scholarly journals DEVELOPMENT OF TRANSFORMATION SYSTEM FOR COFFEA ARABICA USING PARTICLE GUN METHOD

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 661d-661 ◽  
Author(s):  
Chifumi Nagai ◽  
Zhuping Mai ◽  
Jerek Jong
Keyword(s):  
Somatic Embryos ◽  
Coffea Arabica ◽  
Culture Medium ◽  
Gene Transformation ◽  
Ms Medium ◽  
Particle Gun ◽  
Embryogenic Calli ◽  
Blue Mountain ◽  
Secondary Culture ◽  
Leaf Disks

Somatic embryogenesis of coffee has been studied for the purpose of obtaining target tissues for stable genetic transformation through use of a particle gun. Eight cultivars of C. arabica were selected for callus induction from leaves. Primary calli were induced within two weeks in over 98% of the leaf disks explanted on MS medium with 2,4-D and kinetin prior to treatment on a secondary culture medium. Somatic embryos were obtained from `Catuai' and `Blue Mountain' after six months from explanting. Somatic embryos were germinated in MS media and developed into plants. Somatic embryos and embryogenic calli are being used for gene transformation experiments with the helium-driven particle gun.

scholarly journals Somatic embryogenesis of hypocotyl derived calli from an eggplant cultivar

2020 ◽  
Vol 115 (1) ◽  
pp. 151
Author(s):  
Hajar SABET ◽  
Mahmood MALEKI ◽  
Maryam ABDOLI NASAB ◽  
Saeid MIRZAEI
Keyword(s):  
Plant Regeneration ◽  
Culture Medium ◽  
Shoot Elongation ◽  
Gene Transformation ◽  
Ms Medium ◽  
Embryogenic Calli ◽  
Hypocotyl Explants ◽  
Somaclonal Variations ◽  
Micro Propagation ◽  
Embryogenic Callus Induction

<p>Optimization of tissue culture and regeneration conditions of eggplant is necessary for achieving different goals such as gene transformation and the development of somaclonal variations. In this study, hypocotyl explants ware used to produce callus in a medium containing different concentrations of NAA and BAP. Moreover, the concentration of the elements Ca, Mn, Mg, Fe and K were measured and analysed between embryogenic and non-embryogenic calli. For shoot elongation, embryogenic calli were transferred to a new culture medium containing 3.5, 4 and 4.5 mg l<sup>-1</sup> BAP plus 2 mg l<sup>-1</sup> GA3. Finally, produced shoots were rooted in a culture medium containing 1, 1.5 and 2 mg l<sup>-1</sup> NAA. Results showed that the best treatment for the embryogenic callus induction was MS medium containing 0.5 mg l<sup>-1</sup> BAP plus 0.25 mg l<sup>-1</sup> NAA. Two elements, Fe and K, had the highest amount in non-embryogenic calli compare to the embryogenic one. For plant regeneration, MS medium containing 4.5 mg l<sup>-1</sup> BAP plus 2 mg l<sup>-1</sup> GA3 and 2 mg l<sup>-1</sup> NAA were the best treatments for shooting and rooting, respectively. In this study, the best treatments for plant regeneration produced 35 shoots from an explant with 92 % shooting. This regeneration protocol could be useful for gene transformation and micro-propagation studies.</p>

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 AUXIN PULSE IN THE INDUCTION OF SOMATIC EMBRYOS OF Eucalyptus

2019 ◽  
Vol 43 (3) ◽  
Author(s):  
Luciana Coelho de Moura ◽  
Aloisio Xavier ◽  
Ana Cláudia Ferreira da Cruz ◽  
Ricardo Gallo ◽  
Natane Amaral Miranda ◽  
...  
Keyword(s):  
Somatic Embryos ◽  
Culture Medium ◽  
Solid Medium ◽  
Eucalyptus Grandis ◽  
Pulse Treatment ◽  
Embryogenic Competence ◽  
Embryogenic Calli ◽  
Cotyledonary Explants ◽  
Semi Solid ◽  
Initial Source

