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Perbanyakan in vitro tanaman kina (Cinchona ledgeriana Moens) melalui tunas aksiler dan apikal In vitro propagation of cinchona (Cinchona ledgeriana Moens) from axillary and apical buds Imron RIYADI; J. S. TAHARDI TAHARDI
E-Journal Menara Perkebunan Vol 77, No 1: Juni 2009
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractThe use of the appropriate source of nodalbud explants in culture can increase theeffectiveness and efficiency of shootmultiplication. An experiment was conducted todetermine and compare the rate of in vitro shootmultiplication from apical and axillary budsin cinchona (Cinchona ledgeriana Moens) andtheir subsequent growth and development. Theplant material used was Cinchona ledgerianaoriginating from the Indonesian Tea andCinchona Research Institute, Gambung, WestJava. Explants were taken from apical andaxillary nodes from in vitro germinated seedlings.The cultures were incubated at 26 0 C and 60%relative humidity under a 14-h photoperiod withlight intensity of 30 µmol photon/m 2 /sec.provided by cool-white fluorescent tubes (TL40 W) for 4 - 8 weeks. The parameters observedwere shoot multiplication rate, shoot growth anddevelopment such as shoot length, leaf numberand rooting frequency. Apical and axillary nodesproduced shoots at different multiplication rateson Murashige-Skoog (MS) standard mediumcontaining 30 g/L sucrose and supplemented with1 – 5 mg/L BA in combination with 0.1 mg/L IBA.Furthermore, shoots or plantlets of cinchonagrew and developed on the same mediacontaining 5 – 10 mg/L IAA combined with0.5 mg/L IBA. The results showed that shootmultiplication rate was higher in axillary than inapical nodes. The highest multiplication rate inaxillary nodes was 24.6 shootlets with 3 mg/LBA treatment, whereas in apical nodes it was17.2 shootlets with 5 mg/L BA treatment for eightweeks. The highest rooting frequency ofcinchona plantlet was 90%, achieved with 5 mg/LIAA in combination with 0.5 mg/L IBA. Theplantlets were successfully acclimatized andtransplanted to the fieldAbstrakSumber eksplan berupa nodus/tunas padakultur in vitro umum digunakan untuk multi-plikasi tunas. Penelitian ini bertujuan untukmembandingkan tingkat multiplikasi antara tunasapikal dengan tunas aksiler tanaman kina Ledgersecara in vitro. Bahan tanaman yang digunakanadalah kina Ledger (Cinchona ledgeriana Moens)yang berasal dari Pusat Penelitian Teh dan Kina,Gambung, Jawa Barat. Eksplan berupa nodus/tunas apikal dan aksiler asal biji yang dikecam-bahkan secara in vitro. Kultur tersebut diinku-basikan dalam ruang terang pada intensitascahaya 30 μmol foton/m 2 /detik dengan periodepenyinaran 14 jam pada suhu 260 C dankelembaban relatif + 60% selama 4 – 8 minggu.Parameter yang diamati adalah perbandinganmultiplikasi tunas dan pertumbuhan tunas yangmeliputi rata-rata tinggi tunas, jumlah daun danfrekuensi pengakaran. Nodus apikal maupunaksiler menghasilkan tunas dengan tingkatMurashige-Skoog (MS) standar yang me-ngandung sukrosa30 g/L dan ditambahkan BA1 – 5 mg/L dikombinasikan IBA 0,1 mg/L.Selanjutnya tunas/planlet kina tersebut berhasiltumbuhdan berkembang pada medium sama yangdiberi IAA 5 – 10 mg/L dikombinasikan denganIBA 0,5 mg/L. Hasil penelitian menunjukkanbahwa tingkat multiplikasi tunas aksiler lebihtinggi dari pada tunas apikal. Multiplikasi tunasaksiler menghasilkan jumlah tunas rata-ratatertinggi sebesar 24,6 tunas per eksplan padaperlakuan BA 3 mg/L sedangkan multiplikasitunas apikal tertinggi sebesar 17,2 tunas pereksplan pada perlakuan BA 5 mg/L pada umurdelapan minggu. Frekuensi pengakaran planletkina tertinggi mencapai 90% pada perlakuan IAA10 mg/L yang dikombinasikan dengan IBA 0,5mg/L. Planlet yang dihasilkan telah berhasildiaklimatisasi dan dipindahkan ke tempatpersemaian lapang.

