scholarly journals 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

2016 ◽  
Vol 73 (2) ◽  
Author(s):  
Imron RIYADI ◽  
J.S. TAHARDI TAHARDI ◽  
. SUMARYONO

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.

2016 ◽  
Vol 73 (2) ◽  
Author(s):  
Imron RIYADI ◽  
J.S. TAHARDI TAHARDI ◽  
. SUMARYONO

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.


2014 ◽  
Vol 1 (1) ◽  
pp. 21
Author(s):  
Teuku Tajuddin ◽  
Karyanti . ◽  
Tati Sukarnih ◽  
Nadirman Haska

Hutan sagu (Metroxylon sagu Rottb.) dapat ditemukan dalam area yang cukup luas di wilayah Maluku dan Papua. Besarnya keanekaragaman hayati dari pohon sagu dapat dilihat di areal ini. Pohon sagu tumbuh secara alami terutama di daerah dataran atau rawa dengan sumber yang air melimpah. Tanaman sagu dapat diperbanyak dengan metode generatif melalui biji, dan vegetatif melalui tunas anakan. Dalam rangka mendukung perbanyakan pohon induk yang unggul secara in vitro dalam skala besar, perbaikan metode sterilisasi tunas anakan mutlak diperlukan. Tunas anakan muda (15-20 cm) yang diperoleh dari Propinsi Papua digunakan sebagai eksplan. Tujuan percobaan sterilisasi ini dilakukan untuk mendukung perbanyakan pohon sagu secara in vitro. Pada percobaan ini antibiotik digunakan untuk membersihkan jaringan internal eksplan dari jamur dan bakteri. Hasil percobaan ini menunjukkan bahwa campuran alkohol dan antibiotik dapat menekan pertumbuhan kontaminan.Kata kunci: Antibiotik, kontaminan jamur dan bakteri, kultur in vitro, metode sterilisasi, sagu ABSTRACTNatural sago (Metroxylon sagu Rottb.) forest can be found in large area in Maluku and Papua regions. There are wide genetic diversities of sago palm found in these areas. This palm grows along riverbanks and in swampy areas which are not suitable for other crops. Sago palm is propagated generatively by seed and vegetatively by suckers. With the purpose of establishing the in vitro culture method for a large-scale of mass clonally propagation of superior genotypes of sago palm, generating sterilized explants are very important. Young suckers (15-20 cm) obtained from areas of Papua Province were used as explants. The sterilization experiments were carrying out to support the tissue culture of sago palm. Sterilization was conducted using antibiotics in order to get rid of fungi and bacteria from inner part of explants tissues. The results showed that from all sterilization methods tested, the best result was treatment using alcohol and antibiotic as disinfectant agents.Keywords: Antibiotics, fungi and bacteria contaminants, in vitro culture, sterilization method, sago palm


2015 ◽  
Vol 2 (2) ◽  
pp. 73
Author(s):  
Teuku Tajuddin ◽  
Karyanti . ◽  
Tati Sukarnih ◽  
Nadirman Haska

Tanaman sagu (Metroxylon sagu Rottb.) memiliki potensi yang besar sebagai sumber pangan, energi dan bahan baku industri. Kultur jaringan tanaman sagu telah dilakukan di Balai Pengkajian Bioteknologi BPPT dalam rangka perbanyakan genotipe atau aksesi unggul secara massal. Namun demikian, kendala utama yang dihadapi pada perbanyakan in vitro tanaman sagu adalah sulitnya pembentukan akar. Penelitian ini bertujuan untuk mendapatkan kombinasi hormon yang tepat dalam menginduksi perakaran tanaman sagu in vitro. Tunas anakan muda (15-20 cm) yang diperoleh dari daerah Rangkasbitung, Provinsi Banten digunakan sebagai eksplan. Dalam penelitian ini perakaran in vitro diinduksi dengan berbagai perlakuan jenis dan konsentrasi hormon auksin, konsentrasi medium dan jenis agar. Sebagai medium dasar digunakan medium Gamborg. Hasil penelitian menunjukkan bahwa konsentrasi IBA dan NAA yang terbaik adalah pada taraf 35 ppm. Selanjutnya Gelrite memberikan respon yang positif dengan munculnya perakaran pada pangkal eksplan.Kata Kunci: Induksi perakaran,  jenis agar, kultur in vitro, auksin, sagu ABSTRACTSago palm (Metroxylon sagu Rottb) has huge potential as food, energy and industrial bioresources. In vitro culture of sago palm was performed in Biotech Center, BPPT in order to obtain a large-scale of mass clonal propagation of superior genotypes. Nevertheless, the main obstacle for the sago palm in vitro propagation was rooting formation. The purpose of our study was to obtain the best hormones combination for root induction on sago palm shoots in vitro. The young suckers (15-20 cm) obtained from Rangkasbitung area, Banten Province, were used as explants. In our study, in vitro rooting was induced by different types and concentrations of auxin, medium strength and solidifying agents. The shoots were cultured on Gamborg media. The result showed that the best level of both hormones IBA and NAA for root induction was 35 ppm. Moreover the solidifying agent of Gelrite gave positive response by stimulating root at the basal-end.Keywords: Rooting induction, solidifying agent, in vitro cultures, auxin, sago palm


