scholarly journals Keragaman morfologi selama perkembangan embrio somatik sagu (Metroxylon sagu Rottb.) Morphological variations during the development of somatic embryos of sago (Metroxylon sagu Rottb.)

2016 ◽  
Vol 74 (1) ◽  
Author(s):  
Pauline Destinugrainy KASI ◽  
. SUMARYONO
Keyword(s):  
Somatic Embryos ◽  
Developmental Stages ◽  
Morphological Variations ◽  
Embryo Size ◽  
Initial Culture ◽  
Secondary Embryos ◽  
Half Strength ◽  
Metroxylon Sagu ◽  
Average Size

Summary In vitro culture of sago (Metroxylon sagu Rottb.) on an agar-solidified medium consists of somatic embryos of different sizes, colors, and developmental stages.  One gram of mostly globular somatic embryos were cultured on a solid medium to observe their morphological variations with respect to embryo size, color, and developmental stage over one passage of six weeks culture.  The medium was a modified-MS medium with half-strength of macronutrients containing   0.01 mg/L ABA and 2 mg/L kinetin.  At the end of culture passage, fresh weight of embryo increased by 2.3 folds.  The embryo numbers increased by more than two times indicating the formation of secondary embryos.  The average size of sago somatic embryos did not change significantly over the culture period; however, the embryo size was already highly varied at the start and increased gradually as the embryo developed.  At the initial of culture,   33.7 % of the embryos were yellowish, 64.1 % were greenish, and 2.2% were reddish.  By the end of the culture the composition of yellowish embryos increased to 51.2 %, greenish embryo decreased to 42.5 % and red embryos increased to 6.3 %.  At the initial culture, 61 % of the embryos were at the globular, 9 % at heart-shape and 30 % at torpedo stage.  Generally globular embryos developed into later-stage embryos as the culture progressed, although almost 56% of the embryos remained at the globular stage after the sixth week.Ringkasan Kultur in vitro sagu (Metroxylon sagu Rottb.) pada medium padat terdiri dari embrio somatik dalam berbagai ukuran, warna, dan fase perkembangan.  Satu gram embrio somatik yang sebagian besar dalam fase globuler dikulturkan pada medium padat untuk mengamati keragaman morfologi embrio dalam hal ukuran, warna dan fase perkembangan dalam satu periode kultur enam minggu.  Medium kultur adalah MS modifikasi dengan setengah hara makro serta penambahan zat pengatur tumbuh ABA 0,01 mg/L dan kinetin 2 mg/L.  Pada akhir masa kultur bobot embrio segar meningkat 2,3 kali dibandingkan awal masa kultur.  Jumlah embrio juga mengalami peningkatan sebesar lebih dari dua kali yang menunjukkan adanya pembentukan embrio somatik sekunder. Ukuran rata-rata embrio tidak berubah secara signifikan selama masa kultur akan tetapi ukuran embrio telah sangat beragam pada awal kultur dan terus meningkat hingga akhir kultur. Warna embrio mengalami perubahan selama periode kultur.  Pada awal kultur dijumpai 33,7 % embrio berwarna kuning, 64,1 % embrio hijau, dan 2,2 % embrio merah.  Pada akhir kultur presentase embrio kuning meningkat menjadi 51,2 %, embrio hijau menjadi 42,5 %, dan embrio merah 6,3 %.  Pada awal kultur, dijumpai 61 % embrio pada fase globuler, 9 % fase bentuk-hati dan 30 % fase torpedo.  Umumnya embrio globuler berkembang menjadi embrio fase lanjut selama kultur berlangsung, namun 56 % embrio masih tetap dalam fase globuler pada minggu keenam.

scholarly journals Keragaman morfologi selama perkembangan embrio somatik sagu (Metroxylon sagu Rottb.) Morphological variations during the development of somatic embryos of sago (Metroxylon sagu Rottb.)