ABSTRACT The objective of this study was to evaluate the effect of auxin pulse intervals on the induction of somatic embryos of Eucalyptus grandis x E. urophylla and to describe the embryogenic behavior of callus under the effect of auxinic stress. Cotyledons were inoculated in culture medium containing 207.07 µM picloram, a treatment considered as auxin pulse. Explants that were in the auxin pulse treatment were transferred to semisolid or liquid medium containing 20.71 µM picloram after one, two, four or eight days of auxin pulse. In a second experiment, explants that were on auxin pulse treatment were transferred to semi-solid medium containing 20.71 µM picloram after one, two or three days of auxin pulse. Auxiliary picloram pulse treatments (207.02 µM) can be used as an initial source of stress for the acquisition of embryogenic competence. The oxidation of cotyledonary explants may be considered as an indication of the formation of embryogenic calli. The presence of pectins in peripheral regions of somatic pro-embryos can be considered as a marker of somatic embryogenesis in cotyledonary explants of Eucalyptus grandis x E. urophylla.

scholarly journals Plant regeneration from protoplasts of Gentiana straminea Maxim

2016 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Guomin Shi ◽  
Lina Yang ◽  
Tao He
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Protoplast Culture ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Embryogenic Calli ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Solid Liquid

AbstractA protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N6-benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

scholarly journals Plant regeneration through somatic embryogenesis of yacón [smallanthus sonchifolius (poepp. and endl.) H. Robinson]

2009 ◽  
Vol 52 (3) ◽  
pp. 549-554 ◽  
Author(s):  
Cynthia Manyra Corrêa ◽  
Graciele Nicolodi de Oliveira ◽  
Leandro Vieira Astarita ◽  
Eliane Romanato Santarém
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Ms Medium ◽  
Embryogenic Calli ◽  
Efficient System ◽  
Smallanthus Sonchifolius ◽  
Tuberous Roots ◽  
Medicinal Use ◽  
Half Strength

Smallanthus sonchifolius has tuberous roots containing large amounts of fructo-oligosaccharides and its medicinal use has increased due to the hypoglycemic properties reported for this species. An efficient system for propagation via somatic embryogenesis is reported using petiole segments cultivated on MS medium supplemented with combinations of BA, kinetin and 2,4-D, under light and darkness conditions. Embryogenic callus was formed in most of the treatments; however, somatic embryogenesis was promoted by the presence of light. Clusters of somatic embryos appeared on callus surface after 50 days of culture. The highest number of embryos was produced on 0.45 µM BA and 4.5 µM 2,4-D. Embryogenic calli were maintained on MS medium containing 4.5 µM BA and 0.045 µM 2,4-D. Embryos converted on hormone-free half-strength MS medium with 2 g.L-1 activated charcoal and plantlets were transferred to non-sterile conditions for acclimatization, showing 100% of survival.

scholarly journals Somatic Embryogenesis in Four Tree Legumes

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Premananda Das
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Embryogenic Calli ◽  
Friable Embryogenic Callus ◽  
Napthaleneacetic Acid ◽  
Acacia Catechu ◽  
2,4 Dichlorophenoxyacetic Acid

Somatic embryogenesis was achieved in four leguminous tree species, that is, Acacia catechu, Acacia arabica, Hardwickia binata, and Dalbergia sissoo using immature zygotic embryos as explants on Murashige and Skoog (MS) medium supplemented with 0.25–1.0 mg/l Kn (kinetin) and 2.0–3.0 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) or NAA (1-napthaleneacetic acid) and 3% sucrose. MS medium containing 2.0 mg/l 2,4-D and 1.0–1.5 mg/l Kn was noted to be most effective in inducing friable embryogenic callus (FEC); the number of somatic embryos per culture varied in MS medium supplemented with 1.0–2.0 mg/l 2,4-D or NAA and 0.25–1.5 mg/l kinetin. The maximum number of somatic embryos was obtained in MS medium containing 1.5–2.0 mg/l 2,4-D or NAA and 1.0–1.5 mg/l kinetin; proliferation of embryogenic calli was enhanced in cultures having 1.0–2.0 mg/l 2,4-D, 1.0–1.5 mg/l kinetin, and 400–600 mg/l L-Proline. The somatic embryos in various shapes and sizes after the first subculture on MS medium supplemented with 0.1 mg/l IAA and 0.25 mg/l BA; developed shoots and rooted in strength MS medium supplemented with 0.1 mg/l IBA or IAA. The somatic embryo-derived plantlets were transferred to the field after being hardened in the climate-controlled hardening chamber.