2016 ◽  
Vol 2 (1) ◽  
pp. 27
Author(s):  
Teuku Tajuddin ◽  
Karyanti . ◽  
Tati Sukarnih ◽  
Nadirman Haska

Pohon sagu (Metroxylon sagu Rottb.) mempunyai banyak keunggulan dibanding dengan tanaman-tanaman penghasil pati lainnya, khususnya karena memiliki produktivitas yang tinggi, tumbuh di area bantaran sungai dan rawa, yang merupakan lingkungan tidak sesuai bagi pertumbuhan tanaman-tanaman lain. Dalam rangka membangun suatu perkebunan sagu di area yang luas, maka sangat dibutuhkan anakan-anakan sagu yang ukurannya seragam dalam jumlah yang besar. Namun demikian, terbatasnya jumlah anakan yang seragam telah menjadi kendala bagi pengembangan perkebunan sagu. Sebagai alternatif, perbanyakan in vitro dengan induksi tunas langsung dilakukan untuk mendapatkan bibit-bibit sagu dengan genotip unggul secara masal. Anakan sagu yang diperoleh dari Propinsi Maluku digunakan sebagai sumber eksplan. Eksplan dikultur pada media MS dan B5 yang mengandung kombinasi hormon auksin dan sitokinin. Hasil penelitian menunjukkan bahwa perlakuan BAP 2.0 ppm dan NAA 2.0 ppm menghasilkan jumlah tunas terbanyak.Kata kunci: Auksin, sitokinin, in vitro, sagu, inisiasi tunas ABSTRACTSago palm (Metroxylon sagu Rottb.) has many advantages over other starch-producing crops especially for its higher yield, ability to grow along riverbanks and on swampy areas not suitable for other crops. With the purpose of establishing large-scale plantations, a large amount of uniform sago palm suckers are required. However, limited availability of uniform suckers has hindered the mass propagation and development of cultivated Sago palm. Alternatively, in vitro cultures were performed in order to obtain a large-scale of mass clonally propagation of superior genotypes of sago palm. The young suckers obtained from areas of Maluku Province were used as explants. In vitro culture was carried out through direct shooting. The explants were cultured on two kinds of media, which were MS and B5 media containing various growth hormones of auxins and cytokinins. The results showed that the treatment with BAP 2.0 ppm and NAA 2.0 ppm produced the highest number of shoots.Keywords: Auxin, cytokinin, in vitro, sago palm, shoot initiation


1970 ◽  
Vol 20 (2) ◽  
pp. 157-170 ◽  
Author(s):  
Richard M.S. Mulwa ◽  
Margaret M.A. Norton ◽  
Robert M. Skirvin

