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

2018 ◽  
pp. 7-20
Author(s):  
Maria Daniela Artigas Ramirez ◽  
Rafael Fernandez Da Silva
Keyword(s):  
Somatic Embryogenesis ◽  
Azadirachta Indica ◽  
Somatic Embryos ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Secondary Somatic Embryogenesis ◽  
Efficient System ◽  
Whole Process ◽  
Secondary Somatic Embryos

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.

scholarly journals ESTABLECIMIENTO DEL SISTEMA DE REGENERACION POR EMBRIOGÉNESIS SOMÁTICA DE Azadirachta indica A. Juss.

2014 ◽  
Vol 20 (2) ◽  
Author(s):  
Maria D. Artigas R ◽  
Rafael Fernandez Da Silva
Keyword(s):  
Somatic Embryogenesis ◽  
Azadirachta Indica ◽  
Culture System ◽  
Plant Metabolites ◽  
Secondary Somatic Embryogenesis ◽  
Short Term ◽  
Efficient System ◽  
Low Concentrations ◽  
Whole Plants

<p><em>Azadirachta indica</em>, es una planta con múltiples aplicaciones tanto forestal como farmacológica. Por ende, el establecimiento del sistema de cultivo <em>in vitro</em> por embriogénesis somática ofrece diversas y variadas ventajas, tales como obtener plantas altamente productivas en metabolitos. En este estudio, se utilizaron secciones foliares y cotiledonares, inducidas en medios MS (1962) suplementados con: BAP sólo y combinado con ANA/2,4-D, TDZ sólo y con ABA. La regeneración fue con MS sólo o con K + AIA y BAP + AIA. Como resultado se estableció un sistema eficiente con secciones de cotiledones, observándose organogénesis a bajas concentraciones de BAP, mientras a altos niveles de BAP (2,5 mg.L<sup>-1</sup>), así como  con TDZ + ABA (0,02 + 1mg.L<sup>-1</sup>) respectivamente favorecieron la embriogénesis somática primaria y secundaria en un 96 % y 71 % respectivamente. La regeneración fue 71 % con MS, mientras que el enraizamiento fue de 86,67 % con MS½, obteniéndose plantas completas a corto plazo. </p><p><strong>ABSTRACT</strong></p><p><em>Azadirachta indica</em>, is a plant with multiple forest and pharmacological application. Therefore, the establishment of <em>in vitro</em> culture system for somatic embryogenesis offers several distinct advantages such as obtaining highly productive plant metabolites. In this study, were used sections cotyledon and leaf, induced on MS medium (1962) supplemented with: BAP alone and combined with NAA / 2,4-D, TDZ alone and ABA. Regeneration was with MS alone or with K + BAP + IAA and IAA. As a result was established an efficient system with cotyledon sections, being observed organogenesis at low concentrations of BAP, while high levels of BAP (2.5 mg.L<sup>-1</sup>) with 96 % and TDZ + ABA (0.02 + 1mg.L<sup>-1</sup>) with 71 %, favoring the primary and secondary somatic embryogenesis. Regeneration was 71 % with MS; rooting was 86.67 % with MS½, presenting whole plants obtained short term.</p><p><strong>RESUMO</strong></p><p><em>Azadirachta indica, </em>é uma planta com várias aplicaçãos florestas e farmacológicas. Por conseguinte, a criação de sistema de cultur<em>a in vitro </em>para a embriogénese somática apresenta várias vantagens, tais como a obtenção de plantas altamente produtivas de metabolitos. Neste estudo, as seções de cotilédones e folhas, induzidas em meios suplementado en MS (1962) con: BAP sozinho e combinado com ANA / 2,4-D, TDZ e ABA sozinho. A regeneração foi apenas com MS ou com K + BAP + IAA e IAA. Como resultado, foi eficiente con seções cotiledones, observados organogênese em baixas concentrações de BAP, en quanto altos níveis de BAP (2,5 mg L-1) e com TDZ + ABA (0,02 + 1mg.L-1) favoreceu a embriogênese somática primária e secundária até 96 % e 71%, respectivamente. Regeneração estava com MS 71 %, en quanto que o enraizamento foi de 86,67 %, com ½ MS produzindo plantas inteiras curto prazo.</p><p><strong><br /></strong></p><p> </p>

scholarly journals High Efficiency Secondary Somatic Embryogenesis inHovenia dulcisThunb. through Solid and Liquid Cultures

