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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
E-Journal Menara Perkebunan Vol 79, No 2: Desember 2011
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (439.221 KB) | 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.
Identifikasi dan pencegahan kontaminasi pada kultur cair sistem perendaman sesaat Identification and prevention of contamination in liquid culture of temporary immersion system Masna Maya SINTA; Imron RIYADI; . SUMARYONO
E-Journal Menara Perkebunan Vol 82, No 2: Desember 2014
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (230.566 KB) | DOI: 10.22302/iribb.jur.mp.v82i2.21

Abstract

AbstractLiquid culture is commonly used to scale up in vitro culture production as well as to optimize the developmental phase of plant in vitro culture. One of the liquid cultures that has been used widely is temporary immersion system (TIS). The main problem of liquid culture is contamination. The use of antibiotics sometimes controls the contaminants less effectively and hinders the growth of plant culture. The purpose of this research was to determine sources of contaminant on whole sequence of TIS to identify and to prevent the emergence of the contaminants. Sampling method was applied to each section and stage of TIS culture and the contaminants found were identified. The results revealed that compartment of TIS was the main source of contaminant (100%). Furthermore, from all components of TIS compartment, washer (a small ring seal connecting screen disc and basket) was the main source of TIS contaminant (41.2%). Four contaminants found were identified as Bacillus macerans, Bacillus megaterium, Bacillus sphaericus and Bacillus firmus. Two times sterilization of washer in an autoclave at temperature of 121 oC and air pressure of 1 kg/cm2 for 20 minutes before and after being installed reduced the contamination level on TIS culture significantly.AbstrakKultur cair umumnya digunakan untuk meningkatkan skala produksi dan mengoptimalkan fase perkembangan kultur in vitro tanaman. Salah satu jenis kultur cair yang banyak digunakan adalah sistem perendaman sesaat (SPS). Masalah utama dalam kultur cair adalah kontaminasi. Penggunaan antibiotika terkadang kurang efektif dalam me-ngendalikan kontaminan dan menghambat pertumbuhan kultur tanaman. Tujuan dari penelitian ini adalah untuk mengetahui sumber kontaminan pada seluruh rangkaian kultur SPS serta mengidentifikasi dan mencegah munculnya kontaminan tersebut. Metode yang digunakan adalah  pengambilan contoh pada tiap bagian dan fase kultur SPS, serta kontaminan yang ditemukan kemudian diidentifikasi. Hasil penelitian memperlihatkan bahwa kompartemen SPS merupakan sumber utama kontaminan (100%). Selanjutnya, dari seluruh komponen kompartemen SPS, washer (cincin penutup yang menghubungkan penyaring dan keranjang) di dalam rangkaian SPS merupakan sumber utama kontaminan (41,2%).  Empat  kontaminan yang ditemukan diidentifikasi sebagai Bacillus macerans, Bacillus megaterium, Bacillus sphaericus dan Bacillus firmus. Sterilisasi cincin penutup sebanyak  dua  kali  dalam  autoklaf pada suhu 121 oC dan tekanan udara 1 kg/cm2selama 20 menit sebelum dan sesudah dirangkai secara nyata menurunkan tingkat konta-minasi pada kultur SPS. 
Daya hidup planlet karet asal in vitro microcutting pada berbagai periode penutupan sungkup plastik dan komposisi media tumbuh Survival rate of in vitro microcutting-derived rubber plantlets on various plastic cover closed periods and medium compositions . SUMARYONO; Masna Maya SINTA; . NURHAIMI-HARIS
E-Journal Menara Perkebunan Vol 80, No 1: Juni 2012
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (319.119 KB) | DOI: 10.22302/iribb.jur.mp.v80i1.46

