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Embriogenesis Somatik Tidak Langsung pada Tanaman Sagu (Metroxylon sagu Rottb.) Menggunakan Sistem Kultur Suspensi, Perendaman Sesaat, dan Media Padat Imron Riyadi; Darda Efendi; Bambang S. Purwoko; Djoko Santoso
Jurnal AgroBiogen Vol 12, No 1 (2016): Juni
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v12n1.2016.p37-44

Metode kultur in vitro yang tepat akan meningkatkan efektivitas dan efisiensi pada proses penggandaan kalus dan induksiembriogenesis somatik. Penelitian ini bertujuan mengevaluasi efektivitas tiga metode kultur jaringan, yaitu sistem kultursuspensi, sistem perendaman sesaat (SPS) atau temporary immersion system (TIS), dan media padat, untuk proliferasi kalusdan pembentukan embrio somatik secara tidak langsung pada tanaman sagu “Alitir” yang berasal dari Merauke, Papua. Bahantanaman atau eksplan awal yang digunakan adalah kalus remah hasil induksi dari kultur meristem pucuk tunas anakan sagu.Kalus tersebut dikulturkan pada media Murashige dan Skoog (MS) modifikasi dengan penambahan 2,4-D 5,0–15,0 mg/l dikombinasikandengan kinetin 0,1 mg/l menggunakan ketiga metode kultur sehingga terdapat dua belas kombinasi perlakuan.Hasil penelitian menunjukkan bobot segar kalus tertinggi sebesar 12,0 g/bejana dicapai pada metode kultur suspensi denganpenambahan 2,4-D 15,0 mg/l dikombinasikan dengan kinetin 0,1 g/l. Perolehan jumlah embrio somatik tertinggi dicapai padametode kultur suspensi dengan penambahan 2,4-D 5,0 mg/l dikombinasikan dengan kinetin 0,1 g/l sebesar 384,7 buah/bejana.Daya hidup kultur sagu terbaik dan tertinggi (100%) diperoleh pada metode kultur suspensi pada semua perlakuankonsentrasi 2,4-D. Selama proses induksi embrio somatik, terjadi perubahan warna kalus dari sebagian besar kekuninganmenjadi krem dan putih-kekuningan.

Propagasi in vitro tanaman kurma (Phoenix dactylifera L.) pada bioreaktor dengan perendaman sesaat Rizka Tamania SAPTARI; Masna Maya SINTA; Imron RIYADI; . PRIYONO; . SUMARYONO
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.v88i2.394

The cultivation of date palm in Indonesia has increased since the last decade. However, the superior date palm seedlings are still limited and most of them are imported from other countries. The mass supply of superior date palm seedlings can be provided by in vitro propagation in the bioreactor. Therefore, the research was conducted to develop a protocol of date palm in vitro propagation by using Temporary Immersion Bioreactor (TIB). The in vitro propagation was carried out through somatic embryogenesis technique using meristematic tissues isolated from offshoots of date palm female clone cv. Zambli as explants. The explants were sterilized and then cultured to produce embryogenic calli and somatic embryos. Afterwards, somatic embryos germination and plantlets formation were conducted in TIB with treatments of immersion period: 3, 10, and 30 minutes every 6 hours, with 8 replications, The results showed that the optimal somatic embryo germination in TIB was with the immersion period of 30 min every 6 h, resulting in the most formation of shoots and fresh biomass weight increment up to nearly threefold in 6 weeks. Thereafter, plantlets formation in TIB with immersion period of 10 min and 30 min every 6 h exhibited similar performances in producing more plantlets with higher total fresh weight and better vigor than those of 3 min every 6 h. However, there were more rooted plantlets in the TIB with immersion period of 10 min every 6 h. Based on the results, an in vitro propagation protocol via somatic embryogenesis in TIB has been successfully developed for mass propagation of date palm cv. Zambli, which produced plantlets with good vigor and rooting.

