Kultur Embrio Tiga Spesies Kopi pada Umur Buah dan Formulasi Media yang Berbeda

<em>Studying the fruit age and proper media formulation is one of the important stages in embryo culture of coffee. The data is highly benefical, especially in saving embryos generated from intra- and inter-species crosses that fall prematurely or experience problems in germination. The aim of this study was to determine the suitable age and media formulation for embryo culture of Arabica, Robusta, and Liberica coffee. The study was conducted at the Tissue Culture Laboratory, Indonesian Industrial and Beverage Crops Research Institute from January 2019 to November 2020. Murashige and Skoog (MS) media with growth regulators adapted to embryonic development were used in this study. The three types of coffee divided into 5 groups, namely pinhead, immature, early mature, almost mature, mature, and used as planting material. The research was designed in a completely randomized design with 10 replications, and media formulation as a treatment. The results showed that embryo culture of the three coffee species was conducted successfully, except for pinhead fruit. The older the cultured fruit, the higher the percentage of germination. There is a difference in germination time between the three coffee species. The medium for embryo culture should be adjusted with the age of the fruit being cultured. Aside from growing embryos, the cultured mature fruit embryos on MS medium given 0.5 mg/l BA can also be used for propagation by utilizing the secondary somatic embryos formation.</em>

The evaluation of java fine flavor cocoa propagation through somatic embryogenesis technique for germplasm preservation

Somatic embryogenesis is one of the newest technology that applied for the mass production of cocoa. This research aims to evaluate the regeneration rate of somatic embryos through somatic embryogenesis propagation techniques on java fine flavor cocoa. Cultivars in this study are ICCRI 01, ICCRI 02, DR 1, DR 2, DRC 16, DR 38, PNT 16, and PNT 30. Observations include parameters to determine the percentage of primary callus and embryogenic callus formation and the number of somatic embryos produced. Based on data, the ability of callus to produce primary embryos is highly dependent on plant cultivars and explant sources. Five cultivars showed a higher regeneration rate using explants from the petal part, while the rest showed a higher regeneration rate using explants from the staminode section. Embryogenic callus from each cacao cultivar has the same basic structure: a nodular friable structure consisting of many embryonic cells. Some fine flavor cacao cultivars that were able to produce callus and primary somatic embryos could not produce secondary somatic embryos and plantlets. However, two cultivars, which had low potential in producing primary embryos, had the high ability to produce secondary somatic embryos and develop into plantlets.

Morpho-anatomical characterization of secondary somatic embryogenesis in Azadirachta indica (Meliaceae)

Background and Aims: Meliaceae species are extremely recalcitrant during germination and in vitro processes. Therefore, this research focuses on characterization and optimization of a highly efficient system by secondary somatic embryogenesis in Azadirachta indica, which is an important step for enhancing secondary metabolite production and regeneration in recalcitrant species.Material and Methods: Leaf and cotyledon sections were induced in MS medium supplemented with benzylaminopurine (BAP) alone, or combined with 1-naphthaleneacetic acid (NAA) and, abscisic acid (BA) with thidiazuron (TDZ).Key results: Azadirachta indica developed primary somatic embryos with BAP. Shoot and root formation occurred at low concentrations of BAP, while somatic embryogenesis was favored under high levels of BAP or TDZ. Primary and secondary somatic embryos were evidenced continuously and asynchronously. The highest amount of somatic embryos was obtained with cytokinins. However, the concentration might be significant to differentiate between primary and secondary embryos. Moreover, the auxins are key for inducing histodifferentiation in embryos. Shoot induction occurred after transfer of the embryos to hormone-free MS medium. The shoots were rooted in MS1/2.Conclusions: The secondary somatic embryos were distinguished and characterized during the whole process and the efficient system was established with cotyledon sections at short term, which offers several advantages such as the production of metabolites.