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AbstractIn vitro culture through microcutting technology can be used for clonal propagation of rubber (Hevea brasiliensis Muell. Arg.) rootstocks. Acclimatization of in vitro plantlets to ex vitro conditions is a major bottleneck in the micropropagation of many plants.This research was conducted to study the effect of plastic cover closed period and media composition on the survival rate of rubber plantlets. Plantlets derived from microcutting were planted on plastic pots containing a mixture of soil, cocopeat, dung manure, and sand or zeolite. The plantlets were then placed inside a closed transparent plastic cover that opened after 2, 3, 4 and 6 weeks. The cover was placed under tree canopy. The second experiment used the same media composition with or without cocopeat and with sand or zeolite. At 1.5 month after culture, observation was done on the number of survived plantlets, plantlet height and the percentage of rooted plantlets. The results show that the best coverclosed period was six weeks and the best growing medium was a mixture of soil, cocopeat, dung manure, and zeolite (6:2:1:1v/v). On the two combined treatments, the survival rate was 73.3% after 1.5 month of acclimatization. The use of zeolite and a higher soil percentage gave positive influences on rubber plantlet survival rate. The second experiment results confirmed that the use of zeolite was better than sand and the use of cocopeat was definitely needed. It can be concluded that the best of acclimatization of rubber plantlets from microcutting was on a medium mixture of soil, cocopeat, dung manure, and zeolite (6:2:1:1) and placed inside a closed plastic cover for six weeks before the cover was opened gradually. AbstrakKultur in vitro melalui teknologi microcutting dapat digunakan untuk perbanyakan klonal batang bawah tanaman karet (Hevea brasiliensis Muell. Arg.). Aklimatisasi planlet in vitro ke kondisi ex vitro merupakan hambatan utama pada mikropropagasi berbagai jenis tanaman. Penelitian ini dilakukan untuk mempelajari pengaruh lama penutupan sungkup plastik dan komposisi media tumbuh terhadap daya hidup planlet karet. Planlet karet asal microcutting ditanam pada pot plastik berisi media dengan berbagai campuran tanah, cocopeat, pupuk kandang, dan pasir atau zeolit. Planlet selanjutnya diletakkan di dalam sungkup plastik transparan tertutup rapat yang dibuka setelah 2, 3, 4 dan 6 minggu. Sungkup plastik diletakkan di bawah tajuk pepohonan. Percobaan kedua menggunakan komposisi media serupa dengan atau tanpa cocopeat dan dengan pasir atau zeolit. Pada umur 1,5 bulan, pengamatan dilakukan terhadap jumlah planlet yang hidup, tinggi planlet, dan persentase planlet yang berakar. Hasil penelitian menunjukkan bahwa lama penyungkupan terbaik adalah enam minggu dan media tumbuh terbaik adalah campuran tanah, cocopeat, pupuk kandang, dan zeolit (6:2:1:1 v/v). Pada kombinasi kedua perlakuan tersebut, daya hidup planlet karet mencapai 73,3% setelah 1,5 bulan aklimatisasi. Penggunaan zeolit dan persentase tanah yang lebih tinggi berpengaruh positif terhadap daya hidup planlet karet. Hasil percobaan kedua menegaskan bahwa penggunaan zeolit lebih baik daripada pasir dan penggunaan cocopeat mutlak diperlukan. Dapat disimpulkan bahwa aklimatisasi planlet karet asal microcutting terbaik dilakukan pada media campuran tanah, cocopeat, pupuk kandang, zeolit (6:2:1:1) dan diletakkan di dalam sungkup plastik tertutup selama enam minggu sebelum sungkup dibuka secara bertahap.
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
E-Journal Menara Perkebunan Vol 75, No 1: Juni 2007
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (231.615 KB) | DOI: 10.22302/iribb.jur.mp.v75i1.152

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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (448.502 KB) | DOI: 10.22302/iribb.jur.mp.v81i1.53

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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.
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
E-Journal Menara Perkebunan Vol 73, No 1: Juni 2005
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (819.279 KB) | DOI: 10.22302/iribb.jur.mp.v73i1.158

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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.
Pengaruh matriks kapsul terhadap perkecambahan benih sintetik teh (Camellia sinensis L.) Effect of capsule matrix on germination of synthetic seeds of tea (Camellia sinensis L.) . SUMARYONO; Rizka T SAPTARI
Menara Perkebunan Vol. 83 No. 2: 83 (2), 2015
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v83i2.1

