontaminasi.
Keragaman morfologi selama perkembangan embrio somatik sagu (Metroxylon sagu Rottb.) Morphological variations during the development of somatic embryos of sago (Metroxylon sagu Rottb.) Pauline Destinugrainy KASI; . SUMARYONO
Menara Perkebunan Vol. 74 No. 1: 74 (1), 2006
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v74i1.120

Abstract

Summary In vitro culture of sago (Metroxylon sagu Rottb.) on an agar-solidified medium consists of somatic embryos of different sizes, colors, and developmental stages.  One gram of mostly globular somatic embryos were cultured on a solid medium to observe their morphological variations with respect to embryo size, color, and developmental stage over one passage of six weeks culture.  The medium was a modified-MS medium with half-strength of macronutrients containing   0.01 mg/L ABA and 2 mg/L kinetin.  At the end of culture passage, fresh weight of embryo increased by 2.3 folds.  The embryo numbers increased by more than two times indicating the formation of secondary embryos.  The average size of sago somatic embryos did not change significantly over the culture period; however, the embryo size was already highly varied at the start and increased gradually as the embryo developed.  At the initial of culture,   33.7 % of the embryos were yellowish, 64.1 % were greenish, and 2.2% were reddish.  By the end of the culture the composition of yellowish embryos increased to 51.2 %, greenish embryo decreased to 42.5 % and red embryos increased to 6.3 %.  At the initial culture, 61 % of the embryos were at the globular, 9 % at heart-shape and 30 % at torpedo stage.  Generally globular embryos developed into later-stage embryos as the culture progressed, although almost 56% of the embryos remained at the globular stage after the sixth week.Ringkasan Kultur in vitro sagu (Metroxylon sagu Rottb.) pada medium padat terdiri dari embrio somatik dalam berbagai ukuran, warna, dan fase perkembangan.  Satu gram embrio somatik yang sebagian besar dalam fase globuler dikulturkan pada medium padat untuk mengamati keragaman morfologi embrio dalam hal ukuran, warna dan fase perkembangan dalam satu periode kultur enam minggu.  Medium kultur adalah MS modifikasi dengan setengah hara makro serta penambahan zat pengatur tumbuh ABA 0,01 mg/L dan kinetin 2 mg/L.  Pada akhir masa kultur bobot embrio segar meningkat 2,3 kali dibandingkan awal masa kultur.  Jumlah embrio juga mengalami peningkatan sebesar lebih dari dua kali yang menunjukkan adanya pembentukan embrio somatik sekunder. Ukuran rata-rata embrio tidak berubah secara signifikan selama masa kultur akan tetapi ukuran embrio telah sangat beragam pada awal kultur dan terus meningkat hingga akhir kultur. Warna embrio mengalami perubahan selama periode kultur.  Pada awal kultur dijumpai 33,7 % embrio berwarna kuning, 64,1 % embrio hijau, dan 2,2 % embrio merah.  Pada akhir kultur presentase embrio kuning meningkat menjadi 51,2 %, embrio hijau menjadi 42,5 %, dan embrio merah 6,3 %.  Pada awal kultur, dijumpai 61 % embrio pada fase globuler, 9 % fase bentuk-hati dan 30 % fase torpedo.  Umumnya embrio globuler berkembang menjadi embrio fase lanjut selama kultur berlangsung, namun 56 % embrio masih tetap dalam fase globuler pada minggu keenam.
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.