scholarly journals In vitro Regeneration of Clematis Plants in the Nikita Botanical Garden via Somatic Embryogenesis and Organogenesis

2021 ◽  
Vol 12 ◽  
Author(s):  
Irina Mitrofanova ◽  
Natalia Ivanova ◽  
Tatyana Kuzmina ◽  
Olga Mitrofanova ◽  
Natalya Zubkova

The effects of growth regulators, namely, 6-benzylaminopurine (BAP) and thidiazuron (TDZ), on the morphogenic capacity of 13 cultivars of clematis plants, in terms of their morphological structure formation, shoot regeneration, and somatic embryo development, are presented. The clematis cultivars ‘Alpinist,’ ‘Ay-Nor,’ ‘Bal Tsvetov,’ ‘Crimson Star,’ ‘Crystal Fountain,’ ‘Kosmicheskaya Melodiya,’ ‘Lesnaya Opera,’ ‘Madame Julia Correvon,’ ‘Nevesta,’ ‘Nikitsky Rosovyi,’ ‘Nikolay Rubtsov,’ ‘Serenada Kryma,’ and ‘Vechniy Zov’ were taken in collection plots of the Nikita Botanical Gardens for use in study. After explant sterilization with 70% ethanol (1 min), 0.3–0.4% Cl2 (15 min), and 1% thimerosal (10 min), 1-cm long segments with a single node were introduced to an in vitro culture. The explants were established on the basal MS medium supplemented with BAP (2.20–8.90 μM) and 0.049 μM NAA, or TDZ (3.0; 6.0, and 9.0 μM) with 30 g/L sucrose and 9 g/L agar. The medium with 0.89 μM BAP served as the control. Culture vessels and test tubes with the explants were maintained in plant growth chamber-controlled conditions: with a 16-h photoperiod, under cool-white light fluorescent lamps with a light intensity of 37.5 μmol m–2 s–1, at a temperature of 24 ± 1°C. Histological analysis demonstrated that adventitious bud and somatic embryo formation in studied clematis cultivars occurred at numerous areas of active meristematic cell zones. The main role of plant growth regulators and its concentrations were demonstrated. It was determined that maximum adventitious microshoot regeneration without any morphological abnormalities formed on the media supplemented with BAP or TDZ. 4.40 μM BAP, or 6.0 μM TDZ were optimal cytokinin concentrations for micropropagation. The explants of ‘Alpinist,’ ‘Ay-Nor,’ ‘Crimson Star,’ ‘Crystal Fountain,’ ‘Nevesta,’ and ‘Serenada Kryma’ cultivars displayed high morphogenetic capacity under in vitro culturing. During indirect somatic embryogenesis, light intensity 37.5 μmol m–2 s–1 stimulated a higher-number somatic embryo formation and a temperature of 26°C affected somatic embryo development. Active formation of primary and secondary somatic embryos was also demonstrated. 2.20 μM BAP with 0.09 μM IBA affected the high-number somatic embryo formation for eight cultivars. Secondary somatic embryogenesis by the same concentration of BAP was induced. The frequency of secondary somatic embryogenesis was higher in ‘Crystal Fountain’ (100%), ‘Crimson Star’ (100%), ‘Nevesta’ (97%), and ‘Ay-Nor’ (92%) cultivars. Based on these results, the methodology for direct somatic embryogenesis and organogenesis of studied clematis cultivars has been developed.

HortScience ◽  
2000 ◽  
Vol 35 (4) ◽  
pp. 568B-568a
Author(s):  
Lianghong Chen ◽  
Ajmer S. Bhagsari ◽  
Soon O. Park ◽  
Sarwan Dhir

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.


2004 ◽  
Vol 161 (2) ◽  
pp. 229-236 ◽  
Author(s):  
Dunja Leljak-Levanić ◽  
Nataša Bauer ◽  
Snježana Mihaljević ◽  
Sibila Jelaska

2018 ◽  
Vol 8 ◽  
pp. 1293-1308
Author(s):  
D. K. Isutsa ◽  
R. N. Mayoli ◽  
A. B. Nyende ◽  
C. M. Mweu

Coffee is one of the most important crops cultivated in the world for use in beverages and confectionaries. Embryogenesis is a complex process that begins with a single cell and ends with the formation of mature embryos. Somatic embryo development involves accumulation of complex metabolites and storage reserves. This present experiment identified and quantified endogenous phytocomponents and amino acids present during somatic embryogenesis of ‘Ruiru 11’. Laboratory experiments for this study were set up in the Coffee Research Institute, Kenya at Ruiru. Third leaf pair explants were excised from 8-month-old greenhouse-grown mother plants sterilized and cultured in half strength Murashige and Skoog basal salts augmented with Thidiazuron. Once embryos had developed, the cultures were analysed for phytocomponents using GCMS and HPLC. The results showed that palmitoleic and stearic acids were highest (23.3 µg/g and 69.9 µg/g respectively) in brown embryogenic cultures. Cis 7,8 epoxy-2-methyl octadecane was highest (253 µg/g) in green embryogenic cultures. (Z)-3-Tetradecene was highest (25 µg/g) in brown non-embryogenic cultures. Z, Z-3,13- Octadecedien-1-ol and (Z)-7-Hexadecenal were highest (32.1 µg/g and 70.2 µg/g respectively) in green embryogenic cultures. Alanine content was highest (4.4 µg/g) in embryos of brown cultures. Amino acids, fatty acids and their derivatives are potential biomarkers for embryogenesis. Other phytocomponents should be identified and their role in coffee somatic embryogenesis determined. Further studies regarding the status of the phytocomponents identified in the present study, especially in particular stages of embryo development are needed to propose treatments to improve coffee somatic embryo development.


2001 ◽  
Vol 13 (2) ◽  
pp. 196-223 ◽  
Author(s):  
VÍCTOR M. JIMÉNEZ

Different aspects of the in vitro somatic embryogenesis regulation are reviewed in this paper.work. A description of g General aspects, such as terminology, uses, stages of development and factors associated with the somatic embryogenesis, are described. is carried out. Although a brief description ofn the effects of the addition of different plant growth regulators to the culture medium wasis given, the article is centereds itself on the effect that the endogenous hormone concentrations in the initial explants and in the tissue cultures derived from them could play oin the induction and expression of somatic embryogenesis. It is significant that few to emphasize the low amount of systematic studies have been conducted, in this subject, in which different species and hormone groups were compared in cultures with and without embryogenic capacity. Moreover, the lack of correlation between the results presented in different studies the distinct works indicates that the hormone content of the cultures is not the only factor involved.


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