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Jingli Yang ◽  
Songquan Wu ◽  
Chenghao Li
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Large Scale ◽  
High Efficiency ◽  
Secondary Embryogenesis ◽  
Secondary Somatic Embryogenesis ◽  
Liquid Cultures ◽  
Medicinal Tree ◽  
Plant Conversion

Embryogenic callus was obtained from mature seed explants on medium supplemented with 2,4-dichlorophenoxyacetic acid. Primary somatic embryos (SEs) can only develop into abnormal plants. Well-developed SEs could be obtained through secondary somatic embryogenesis both in solid and liquid cultures. Temperature strongly affected induction frequency of secondary embryogenesis. Relatively high temperature (30∘C) and germinated SEs explants were effective for induction of secondary somatic embryos, and low temperature (20∘C) was more suitable for further embryo development, plantlet conversion, and transplant survival. Somatic embryos formed on agar medium had larger cotyledons than those of embryos formed in liquid medium. Supplementing 0.1 mg L−16-benzyladenine (BA) was effective for plant conversion; the rate of plant conversion was 43.3% in somatic embryos from solid culture and 36.5% in embryos from liquid culture.In vitroplants were successfully acclimatized in the greenhouse. The protocol established in this study will be helpful for large-scale vegetative propagation of this medicinal tree.

scholarly journals Primary and secondary somatic embryogenesis in Jatropha curcas L. From leaf transverse thin cell layers

2018 ◽  
Vol 14 (4) ◽  
pp. 661-671
Author(s):  
Nguyen Thi Kim Loan ◽  
Do Dang Giap ◽  
Tran Trong Tuan ◽  
Nguyen Thi Thanh Hien ◽  
Nguyen Phuc Huy ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Jatropha Curcas ◽  
Somatic Embryos ◽  
Formation Rate ◽  
Ms Medium ◽  
Secondary Somatic Embryogenesis ◽  
Thin Cell Layers ◽  
Cell Layers ◽  
Secondary Embryos ◽  
Transverse Thin Cell Layers

An efficient method for plant regeneration in Jatropha curcas L. via primary and secondary somatic embryogenesis culture from ex vitro leaves of 6-month-old plants was presented in this study. Leaves were cut into transverse thin cell layers (tTCLs) and cultured on MS medium supplemented with kinetin (KIN) at 0.5, 1.0, 1.5, and 2.0 mg/l in combination with indole-3-butyric acid (IBA) at 0.1, 0.5, and 1.0 mg/l or 2,4-dichlorophenoxyacetic acid (2,4-D) at 1.0, 1.5 and 2.0 mg/l . The highest embryogenic callus formation rate (89.3%) was obtained on medium supplemented with 1.0 mg/l KIN and 1.5 mg/l 2,4-D. The calli were selected for the study of primary somatic embryogenesis on MS medium containing 2,4-D (0.01, 0.03, 0.05, and 0.07 mg/l) or KIN (0.5, 1.0, 1.5, and 2.0 mg/l). The highest primary somatic embryos formation rate (76.67%) was achieved on MS medium supplemented with 1.0 mg/l KIN. The primary embryos were cultured on medium supplemented with KIN (0.1, 0.5, 1.0, 1.5, and 2.0 mg/l) combined with 0.2 mg/l indole-3-butyric acid (IBA) or 0.05 mg/l 2,4-D. The combination of 1.5 mg/l KIN and 0.05 mg/l 2,4-D was suitable for secondary embryos formation. Embryos proliferated rapidly, and the highest number of secondary embryos (77.5 embryos) wasobtained from a single primary embryos inoculated. Results also showed that the addition of proline (0.75 g/l) or spermidine (0.15 mM) to the culture medium increased the number of secondary embryos considerably. The fully developed plantlets exhibiting healthy roots and shoots were obtained when somatic embryos were sub-cultured onto B5 medium containing 1.5 mg/l IBA.

