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 ◽  
...  

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.

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Jingli Yang ◽  
Songquan Wu ◽  
Chenghao Li

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.


Author(s):  
T.T.B. Phuong ◽  
V.P. Trung ◽  
N.H. An ◽  
N.D. Tuan ◽  
P.T.T. Nguyen

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.


HortScience ◽  
2004 ◽  
Vol 39 (2) ◽  
pp. 321-323 ◽  
Author(s):  
Hamid Elhag ◽  
Mahmoud M. El-Olemy ◽  
Mansour S. Al-Said

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).


2019 ◽  
Vol 6 (1) ◽  
pp. 83
Author(s):  
Sulastri Sulastri ◽  
Winda Nawfetrias ◽  
Djatmiko Pinardi ◽  
Henti Rosdayanti

In Vitro Somatic Embryogenesis and Plantlet Regeneration of Three Varieties of Alfalfa (Medicago sativa L.)ABSTRACTAlfalfa (Medicago sativa L.) is a valuable plant as a source of food for animal, forage, pharmaceutical, medicine, food supplement, and human consumption.  In vitro selection technology combined with induction or spontaneous mutagenesis has been effective in altering or isolating genetic variability for desirable characters.  Consequently, a reproducible in vitro propagation technique of that plant is mandatory. The aim of the research was to obtain information on the embryogenic callus induction, somatic embryogenesis, and plantlet regeneration of three varieties of alfalfa. The results showed that an optimum embryogenic callus induction (82%) was obtained on Murashige & Skoog (MS) basal medium containing 2 ppm 2,4-dichlorophenoxyacetic acid (2,4-D), 2 ppm kinetin and 2 ppm a-naphthaleneacetic acid (NAA). Those embryogenic calli could subsequently develop into somatic embryos, which germinated and regenerated into normal plantlets on R1 medium consisting of MS nutrients without the addition of plant growth regulator.Keywords: alfalfa, callus, embryogenic, plantlets, regeneration ABSTRAKAlfalfa (Medicago sativa) adalah tanaman berharga sebagai sumber makanan untuk hewan, yaitu hijauan pakan ternak, farmasi, obat-obatan, suplemen makanan dan konsumsi manusia. Teknologi seleksi in vitro yang dikombinasikan dengan induksi atau mutagenesis spontan telah terbukti efektif dalam mengubah atau mengisolasi variabilitas genetik untuk karakter yang diinginkan. Oleh sebab itu, keberhasilan teknik perbanyakan in vitro yang telah terbukti dapat direproduksi dari tanaman tersebut menjadi syarat yang harus terpenuhi. Tujuan dari penelitian ini adalah untuk mendapatkan informasi mengenai induksi kalus embriogenik, embriogenesis somatik dan regenerasi planlet dari tiga varietas alfalfa. Hasil penelitian menunjukkan bahwa induksi kalus embriogenik optimal (82%) didapat pada media Murashige & Skoog (MS) dengan  2 ppm 2,4-dichlorophenoxyacetic acid (2,4-D), 2 ppm kinetin dan 2 ppm a-naphthaleneacetic acid (NAA). Kalus embriogenik tersebut dapat membentuk embrio somatik, embrionya berkecambah dan beregenerasi membentuk planlet normal pada perlakuan media R1 yaitu nutrisi MS tanpa penambahan zat pengatur tumbuh.Kata Kunci: alfalfa, embriogenik, kalus, planlet, regenerasi


HortScience ◽  
2017 ◽  
Vol 52 (12) ◽  
pp. 1755-1758
Author(s):  
Ya-Long Qin ◽  
Xiao-Chun Shu ◽  
Wei-Bing Zhuang ◽  
Feng Peng ◽  
Zhong Wang

Callus induction and plant regeneration play a key role in transgenic technology. Although much progress has been made with respect to eggplant, this type of research is insufficiently developed in Solanum torvum (a wild relative of eggplant), which contains a large number of resistance genes. Here, a high-efficiency regeneration system of S. torvum was established. Stem segments and leaves were cultured on Murashige and Skoog (MS) medium supplemented with 0.5–3.0 mg·L−1 6-benzyladenine (6-BA) and 0.1–0.6 mg·L−1 α-naphthaleneacetic acid (NAA). The highest callus induction ratio (100%) was produced on MS + 1.0 mg·L−1 6-BA + 0.5 mg·L−1 NAA. The combination of 0.5 mg·L−1 BA and 1.0 mg·L−1 2,4-dichlorophenoxyacetic acid in MS medium (double microelement) was the best for plant regeneration. Well-developed shoots rooted on half-strength MS medium supplemented with 0.1 mg·L−1 indole-3-acetic acid (IAA). These results will be helpful for functional verification of resistance genes from S. torvum and may be useful to those working in the field of eggplant breeding.