Kriteria planlet kelapa kopyor yang siap untuk diaklimatisasi [Criteria of kopyor coconut plantlets ready to be acclimatized] . SUMARYONO; Imron RIYADI
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Kopyor coconut is a special coconut grown in Indonesia. Nuts of kopyor can not be used as a source of planting material due to its endosperm damage; therefore in vitro embryo rescue technique is applied to propagate kopyor coconuts. Acclimatization is a critical stage during in vitro culture of kopyor coconut. Experiments were conducted to determine the effect of plantlet initial conditions prior to acclimatization on survival and growth in ex vitro conditions. Five replications of 50 plantlets of tall variety of kopyor coconut with different shoot and root conditions were used in the acclimatization process. The coconut plantlets were planted on plastic pots containing a mixture of soil, sand and dung manure, and then placed inside a closed plastic tunnel. The tunnel was opened gradually after 3 months and the plants were transferred to the nursery after 4.5 months. Survival frequency and growth (plant height, leaf number and shoot diameter) of the plantlets were observed after 6 months of acclimatization period. Research results show that the initial plantlet height and initial root length affected significantly the survival rate and growth of the plantlets of kopyor coconut during acclimatization. Other parameters of plantlet initial conditions such as leaf number, stem diameter, primary root number, and the existence of secondary roots did not influence the survival rate and growth of the plantlets. In order to obtain high survival rate (84.7%) and good growth of plantlets during acclimatization, the plantlet height must be at least 20 cm prior to acclimatization. Differences in root length on plantlets with the height of 20 cm or more, did not affect survival percentage of the vitroplants.

Pengaruh TDZ terhadap induksi embrio somatik sagu (Metroxylon sagu Rottb.) pada tiga metode kultur berbeda (Effect of TDZ on the somatic embryo induction of sago palm (Metroxylon sagu Rottb.) in three different culture methods) Imron Riyadi; Darda EFENDI; Bambang S PURWOKO; Djoko SANTOSO
E-Journal Menara Perkebunan Vol 86, No 1 (2018): April, 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractA right combination of cytokinin is able to support the process of callus differentiation to somatic embryo formation in plant somatic embryogenesis. Liquid culture application could increase the efficiency of in vitro culture process on plants. This research aimed to determine the best concentration of TDZ combined with kinetin for callus differentiation to somatic embryo of sago palm on three culture methods. Plant material used was embryogenic callus derived from tips meristem culture from sucker of Alitir sago palm. Callus was cultured on modified MS media added with: 0.0, 0.1, 0.5 and 1.0 mg/L TDZ combined with 0.5 mg/L kinetin for 12 weeks with subcultures every 6 weeks. Three culture methods used were suspension, temporary immersion system (TIS), and solid media. There were 12 treatments with 4 replicates. The results showed that the highest number of somatic embryos was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in 6 weeks (167.3 embryos/flask) and 12 weeks (389.2 embryos/flask) with its fresh weight of 18.4 g and 29.1 g, respectively. The highset survival rate in final culture (12 weeks) was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin (100%). The shortest time for somatic embryos expression was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in two weeks after culture. Histological analysis of early-stage somatic embryos showed the presence of dense and compact cellular arrangements which formed growth spot axis for shoot or SAM (shoot apical meristem) and root or RAM (root apical meristem) that connected each other. [Key words: culture method, embryogenic callus, Metroxylon sagu Rottb., kinetin, sago palm, TDZ] AbstrakAplikasi kombinasi sitokinin yang tepat dapat mendorong proses diferensiasi kalus membentuk embrio somatik pada proses embriogenesis somatik tanaman. Penggunaan metode kultur cair dapat meningkatkan efisiensi proses kultur in vitro tanaman. Penelitian ini bertujuan untuk menentukan konsentrasi TDZ terbaik dikombinasikan dengan kinetin dalam proses diferensiasi kalus membentuk embrio somatik tanaman sagu pada tiga metode kultur. Bahan tanam penelitian berupa kalus embriogenik tanaman sagu asal kultur meristem pucuk dari anakan sagu jenis Alitir. Kalus dikulturkan pada media modifikasi dengan penambahan TDZ dengan konsentrasi 0,1; 0,5; dan 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L selama 12 minggu yang disubkultur pada umur 6 minggu. Metode kultur yang digunakan terdiri atas tiga macam yaitu: kultur suspensi, sistem perendaman sesaat (SPS) dan media padat. Perlakuan terdiri atas 12 kombinasi perlakuan dengan empat ulangan. Hasil penelitian menunjukkan bahwa rerata jumlah embrio somatik tertinggi dicapai pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L baik pada umur kultur 6 minggu (167,3 buah) maupun umur 12 minggu (389,2 buah). Rerata bobot segar tertinggi juga diperoleh pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L pada umur kultur 6 minggu (18,4 g) dan 12 minggu (29,1 g). Rerata daya hidup kultur akhir (12 minggu) tertinggi sebesar 100% diperoleh pada perlakuan SPS. Induksi embrio somatik tercepat yakni setelah dua minggu diperoleh pada metode kultur SPS dengan TDZ 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L. Analisis histologi embrio somatik stadium awal menunjukkan adanya susunan sel yang rapat dan kompak yang menyusun semacam poros atau berkas titik tumbuh tunas atau SAM (shoot apical meristem) maupun akar atau RAM (root apical mersitem) yang saling terhubung.[Kata kunci: kalus embriogenik, metode kultur, kinetin, TDZ, sagu, Metroxylon sagu]