Abundant embryogenic callus was obtained from leaf and floral explants of "Chancellor" grape by continuous culture for 12 weeks on Nitsch and Nitsch basal medium supplemented with 9 μM 2, 4-D + 17 μM IASP + either 1 μM BA or 1 μM TDZ (ECIM) in darkness. They were successfully maintained by a five to six week subculture interval on NN medium containing 2 μM 2, 4-D + 0.2 μM TDZ + 4 μM IASP (LTMM). Near synchronous embryo developed from embryogenic callus on medium containing 10 μM IASP + 8 μM NOA + 1 μM TDZ + 1 μM ABA + 2.5 g/l AC (EDMM).  Individually separated somatic embryos were germinated on both NN and half strength of MS containing 0.5 μM BA + 0.025 μM NAA, respectively; normal plantlet conversion from embryos was low (35%).  Whole fruiting plants were obtained. Aberrant embryo development was characterized by failure to form functional shoot meristems following the initial cotyledon expansion during germination. These observations indicate that the embryo conversion stage of the regeneration is difficult and remains a limiting factor requiring more empirical experimentation for improvement in grape tissue culture.   Key words: Chancellor grape, Regeneration, Somatic embryogenesis   D.O.I. 10.3329/ptcb.v20i2.6895   Plant Tissue Cult. & Biotech. 20(2): 157-170, 2010 (December)


2016 ◽  
Vol 3 (2) ◽  
pp. 71
Author(s):  
Nur Ajijah ◽  
RR. Sri Hartati

<p><em>Information on the effect of cytokinins on cacao (</em>Theobroma cacao<em> L.) primary somatic embryogenesis and its interaction with explant types and genotypes is not yet known. This study aimed to evaluate the effect of cytokinins and its interaction with explant types and genotypes on cacao somatic embryogenesis. The study was conducted at tissue culture laboratory of IAARD, Bogor from April until December 2012 and October 2014 until February 2016. Three types of cytokinins i.e. kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M), thidiazuron (0.01, 0.02, and 0.04 </em><em>μ</em><em>M) and benzylaminopurine (0.55, 1.11, and 2.22 </em><em>μ</em><em>M) in combination with 9 </em><em>μ</em><em>M 2,4-D were tested for their effectiveness in inducing somatic embryogenesis from petals and staminoid explants of Cimanggu 1 genotype. Furthermore, three levels of kinetin (0.58, 1.16, and 2.32 </em><em>μ</em><em>M</em><em>) also in combination with 9 </em><em>μ</em><em>M 2,4-D were evaluated for their influences on the somatic embryogenesis from petals and staminoid explants of three cacao genotypes i.e. Sulawesi 02, ICCRI 04 and Cimanggu 3. The result demonstrated that 2.32 </em><em>μ</em><em>M kinetin and staminoids explant were more effective to induce cacao somatic embryogenesis of Cimanggu 1 genotype (7%, 0.23 embryos/explant). Additionally, there were interaction effects between the level of kinetin with explant types and genotype on the percentage of explants forming embryo at 12 weeks after culture. The highest percentage of somatic embryo formation was shown by ICCRI 04 genotype with the use of petals explant and a kinetin level of 1.16 </em><em>μ</em><em>M (31.85%), but not significantly different from the level of kinetin 2.23 </em><em>μ</em><em>M (25.55%). The formation of primary somatic embryos of cacao is largely determined by the type and level of cytokinins, type of explant, and genotype.</em></p>


2018 ◽  
Vol 41 (4) ◽  
Author(s):  
Marlúcia Souza Pádua ◽  
Raíssa Silveira Santos ◽  
Luciano Vilela Paiva ◽  
Vanessa Cristina Stein ◽  
Luciano Coutinho Silva

ABSTRACT Oil palm is a woody monocot of economic importance due to high oil production from its fruits. Currently, the conventional method most used to propagate oil palm is seed germination, but success is limited by long time requirements and low germination percentage. An alternative for large-scale propagation of oil palm is the biotechnological technique of somatic embryogenesis. The rooting of plants germinated from somatic embryos is a difficult step, yet it is of great importance for later acclimatization and success in propagation. The aim of this study was to evaluate the effect of the auxins indole acetic acid (IAA) and indole butyric acid (IBA) on the rooting of somatic embryos of Tenera hybrid oil palm. Plants obtained by somatic embryogenesis were inoculated in modified MS medium with 10% sucrose and 0.6% agar and supplemented with IAA or IBA at concentrations of 5 µM, 10 µM, and 15 µM, and the absence of growth regulators. After 120 days, the presence of roots, root type, length of the longest root, number of roots, number of leaves, and shoot length were analyzed. Growth regulators were favorable to rooting; plants cultivated with IBA growth regulator at 15 µM showed higher rooting percentage (87%) and better results for the parameters of number of roots (1.33) and shoot length (9.83).