scholarly journals The Effects of 2,4-Dichlorophenoxyacetic Acid and α-Naphthaleneacetic Acid on Biomass Increment, Rhizogenesis and Somatic Embryogenesis of Suspension-cultured Dinh Lang Cells [Polyscias fruticosa (L.) Harms]

2021 ◽  
Author(s):  
T.T.B. Phuong ◽  
V.P. Trung ◽  
N.H. An ◽  
N.D. Tuan ◽  
P.T.T. Nguyen
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Bioactive Compound ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Triterpenoid Saponins ◽  
Cell Biomass ◽  
2,4 Dichlorophenoxyacetic Acid

Background: Dinh Lang [Polyscias fruticosa (L.) Harms] is a medicinal plant widely grown in Vietnam, with proven note-worthy health benefits. However, Dinh Lang’s amounts of triterpenoid saponins could not meet the need of the pharmaceutical industry. Thus, this study’s purpose is to figure out the optimal condition for raising Dinh Lang’s cell biomass, rhizogenesis and somatic embryogenesis to provide materials for bioactive compound productions. Methods: Different 2,4-dichlorophenoxyacetic acid and α-naphthaleneacetic acid concentrations (0.5, 1.0, 1.5 and 2.0 mg/L) were examined to determine the best amount of each plant growth regulator for raising cells’ biomass, rhizogenesis and somatic embryogenesis. In each treatment, two grams of eight-week-old calli were cultured in 50 mL of liquid MS medium. Result: It is demonstrated by the results that liquid MS medium containing 1.5 mg/L α-naphthaleneacetic acid has the capacity of producing the highest numbers of somatic embryos (489 embryos per flask) and rooted cells (259.5 cells per flask), while the fresh weight of cells cultured in the medium given 1.5 mg/L 2,4-dichlorophenoxyacetic acid reached its peak of 5.7 g.

scholarly journals Development and regeneration of somatic embryos from leaves-derived calli of Coffea liberica

2020 ◽  
Vol 21 (12) ◽  
Author(s):  
FITRIA ARDIYANI ◽  
Edy Setiti Wida Utami ◽  
HERY PURNOBASUKI ◽  
SENJA APRILIA PARAMITA
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Somatic Embryos ◽  
Economic Value ◽  
Genetic Material ◽  
Clonal Plants ◽  
Ms Medium ◽  
Cotyledonary Stage ◽  
Coffea Liberica

Abstract. Ardiyani F, Utami ESW, Purnobasuki H, Paramita SA. 2020. Development and regeneration of somatic embryos from leaves-derived calli of Coffea liberica. Biodiversitas 21: 5829-5834. Coffea liberica is an important and potentially commercial plant with a high economic value from the Coffea genus. Therefore, the availability of planting material is needed to increase productivity and ensure the sustainability of its farming. Somatic embryogenesis is a powerful propagation method used to produce clonal plants from limited genetic material. In the present research, we have shown that C. liberica could be successfully regenerated in vitro via somatic embryogenesis from leaves derived embryogenic callus. These calli were cultured on Murashige Skoog (MS) medium added with 1 mgL-1 BAP or in combination with 2.4 D (0.5, 1.0, 1.5 and 2 mgL-1) for embryo development induction. Furthermore, the medium containing only BAP was best for embryo development induction after culturing for 12 weeks, with the highest number of cotyledonary stage embryos (17.8%) and producing a total of embryo (20.2). Following cotyledonary stage embryo were cultured on new MS medium containing 0.5 mgL-1 BAP, 0.5 mgL-1 IAA, 0.5 mgL-1 NAA only, and 0.5 mgL-1 BAP in combination with 0.5 mgL-1 IAA or 0.5 mgL-1 NAA. Interestingly, the results showed that cotyledonary stage embryos were converted into complete plants at all treatment, but the MS medium containing 0.5 mgL-1 BAP was found to be the most effective in promoting regeneration with 2.6 leaves per-plantlet and height of 5.2 mm. Based morphological analysis confirm that the development of somatic embryo from leaves-derived calli of Coffea liberica started with the formation of embryo globular, heart, torpedo, cotyledonary stages, and finally conversion of cotyledonary embryo into complete plant.