2016 ◽  
Vol 74 (1) ◽  
Author(s):  
Pauline Destinugrainy KASI ◽  
. SUMARYONO
Keyword(s):  
Somatic Embryos ◽  
Developmental Stages ◽  
Morphological Variations ◽  
Embryo Size ◽  
Initial Culture ◽  
Secondary Embryos ◽  
Half Strength ◽  
Metroxylon Sagu ◽  
Average Size

Summary In vitro culture of sago (Metroxylon sagu Rottb.) on an agar-solidified medium consists of somatic embryos of different sizes, colors, and developmental stages.  One gram of mostly globular somatic embryos were cultured on a solid medium to observe their morphological variations with respect to embryo size, color, and developmental stage over one passage of six weeks culture.  The medium was a modified-MS medium with half-strength of macronutrients containing   0.01 mg/L ABA and 2 mg/L kinetin.  At the end of culture passage, fresh weight of embryo increased by 2.3 folds.  The embryo numbers increased by more than two times indicating the formation of secondary embryos.  The average size of sago somatic embryos did not change significantly over the culture period; however, the embryo size was already highly varied at the start and increased gradually as the embryo developed.  At the initial of culture,   33.7 % of the embryos were yellowish, 64.1 % were greenish, and 2.2% were reddish.  By the end of the culture the composition of yellowish embryos increased to 51.2 %, greenish embryo decreased to 42.5 % and red embryos increased to 6.3 %.  At the initial culture, 61 % of the embryos were at the globular, 9 % at heart-shape and 30 % at torpedo stage.  Generally globular embryos developed into later-stage embryos as the culture progressed, although almost 56% of the embryos remained at the globular stage after the sixth week.Ringkasan Kultur in vitro sagu (Metroxylon sagu Rottb.) pada medium padat terdiri dari embrio somatik dalam berbagai ukuran, warna, dan fase perkembangan.  Satu gram embrio somatik yang sebagian besar dalam fase globuler dikulturkan pada medium padat untuk mengamati keragaman morfologi embrio dalam hal ukuran, warna dan fase perkembangan dalam satu periode kultur enam minggu.  Medium kultur adalah MS modifikasi dengan setengah hara makro serta penambahan zat pengatur tumbuh ABA 0,01 mg/L dan kinetin 2 mg/L.  Pada akhir masa kultur bobot embrio segar meningkat 2,3 kali dibandingkan awal masa kultur.  Jumlah embrio juga mengalami peningkatan sebesar lebih dari dua kali yang menunjukkan adanya pembentukan embrio somatik sekunder. Ukuran rata-rata embrio tidak berubah secara signifikan selama masa kultur akan tetapi ukuran embrio telah sangat beragam pada awal kultur dan terus meningkat hingga akhir kultur. Warna embrio mengalami perubahan selama periode kultur.  Pada awal kultur dijumpai 33,7 % embrio berwarna kuning, 64,1 % embrio hijau, dan 2,2 % embrio merah.  Pada akhir kultur presentase embrio kuning meningkat menjadi 51,2 %, embrio hijau menjadi 42,5 %, dan embrio merah 6,3 %.  Pada awal kultur, dijumpai 61 % embrio pada fase globuler, 9 % fase bentuk-hati dan 30 % fase torpedo.  Umumnya embrio globuler berkembang menjadi embrio fase lanjut selama kultur berlangsung, namun 56 % embrio masih tetap dalam fase globuler pada minggu keenam.

scholarly journals 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

2016 ◽  
Vol 69 (2) ◽  
Author(s):  
. SUMARYONO ◽  
Imron RIYADI ◽  
J.S. TAHARDI
Keyword(s):  
Developmental Stage ◽  
Camellia Sinensis ◽  
Embryo Culture ◽  
Somatic Embryos ◽  
Color Change ◽  
Fresh Weight ◽  
Morphological Variations ◽  
Initial Culture ◽  
Average Size

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. 