Regeneration of oil palm plantlets introduced by P5CS gene using Agrobacterium-mediated transformation Asmini BUDIANI; Imam Bagus NUGROHO; Hayati MINARSIH; Imron RIYADI
E-Journal Menara Perkebunan Vol 87, No 2 (2019): OKTOBER, 2019
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstrakCekaman kekeringan dapat mempengaruhi produktivitas tanaman perkebunan. Rekayasa genetika merupakan salah satu cara untuk meningkatkan produktivitas tanaman perkebunan penting seperti kelapa sawit. Tujuan dari penelitian ini adalah melakukan perekayasaan kelapa sawit melalui introduksi gen P5CS dengan transformasi berbasis Agrobacterium untuk meningkatkan ketahanan tanaman terhadap cekaman kekeringan. Pada penelitian ini perakitan kelapa sawit transgenik yang tahan terhadap cekaman kekeringan dilakukan melalui transformasi gen P5CS (Δ1-pyrroline-5-carboxylate synthetase) ke dalam kalus embriogenik (embryogenic calli – EC) menggunakan Agrobacterium. Plasmid pBI_P5CS yang membawa gen P5CS ditransfer dari Escherichia coli XL1 Blue ke Agrobacterium tumefaciens AGL1 melalui konjugasi. Selanjutnya klon Agrobacterium yang membawa plasmid pBI_P5CS digunakan untuk menginfeksi kalus embriogenik kelapa sawit dengan perlakuan 100 ppm asetosiringon. Kalus transforman diregenerasi pada media de Fossard (DF) yang ditambahkan 50 ppm kanamisin dan 250 ppm sefotaksim. Kalus transforman diseleksi melalui uji GUS dan metode PCR menggunakan primer NPTII dan P5CS1. Uji GUS dilakukan untuk menyeleksi kalus transforman yang ditunjukkan dengan reaksi positif pembentukan warna biru pada kalus yang berhasil ditransformasi dengan konstruk pBI_P5CS. Pengujian dengan menggunakan PCR memberikan hasil positif dengan adanya profil pita PCR pada visualisasi menggunakan pewarnaan SYBR Green, yang menunjukkan amplikon berukuran ~ 0,7 kb untuk gen NPTII dan ~ 0,4 kb untuk gen P5CS pada elektroforesis dengan gel agarosa. Hasil dari penelitian ini adalah diperolehnya kalus transforman terseleksi yang telah diregenerasi dan tumbuh menjadi planlet.[Kata kunci: cekaman kekeringan, Elaeis guineensis Jacq., rekayasa genetika, planlet]Abstract Environmental abiotic stressors particularly drought has detrimental effects upon the productivity of estate crops. Increasing the crop tolerance towards drought stress through genetic engineering is one of the strategies employed to maintain steady productivity of valuable crop, i.e. oil palm. The aim of this study was to engineer oil palm with a better tolerance towards drought by introducing P5CS (Δ1-pyrroline-5-carboxylate synthetase) gene via Agrobacterium–mediated transformation into embryogenic calli (EC). The pBI_P5CS plasmid harboring P5CS gene was transferred from Escherichia coli XL1 Blue to Agrobacterium tumefaciens AGL1 by conjugation. The positive clone of transformed Agrobacterium was then used to infect oil palm EC by the addition of 100 ppm acetosyringone. The transformed ECs were regenerated in the de Fossard (DF) media supplemented by 50 ppm kanamycin and 250 ppm cefotaxime followed by GUS assay and PCR-based screening using NPTII and P5CS1 primers. The positive EC clones were confirmed by GUS assay, which produced blue coloration on positive transformed oil palm EC. A positive result of PCR screenings was depicted by PCR products in SYBR Green staining gel agarose electrophoresis with the expected band size of ~ 0.7 kb for the NPTII gene and ~ 0.4 kb for the P5CS gene. This study has successfully selected and regenerated pBI_P5CS transformed oil palm embryogenic calli into plantlets.[Keywords: drought tolerance, Elaeis guineensis Jacq., genetic engineering, plantlets]

