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Kloning dan karakterisasi daerah promoter gen penyandi ADP glucose pyrophosphorylase dari Metroxylon sagu rendemen pati-tinggi dan -rendah [Cloning and characterization of promoter region of ADP glucose pyrophosphorylase-encoding gene from Metroxylon sagu with high- and low-starch content] Asmini BUDIANI; Riza Arief PUTRANTO; Hayati MINARSIH; Imron RIYADI; . SUMARYONO; Barahima ABBAS
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v84i1.200

ADP-glucose pyrophosphorylase (AGPase) is one of the key enzymes in the starch biosynthesis. In many plants, the activity of this enzyme was reported to affect the yield and composition of the produced starch. This research is a part of an effort to develop molecular markers for early selection of high starch-yielding of sago palm. The purpose of the research was to isolate promoters of AGP gene and to analyze the differences in their DNA sequences between sago palm with high starch content (MsHS) and low starch content (MsLS). DNA was isolated and purified from the leaves of the two sago palm. The promoter region of AGP was amplified by Genome Walking technique. The specific primers were designed by Primer3 program based on the information of DNA sequence of AGP genes of sago palm from previous studies. Selected DNA fragments resulted from Genome Walking were isolated from the gel, cloned into E. coli, and analyzed its DNA sequence. DNA sequence analysis showed that one DNA fragment from MsHS (± 1500 bp) and one DNA fragment from MsLS (> 2000 bp) were confirmed as a 5’ upstream of the AGP gene. Further in silico analysis using MEME program identified various DNA motifs of cis-acting elements, which confirmed that those DNA fragment were promoter region of the gene. Preliminary analysis showed the differences in DNA sequences and motives of cis-acting elements in the promoter region of the two samples which might influence or indirectly associated with the character of the starch yield in sago palm.

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
E-Journal Menara Perkebunan Vol 76, No 1: Juni 2008
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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.

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
E-Journal Menara Perkebunan Vol 78, No 1: Juni 2010
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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 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
E-Journal Menara Perkebunan Vol 77, No 2: Desember 2009
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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.

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
E-Journal Menara Perkebunan Vol 81, No 1: Juni 2013
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractAcclimatization of plantlets is a critical stage in themicropropagation of many plants. An experiment wasconducted to determine the effect of pre-condition periodand vessel closure on the growth and survival rate ofrubber (Hevea brasiliensis Muell.Arg.) plantlets derivedfrom in vitro microcutting during acclimatization.Plantlets were planted in plastic pots containing mixedgrowing media after being conditioned in ex vitroenvironment for 0, 3 and 6 days. Five closure vesseltreatments were closed pots placed in opened container,opened pots in closed glass container, closed pots inclosed glass container, opened pots in closed plasticcontainer, and closed pots in closed plastic container.Observation on leaf conditions, rooting frequency, andplant height were conducted at 1.5 months and on thepercentage of survive plantlets at 1.5 and 3 months afteracclimatization. The results showed that pre-conditionwas required to increase survival rate and growth of theplantlets. Pre-condition period of six days gave a highersurvival rate than 0 and 3 days which reached 100% and93% on opened pot in closed plastic container and closedpot in opened container, respectively after 1.5 monthsand was reduced to 80% after three months ofacclimatization. The highest formation of new leaves androots were also obtained on six days pre-conditionperiod. Plantlets with pre-condition for six days and wereplanted on closed pots in an opened container had thebest rooting frequency which was 90%. The resultshowed that the highest survival rate (80%) of rubberplantlets after three months was obtained when theplantlets were pre-conditioned in ex vitro conditions forsix days before acclimatization and planted on openedpots in a closed plastic container or closed pots in anopened container.AbstrakAklimatisasi planlet merupakan tahap kritis dalammikropopagasi tanaman. Penelitian dilakukan untuk me-nentukan pengaruh periode pra-kondisi dan penyung-kupan terhadap pertumbuhan dan daya hidup planletkaret (Hevea brasiliensis Muell.Arg.) asal stek mikro (invitro microcutting) selama aklimatisasi. Planlet ditanampada pot plastik berisi campuran media tanam setelahdikondisikan lebih dahulu pada lingkungan luar selama 0,3 dan 6 hari. Lima perlakuan penyungkupan adalahpenanaman planlet pada pot tertutup diletakkan dalamwadah terbuka, pot terbuka dalam wadah kaca tertutup,pot tertutup dalam wadah kaca tertutup, pot terbukadalam wadah plastik tertutup dan pot tertutup dalamwadah plastik tertutup. Pengamatan keadaan daun, pem-bentukan akar dan tinggi tanaman dilakukan pada 1,5bulan, sedangkan persentase planlet yang hidup diamatipada 1,5 dan 3 bulan setelah aklimatisasi. Hasil penelitianmenunjukkan bahwa periode pra-kondisi diperlukanuntuk meningkatkan daya hidup dan pertumbuhanplanlet. Pra-kondisi selama enam hari memberikan dayahidup planlet lebih tinggi dibandingkan dengan 0 dan 3hari yaitu 100% dan 93% pada perlakuan penanamanpada pot terbuka dalam wadah plastik tertutup dan pottertutup dalam wadah terbuka setelah 1,5 bulan danmenjadi 80% setelah tiga bulan. Penambahan daun barudan pembentukan akar tertinggi juga terdapat padaperlakuan pra-kondisi enam hari. Perlakuan pra-kondisienam hari dengan pot tertutup yang diletakkan dalamwadah terbuka memperlihatkan persentase pembentukanakar yang paling baik yakni 90%. Hasil peneltian me-nunjukkan bahwa daya hidup planlet karet tertinggi(80%) pada umur tiga bulan diperoleh apabila planletdipra-kondisi pada lingkungan ex vitro selama enam hari,kemudian ditanam pada pot terbuka dalam wadah plastiktertutup atau pot tertutup dalam wadah terbuka.

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
E-Journal Menara Perkebunan Vol 74, No 1: Juni 2006
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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.

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
E-Journal Menara Perkebunan Vol 83, No 2: Desember 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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.

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
E-Journal Menara Perkebunan Vol 69, No 2: Desember 2001
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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.

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
E-Journal Menara Perkebunan Vol 73, No 2: Desember 2005
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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.

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
E-Journal Menara Perkebunan Vol 83, No 2: Desember 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

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.

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