Long-term in vitro Storage Technique of Valuable Birch Genotypes and Plant Production on its Basis

2019 ◽  
pp. 57-67
Author(s):  
T.M. Tabatskaya ◽  
N.I. Vnukova
Keyword(s):  
Plant Production ◽  
High Survival ◽  
Ex Vitro ◽  
Regenerative Capacity ◽  
In Vitro Storage ◽  
Interspecific Differences ◽  
Regenerated Plants ◽  
Growing Conditions

A technique for the long-term (up to 27 years) in vitro storage of valuable birch genotypes under normal (25 °C, 2.0 klx, 16-h day and 8-h night) and low temperature (4 °C, 0.5 klx, 6-h day and 18-h night) growing conditions on hormone-free media has been described. The study explored for the first time the influence of different strategies to store the clones of Betula pubescens and B. pendula var. сarelica (6 genotypes) on the regenerative capacity of collection samples, adaptive potential of regenerated plants and plant production by the in vitro and ex vitro techniques. It was established that both storage strategies provided a persistently high survival rate (82-100%) and regenerative capacity of in vitro shoots (the multiplication coefficient of 4.2-6.3 and rhizogenic activity of 90-100%). The clones retained their characteristics of height growth under the in vitro and ex vitro conditions, and demonstrated intraclonal homogeneity and lack of signs of somaclonal variability. The plants showed substantial interspecific differences at the stage of multiplication and transfer to the greenhouse. The highest percentage of acclimated plants (75-98% depending on the clone genotype) was obtained after planting of micro plants straight in the greenhouse, which simplified the technology and made plant production less costly. long-term in vitro storage, birch, species, genotype, micropropagation, ex vitro adaptation, plant material

scholarly journals Organogenesis and long-term micropropagatlon of Polish pea cultivars

2011 ◽  
Vol 72 (4) ◽  
pp. 295-302 ◽  
Author(s):  
Tomasz Pniewski ◽  
Joanna Wachowiak ◽  
Józef Kapusta ◽  
Andrzej B. Legocki
Keyword(s):  
De Novo ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Ex Vitro ◽  
Regenerated Plants ◽  
Axillary Meristems ◽  
Half Strength

The complete protocol for regeneration and long-term micropropagation of several Polish cultivars of pea (<em>Pisum sativum </em>L.) has been elaborated. The shoots were the most likely regenerated via de novo organogenesis. The adventitious buds formed in callus derived from cotyledons tissue adjacent to the axillary meristems of immature embryos. All cultivars' calli regenerated several shoots per explant on the MS medium supplemented with B5 vitamins and 4.5 mgl<sup>-1</sup> of BAP, however some differences in regeneration capacity among cultivars were observed. The plantlets were subsequently micropropagated with slightly higher efficiency and preserving a good viability over the long-term culture on a medium containing 2.0 mgl<sup>-1</sup> than one with 4.5 mgl<sup>-1</sup> of BAP. The additional step of the pre-conditioning culture of multiplicated shoots on a medium with very low BAP concentration i.e. 0.02 mgl<sup>-1</sup> was applied and appeared to be beneficial before rooting in vitro or grafting. The modified MS-derived medium with the half-strength of MS macroelements but with the full original dose of calcium and supplemented with B5 vitamins and 1.0 mgl<sup>-1</sup> of NAA was developed for effective rooting. The shoots were also sufficiently transferred into ex vitro conditions using grafting. The majority of the regenerated plants had adapted to in vivo conditions in a greenhouse and subsequently has set seeds. The presented protocol provides relatively efficient rate of de novo pea regeneration and would be useful for <em>Agrobacterium</em>-mediated transformation purposes.

scholarly journals Micropropagation of Tarenaya rosea (Cleomaceae) from leaf explants

Rodriguésia ◽  
2021 ◽  
Vol 72 ◽  
Author(s):  
Claudia Simões-Gurgel ◽  
Tatiana Carvalho de Castro ◽  
Cátia Henriques Callado ◽  
Lívia da Silva Cordeiro ◽  
Norma Albarello
Keyword(s):  
Large Scale ◽  
Plant Conservation ◽  
Leaf Explants ◽  
Plant Production ◽  
Anthocyanin Content ◽  
Alternative Source ◽  
Regeneration Rate ◽  
Culture Techniques ◽  
Regenerated Plants