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Abstract         Synthetic seed technology has been developed to combine the practical use of ordinary seeds and clonal plant materials. Single somatic embryos of tea (Camellia sinensis L.) clone GMB 9 were encap-sulated using sodium alginate and CaCl2 at different concentrations to produce synthetic seeds.  Encapsulating matrices with and without somatic embryos were made of WP medium and 20 g/L sucrose with   1, 2 or 3%  sodium alginate and 50 or 100 mM CaCl2. Encapsulating matrices without somatic embryos were then tested its physical characteristics, whereas the capsules with somatic embryos were observed its germination rate and secondary embryo formation every week up to six weeks. The results showed that the concentrations of sodium alginate and CaCl2 were affected significantly the physical characteristics of encap-sulating matrix produced. The sufficient level of hardness and highest germination rate was obtained from 2% sodium alginate and 50 mM CaCl2. Sodium alginate at 1% or less produced soft, leaky and oval encapsulating matrices which were not suitable for synthetic seeds. Sodium alginate 3% and 100 mM CaCl2  produced rounded and very hard encapsulating matrices and inhibited the germination of somatic embryos. Germination rates of tea synthetic seeds and somatic embryos without encapsulation were 5 to 20% after six weeks. Abstrak         Teknologi benih sintetik dikembangkan untuk memadukan kepraktisan penggunaan benih biasa dengan bahan tanam klonal. Embrio somatik tunggal dari tanaman teh (Camellia sinensis L.) klon GMB 9 dienkapsulasi menggunakan natrium alginat   dan CaCl2 untuk membuat benih sintetik. Matriks kapsul dengan dan tanpa embrio somatik dibuat dari medium WP dengan sukrosa 20 g/L dicampur dengan natrium alginat 1, 2, atau 3% dan CaCl2 50  atau 100 mM.  Matriks  kapsul  tanpa embrio somatik kemudian diuji sifat fisiknya, sedangkan matriks kapsul dengan embrio somatik diamati daya kecambah dan pembentukan embrio somatik sekunder setiap minggu sampai dengan enam minggu. Hasil penelitian menunjukkan bahwa konsentrasi natrium alginat dan CaCl2 sangat berpengaruh terhadap sifat fisik matriks kapsul yang dihasilkan. Tingkat kekerasan yang memadai dan menghasilkan persentase perkecambahan yang tinggi diperoleh dengan alginat 2% dan CaCl2 50 mM. Natrium alginat pada konsentrasi 1% atau kurang menghasilkan matriks kapsul yang  lunak, mudah bocor dan berbentuk lonjong sehingga tidak sesuai untuk benih sintetik. Natrium alginat konsentrasi 3% pada larutan CaCl2 100 mM menghasilkan benih sintetik yang bulat, sangat keras dan menghambat perkecambahan embrio somatik teh.  Daya kecambah benih sintetik dan embrio somatik teh tanpa enkapsulasi berkisar  5 - 20% setelah enam minggu. 
Ekspresi dan kloning gen penyandi ADP-Glucose Phyrophosphorylase dari tanaman sagu (Metroxylon sagu Rottb.) Expression and cloning of gene encoding ADP-Glucose Phyrophosphorylase from sago palm (Metroxylon sagu Rottb.) Asmini BUDIANI1; Riza Arief PUTRANTO; Hayati MINARSIH; Imron RIYADI; . SUMARYONO; Barahima ABBAS
Menara Perkebunan Vol. 83 No. 2: 83 (2), 2015
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v83i2.4

