scholarly journals Regeneration of Pharbitis nil from immature embryos by somatic embryogenesis

2014 ◽  
Vol 66 (2) ◽  
pp. 159-163 ◽  
Author(s):  
Alina Trejgell ◽  
Andrzej Tretyn
Keyword(s):  
Somatic Embryogenesis ◽  
Gibberellic Acid ◽  
Growth Medium ◽  
Somatic Embryos ◽  
Immature Embryos ◽  
Pharbitis Nil ◽  
Naphthaleneacetic Acid ◽  
Photoperiodic Induction ◽  
Root Region

Immature embryos of <em>Pharbitis nil</em> Chois. were used for the study. They were isolated from previously sterilized fruits and afterwards transferred to Murashige and Skoog (MS) growth medium with 0.8% agar. Immature embryos were cut across their axis. After 6-8 weeks of cultivation in the injury place (hypocotyl-root region) somatic embryos were formed. These embryos were isolated and each of them was transferred into a separate tube containing MS supplemented with naphthaleneacetic acid (NAA; in concentration 0.1 mg•dm<sup>-3</sup> ) and gibberellic acid (GA<sub>3</sub>; in concentration 0.5 mg•dm<sup>-3</sup>). Under these conditions about 26% of somatic embryos regenerated into complete plants. Two-three weeks after the photoperiodic induction flowers appeared on these plants. A few weeks after pollination normal seeds were developed from these flowers.

scholarly journals Efficient Somatic Embryogenesis and Plant Regeneration from Immature Embryos of Tapiscia sinensis Oliv., an Endemic and Endangered Species in China

HortScience ◽  
2014 ◽  
Vol 49 (12) ◽  
pp. 1558-1562 ◽  
Author(s):  
Yuyu Wang ◽  
Faju Chen ◽  
Yubing Wang ◽  
Xiaoling Li ◽  
Hongwei Liang
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Activated Charcoal ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Immature Embryos ◽  
Zygotic Embryos ◽  
Naphthaleneacetic Acid ◽  
Embryogenic Calli ◽  
Induction Rate

High-frequency somatic embryogenesis and plant regeneration were achieved from immature cotyledonary-stage embryos in the endangered plant, Tapiscia sinensis Oliv. Plant growth regulators with different concentrations and combinations on embryogenesis capacity were studied. The optimal explants for in vitro somatic embryogenesis were immature embryos in T. sinensis. A high callus induction rate of 100% was achieved on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg·Ll−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5% (w/v) activated charcoal. Alternatively, a high induction rate (96.16%) of somatic embryogenesis was obtained on MS basal medium supplemented with the combination of 0.05 mg·L−1 α-naphthaleneacetic acid (NAA) and 0.2 mg·L−1 6-benzylaminopurine (6-BA), and somatic embryos proliferated fastest on the mentioned medium supplemented with 0.5% (w/v) activated charcoal and 3% (w/v) sucrose, inoculation of explants proliferating 21 times in the 23-day subculture. Of the 100 plantlets transferred to field after the acclimation, 95 (95%) survived. Based on the histocytological observations, the development of somatic embryos was similar to that of zygotic embryos. There were two accumulation peaks of starch grains in the embryogenic calli and in the globular-stage embryos, both closely related to the energy supply, and the embryoids were of multicelluar origin.

Effects of polyamines, polyamine precursors, and polyamine biosynthetic inhibitors on somatic embryogenesis from eggplant (Solarium melongena) cotyledons

1988 ◽  
Vol 66 (9) ◽  
pp. 1734-1742 ◽  
Author(s):  
Pierre R. Fobert ◽  
David T. Webb
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulator ◽  
Somatic Embryos ◽  
Second Messengers ◽  
Naphthaleneacetic Acid ◽  
Polyamine Synthesis ◽  
Nonspecific Effects ◽  
Competitive Inhibitors ◽  
Murashige And Skoog Medium ◽  
Low Levels