Evaluasi varietas, sumber eksplan dan strain Agrobacterium terhadap keberhasilan transformasi tebu dengan gen P5CS Evaluation of varieties, explant sources, and Agrobacterium strains for successful sugarcane transformation using P5CS gene Hayati MINARSIH; Dwi SUBIYARTI; Imron RIYADI; Soekarno Mismana PUTRA; Laksmi AMBARSARI
E-Journal Menara Perkebunan Vol 83, No 1: Juni 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Abstract Genetic transformation can be used as an alter-native to develop sugarcane (Saccharum officinarum L.) tolerant to drought stress. P5CS gene has a role in biosynthesis of proline, an amino acid that accumulated under drought stress conditions. Transfer of a P5CS gene construct into plant cells in conjunction with regeneration of transgenic plantlets may develop sugarcane tolerant to drought stress. The aim of this research was to obtain an optimal transformation method which includes a suitable strain of Agrobacterium tumefaciens, and the best sugarcane explant and variety. The results showed that transfer of P5CS gene has been successfully carried out on sugarcane explants from solid media-derived calli, embryogenic calli and somatic embryos derived from temporary immersion system (TIS) culture. Whilst Agrobacterium strain LBA4404 was indicated as the most effective transformation vector. The regeneration of Kidang Kencana variety transformants from calli and somatic embryos was better than those of PS 881 and PS 891. The best performance of transformants based on the source of explants obtained from somatic embryos from TIS culture. Moreover, a succesfull Agrobacterium mediated transformation on sugarcane was indicated by transient expression of Gus gene and the ability of the transformants grew in a selection medium containing 50 ppm of kanamycin.Abstrak Transformasi genetik dapat digunakan sebagai upaya untuk merakit tebu (Saccharum officinarum L.) toleran terhadap cekaman kekeringan. Gen P5CS diketahui berperan dalam biosintesis prolin, yaitu asam amino yang umumnya terakumulasi ketika tanaman mengalami cekaman kekeringan. Transfor-masi gen P5CS dan regenerasi transgeniknya mungkin dapat menghasilkan tanaman tebu trans-genik yang toleran terhadap cekaman kekeringan. Tujuan penelitian ini adalah untuk mendapatkan metode transformasi yang optimum yang mencakup strain Agrobacterium tumefaciens yang sesuai, sumber eksplan dan varietas tebu terbaik sebagai target transformasi. Hasil penelitian menunjukkan bahwa transformasi gen P5CS telah berhasil dilakukan ke eksplan tebu baik yang berupa kalus asal media padat maupun kalus embriogenik dan embrio somatik asal kultur sistem perendaman sesaat (SPS). Sementara itu strain A. tumefaciens LBA4404 menunjukkan hasil yang paling efektif sebagai vektor transformasi. Pertumbuhan transforman baik pada kalus maupun embrio somatik pada varietas Kidang Kencana terlihat paling baik dibandingkan dengan varietas PS 881 dan PS 891. Sumber eksplan yang paling efektif adalah embrio somatik yang diperoleh dari kultur SPS. Keberhasilan transformasi tebu me-lalui Agrobacterium ditunjukkan oleh ekspresi transien dari gen GUS dan kemampuan dari trans-forman untuk tumbuh di media yang mengandung 50 ppm kanamisin.

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.