Abstract In vitro culture techniques are recognized as efficient strategies for large-scale plant production, as well as providing alternatives for plant conservation. In this study the micropropagation of Tarenaya rosea was established using petiole and foliar blade segments cultivated on MS medium with 6-benzyladenine (BA) and/or 6-furfurylaminopurine (KIN). The regeneration rate from explants was evaluated after 30-days in culture, as well as the proliferation rate from explant-derived shoots, reached after four subcultures performed at 30-days in culture. In vitro propagation occurred by both direct (DO) and indirect (IO) organogenesis. The highest regeneration rates by DO (50% to 100%) were reached on media containing only BA, while morphogenic calluses (IO) were mainly formed with BA+KIN. Explants on media with BA showed the presence of small black nodules on their surface, and histological analysis revealed the presence of trichomes with anthocyanin content. Elongation and rooting were reached on growth regulator-free MS. Acclimatization rates around 80% were achieved and the in vitro-regenerated plants were successfully maintained under field conditions. Results show significant morphogenetic potential of T. rosea from leaf explants, mainly when cultivated in the presence of 4.4 µM BA, providing a new alternative source of plant material for biotechnological and in vitro conservation studies.

scholarly journals Ex Vitro Rooting of American Chestnut Improves Acclimatization Survival and Plantlet Quality

2016 ◽  
Vol 34 (3) ◽  
pp. 75-79 ◽  
Author(s):  
Allison D Oakes ◽  
Tyler R. Desmarais ◽  
William A. Powell ◽  
Charles A. Maynard
Keyword(s):  
Tissue Culture ◽  
Keystone Species ◽  
Ex Vitro Rooting ◽  
Plant Production ◽  
Shoot Cultures ◽  
Ex Vitro ◽  
Regulatory Review ◽  
Survival Percentage ◽  
Virus Indexing

Tissue culture of plants has many applications, from producing genetically identical horticultural varieties, to production of secondary metabolites, to virus indexing, and most relevantly, developing novel traits by genetic transformation. Using Agrobacterium-mediated transformation on somatic embryos, blight-resistant American chestnuts [Castanea dentata (Marsh.) Borkh.] have been developed as shoot cultures in plant tissue culture. Rooting tissue-cultured shoots and acclimatizing the rooted plantlets are key steps in tree production. In this study, in vitro and ex vitro rooting methods were compared. The ex vitro method resulted in a lower initial rooting percentage but an overall higher survival percentage, resulting in higher potted plant production. The higher survival was likely due to partial acclimatization taking place before the plantlets were transplanted into potting mix. After 8 weeks, plantlets rooted via the ex vitro method were taller, and had more, and larger, leaves than the in vitro-rooted plantlets. These trees are currently in high demand for inoculation studies for federal regulatory review and eventually for restoration of this keystone species to its native habitat.

scholarly journals Preliminary results of in vitro culture of pea and lupin embryos for the reduction of generation cycles in single seed descent technique

2013 ◽  
Vol 82 (3) ◽  
pp. 231-236 ◽  
Author(s):  
Maria Surma ◽  
Tadeusz Adamski ◽  
Wojciech Święcicki ◽  
Paweł Barzyk ◽  
Zygmunt Kaczmarek ◽  
...  
Keyword(s):  
Root Development ◽  
Ms Medium ◽  
Ex Vitro ◽  
Single Seed Descent ◽  
Yellow Lupin ◽  
Single Seed ◽  
Temperature Regimes ◽  
Regenerated Plants ◽  
Almost All