1986 ◽  
Vol 64 (2) ◽  
pp. 355-361 ◽  
Author(s):  
S. Gleddie ◽  
W. A. Keller ◽  
G. Setterfield

Cell suspension cultures of eggplant (Solanum melongena L.) were initiated from embryogenic callus cultures and maintained in medium supplemented with either 1-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Higher yields of protoplasts were obtained from cells grown in 2,4-D than in NAA. The efficiency of cell division was also greater in protoplast cultures derived from cells grown in the presence of 2,4-D. Protoplast-derived cells formed somatic embryos in modified Kao or Nagata and Takebe media which were supplemented with 1 mg/L 2,4-D. Early stages of embryogenesis were observed in liquid medium; however, these embryos and associated cell colonies were transferred onto agar-solidified medium for subsequent regeneration. Mature plants were established in soil in the greenhouse.


2018 ◽  
Vol 41 (4) ◽  
Author(s):  
Marlúcia Souza Pádua ◽  
Raíssa Silveira Santos ◽  
Luciano Vilela Paiva ◽  
Vanessa Cristina Stein ◽  
Luciano Coutinho Silva

ABSTRACT Oil palm is a woody monocot of economic importance due to high oil production from its fruits. Currently, the conventional method most used to propagate oil palm is seed germination, but success is limited by long time requirements and low germination percentage. An alternative for large-scale propagation of oil palm is the biotechnological technique of somatic embryogenesis. The rooting of plants germinated from somatic embryos is a difficult step, yet it is of great importance for later acclimatization and success in propagation. The aim of this study was to evaluate the effect of the auxins indole acetic acid (IAA) and indole butyric acid (IBA) on the rooting of somatic embryos of Tenera hybrid oil palm. Plants obtained by somatic embryogenesis were inoculated in modified MS medium with 10% sucrose and 0.6% agar and supplemented with IAA or IBA at concentrations of 5 µM, 10 µM, and 15 µM, and the absence of growth regulators. After 120 days, the presence of roots, root type, length of the longest root, number of roots, number of leaves, and shoot length were analyzed. Growth regulators were favorable to rooting; plants cultivated with IBA growth regulator at 15 µM showed higher rooting percentage (87%) and better results for the parameters of number of roots (1.33) and shoot length (9.83).


1988 ◽  
Vol 66 (9) ◽  
pp. 1734-1742 ◽  
Author(s):  
Pierre R. Fobert ◽  
David T. Webb

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.


1970 ◽  
Vol 14 ◽  
pp. 31-38 ◽  
Author(s):  
M Rahman ◽  
M Asaduzzaman ◽  
N Nahar ◽  
MA Bari

Somatic embryos were obtained from cotyledon and midrib explants of Solanum melongena L., cultivar Loda. For callus induction, medium was supplemented with different concentrations of auxin singly or in combination with BAP. The best callusing 83-85% was obtained from both of the explants cultured on MS medium containing 2.0 mgl-1NAA + 0.05 mgl-1BAP. Somatic embryogenesis and shoot regeneration was achieved after transferring the calli to MS medium supplemented with BAP, GA3, NAA and Zeatin. Cotyledon derived calli showed better performance (87%) for regeneration than that of midrib (82%) when sub cultured on MS medium having 2.0 mgl-1 Zeatin + 1.0 mgl-1 BAP. For root induction, MS + 3.0 mgl-1 IBA was proved to be better treatment for average number (14-15) and mean length (12 cm) of roots than those of other treatments. Key words: Eggplant; cotyledon; midrib; callus induction; somatic embryo J. bio-sci. 14: 1-9, 2006


1989 ◽  
Vol 19 (2) ◽  
pp. 285-288 ◽  
Author(s):  
S. A. Merkle ◽  
A. T. Wiecko

Tissue cultures were initiated from developing seeds of black locust (Robiniapseudoacacia L.) collected from three trees at weekly intervals from 1 week following anthesis until early fruit maturity. Explants were cultured on media containing 0, 2, or 4 mg/L 2,4-dichlorophenoxyacetic acid and 0 or 0.25 mg/L 6-benzyladenine. Seeds explanted onto hormone-supplemented media remained on these media for 1 or 3 weeks before being placed on hormone-free media, or were maintained on hormone-supplemented media for the entire study. Direct somatic embryogenesis was observed in a single culture, initiated from a seed collected 4 weeks after anthesis and cultured for 1 week on a medium supplemented with 4 mg/L 2,4-dichlorophenoxyacetic acid and 0.25 mg/L 6-benzyladenine before transfer to basal medium. Although it could not be discerned from which part of the explant somatic embryos were derived, secondary embryogenesis continued from the radicles of cotyledonary-stage somatic embryos. Most somatic embryos were well formed, with two distinct cotyledons. Embryos germinated precociously, producing plantlets that were initially weak but later gained vigor and resembled seedlings.


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