Pengaruh periode perendaman air dan komposisi media tumbuh terhadap keberhasilan aklimatisasi planlet sagu (Effect of water immersion period and growing media composition on acclimatization success of sago palm plantlets ) . SUMARYONO; Imron RIYADI
E-Journal Menara Perkebunan Vol 85, No 2 (2017): Oktober 2017
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Sago palm (Metroxylon sagu Rottb.) is a carbohydrate-producing crop, commonly propagated by suckers. The availability of planting materials in a large quantity hinders the development of commercial sago plantations. Sago propagation by tissue culture via somatic embryogenesis has been developed to provide superior planting materials of sago palm. One of the major problems faced in tissue culture of sago palm is the low survival rate of plantlets during acclimatization period. The objective of this research was to increase the acclimatization success of sago plantlets in term of survival rate and growth at ex vitro conditions. The experiments were conducted using a randomized block design with two factors i.e. water immersion period and growing media composition. The water immersion periods used were without immersion, immersion for 2 days with 1 day intermittent period, immersion for 1 day with 1 day intermittent period, immersion for 1 day with 2 days intermittent period, and continuous immersion. The growing media used were consisted of top soil, sand, dung manure, and cocopeat at different compositions. Sago plantlets were planted on small plastic pots and placed inside a closed plastic tunnel for 12 weeks. Research results showed that continuous water immersion and mixed composition of soil, sand, and cocopeat (1:1:2 v/v) was the best conditions for acclimatization of sago plantlets with the survival rate of 70% after 12 weeks. The survived plants had good leaves and roots, ready to be transferred to big plastic bags in the main nursery. [Keywords: Metroxylon sagu, sago palm, acclimatization, immersion, media composition] AbstrakTanaman palma sagu (Metroxylon sagu Rottb.) termasuk tanaman penghasil karbohidrat yang umumnya diperbanyak dengan anakan (suckers). Ketersediaan bahan tanam dalam jumlah besar merupakan hambatan pengembangan perkebunan sagu komersial. Kultur jaringan tanaman sagu telah dikembangkan melalui teknik embriogenesis somatik untuk memenuhi kebutuhan bahan tanam unggul sagu. Salah satu masalah utama dalam kultur jaringan sagu adalah rendahnya daya hidup planlet pada tahap aklimatisasi. Penelitian ini bertujuan meningkatkan keberhasilan aklimatisasi planlet sagu yang meliputi daya hidup dan pertumbuhan bibit pada lingkungan ex vitro. Percobaan dilakukan menggunakan rancangan acak kelompok dengan dua faktor yaitu perendaman air dan komposisi media. Perlakuan perendaman air adalah tanpa perendaman, perendaman 2 hari diselang tanpa perendaman 1 hari, perendaman 1 hari diselang tanpa perendaman 1 hari, perendaman 1 hari diselang tanpa perendaman 2 hari, dan perendaman terus menerus. Komposisi media tumbuh yang digunakan berupa perbandingan volume penyusun yaitu tanah, pasir, pupuk kandang dan cocopeat. Planlet ditanam di pot kecil dan diletakkan di dalam sungkup plastik tertutup selama 12 minggu. Hasil penelitian menunjukkan bahwa perlakuan perendaman terus menerus dan media tumbuh campuran tanah, pasir, cocopeat (1:1:2 v/v) merupakan kondisi terbaik pada aklimatisasi planlet sagu dengan daya hidup mencapai 70% setelah 12 minggu. Bibit yang dihasilkan memiliki daun dan perakaran yang baik, siap untuk dipindahkan ke pot plastik besar di persemaian utama.[Kata kunci: Metroxylon sagu, sagu, aklimatisasi, perendaman, komposisi media]

Determination of the optimum initial callus weight for the efficient propagation of sugarcane in temporary immersion bioreactor Rizka Tamania SAPTARI; Imron RIYADI; Masna Maya SINTA; M Eko Riyo Bayu PRASETYO; Sylvia LINDAWATI; Sumaryono SUMARYONO
E-Journal Menara Perkebunan Vol 90, No 2 (2022): Oktober, 2022
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v90i2.505