Somatic Embryogenesis From Styles of Different Cultivars of Citrus limon (L.) Burm

1994 ◽  
Vol 42 (5) ◽  
pp. 587 ◽  
Author(s):  
FD Pasquale ◽  
F Carimi ◽  
FG Crescimanno
Keyword(s):  
Somatic Embryos ◽  
Culture Medium ◽  
Callus Formation ◽  
Basal Medium ◽  
Citrus Limon ◽  
Naphthaleneacetic Acid ◽  
Malt Extract ◽  
Ms Medium ◽  
Culture Initiation ◽  
Embryogenic Response

Plants were regenerated from styles of three cultivars of lemon (Citrus limon (L.) Burm.), 'Monachello', 'Lunario' and 'Femminello Santa Teresa'. Styles and stigmas were excised from flowers, and cultured on Murashige and Skoog (MS) basal medium. Callus formation occurred from the style base 2 weeks after culture initiation and was significantly (P < 0.01) greater in immature flowers growing on BAP supplemented medium. Somatic embryos appeared 8 weeks later. The addition of BAP in the culture medium increased significantly (P < 0.01) the embryogenic response of the style, and the best regeneration occurred on MS basal medium supplemented with 13.3 μM 6-benzylaminopurine (BAP) and 146 mM sucrose. Once embryos were transferred to MS medium supplemented with 500 mg L-1 malt extract and 0.27 μM α-naphthaleneacetic acid (NAA), they developed into plantlets. After 5 or 6 months of continuous subculturing, the embryogenic callus became habituated and new embryos were still arising.

scholarly journals Somatic Embryogenesis and Plant Development in Centella asiatica L., a Highly Prized Medicinal Plant of the Tropics

HortScience ◽  
2007 ◽  
Vol 42 (3) ◽  
pp. 633-637 ◽  
Author(s):  
Nirmal Joshee ◽  
Bipul K. Biswas ◽  
Anand K. Yadav
Keyword(s):  
Somatic Embryogenesis ◽  
Sodium Alginate ◽  
Somatic Embryos ◽  
Centella Asiatica ◽  
High Rate ◽  
Treatment Withdrawal ◽  
Induction Medium ◽  
Ms Medium ◽  
Research Experience ◽  
Embryogenic Calli

Past research experience with Centella asiatica micropropagation suggests a very high rate of contamination during the culture establishment stage. We demonstrate protocols for successful sterilization of Centella explants prepared from field-grown plants with an abundance of fungal and bacterial contamination. Sequential steps during sterilization and explant preparation process included a dip for 30 s in 70% ethyl alcohol, weak bleach treatment for 12 min, and a 60-min soak in plant preservative mixture before establishing cultures. We also report a reproducible system for somatic embryogenesis in Centella using leaf and stolon tip explants collected from naturally growing populations. Somatic embryos were induced within 3 to 4 weeks of culture in the dark on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D). Initial embryogenic mass appeared as nodular callus, which eventually developed into actual somatic embryos exhibiting globular, heart-shaped, and cotyledonary stages. Leaves produced embryogenic calli at 2.26 and 4.52 μm 2,4-D, whereas stolon tips were responsive only in the 9.04 μm 2,4-D treatment. Withdrawal of 2,4-D/growth regulators from the induction medium resulted in the maturation and further development of the embryos into plantlets. Regular subculturing of the embryogenic calli into MS medium sustained their regenarability for more than 1 year. Somatic embryos were individually encapsulated in sodium alginate and calcium chloride-based encapsulation matrix to produce artificial or synthetic seeds (synseeds). Synseeds with 2% sodium alginate were found best for the survival and germination recorded after their storage at 5 to 8 °C for 30 and 60 days. We report protocols for C. asiatica to reduce explant contamination before establishment of cultures on somatic embryo induction medium and efficient somatic embryogenesis to facilitate conservation and mass production of elite germplasm. This may further assist rapid dissemination of superior clones needed for research and commercial production.

scholarly journals A Comparative Study on Plant Regeneration from Protoplasts of Two Genotypes of Asparagus officinalis L.