scholarly journals 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

2016 ◽  
Vol 69 (2) ◽  
Author(s):  
. SUMARYONO ◽  
Imron RIYADI ◽  
J.S. TAHARDI
Keyword(s):  
Developmental Stage ◽  
Camellia Sinensis ◽  
Embryo Culture ◽  
Somatic Embryos ◽  
Color Change ◽  
Fresh Weight ◽  
Morphological Variations ◽  
Initial Culture ◽  
Average Size

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. 

scholarly journals MORPHOLOGICAL CHANGES DURING THE DEVELOPMENT OF SOMATIC EMBRYOS OF SAGO (Metroxylon sagu Rottb.)

2016 ◽  
Vol 8 (2) ◽  
pp. 43 ◽  
Author(s):  
Pauline D. Kasi ◽  
Sumaryono Sumaryono
Keyword(s):  
Developmental Stage ◽  
Somatic Embryos ◽  
Large Scale ◽  
Developmental Stages ◽  
Morphological Changes ◽  
Petri Dish ◽  
Synthetic Seed ◽  
Globular Embryos ◽  
Metroxylon Sagu

Development of somatic embryos of sago (Metroxylon sagu Rottb.) on agar-solidified medium are highly varied producing heterogeneous seedlings. Understanding of this phenomenon may help in improving the cultural procedures and conditions of sago<br />somatic embryogenesis to obtain uniform seedlings in a large scale. This experiment was conducted at the laboratory for plant cell culture and micropropagation, Indonesian Biotechnology Research Institute for Estate Crops from January to March 2006 to examine morphological changes i.e. color and development stages of sago during their somatic embryo development on an agar-solidified medium. Twenty single globular somatic embryos of sago with specific color (yellowish, greenish, and reddish) were cultured in a Petri dish supplemented with a solid medium. The medium was a micronutrients-modified MS (MMS) with half strength of macronutrients containing 0.01 mg l-1 ABA, 2 mg l-1 kinetin, 20 g l-1 sucrose, 0.5 g l-1 activated charcoal, and 2 g l-1 gelrite. Parameter observed was the percentage of embryo’s number based on color and developmental stage. The result showed that at the end of 6-week culture passage, most originally greenish (80.8%) and reddish (95.8%) embryos remained unchanged in their colors, whereas almost half of the originally yellowish embryos turned to greenish and only 30%<br />remained yellowish. At the same time, single globular embryos have changed gradually into the next developmental stages, although not all of the embryos were germinated. The initial color of embryo affected the rate of the developmental stage changes. Yellowish and greenish globular embryos developed more rapidly into cotyledon or germinant stages at 58% and 55% respectively, in 6 weeks than the reddish ones (41%). Therefore, the yellowish and greenish embryos are the best sources of material for in vitro mass propagation and synthetic seed production of sago.

scholarly journals MORPHOLOGICAL CHANGES DURING THE DEVELOPMENT OF SOMATIC EMBRYOS OF SAGO (Metroxylon sagu Rottb.)

2016 ◽  
Vol 8 (2) ◽  
pp. 43
Author(s):  
Pauline D. Kasi ◽  
Sumaryono Sumaryono
Keyword(s):  
Developmental Stage ◽  
Somatic Embryos ◽  
Large Scale ◽  
Developmental Stages ◽  
Morphological Changes ◽  
Petri Dish ◽  
Synthetic Seed ◽  
Globular Embryos ◽  
Metroxylon Sagu