AbstrakBioreaktor perendaman sesaat (BPS) telah digunakan secara luas untuk propagasi skala massal berbagai tanaman penting, termasuk tanaman tebu. BPS menyediakan sistem kultur semi-otomatis dan kondisi optimal bagi pertumbuhan tanaman. Beberapa faktor menentukan pertumbuhan tanaman pada BPS, salah satunya densitas dari eksplan. Oleh karena itu, penelitian dilakukan untuk menentukan bobot awal yang optimal untuk kalus tebu yang dikulturkan pada BPS, serta mengevaluasi pengaruh perbedaan bobot awal kalus tersebut terhadap proliferasi dan regenerasi kalus tebu. Kalus tebu diinduksi dari daun muda yang masih menggulung dari empat varietas tebu unggul Indonesia. Bobot awal kalus yang dikultur ke dalam bejana TIB yaitu 0,05 g; 0,1 g; 0,2 g; 0,5 g; dan 1,0 g untuk setiap bejana. Kalus kemudian melalui tahap proliferasi pada BPS sebanyak tiga siklus, kemudian kalus diregenerasi pada BPS dengan perlakuan auksin dan sitokinin. Hasil penelitian menunjukkan bahwa 0,2 g merupakan bobot awal kalus yang efisien untuk proliferasi kalus tebu pada TIB, dimana eksponensial multiplikasi kalus tercapai pada bobot awal tersebut, yaitu untuk masing-masing varietas 130,3 kali (PSKA 942), 136,8 kali (PS 094), 21,3 (PS 881), dan 12,9 kali (PS 091) setelah 12 minggu. Densitas kalus pada TIB berkorelasi negatif dengan karakteristik fisikokimia medium. Hal ini menggambarkan variasi intensitas pertumbuhan dan metabolisme kalus dengan adanya perbedaan densitas pada BPS. Penggunaan BAP 0,2 mg L-1 bersama kinetin 0,2 mg L-1 paling sesuai untuk memacu regenerasi kalus tebu dengan menghasilkan jumlah tunas terbanyak dalam waktu relatif lebih cepat (1 – 2 minggu lebih cepat) dibandingkan perlakuan lainnya dan dengan tingkat kejadian pencoklatan yang rendah.[Kata kunci: kultur in vitro, kultur cair, proliferasi]AbstractTemporary immersion bioreactor (TIB) has been utilized for the mass-scale propagation of many important plants, including sugarcane. TIB facilitates a semiautomated culture system and provides optimal conditions for plant growth. Several factors determine plant growth in the TIB, such as explant density. Therefore, an experiment was carried out to determine the optimal initial weight of sugarcane calli and to evaluate its effect on the proliferation and regeneration in TIB. Sugarcane calli were induced from spindle leaves isolated from four Indonesian prime sugarcane varieties. The initial weights of the calli cultured in the TIB flasks were 0.05 g, 0.1 g, 0.2 g, 0.5 g and 1.0 g per flask. The calli were proliferated through three cycles in TIB, and subsequently regenerated in TIB with auxin and cytokinin treatments. The results of the experiments showed that 0.2 g was the most efficient initial weight for sugarcane callus proliferation in the TIB, resulting in an exponential multiplication rate of 130.3-fold (PSKA 942), 136.8-fold (PS 094), 21.3-fold (PS 881), and 12.9-fold (PS 091) within 12 weeks. In the TIB, callus density showed a negative correlation with the physicochemical properties of the medium, demonstrating various growth intensities or metabolic activities of calli at different densities in the TIB. The use of 0.2 mg L-1 BAP along with 0.2 mg L-1 kinetin was suitable for promoting the regeneration of sugarcane calli and producing the highest number of shoots in a relatively short amount of time (1 – 2 weeks faster) with low incidences of browning.[Keywords: in vitro culture, liquid culture, proliferation]

Kriteria planlet kelapa kopyor yang siap untuk diaklimatisasi [Criteria of kopyor coconut plantlets ready to be acclimatized] . SUMARYONO; Imron RIYADI
Menara Perkebunan Vol. 84 No. 1 (2016): 84 (1), 2016
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v84i1.203

