scholarly journals Abscisic Acid Effect on Improving Horse Chestnut Secondary Somatic Embryogenesis

HortScience ◽  
2012 ◽  
Vol 47 (12) ◽  
pp. 1741-1744 ◽  
Author(s):  
Dušica Ćalić ◽  
Nina Devrnja ◽  
Jelena Milojević ◽  
Igor Kostić ◽  
Dušica Janošević ◽  
...  

The effect of abscisic acid on the development of primary androgenic embryo and secondary somatic embryogenesis was investigated with the aim of improving multiplication rates and secondary somatic embryo quality in horse chestnut microspore and anther culture. The early embryo stage (globular) had a better response than late stages (heart, torpedo, and cotyledonary) in both types of cultures. Also, microspore culture had a high potential for mass secondary embryo production. The number of secondary somatic embryos was three times higher on hormone-free medium than on medium enriched with 0.01 mg·L−1 abscisic acid. However, most of the embryos on hormone-free medium had abnormal morphology. For this reason, abscisic acid was added to the media to improve embryo quality. The morphology of abscisic acid treated embryos was better than abscisic acid non-treated embryos. The optimal abscisic acid concentration for secondary somatic embryo induction and production of high-quality embryos was 0.01 mg·L−1. Overall, the effect of abscisic acid on the induction of secondary somatic embryogenesis and plant regeneration of androgenic embryos of this species may be helpful for the further synthesis of secondary metabolites in vitro and their application in the pharmaceutical industry.

1998 ◽  
Vol 25 (8) ◽  
pp. 909 ◽  
Author(s):  
F. Bagnoli ◽  
M. Capuana ◽  
M. L. Racchi

Catalase (CAT) and superoxide dismutase (SOD), two of the major antioxidant enzyme systems, were examined by native PAGE at different stages of zygotic and somatic embryogenesis of horse chestnut (Aesculus hippocastanum L.). During both zygotic and somatic embryogenesis, CAT and SOD specific activities increased, but electrophoretic analysis revealed remarkable differences in the isoenzyme patterns. Two CAT isoforms were differentially present during zygotic embryogenesis. The transition from the fast to the slow migrating form occurred in July, approximately 2 months after pollination. In contrast to zygotic, the two isoforms were continuously detectable during somatic embryo-genesis. In fact, with the exception of the callus stage, in which only one form was present, both of the CAT isoforms are equally active during the somatic embryo development. Unlike CAT, all SOD isoenzymes, one Mn-SOD and five Cu/Zn-SODs, were present during all the stages of zygotic embryo formation, but only Mn-SOD and an Fe-SOD were detected during somatic embryogenesis. These results suggest the occurrence of oxidative stress conditions during in vitro culture which, in horse chestnut, could account for the difficulties observed in the development of the somatic embryo into a plantlet.


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.


2014 ◽  
Vol 20 (2) ◽  
Author(s):  
Maria D. Artigas R ◽  
Rafael Fernandez Da Silva

<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>


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.


Plant Science ◽  
2016 ◽  
Vol 253 ◽  
pp. 98-106 ◽  
Author(s):  
Hugo Pacheco de Freitas Fraga ◽  
Leila do Nascimento Vieira ◽  
Catarina Corrêa Puttkammer ◽  
Henrique Pessoa dos Santos ◽  
Julio de Andrade Garighan ◽  
...  

1990 ◽  
Vol 68 (12) ◽  
pp. 2583-2589 ◽  
Author(s):  
S. M. Attree ◽  
T. E. Tautorus ◽  
D. I. Dunstan ◽  
L. C. Fowke

Somatic embryo maturation, germination, and soil establishment frequencies were compared for two conifer species, white and black spruce (Picea glauca and Picea mariana). The comparison of the two species regenerated and established in soil under the same conditions showed black spruce to be the most responsive. Shorter exposure times to 32 μM abscisic acid were not as effective as maturation on a medium containing 16 μM abscisic acid for 28 days. This gave similar maturation frequencies for the two species (6–8%), and germination frequencies of 64% for white spruce and over 73% for black spruce. Over 1800 black and white spruce plantlets were recovered, and more than 400 were transferred from in vitro to nonsterile conditions. Sixty percent (160) of the black spruce plantlets survived transfer and continued to grow vigorously. By comparison only 18% (29) of the white spruce plantlets survived, and half of these rapidly produced dormant buds and underwent no further shoot growth. White spruce plants that did not produce dormant buds grew vigorously. These results indicate that there are large differences in the ability of these closely related species to respond to plantlet establishment following regeneration from somatic embryos, and that black spruce is highly responsive to micropropagation by this method. Key words: Picea glauca, Picea mariana, somatic embryogenesis, maturation, germination, soil establishment.


2013 ◽  
Vol 8 (6) ◽  
pp. 591-599 ◽  
Author(s):  
Agata Ptak ◽  
Anna Tahchy ◽  
Edyta Skrzypek ◽  
Tomasz Wójtowicz ◽  
Dominique Laurain-Mattar

AbstractIn vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).


2000 ◽  
Vol 80 (2) ◽  
pp. 271-276 ◽  
Author(s):  
Lining Tian ◽  
Daniel C. W. Brown

Recovery of tissue culture-derived plants through somatic embryogenesis is a useful system for genetic engineering of soybean. The effect of abscisic acid (ABA) on soybean somatic embryogenesis, development, and maturation was investigated. ABA at 1, 10, 50, 100, and 500 µM were applied at different stages of embryo development; namely, at the globular stage in suspension culture, at the development stage and at the maturation stage on solid media. ABA promoted embryo growth and development when applied at the globular stage. Embryo size, after 15 d and after 1 mo on development medium, was significantly greater than that without exposure to ABA. ABA promoted normal embryo morphogenesis and 62% more normal embryos developed when embryos were treated with ABA at the globular stage. ABA treated-embryos showed an increased tolerance to partial desiccation (from 24% to 78%) and exhibited an increased germination capability relative to non-ABA-treated controls (54% versus 8%). Somatic embryos appeared to undergo a decreasing sensitivity to ABA during maturation. ABA did not show an effect when applied during embryo development and maturation stages. A protocol for more normal embryo formation and improved embryo germination is reported. Key words: Glycine max, somatic embryogenesis, in vitro culture


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