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 872E-872
Author(s):  
Sandip Mukhopadhyay ◽  
Yves Desjardins
Keyword(s):  
Somatic Embryos ◽  
Culture System ◽  
Culture Media ◽  
Carbon Sources ◽  
Media Culture ◽  
Asparagus Officinalis ◽  
Ms Medium ◽  
Embryogenic Calli ◽  
Half Strength ◽  
Different Levels

The effects of culture media, culture modes, and carbon sources on plating efficiencies of protoplasts of two genotypes of Asparagus officinalis L. were investigated. Protoplasts grew best in a semisolid culture system containing half-strength MS medium with 1 mg NAA/liter, 0.5 mg zeatin/liter, 0.6 M glucose, and 0.1% Gelrite. The plating efficiencies were 12.5% and 8.1% for genotypes G 203 and G 171, respectively. Embryogenic calli were produced from protoplast-derived microcalli after culturing on MS medium with 1 mg 2,4-D, 3% sucrose, and 0.2% Gelrite. The somatic embryos were initiated, matured, and then germinated to plantlets in MS medium containing 0.1 mg NAA/liter, 0.3 mg 2-iP/liter (EMM), and different levels of carbohydrates. Transfer of somatic embryos from EMM with 10% glucose to EMM containing 2% sucrose produced the highest number of bipolar embryos and plantlets. The plantlets regenerated shoots and roots in MS medium with 3% sucrose, 0.1 mg NAA/liter, 0.1 mg kinetin/liter, and 1.28 mg ancymidol/liter. Cytological analysis of these plants revealed 2n = 20 chromosomes.

scholarly journals Pengaruh 2,4-D terhadap Induksi Embrio Somatik Kopi Arabika

2017 ◽  
Vol 10 (2) ◽  
pp. 82 ◽  
Author(s):  
Imron Riyadi ◽  
NFN Tirtoboma
Keyword(s):  
Somatic Embryos ◽  
Coffea Arabica ◽  
Plant Material ◽  
Culture Medium ◽  
Standard Medium ◽  
Arabica Coffee ◽  
Murashige Skoog ◽  
Young Leaves ◽  
Coffea Arabica L ◽  
Direct Induction

<p><strong>Abstract</strong></p><p>Direct induction of somatic embryos in Arabica coffee (Coffea arabica L.) using plant groth regulators (PGR's) has been successful. The concentration and combination of different kinds of PGR's can influence the response and success in embryo induction. An experiment was conducted to determine the optimal concentration of 2.4-D in combination with kinetin for direct induction and proliferation of somatic embryos. The plant material used was Arabica coffee var. Kartika-l originating from The Indonesian Coffee and Cacao Research Institute, Jember. Explants were taken from young leaves of reddish-green in color. Somatic embryos were induced directly on a Murashige-Skoog (MS) standard medium containing 30 g/l sucrose and supplemented with 0, 1, 2, 4, and 8 mg/l 2.4-D in combination with 0.1 mg/l kinetin each. The cultures were incubated in the dark at temperature 26oC and RH +60% for 6 weeks with 10 replications. The results showed that somatic embryogenesis in Arabica coffee was best induced in a culture medium wiyh 2.4-D at 4 mg/l, combination with 0.1 mg/l kinetin. Induction of somatic embryos was achieved at 100% 4 weeks after culture. Three morphological stages of embryo development were identified: globular, early heart, and middle heart. The embryos were of three distinct colors such as, yellowish, yellowish-white, and white. The highest rate of proliferation of somatic embryos was achieved at 2 mg/l, 2.4-D in combination with 0.1 mg/l kinetin averaging 68.53 embryos per explant 6 weeks after subculture.</p>

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