Physiological responses and P5CS gene expression of transgenic oil palm plantlet induced by drought stress Turhadi TURHADI; Hayati MINARSIH; Imron RIYADI; . PRIYONO; Asmini BUDIANI
E-Journal Menara Perkebunan Vol 88, No 2 (2020): Oktober,2020
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v88i2.386

Drought is one of the limiting factors in crop cultivation, such as in oil palm (Elaeis guineensis Jacq.). The transgenic approaches are expected to increase plant tolerance to drought stress and minimize low productivity when drought occurs. Proline is an osmoprotectant compound in plants which its biosynthesis involved the P5CS gene. The objective of this study was to evaluate the tolerance level of P5CS-transgenic oil palm to drought stress induced by polyethylene glycol 6000 (PEG-6000). In this present study, the transgenic and non-transgenic oil palms were treated by 0, 2, and 4% PEG-6000 under in vitro conditions. The experiment was arranged as a factorial completely randomized design with three replications. The drought level score, total chlorophyll content, carotenoids, and proline content, as well as P5CS gene expression in leaf tissues were observed at 7 and 14 days after stress treatments. The result showed that transgenic plantlets had a lower drought level score than those of non-transgenic lines. A concentration of 4% PEG-6000 treatment reduced the total chlorophyll and carotenoids contents than that of 2% concentration in non-transgenic plantlets at 7 and 14 day after treatments (DAT). In addition, proline content and P5CS gene expression level in transgenic had been significantly increased during stress treatment. Based on these results, it can be concluded that the P5CS transgene increased the drought stress tolerance of oil palm.

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.

Direct somatic embryogenesis and plant regeneration in tea by temporary liquid immersion Embriogenesis somatik langsung dan regenerasi tanaman teh melalui perendaman sesaat J S TAHARDI; Tatik RAISAWATI; Imron RIYADI; W A DODD
E-Journal Menara Perkebunan Vol 68, No 1: Juni 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Ringkasan Perbanyakan tanaman teh [Camellia sinensis (L.) O. Kuntze] melalui stek tunas berdaun tunggal hanya dapat menghasilkan klon unggul dalam jumlah terbatas. Oleh sebab itu diperlukan metode alternatif dengan teknik kultur sel dan jaringan untuk perbanyakan klonal secara cepat. Dalam penelitian ini dikembangkan metode yang lebih efektif untuk regenerasi tanaman teh melalui embriogenesis somatik langsung. Massa proembriogenik dari eksplan kotiledon dihasilkan dengan frekuensi 56,7% dalam media MS padat setengah konsentrasi yang mengandung BAP 2 mg1L. Proliferasi, perkembangan, pendewasaan dan perkecambahan embrio somatik diperoleh dengan sistem perendaman sesaat (SPS) yang menggunakan media MS cair setengah konsentrasi, yang diperkaya dengan zat pengatur tumbuh dengan berbagai konsentrasi. Proliferasi embrio meningkat 4,3 kali dalam media yang diberi BAP 2 mglL; perkembangan dan pendewasaannya meningkat dengan penambahan kinetin dan ABA masing-masing pada konsentrasi 0,1 mg1L yang 30% diantaranya berkecambah dan membentuk planlet tanpa penambahan zat pengatur tumbuh. Protokol SPS tersebut merupakan sistem in vitro yang berpotensi bagi proliferasi dan perkembangan embrio somatik tanaman teh yang cepat dan sinkron dari kultur kotiledon, serta regenerasinya menjadi planlet tanpa melalui fase kalus.Summary Tea propagation by single-leaf bud cuttings has limited applications for rapid dissemination of planting materials from new elite clones. An alternative method for rapid cloning by cell and tissue culture technique is necessary. In this study we have established an improved method for tea [Camellia sinensis (L.) O. Kuntze] plant regeneration via direct somatic embryogenesis. Clumps of proembryogenic masses were initiated at a frequency of 56.7% from cotyledonary slices cultured on a half-strength MS agar-gelled medium supplemented with 2 mg/L BAP. Proliferation, development, maturation and germination of somatic embryos were achieved using the temporary immersion system (TIS) provided with halfstrength MS liquid media supplemented with varying concentrations of growth regulators. Embryo proliferation increased by 4.3-fold in medium provided with 2 mg/L BAP; their development and maturation were enhanced by the presence of both kinetin and ABA at 0.1 mg/L each. Germination and plant recovery were achieved at a frequency of about 30% without the use of growth regulators. The TIS protocol described above represents an in vitro system potential for rapid proliferation and synchronized development of tea somatic embryos from cotyledon cultures, and their regeneration into plantlets without an intervening callus phase.