Development of somatic embryos of sago (Metroxylon sagu Rottb.) on agar-solidified medium are highly varied producing heterogeneous seedlings. Understanding of this phenomenon may help in improving the cultural procedures and conditions of sago<br />somatic embryogenesis to obtain uniform seedlings in a large scale. This experiment was conducted at the laboratory for plant cell culture and micropropagation, Indonesian Biotechnology Research Institute for Estate Crops from January to March 2006 to examine morphological changes i.e. color and development stages of sago during their somatic embryo development on an agar-solidified medium. Twenty single globular somatic embryos of sago with specific color (yellowish, greenish, and reddish) were cultured in a Petri dish supplemented with a solid medium. The medium was a micronutrients-modified MS (MMS) with half strength of macronutrients containing 0.01 mg l-1 ABA, 2 mg l-1 kinetin, 20 g l-1 sucrose, 0.5 g l-1 activated charcoal, and 2 g l-1 gelrite. Parameter observed was the percentage of embryo’s number based on color and developmental stage. The result showed that at the end of 6-week culture passage, most originally greenish (80.8%) and reddish (95.8%) embryos remained unchanged in their colors, whereas almost half of the originally yellowish embryos turned to greenish and only 30%<br />remained yellowish. At the same time, single globular embryos have changed gradually into the next developmental stages, although not all of the embryos were germinated. The initial color of embryo affected the rate of the developmental stage changes. Yellowish and greenish globular embryos developed more rapidly into cotyledon or germinant stages at 58% and 55% respectively, in 6 weeks than the reddish ones (41%). Therefore, the yellowish and greenish embryos are the best sources of material for in vitro mass propagation and synthetic seed production of sago.

scholarly journals In vitro Embryo Morphogenesis and Micropropagation of Dendrobium aggregatum Roxb.

2014 ◽  
Vol 23 (2) ◽  
pp. 241-249 ◽  
Author(s):  
Mohammad Musharof Hossain
Keyword(s):  
Seed Germination ◽  
Plant Growth ◽  
Developmental Stages ◽  
Cell Mass ◽  
Multiple Shoot ◽  
Seedling Development ◽  
Ms Medium ◽  
Multiple Shoot Buds ◽  
Half Strength

In vitro embryo morphogenesis and micropropagation of Dendrobium aggregatum Roxb. were described. The gradual developmental stages of embryos to seedlings were traced out. Within two weeks of culture the cells of undifferented embryos underwent repeated aniclinal and periclinal division producing a compact, green parenchymatous cell mass called spherule that emerged out by rupturing the testa. The spherules subsequently differentiated into greenish protocorms were considered as typical seed germination. Germination occurred on both (MS and Phytamax (PM) medium but MS medium proved to be more efficient. The primary protocorms underwent profuse proliferation through production of secondary (2º) protocorms when transferred to different plant growth regulators (PGRs) supplemented MS; the medium fortified with 2.0 mg/l BAP and 1.0 mg/l NAA proved to be most effective for induction of 2º protocorms and seedling development. Multiple shoot buds (MSBs) were induced in pseudobulb segments of the in vitro grown seedlings when cultured on different PGRs supplemented media; and the maximum number of MSBs were obtained MS + 2.0 mg/l BAP + 0.5 mg/l picloram. The MSBs underwent elongation and then they rooted when they were transferred to half strength of MS + 0.5 mg/l IAA. The well rooted plantlets were finally transferred to outside natural environment with 80% survival. D. O. I. http://dx.doi.org/10.3329/ptcb.v23i2.17525 Plant Tissue Cult. & Biotech. 23(2): 241-249, 2013  (December)

scholarly journals High-frequency Direct Somatic Embryogenesis and Plantlet Regeneration from Leaves Derived from In Vitro-Germinated Seedlings of a Coffea arabica Hybrid Cultivar

HortScience ◽  
2016 ◽  
Vol 51 (9) ◽  
pp. 1148-1152 ◽  
Author(s):  
Jane Kahia ◽  
Margaret Kirika ◽  
Hudson Lubabali ◽  
Sinclair Mantell
Keyword(s):  
Tissue Culture ◽  
High Frequency ◽  
Somatic Embryos ◽  
Coffea Arabica ◽  
Alternative Methods ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Embryogenic Cultures ◽  
Half Strength

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.

scholarly journals 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)

