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.
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.
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.
The 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.
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