Pengaruh interval dan lama perendaman terhadap pertumbuhan dan pendewasaan embrio somatik tanaman sagu (Metroxylon sagu Rottb.) Effect of immersion interval and duration on the growth and maturation of somatic embryos of sago palm ( Metroxylon sagu Rottb.) Imron RIYADI; . SUMARYON
E-Journal Menara Perkebunan Vol 77, No 2: Desember 2009
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractLiquid culture via temporary immersionsystem (TIS) has a potency for enhancingmaturity and uniformity of plant somatic embryos(SE). An experiment was conducted to determinethe effect of medium immersion interval andduration on the growth and maturation of sagoSE in TIS. A clump of SE at globular stagederived from sucker’s tip meristem culture wasused as material source. The SE were cultured ona modified Murashige and Skoog medium addedwith 0.01 mg/L ABA, 1.0 mg/L kinetin and0.1 mg/L GA 3 . The treatments used were TISwith immersion interval of 3, 6 and 12 hourswith duration of 1 and 3 minutes. Solid mediumwas used as a control. The results show that TISwith immersion interval 12 hours for threeminutes produced the highest SE biomass(14.6 g/flask) which had increased by 9.8-foldwithin six weeks. The longer of immersioninterval (less frequent) and the longer ofimmersion duration (three minutes) increasedsignificantly biomass fresh weight of of sago SE.SE biomass of sago on solid medium wassignificantly lower than those of in liquid mediaof TIS. The highest number of advanced stageembryos (torpedo, cotyledonary and earlygerminant) of 643 or 48.2% from the totalnumber of SE was achieved in TIS with 12 hoursinterval for three minutes. During the maturationof sago SE, the color of embryos has changedfrom mostly yellowish to greenish and reddish.AbstrakKultur cair dengan sistem perendaman sesaat(SPS) berpotensi untuk meningkatkanpendewasaan dan keseragaman embrio somatik(ES) tanaman. Penelitian ini bertujuan untukmenentukan pengaruh interval dan lamaperendaman terhadap proses pendewasaan ESsagu dalam SPS. Bahan yang digunakan berupaES fase globuler asal kultur pucuk tunas anakansagu. ES sagu dikulturkan dalam mediumMurashige dan Skoog yang dimodifikasiditambah ABA 0,01 mg/L; kinetin 1 mg/L danGA 3 0,1 mg/L. Perlakuan yang digunakan adalahkultur SPS dengan interval perendaman 3, 6 dan12 jam dengan lama perendaman 1 dan 3 menitserta kultur padat sebagai pembanding. Hasilpenelitian menunjukkan bahwa perlakuan SPSdengan interval perendaman 12 jam selama tigamenit menghasilkan bobot biomassa ES tertinggiyaitu 14,6 g/bejana yang meningkat 9,8 kalidalam waktu enam minggu. Interval peren-daman lebih lama (lebih jarang) dan lama peren-daman lebih panjang (tiga menit) meningkatkansecara nyata bobot segar biomassa ES sagu.Biomassa ES sagu pada medium padat secaranyata lebih rendah dibandingkan dengan kulturkotiledon dan kecambah dini) tertinggi yaitu 643atau 48,2% dari jumlah total ES diperoleh pada perlakuan SPS interval 12 jam dengan lama tigamenit. Seiring dengan pendewasaan ES sagu, terjadi perubahan warna dari sebagian besar kuning menjadi hijau dan merah.

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.