Kopyor coconut is a special coconut grown in Indonesia. Nuts of kopyor can not be used as a source of planting material due to its endosperm damage; therefore in vitro embryo rescue technique is applied to propagate kopyor coconuts. Acclimatization is a critical stage during in vitro culture of kopyor coconut. Experiments were conducted to determine the effect of plantlet initial conditions prior to acclimatization on survival and growth in ex vitro conditions. Five replications of 50 plantlets of tall variety of kopyor coconut with different shoot and root conditions were used in the acclimatization process. The coconut plantlets were planted on plastic pots containing a mixture of soil, sand and dung manure, and then placed inside a closed plastic tunnel. The tunnel was opened gradually after 3 months and the plants were transferred to the nursery after 4.5 months. Survival frequency and growth (plant height, leaf number and shoot diameter) of the plantlets were observed after 6 months of acclimatization period. Research results show that the initial plantlet height and initial root length affected significantly the survival rate and growth of the plantlets of kopyor coconut during acclimatization. Other parameters of plantlet initial conditions such as leaf number, stem diameter, primary root number, and the existence of secondary roots did not influence the survival rate and growth of the plantlets. In order to obtain high survival rate (84.7%) and good growth of plantlets during acclimatization, the plantlet height must be at least 20 cm prior to acclimatization. Differences in root length on plantlets with the height of 20 cm or more, did not affect survival percentage of the vitroplants.

Pengaruh periode perendaman air dan komposisi media tumbuh terhadap keberhasilan aklimatisasi planlet sagu (Effect of water immersion period and growing media composition on acclimatization success of sago palm plantlets ) . SUMARYONO; Imron RIYADI
Menara Perkebunan Vol. 85 No. 2 (2017): 85 (2), 2017
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v85i2.242

Sago palm (Metroxylon sagu Rottb.) is a carbohydrate-producing crop, commonly propagated by suckers. The availability of planting materials in a large quantity hinders the development of commercial sago plantations. Sago propagation by tissue culture via somatic embryogenesis has been developed to provide superior planting materials of sago palm. One of the major problems faced in tissue culture of sago palm is the low survival rate of plantlets during acclimatization period. The objective of this research was to increase the acclimatization success of sago plantlets in term of survival rate and growth at ex vitro conditions. The experiments were conducted using a randomized block design with two factors i.e. water immersion period and growing media composition. The water immersion periods used were without immersion, immersion for 2 days with 1 day intermittent period, immersion for 1 day with 1 day intermittent period, immersion for 1 day with 2 days intermittent period, and continuous immersion. The growing media used were consisted of top soil, sand, dung manure, and cocopeat at different compositions. Sago plantlets were planted on small plastic pots and placed inside a closed plastic tunnel for 12 weeks. Research results showed that continuous water immersion and mixed composition of soil, sand, and cocopeat (1:1:2 v/v) was the best conditions for acclimatization of sago plantlets with the survival rate of 70% after 12 weeks. The survived plants had good leaves and roots, ready to be transferred to big plastic bags in the main nursery. [Keywords: Metroxylon sagu, sago palm, acclimatization, immersion, media composition] AbstrakTanaman palma sagu (Metroxylon sagu Rottb.) termasuk tanaman penghasil karbohidrat yang umumnya diperbanyak dengan anakan (suckers). Ketersediaan bahan tanam dalam jumlah besar merupakan hambatan pengembangan perkebunan sagu komersial. Kultur jaringan tanaman sagu telah dikembangkan melalui teknik embriogenesis somatik untuk memenuhi kebutuhan bahan tanam unggul sagu. Salah satu masalah utama dalam kultur jaringan sagu adalah rendahnya daya hidup planlet pada tahap aklimatisasi. Penelitian ini bertujuan meningkatkan keberhasilan aklimatisasi planlet sagu yang meliputi daya hidup dan pertumbuhan bibit pada lingkungan ex vitro. Percobaan dilakukan menggunakan rancangan acak kelompok dengan dua faktor yaitu perendaman air dan komposisi media. Perlakuan perendaman air adalah tanpa perendaman, perendaman 2 hari diselang tanpa perendaman 1 hari, perendaman 1 hari diselang tanpa perendaman 1 hari, perendaman 1 hari diselang tanpa perendaman 2 hari, dan perendaman terus menerus. Komposisi media tumbuh yang digunakan berupa perbandingan volume penyusun yaitu tanah, pasir, pupuk kandang dan cocopeat. Planlet ditanam di pot kecil dan diletakkan di dalam sungkup plastik tertutup selama 12 minggu. Hasil penelitian menunjukkan bahwa perlakuan perendaman terus menerus dan media tumbuh campuran tanah, pasir, cocopeat (1:1:2 v/v) merupakan kondisi terbaik pada aklimatisasi planlet sagu dengan daya hidup mencapai 70% setelah 12 minggu. Bibit yang dihasilkan memiliki daun dan perakaran yang baik, siap untuk dipindahkan ke pot plastik besar di persemaian utama.[Kata kunci: Metroxylon sagu, sagu, aklimatisasi, perendaman, komposisi media]