Eggplant (Solarium melongena L.) cotyledons grown on Murashige and Skoog medium with naphthaleneacetic acid formed callus, roots, and somatic embryos. Low levels of naphthaleneacetic acid (0.1 – 0.5 mg L−1) favoored rhizogenesis, intermediate levels (1.0 – 5.0 mg L−1) favoured embryogenesis, and high levels (10 – 50 mg L−1) favoured callogenesis. Addition of polyamines or their precursors did not induce morphogenesis on medium containing no growth regulator, nor did it affect embryogenesis on medium containing naphthaleneacetic acid, except at the highest concentrations tested, which were inhibitory. Enzyme-activated inhibitors of putrescine synthesis significantly reduced embryogenesis and stimulated rhizogenesis. α-Difluoromethylornithine was more potent in inhibiting embryogenesis and stimulating rhizogenesis than was α-difluoromethylarginine. α-Difluoromethylarginine did not inhibit growth and α-difluoromethylornithine stimulated growth. Addition of putrescine with α-difluoromethylornithine restored embryogenesis to control levels and reduced rhizogenesis. Competitive inhibitors of polyamine synthesis had nonspecific effects. Compared with seedling cotyledons, expiants grown on 5.0 mg naphthaleneacetic acid per litre contained slightly less free soluble putrescine and about the same amount of spermidine. At day 8, free putrescine and spermidine levels were higher in explants grown on naphthaleneacetic acid than in those grown on medium containing no growth regulator. Addition of α-difluoromethylornithine greatly reduced the putrescine and spermidine titres of the explants. Application of putrescine with α-difluoromethylornithine dramatically increased putrescine titres but not spermidine titres. Although the results suggest a role for polyamines in eggplant somatic embryogenesis, they do not support the hypothesis that polyamines act as auxin- or cytokinin-like growth regulators or as second messengers for auxin in this system.

scholarly journals Somatic Embryos Derived from Cotyledons of Cucumber

1990 ◽  
Vol 115 (4) ◽  
pp. 691-696 ◽  
Author(s):  
Rebecca M. Cade ◽  
Todd C. Wehner ◽  
Frank A. Blazich
Keyword(s):  
Tissue Culture ◽  
Acetic Acid ◽  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Culture Media ◽  
Naphthaleneacetic Acid ◽  
Histological Evidence ◽  
Tissue Culture Media ◽  
Maturation Medium ◽  
Dichlorophenoxy Acetic Acid

Two studies were conducted to test the effects of various tissue culture media on somatic embryogenesis from cotyledon tissue of cucumber (Cucumis sativus L.). The two best media for embryo initiation were Murashige and Skoog (MS) salts and vitamins containing either 1 or 2 mg 2,4-D/liter and 0.5 mg kinetin/liter. In the second study, embryos developed more normally. More plantlets developed when tissue was removed from the initiation medium after 3 weeks and transferred to MS containing 1 mg NAA/liter and 0.5 mg kinetin/liter for 3 weeks, rather than leaving the embryos on a medium containing 2,4-D. Histological evidence indicated that the embryos were multicellular in origin. Charcoal in the maturation medium inhibited embryo development. Chemical names used: (2,4-dichlorophenoxy) -acetic acid (2,4-D); N-(2-furanylmethyl)-lH-purine-6-amine (kinetin); 1-naphthaleneacetic acid (NAA).

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 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 Effect of Explant Resource and Growth Regulators on Somatic Embryogenesis and Plant Regeneration in Sweetpotato

HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 568B-568a
Author(s):  
Lianghong Chen ◽  
Ajmer S. Bhagsari ◽  
Soon O. Park ◽  
Sarwan Dhir
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryo ◽  
Embryo Development ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Shoot Tip ◽  
Naphthaleneacetic Acid ◽  
Somatic Embryo Development