EFFECTS OF EXPLANT TYPE, CULTURE MEDIA AND PICLORAM AND DICAMBA GROWTH REGULATORS ON INDUCTION AND PROLIFERATION OF SOMATIC EMBRYOS IN EUCALYPTUS GRANDIS X E. UROPHYLLA1

ABSTRACT The objective of the present study was to test the effects of explant type, auxin concentrations, culture media, and auxin concentrations on the induction and proliferation of somatic embryos of Eucalyptus grandis x E. urophylla. Seeds and cotyledons were used as explants and inoculated in culture media containing 1.13, 2.26, 3.39 and 4.52 µM dicamba or 4.14, 10.35, 20.71 and 31.06 µM picloram. Embryogenic calli induced in the picloram treatments were used as explants and inoculated in semisolid or liquid media containing 4.14, 10.35, 20.71 and 31.06 µM picloram and keeping the origin of the embryogenic callus (seeds or cotyledons) and the concentration of picloram in those who were in the induction phase. Statistical, descriptive and anatomical analyses were performed. Induction of somatic pro-embryos into the juvenile plant material of Eucalyptus grandis x E. urophylla was performed using seeds or cotyledons as the source of explants, with the addition of dicamba and picloram as growth regulators. The use of cotyledons as a source of explants and the concentration of 4.1 µM picloram added to the culture media resulted in a higher induction of somatic pro-embryos. Proliferation of secondary somatic embryos was achieved using liquid medium added with picloram.

The Induction of Primary and Secondary Somatic Embryo to Support Arabica Coffee Propagation

The primary and secondary somatic embryogenesis can be used to propagate Coffea arabica L clonally. However, the success of this propagation was depended on plant growth regulator and varieties. This study aimed to examine the possibility of 2,4-D and thidiazuron application to form primary and secondary somatic embryo to support Arabica coffee clonal propagation. The study consisted of two activities (1) 2,4-D and thidiazuron Application to Induce Primary Somatic Embryogenesis of Arabica Coffee and (2) The Application of thidiazuron in Solid and Semi-Solid Media to Induce Secondary Somatic Embryos. The results indicated significant effect of varieties and plant growth regulator on fresh weight, number of torpedo and germinated embryo. However, it showed no significant effect on callus formation percentage. The best medium to induce primary somatic embryogenesis depending on variety, on the treatment of 4.52 μM 2,4 -D +18.16 μM thidiazuron was the best for AS2K and Sigarar Utang varieties, S 795 at 4.52 μM 2,4-D + 9.08 μM thidiazuron, whereas Kartika at 4.52 μM 2.4-D + 13.62 μM thidiazuron. The morphology of coffee somatic embryo was normal. Primary somatic embryo was developed indirectly, whereas the secondary somatic embryo was directly. The application of 9.08 μM thidiazuron increased the percentage and number of secondary somatic embryos, hence enhancing number of Arabica coffee planlet. Keywords : Coffea arabica L, 2,4-D, thidiazuron, semi-solid media, Indirect somatic embryogenesis

Somatic Embryogenesis and Genetic Uniformity of Cassava Plants Regenerated from Secondary Somatic Cotyledons Preserved in Osmotic Agents

Somatic embryogenesis, plant regeneration and genetic stability of regenerants grown from cassava secondary somatic cotyledon preserved at 160C on medium containing mannitol or sorbitol alone and their combinations were investigated. Irrespective of osmotic agents in the medium, survival of cotyledon explant, frequency of somatic embryos, shoot induction, number of somatic embryo per explant, shoot elongation and rooting decreased as preservation period increased. The highest survival rate of cotyledon explants, frequency of somatic embryos, shoot induction and shoot elongation were observed on media containing 2% mannitol. However, the highest per cent rooting occurred on medium containing mannitol alone at 8 months after storage (MAS) and on media containing mannitol or sorbitol alone at 16 MAS. RAPD analysis suggested genetic uniformity among regenerants and their control plant. Osmotic preservation of secondary somatic embryos of cassava on 2% mannitol at 160C is the best slow?growth method.Plant Tissue Cult. & Biotech. 26(1): 47-54, 2016 (June)