scholarly journals SOMATIC EMBRYOGENESIS OF CASSAVA (MANIHOT ESCULENTA CRANTZ) VAR. KM297

2011 ◽  
Vol 14 (3) ◽  
pp. 14-22
Author(s):  
Kien Van Vu ◽  
Sanh Du Nguyen
Keyword(s):  
Somatic Embryogenesis ◽  
Manihot Esculenta ◽  
Somatic Embryos ◽  
Light Condition ◽  
Ms Medium ◽  
Dark Condition ◽  
Manihot Esculenta Crantz ◽  
Immature Leaf

Somatic embryos of cassava var. KM297 received from pieces of in vitro immature leaf lobes or cotyledon of somatic embryos, were induced on the MS medium supplemented with 8mg/l picloram after 13 days inoculation in the dark condition. Different states of embryo were obtained after 10 days cultured on MS medium supplemented with 0.1 mg/l BA and 0.01mg/l NAA, in the light condition. Role of endogenous AIA and Zeatin of the globular state of embryos was studied.

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 Effect of Plant Growth Regulators on Somatic Embryogenesis and Plantlet Development of Turkey Berry (Solanum torvum SW)

2021 ◽  
pp. 1-8
Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Solanum Torvum ◽  
Plantlet Formation ◽  
Regenerated Plants

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18)  along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted  on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.

scholarly journals Somatic embryogenesis from Hoan ngoc(Pseuderanthemum palatiferum (Ness) Radlk) leaf callus

2014 ◽  
Vol 17 (2) ◽  
pp. 100-107
Author(s):  
Nhung Thi Tuyet Tran ◽  
Hoang Ngo Phan ◽  
Sanh Du Nguyen
Keyword(s):  
Somatic Embryogenesis ◽  
Biological Activity ◽  
Plant Growth ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Leaf Tissue ◽  
Ms Medium ◽  
The Mean ◽  
Endogenous Plant

Pseuderanthemum palatiferum contains many compounds which have pharmaceutical and medicinal values. In this study, the in vitro somatic embryogenesis of callus formed from Pseuderanthemum palatiferum’s leaves was performed and analyzed. The calli were induced in three weeks in MS medium with 1 mg.l-1 or 2 mg.l-1 of 2,4-D. The somatic embryos were developed as follows: the globular embryos formed after 9 days, the heart embryos formed after 13 days, the torpedo embryos formed after 15 days and the cotyledonary embryos formed after 17 days. The mean number of obtained embryos was 28.5 embryos/cm2 of leaf tissue. The intensity of respiration and biological activity of endogenous plant growth regulators of leaf tissue during culture were recorded.

scholarly journals Somatic embryogenesis from zygotic embryos of Euterpe oleracea Mart.

2002 ◽  
Vol 24 (3) ◽  
pp. 601-603 ◽  
Author(s):  
Ana da Silva Ledo ◽  
Osmar Alves Lameira ◽  
Abdellatif Kemaleddine Benbadis ◽  
Ilmarina Campos de Menezes ◽  
Maria do Socorro Padilha de Oliveira ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Zygotic Embryos ◽  
Ms Medium ◽  
Euterpe Oleracea ◽  
Mineral Salts ◽  
Vegetable Material ◽  
Morphogenetic Responses ◽  
Euterpe Oleracea Mart

The aim of this work was to study the morphogenetic responses of zygotic embryos of açai palm (Euterpe oleracea Mart.) submitted to several conditions of in vitro culture. Several research experiments were conducted, in laboratory, using vegetable material collected from açai palm plants at Embrapa Amazon Oriental, Belém-PA, Brazil. It was possible to verify the expression of a direct, repetitive and no-synchronized model of somatic embryogenesis in mature zygotic embryos cultivated in primary MS medium supplemented with 2,4-D (339.36 muM) and transferred to a secondary MS medium in the presence of NAA (0.537 muM) and 2iP (12.30 muM). The conversion of somatic embryos into seedlings was reached after 210 days with the transfer of the cultures to a third medium with sucrose and mineral salts concentrations reduced to a half, without growth regulators.

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