Abstract

AbstractSago palm (Metroxylon sagu Rottb.) is a potential food and energy resources becouse it is the highest starch producing plant.  Breeding of sago palm should be directed to produce elite genotype with superior characters such as high starch content, wider pith diameter, without spine and high starch quality. However, research on sago palm in Indonesia so far is limited espescially in the field of cultivation and breeding, and attempt to produce such elite would take long time. Availability of molecular marker for starch content would be beneficial to shorten the length period of breeding. ADP-Glucose Phyrophosphorylase is one of the important enzymes in starch biosynthesis. Therefore its gene is an interesting subject in order to develope molecular marker of high starch content.  This research was aimed to study the expression of gene encoding AGP in the sago palm with high starch content versus low starch content, and to clone the full cds of the gene. RNA was isolated from leaf and pith of both palms. Exspression analysis and amplify-cation of full cds were conducted by Reverse Transcryptase-Polymerase Chain Reaction (RT-PCR) using specific primers. The results showed that sago palm with higher starch content expressed AGP higher than that of sago palm with lower  starch content. Expression of AGP in the full developing leaf was higher than in the young leaf, and there was no expression detected in the pith. The full cds of AGP was successfully amplified and cloned. Even though the DNA sequence showed high homology with DNA sequence of the same gene that has been deposited in GenBank, there were differences in severall nucleotide including that in the active domain of the enzyme.AbstrakTanaman sagu merupakan sumber pangan dan energi yang sangat potensial untuk dikembangkan karena merupakan tanaman penghasil karbihidrat tertinggi. Pemuliaan tanaman sagu mestinya diarah-kan untuk menghasilkan bibit sagu yang selain memiliki rendemen pati tinggi, juga memiliki diameter empulur besar, tidak berduri dan memiliki cita rasa pati yang enak. Namun, sampai saat ini riset mengenai sagu di Indonesia masih sangat terbatas, sehingga pemuliaan sagu untuk menghasilkan bibit unggul demikian akan memerlukan waktu lama. Ketersediaan penanda rendemen pati akan sangat membantu mempercepat pemuliaan tanaman tersebut. ADP-Glucose Pyrophosphorylase adalah salah satu enzim yang berperan penting dalam biosintesis pati, sehingga gene penyandinya merupakan subjek yang menarik dalam pengembangan marka kandungan pati tinggi.  Sebagai bagian dari upaya untuk mendapat-kan penanda rendemen pati tinggi pada tanaman sagu, penelitian ini bertujuan untuk mempelajari ekspresi gen penyandi AGP. RNA diisolasi dari daun tanaman sagu rendemen pati rendah dan tanaman sagu rendemen pati tinggi. Perbedaan tingkat ekspresi gen penyandi AGP dari tanaman sagu rendemen pati tinggi vs rendemen pati rendah, dianalisis dengan teknik Reverse-Transcryptase PCR menggunakan primer spesifik. Hasil penelitian menunjukkan bahwa tanaman sagu rendemen pati tinggi mengekspresikan AGP lebih tinggi dibandingkan dengan tanaman sagu rendemen pati rendah. Ekspresi gen tersebut pada daun tua (full developing leaf) lebih tinggi di-bandingkan dengan pada daun muda, dan pada empulur tidak dideteksi ekspresi gen tersebut. Daerah penyandi lengkap AGP subunit kecil telah diklon. Meskipun memiliki homologi yang tinggi dengan sekuen DNA gen yang sama yang telah dideposit pada  GenBank,  namun terdapat perbedaan beberapa nukleotida termasuk pada daerah domain aktif dari enzim tersebut. 
Identifikasi dan pencegahan kontaminasi pada kultur cair sistem perendaman sesaat Identification and prevention of contamination in liquid culture of temporary immersion system Masna Maya SINTA; Imron RIYADI; . SUMARYONO
Menara Perkebunan Vol. 82 No. 2: 82 (2), 2014
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v82i2.21

Abstract

AbstractLiquid culture is commonly used to scale up in vitro culture production as well as to optimize the developmental phase of plant in vitro culture. One of the liquid cultures that has been used widely is temporary immersion system (TIS). The main problem of liquid culture is contamination. The use of antibiotics sometimes controls the contaminants less effectively and hinders the growth of plant culture. The purpose of this research was to determine sources of contaminant on whole sequence of TIS to identify and to prevent the emergence of the contaminants. Sampling method was applied to each section and stage of TIS culture and the contaminants found were identified. The results revealed that compartment of TIS was the main source of contaminant (100%). Furthermore, from all components of TIS compartment, washer (a small ring seal connecting screen disc and basket) was the main source of TIS contaminant (41.2%). Four contaminants found were identified as Bacillus macerans, Bacillus megaterium, Bacillus sphaericus and Bacillus firmus. Two times sterilization of washer in an autoclave at temperature of 121 oC and air pressure of 1 kg/cm2 for 20 minutes before and after being installed reduced the contamination level on TIS culture significantly.AbstrakKultur cair umumnya digunakan untuk meningkatkan skala produksi dan mengoptimalkan fase perkembangan kultur in vitro tanaman. Salah satu jenis kultur cair yang banyak digunakan adalah sistem perendaman sesaat (SPS). Masalah utama dalam kultur cair adalah kontaminasi. Penggunaan antibiotika terkadang kurang efektif dalam me-ngendalikan kontaminan dan menghambat pertumbuhan kultur tanaman. Tujuan dari penelitian ini adalah untuk mengetahui sumber kontaminan pada seluruh rangkaian kultur SPS serta mengidentifikasi dan mencegah munculnya kontaminan tersebut. Metode yang digunakan adalah  pengambilan contoh pada tiap bagian dan fase kultur SPS, serta kontaminan yang ditemukan kemudian diidentifikasi. Hasil penelitian memperlihatkan bahwa kompartemen SPS merupakan sumber utama kontaminan (100%). Selanjutnya, dari seluruh komponen kompartemen SPS, washer (cincin penutup yang menghubungkan penyaring dan keranjang) di dalam rangkaian SPS merupakan sumber utama kontaminan (41,2%).  Empat  kontaminan yang ditemukan diidentifikasi sebagai Bacillus macerans, Bacillus megaterium, Bacillus sphaericus dan Bacillus firmus. Sterilisasi cincin penutup sebanyak  dua  kali  dalam  autoklaf pada suhu 121 oC dan tekanan udara 1 kg/cm2selama 20 menit sebelum dan sesudah dirangkai secara nyata menurunkan tingkat konta-minasi pada kultur SPS. 
Daya hidup planlet karet asal in vitro microcutting pada berbagai periode penutupan sungkup plastik dan komposisi media tumbuh Survival rate of in vitro microcutting-derived rubber plantlets on various plastic cover closed periods and medium compositions . SUMARYONO; Masna Maya SINTA; . NURHAIMI-HARIS
Menara Perkebunan Vol. 80 No. 1: 80 (1), 2012
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v80i1.46