Pengaruh jenis penutup botol kultur terhadap pertumbuhan planlet kelapa sawit (Elaeis guineensis Jacq.) Effect of different culture vessel closures on the growth of oil palm (Elaeis guineensis Jacq.) plantlets Masna Maya SINTA; Imron RIYADI; . UMARYONO
E-Journal Menara Perkebunan Vol 79, No 1: Juni 2011
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractMicroenvironment inside the culture vessel such astemperature, light intensity, relative humidity, and aerationaffect growth and development of plantlets. This experimentwas conducted to determine the effect of different culturevessel closures on microenvironmental conditions inside thevessel and on growth of plantlets of oil palm. Shoots of oilpalm derived from somatic embryos were cultured on DFmedium for eight weeks in transparent culture bottlescovered with five different vessel closures e.i. screw cap withplastic wrap, screw cap, plastic wrap, aluminum foil, andautoclavable plastic. The culture vessels were placed in theculture room with light intensity 20 µmol/m 2 /sec for 12 hoursphotoperiod, at room temperature 26°C. Parametersobserved on plantlet growth were shoot height, biomass freshweight, leaf number, and leaf color grade, while onmicroenvironment were temperature and light intensity. Atthe end of experiment, the volume and fresh weight of theremaining medium were measured to determine evaporationrate of each treatment. Results show that the use of differentculture vessel closures affected the microenvironment insidethe vessel, the volume of the remaining medium, and thegrowth of the plantlets. The closure increased thetemperature by 1.6 – 2.6°C and decreased the light intensityby 1.7 – 8.7 µmol/m 2 /sec inside the culture vessels dependson the culture vessel closures. Culture vessels with aluminumfoil closure had the lowest temperature (28.9°C) and thelowest light intensity (10.8 µmol/m 2 /sec) gave the best resultin the growth of the plantlets. Better plantlets growth wasalso observed in the culture vessel with autoclavable plasticclosure that less expensive, therefore it can be used as analternative vessel closure for the growth of oil palm plantlets.AbstrakLingkungan mikro di dalam botol kultur seperti suhu,intensitas cahaya, kelembaban nisbi dan aerasi mem-pengaruhi pertumbuhan dan perkembangan planlet.Penelitian ini dilakukan untuk mengetahui pengaruhpenggunaan penutup botol kultur yang berbeda terhadapkondisi lingkungan mikro di dalam botol kultur danpertumbuhan planlet kelapa sawit. Planlet kelapa sawit asalembrio somatik dikulturkan dalam botol kultur bening berisimedium DF selama delapan minggu dan ditutup mengguna-kan lima jenis penutup botol yang berbeda yaitu tutup ulirdengan plastik wrap, tutup ulir, plastik wrap, aluminium foildan plastik tahan diautoklaf. Kultur diletakkan dalam ruangkultur, di bawah lampu TL dengan intensitas cahaya20 µmol/m 2 /detik dan suhu ruang 26 o C. Parameterpertumbuhan planlet yang diamati adalah tinggi planlet,bobot basah, jumlah daun dan kelas warna daun, sedangkanlingkungan mikro adalah suhu dan intensitas cahaya. Padaakhir eksperimen, volume dan bobot basah medium yangtersisa diukur untuk mengetahui tingkat penguapan padasetiap perlakuan. Hasil penelitian menunjukkan bahwapenggunaan penutup botol yang berbeda berpengaruhterhadap lingkungan mikro, volume medium tersisa dalambotol kultur dan pertumbuhan planlet. Penutup botolmeningkatkan suhu 1,6 – 2,6 o C dan menurunkan intensitascahaya 1,7 – 8,7 µmol/m 2 /detik di dalam botol tergantungpada jenis penutup botol yang digunakan. Botol kulturdengan penutup berbahan aluminium foil mempunyaiintensitas cahaya terendah (10,8 µmol/m 2 /detik) dan suhuterendah (28,9 o C) memberikan hasil terbaik pada pembesaranplanlet kelapa sawit. Pertumbuhan planlet yang baik jugaterdapat pada botol kultur dengan penutup plastik tahandiautoklaf yang lebih murah, sehingga penutup ini dapatdigunakan sebagai pilihan untuk pembesaran planlet kelapasawit.

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