Pengaruh TDZ terhadap induksi embrio somatik sagu (Metroxylon sagu Rottb.) pada tiga metode kultur berbeda (Effect of TDZ on the somatic embryo induction of sago palm (Metroxylon sagu Rottb.) in three different culture methods) Imron Riyadi; Darda EFENDI; Bambang S PURWOKO; Djoko SANTOSO
Menara Perkebunan Vol. 86 No. 1 (2018): 86 (1), 2018
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v1i1.258

AbstractA right combination of cytokinin is able to support the process of callus differentiation to somatic embryo formation in plant somatic embryogenesis. Liquid culture application could increase the efficiency of in vitro culture process on plants. This research aimed to determine the best concentration of TDZ combined with kinetin for callus differentiation to somatic embryo of sago palm on three culture methods. Plant material used was embryogenic callus derived from tips meristem culture from sucker of Alitir sago palm. Callus was cultured on modified MS media added with: 0.0, 0.1, 0.5 and 1.0 mg/L TDZ combined with 0.5 mg/L kinetin for 12 weeks with subcultures every 6 weeks. Three culture methods used were suspension, temporary immersion system (TIS), and solid media. There were 12 treatments with 4 replicates. The results showed that the highest number of somatic embryos was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in 6 weeks (167.3 embryos/flask) and 12 weeks (389.2 embryos/flask) with its fresh weight of 18.4 g and 29.1 g, respectively. The highset survival rate in final culture (12 weeks) was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin (100%). The shortest time for somatic embryos expression was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in two weeks after culture. Histological analysis of early-stage somatic embryos showed the presence of dense and compact cellular arrangements which formed growth spot axis for shoot or SAM (shoot apical meristem) and root or RAM (root apical meristem) that connected each other. [Key words: culture method, embryogenic callus, Metroxylon sagu Rottb., kinetin, sago palm, TDZ] AbstrakAplikasi kombinasi sitokinin yang tepat dapat mendorong proses diferensiasi kalus membentuk embrio somatik pada proses embriogenesis somatik tanaman. Penggunaan metode kultur cair dapat meningkatkan efisiensi proses kultur in vitro tanaman. Penelitian ini bertujuan untuk menentukan konsentrasi TDZ terbaik dikombinasikan dengan kinetin dalam proses diferensiasi kalus membentuk embrio somatik tanaman sagu pada tiga metode kultur. Bahan tanam penelitian berupa kalus embriogenik tanaman sagu asal kultur meristem pucuk dari anakan sagu jenis Alitir. Kalus dikulturkan pada media modifikasi dengan penambahan TDZ dengan konsentrasi 0,1; 0,5; dan 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L selama 12 minggu yang disubkultur pada umur 6 minggu. Metode kultur yang digunakan terdiri atas tiga macam yaitu: kultur suspensi, sistem perendaman sesaat (SPS) dan media padat. Perlakuan terdiri atas 12 kombinasi perlakuan dengan empat ulangan. Hasil penelitian menunjukkan bahwa rerata jumlah embrio somatik tertinggi dicapai pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L baik pada umur kultur 6 minggu (167,3 buah) maupun umur 12 minggu (389,2 buah). Rerata bobot segar tertinggi juga diperoleh pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L pada umur kultur 6 minggu (18,4 g) dan 12 minggu (29,1 g). Rerata daya hidup kultur akhir (12 minggu) tertinggi sebesar 100% diperoleh pada perlakuan SPS. Induksi embrio somatik tercepat yakni setelah dua minggu diperoleh pada metode kultur SPS dengan TDZ 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L. Analisis histologi embrio somatik stadium awal menunjukkan adanya susunan sel yang rapat dan kompak yang menyusun semacam poros atau berkas titik tumbuh tunas atau SAM (shoot apical meristem) maupun akar atau RAM (root apical mersitem) yang saling terhubung.[Kata kunci: kalus embriogenik, metode kultur, kinetin, TDZ, sagu, Metroxylon sagu]

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