Abstract

AbstractIn vitro culture through microcutting technology can be used for clonal propagation of rubber (Hevea brasiliensis Muell. Arg.) rootstocks. Acclimatization of in vitro plantlets to ex vitro conditions is a major bottleneck in the micropropagation of many plants.This research was conducted to study the effect of plastic cover closed period and media composition on the survival rate of rubber plantlets. Plantlets derived from microcutting were planted on plastic pots containing a mixture of soil, cocopeat, dung manure, and sand or zeolite. The plantlets were then placed inside a closed transparent plastic cover that opened after 2, 3, 4 and 6 weeks. The cover was placed under tree canopy. The second experiment used the same media composition with or without cocopeat and with sand or zeolite. At 1.5 month after culture, observation was done on the number of survived plantlets, plantlet height and the percentage of rooted plantlets. The results show that the best coverclosed period was six weeks and the best growing medium was a mixture of soil, cocopeat, dung manure, and zeolite (6:2:1:1v/v). On the two combined treatments, the survival rate was 73.3% after 1.5 month of acclimatization. The use of zeolite and a higher soil percentage gave positive influences on rubber plantlet survival rate. The second experiment results confirmed that the use of zeolite was better than sand and the use of cocopeat was definitely needed. It can be concluded that the best of acclimatization of rubber plantlets from microcutting was on a medium mixture of soil, cocopeat, dung manure, and zeolite (6:2:1:1) and placed inside a closed plastic cover for six weeks before the cover was opened gradually. AbstrakKultur in vitro melalui teknologi microcutting dapat digunakan untuk perbanyakan klonal batang bawah tanaman karet (Hevea brasiliensis Muell. Arg.). Aklimatisasi planlet in vitro ke kondisi ex vitro merupakan hambatan utama pada mikropropagasi berbagai jenis tanaman. Penelitian ini dilakukan untuk mempelajari pengaruh lama penutupan sungkup plastik dan komposisi media tumbuh terhadap daya hidup planlet karet. Planlet karet asal microcutting ditanam pada pot plastik berisi media dengan berbagai campuran tanah, cocopeat, pupuk kandang, dan pasir atau zeolit. Planlet selanjutnya diletakkan di dalam sungkup plastik transparan tertutup rapat yang dibuka setelah 2, 3, 4 dan 6 minggu. Sungkup plastik diletakkan di bawah tajuk pepohonan. Percobaan kedua menggunakan komposisi media serupa dengan atau tanpa cocopeat dan dengan pasir atau zeolit. Pada umur 1,5 bulan, pengamatan dilakukan terhadap jumlah planlet yang hidup, tinggi planlet, dan persentase planlet yang berakar. Hasil penelitian menunjukkan bahwa lama penyungkupan terbaik adalah enam minggu dan media tumbuh terbaik adalah campuran tanah, cocopeat, pupuk kandang, dan zeolit (6:2:1:1 v/v). Pada kombinasi kedua perlakuan tersebut, daya hidup planlet karet mencapai 73,3% setelah 1,5 bulan aklimatisasi. Penggunaan zeolit dan persentase tanah yang lebih tinggi berpengaruh positif terhadap daya hidup planlet karet. Hasil percobaan kedua menegaskan bahwa penggunaan zeolit lebih baik daripada pasir dan penggunaan cocopeat mutlak diperlukan. Dapat disimpulkan bahwa aklimatisasi planlet karet asal microcutting terbaik dilakukan pada media campuran tanah, cocopeat, pupuk kandang, zeolit (6:2:1:1) dan diletakkan di dalam sungkup plastik tertutup selama enam minggu sebelum sungkup dibuka secara bertahap.