This study was carried out to optimize conditions for plant regeneration of sweetpotato [Ipomoea batatas (L.) Lam] using shoot tips, petioles, and leaves of Selection 75-96-1 as explants in Murashige and Skoog (MS) with several growth regulators at different levels. Callus initiation and callus proliferation media were 9.0 μm 2,4-dichlorophenoxyacetic acid (2,4-D) and 9.0 μm 2,4-D + 1.1 μm N6-benzyladenine (6-BA) in protocol I; 8.1 μm α-naphthaleneacetic acid (NAA) + 1.2 μm kinetin (KIN) and 5.4 μm NAA + 4.6 μm KIN in protocol II; 0.9 μm 2,4-D, and 0.9 μm 2,4-D + 1.2 μm N-isopenylamino purine (2iP) in protocol III; NAA (8.1 μm) + KIN (1.2 μm) and 2,4-D (0.9 μm) + 2ip (1.2 μm) in protocol IV, respectively. In protocol I and II, shoot tip, petiole, and leaf were used, but only petiole and leaf in protocol III and IV. In the protocol I and II, somatic embryos were obtained only from shoot tip explants; in protocol III and IV, only from petioles. The frequencies of somatic embryo development were 33.3% in protocol I, 42.1% in protocol II, 21.2% in protocol III, and 10.3% in protocol IV, respectively. The leaf explants failed to produce somatic embryos in all the experiments. In protocol I, somatic embryogenesis occurred through the well-known sequence of globular-, heart-shaped-, torpedo-, and cotyledon-type embryos. However, in protocol II, the structures resembling plumule and radicle were observed before the emergence of torpedo/cotyledon type embryo clusters. The somatic embryogenesis in protocol III and IV was similar to that in protocol I. Growth regulators influenced somatic embryo development. Further, this study showed that explant resource and growth regulators affected the frequency of plant regeneration in sweetpotato.

scholarly journals High-Efficiency Somatic Embryogenesis from Seedlings of Koelreuteria paniculata Laxm.

Forests ◽  
2018 ◽  
Vol 9 (12) ◽  
pp. 769 ◽  
Author(s):  
Xiong Yang ◽  
Xiaoyu Yang ◽  
Ting Guo ◽  
Kai Gao ◽  
Tianyun Zhao ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Callus Induction ◽  
Somatic Embryos ◽  
Large Scale ◽  
High Efficiency ◽  
Naphthaleneacetic Acid ◽  
Clonal Culture ◽  
Embryogenic Calluses ◽  
Koelreuteria Paniculata ◽  
2,4 Dichlorophenoxyacetic Acid

Research Highlights: In the current study, we established a method for plant regeneration via somatic embryogenesis (SE) in Koelreuteria paniculata Laxm. for the first time. Background and Objectives: K. paniculata is an important ornamental and medicinal plant in China. However, the plant has difficulty with asexual reproduction, which imposes a limitation on large-scale propagation. Materials and Methods: Embryogenic calluses were induced from stems of aseptic seedlings on induction media. The effects of different media types and concentrations of N6-benzyladenine (BA), α-naphthaleneacetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D) on callus induction were examined. Embryogenic calluses were then transferred to Driver-Kuniyuki Walnut (DKW) media containing NAA (0.1–0.2 mg L−1) or 2,4-D (0.5–2.0 mg L−1) to develop somatic embryos. Cotyledon embryos were cultured on DKW media containing NAA (0.1–0.2 mg L−1) until maturation, and were then transferred to 1/2 DKW medium supplemented with 1.0 mg L−1 indole-3-butyric acid (IBA) to produce complete plants. The effects of IBA and NAA on rhizogenesis were then examined by clonal culture. Results: The maximum callus induction frequency (80.25%) was obtained on DKW medium supplemented by 0.5 mg L−1 BA, 0.25 mg L−1 NAA, and 1.5 mg L−1 2,4-D. NAA had a more pronounced effect on somatic embryo growth than did 2,4-D, with a maximum SE frequency (54.75%) observed with 0.1 mg L−1 NAA added to DKW medium. For clonal culture, the highest rooting rate (52%) was observed on 1/4 DKW medium containing 1.5 mg L−1 IBA. Histology studies confirmed the presence of embryogenic calluses and somatic embryos in different stages. Conclusions: This protocol provides a novel method for large-scale propagation of K. paniculata, and creates opportunities for genetic engineering in this species.

scholarly journals Enhancement of Somatic Embryogenesis and Production of Developmentally Arrested Embryos in Nigella sativa L.