2018 ◽  
Vol 86 (1) ◽  
Author(s):  
Imron Riyadi ◽  
Darda EFENDI ◽  
Bambang S PURWOKO ◽  
Djoko SANTOSO
Keyword(s):  
Somatic Embryo ◽  
Shoot Apical Meristem ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Apical Meristem ◽  
Culture Methods ◽  
Sago Palm ◽  
Callus Differentiation ◽  
Metroxylon Sagu

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]

scholarly journals Somatic Embryogenesis in Centella asiatica, a Valuable Medicinal Plant

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 859C-859
Author(s):  
Nirmal Joshee* ◽  
Bipul K. Biswas ◽  
Anand K. Yadav
Keyword(s):  
Somatic Embryos ◽  
Bacterial Contamination ◽  
Developmental Stages ◽  
Basal Medium ◽  
Centella Asiatica ◽  
Perennial Herb ◽  
Culture Techniques ◽  
Nodular Callus ◽  
Further Development

Centella asiatica L. (Apiaceae family), also called `Indian Pennywort,' is a prostrate, faintly aromatic, and stoloniferous perennial herb with long petiolated leaves. In the Ayurvedic medicine, it is reputed as a nervine tonic along with antibacterial, antifeedant, antileprotic and wound healing properties. Centella contains glycosides, indocentelloside, brahmoside, and asiaticoside. Its leaves are rich in carotenoids and vitamins B and C. In vitro culture techniques which offer a viable tool for mass propagation of plants have recently become increasingly popular for conservation of rare, endangered and threatened medicinal plants germplasm. Centella tissue culture has been reported to experience high incidences of microbial contamination which drastically reduces survival of explants. Thus, the main purpose of this study was to develop an efficient micropropagation technique for Centella asiatica to reduce explant contamination and rapidly disseminate superior clones for research and production. Here we present induction and further development of somatic embryos, using Centella stolons as explants. Somatic embryos were induced in response to 2,4-D shock on MS medium. Initially, somatic embryos appeared as highly nodular callus and eventually developed into somatic embryos that exhibited globular, heart shaped and cotyledonary stages. After auxin shock, cultures were regularly transferred to MS basal medium where somatic embryos completed various developmental stages and then germinated to give rise to new plantlets. In this presentation, we will demonstrate complete protocols for the successful sterilization of Centella explants prepared from plants that had abundance of fungal and bacterial contamination.

scholarly journals Somatic Embryogenesis using Immature Zygotic Embryos from Developing Fruits of Papaya (Carica papaya)

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 783E-783
Author(s):  
S.K. Dhir ◽  
U.L. Yadava
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Carica Papaya ◽  
Induction Medium ◽  
Zygotic Embryos ◽  
Synthetic Seed ◽  
Factors Affecting ◽  
Secondary Embryos ◽  
Potential Use

An efficient protocol has been developed for the in vitro multiplication of papaya (Carica papaya L.) through somatic embryogenesis utilizing immature zgotic embryos. Somatic embryos were initiated on MS basel media supplemented with 5 mg·liter–1 2,4-D, 400 mg·liter–1 glutamine, and 6% sucrose. After culturing for 2 months, 65% of the explants became highly embryogenic. Each explant produced 50 to 80 embryos in 4 months on culture induction medium. Frequency of embryogenesis was increased (75 to 150 somatic embryos on 80% explants) upon supplementing medium with 4% maltose as a carbon source and 100 mg·liter–1 L-asparagine. The embryogenic callus appeared yellow and embryos at different stages of development were well-organized. On regular subculturing, these cultures continued to produce secondary embryos. Following their transfer to the hormone-free medium supplemented with 4% maltose, these embryos germinated. The somatic embryogenesis system is rapid, repetitive, and highly proliferative. Thus, this system may have a potential use in the development of synthetic seed and transgenic papaya plants. Details of important factors affecting somatic embryogenesis will be discussed.

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