HortScience ◽  
2016 ◽  
Vol 51 (9) ◽  
pp. 1148-1152 ◽  
Author(s):  
Jane Kahia ◽  
Margaret Kirika ◽  
Hudson Lubabali ◽  
Sinclair Mantell

Breeding work carried out during the period 1971–85 by the Coffee Research Institute, Ruiru, Kenya resulted in the release of a new improved hybrid Coffea arabica named Ruiru 11. The cultivar combines resistance to coffee berry disease (CBD) and leaf rust, with high yield and good cup quality attributes. The propagation by F1 hybrid seeds production, cuttings, and tip grafting do not produce enough planting materials. There was a need to explore alternative methods and tissue culture offers potential options. The objective of the study was to evaluate the effect of explant sources and cytokinins on induction and regeneration of somatic embryos. Eight different explants were cultured on half-strength Murashige and Skoog (MS) medium supplemented with 10 µm benzylaminopurine (BAP). The effect of kinetin, N6-(2-isopentyl) adenine (2iP) evaluated at (0, 0.5, 5, or 25 µm) or thidiazuron (TDZ) (0, 0.5, 1.0, or 5 µm) added in separate experiments was also evaluated. The percentage of embryogenic cultures and the numbers of embryos per explant were determined after 3 months’ culture. The explant type had a significant effect (P > 0.05) on the induction of somatic embryos. Explants from in vitro-germinated seedlings produced the highest embryogenic cultures (90%) and the highest mean number of embryos (19.36) per explant. Cytokinins strongly enhanced induction and regeneration of somatic embryos. TDZ at 1 µm produced the highest embryogenic cultures (100%) and the highest mean number of embryos (24.2). The embryos were germinated on half-strength MS medium without any hormones. A high (98%) survival rate of the regenerated plantlets was recorded over all the treatments in the greenhouse. This is the first report on induction of high-frequency direct somatic embryos from coffee juvenile tissues. This is of great significance in tissue culture and indeed molecular biology manipulations because it allows regeneration of coffee from several explants.


1990 ◽  
Vol 20 (11) ◽  
pp. 1759-1765 ◽  
Author(s):  
F. B. Webster ◽  
D. R. Roberts ◽  
S. M. McInnis ◽  
B. C. S. Sutton

To apply somatic embryogenesis to clonal propagation of forest species, the technique must be applicable to a broad range of genotypes and allow efficient regeneration of phenotypically normal plants. Seventy-one lines (genotypes) of embryogenic cultures from six open-pollinated families were obtained by culturing immature embryos of interior spruce. Interior spruce represents a mixture of two closely related species, Piceaglauca (Moench) Voss and Piceaengelmannii Parry, from the interior of British Columbia where they hydridize with one another. The abscisic acid dependent developmental profile (the proportion of rooty embryos, shooty embryos, precociously germinating embryos, and mature embryos over a range of abscisic acid concentrations) differed among genotypes, but in general, production of mature somatic embryos was highest at 40 and 60 μM abscisic acid. Treatment of mature embryos with a high relative humidity treatment resulted in partial drying of the embryos and upon rehydration, markedly enhanced germination of the eight genotypes tested. Within 1 week of being placed under germination conditions, somatic embryos treated with the high relative humidity treatment showed 80–100% germination for 12 of the genotypes, and most genotypes had germination rates of greater than 40%. Survival of "emblings" (germinants from somatic embryos) following transfer to soil, acclimatization, and first season's growth in the nursery was 80% or greater for most genotypes. Over 1200 emblings were tested for nursery performance, representing the first large-scale evaluation of conifer somatic embryos under exvitro conditions. Growth rates, final height, shoot and root morphology, and frost hardiness were similar for emblings and seedlings following the first growing season. These results indicate that somatic embryogenesis can be used for the production of planting stock for a range of interior spruce genotypes.


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