HortScience ◽  
2004 ◽  
Vol 39 (2) ◽  
pp. 321-323 ◽  
Author(s):  
Hamid Elhag ◽  
Mahmoud M. El-Olemy ◽  
Mansour S. Al-Said
Keyword(s):  
Somatic Embryogenesis ◽  
Liquid Medium ◽  
Somatic Embryos ◽  
Nigella Sativa ◽  
Basal Medium ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Root Explants ◽  
Friable Callus ◽  
2,4 Dichlorophenoxyacetic Acid

Somatic embryogenesis of Nigella sativa was investigated with the objective of inducing and isolating somatic embryos for biosynthetic studies. Callus cultures were initiated from leaf, stem, and root explants of axenic seedlings on MSB5 basal medium supplemented with kinetin (0.46 μm) and 2,4-D (4.5 or 13.5 μm) or NAA (5.4 or 16.2 μm) in the dark. Cultures initiated and subcultured on medium containing NAA produced friable callus with numerous roots regardless of explant type. Cultures initiated, subcultured, or both, on medium with low 2,4-D concentration produced shiny embryogenic masses. These cultures differentiated into somatic embryos on medium containing NAA. The embryos developed into leafy structures on basal medium devoid of growth regulators. When the embryogenic callus was transferred to liquid medium containing NAA, numerous embryos and clusters of embryos were released into the liquid medium but, in contrast to solid medium, development remained arrested at the early embryonic stages. The developmentally arrested embryos were tested for production of active constituents of N. sativa oil. Chemical names used: 2,4-dichlorophenoxyacetic acid (2,4-D); α-naphthaleneacetic acid (NAA); kinetin (K).

Indirect somatic embryogenesis and morphohistological analysis in Capsicum chinense

Biologia ◽  
2010 ◽  
Vol 65 (3) ◽  
Author(s):  
Laura Solís-Ramos ◽  
Sara Nahuath-Dzib ◽  
Antonio Andrade-Torres ◽  
Felipe Barredo-Pool ◽  
Tomas González-Estrada ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Zygotic Embryo ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Capsicum Chinense ◽  
Regeneration System ◽  
Vascular Connection ◽  
Indirect Somatic Embryogenesis ◽  
Semi Solid

AbstractCapsicum chinense is recalcitrant in in vitro morphogenesis. No efficient, reproducible somatic embryogenesis regeneration system exists for this species, impeding regeneration from transformed cells. An indirect somatic embryogenesis protocol is developed using mature C. chinense zygotic embryo segments (ZES). The ZES cultured in semi-solid Murashige-Skoog (MS) medium supplemented with 8.9 μM naphthaleneacetic acid, 11.4 μM indoleacetic acid and 8.9 μM 6-benzylaminopurine, developed an embryogenic callus and 8% of the calli developed somatic embryos. Torpedo-stage somatic embryos were detached from the callus and subcultured in semi-solid MS medium without growth regulators, producing a 75% conversion rate to plantlets with well-formed root tissue. Histological analysis showed the developed structures to have no vascular connection with the callus and to be bipolar, confirming that this protocol induced formation of viable somatic embryos from mature C. chinense ZES. All acclimated plantlets survived under greenhouse conditions. This protocol will facilitate regeneration of genetically transformed plants using either biolistics or Agrobacterium tumefaciens approach.

Somatic embryogenesis and plant regeneration from immature embryos of Carica papaya × Carica cauliflora cultured in vitro

1991 ◽  
Vol 69 (9) ◽  
pp. 1913-1918 ◽  
Author(s):  
M. H. Chen ◽  
C. C. Chen ◽  
D. N. Wang ◽  
F. C. Chen
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Interspecific Hybrids ◽  
Carica Papaya ◽  
Immature Embryos ◽  
Embryogenic Cultures ◽  
Regenerated Plants ◽  
Hybrid Embryo

Somatic embryos were induced directly on immature embryos of Carica papaya × Carica cauliflora hybrids cultured on modified Murashige and Skoog's medium. When transferred to medium supplemented with abscisic acid, individual somatic embryos proliferated numerous daughter embryos through repeated embryogenesis. Light microscopic study of the repeatedly embryogenic cultures showed that daughter embryos arose from single superficial cells of parent embryos. Plant regeneration occurred following transfer of somatic embryos to medium devoid of plant growth regulators. Regenerated plants were intermediate between C. papaya and C. cauliflora in several morphological respects and showed isozyme patterns specific to both species as well as some new bands, indicating that they are indeed interspecific hybrids. Key words: Carica, interspecific hybrid, embryo culture, somatic embryogenesis.

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