The aim of the studies was to establish in vitro conditions for the culture of pea and lupin embryos as the first step in the development of an in vitro assisted single seed descent technique for the attainment of homozygous populations. Materials for the study included of pea, and narrow-leafed and yellow lupin cultivars. Embryos dissected from mature but still-green seeds were cultured in vitro on two modified MS media and under three temperature regimes. Shoot and root lengths of regenerated plants were measured after 7, 14 and 21 days of culture. For pea plants full-strength MS medium with 4 g l<sup>−1</sup> agar and temperature 22/ 20°C (day/night) appeared to be the most conducive to shoot and root development, whereas for lupin plants lower temperatures were more propitious: 12°C in the dark for narrow-leafed lupin and 16/ 12°C (day/night) for yellow lupin. Almost all the cultured embryos developed into plants, but not all the regenerated plants survived acclimation to ex vitro conditions.

scholarly journals Inexpensive Screening Method to Validate the Efficacy of Ethanedinitrile Fumigant on the Forest Invasive Nematode Pest Bursaphelenchus xylophilus

2020 ◽  
Vol 12 (11) ◽  
pp. 4765
Author(s):  
Ondřej Douda ◽  
Václav Stejskal ◽  
Marie Manasova ◽  
Miloslav Zouhar ◽  
Jonáš Hnatek
Keyword(s):  
Screening Method ◽  
Global Scale ◽  
Bursaphelenchus Xylophilus ◽  
Laboratory Method ◽  
High Survival ◽  
Initial Screening ◽  
Innovative Method ◽  
International Laws

At a global scale, the sustainability of forests is endangered by multiple invasive species, including the pine wood nematode (Bursaphelenchus xylophilus), a quarantine pest. International laws and standards require that all exported wood coming from countries in which B. xylophilus is present be chemically or physically treated. Since a major fumigant, methyl bromide, was banned, there has been a need to generate data for alternative fumigants, such as ethanedinitrile (EDN), for this purpose. Since the field screening of fumigants (i.e., the application of various doses to and exposure times of naturally infested wood logs) is prohibitively expensive, the aim of this study was to develop a quick and inexpensive laboratory method. Here, we suggest and describe an innovative method based on sawdust cultures for EDN efficacy screening. In the validation part of this study, we demonstrated (i) the high survival of the nematodes in the sawdust and (ii) the high efficacy of EDN against this pest under in vitro conditions; 100% mortality was observed after 6 h of EDN exposure to a dose of 25 g/m3. In particular, our newly developed model system could be used for the initial screening of various doses of and exposure protocols for EDN and similar types of fumigants developed with the intention of regulating B. xylophilus occurrence in exported wood. It is believed that the validated method may help to develop new and effective EDN fumigation procedures and thereby contribute to the long-term protection of forests worldwide.

scholarly journals Influence of Polyethylene Glycol on Leaf Anatomy, Stomatal Behavior, Water Loss, and Some Physiological Traits of Date Palm Plantlets Grown In Vitro and Ex Vitro

Plants ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 1440
Author(s):  
Amal F. M. Zein El Din ◽  
Mohamed F. M. Ibrahim ◽  
Reham Farag ◽  
Hany G. Abd El-Gawad ◽  
Ahmed El-Banhawy ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Photosynthetic Pigments ◽  
Leaf Anatomy ◽  
Water Loss ◽  
Date Palm ◽  
High Survival ◽  
Ex Vitro ◽  
Stomatal Behavior ◽  
Ex Vitro Transfer

Few reports explain the mechanism of PEG action on stomatal behavior and anatomical structure and analyze the photosynthetic pigments of in vitro date palm plantlets for better tolerance to ex vitro exposure. The main challenge for in vitro micropropagation of date palm techniques remains restricted to high survival rates and vigorous growth after ex vitro transplantation. In vitro hardening is induced by Polyethylene glycol PEG (0.0, 10, 20, 30 g L−1) for 4 weeks. Leaf anatomy, stomatal behavior, water loss %, photosynthetic pigments, and reducing sugars were examined in date palm plantlets (Phoenix dactylifera L.) cv. (Sewi) after 4 weeks from in vitro PEG treatment and after 4 weeks from ex vitro transplanting to the greenhouse. Leaf anatomy and the surface ultrastructure of in vitro untreated leaves showed a thin cuticle layer, wide opened malfunctioning stomata, and abnormal leaf anatomy. Furthermore, addition of PEG resulted in increasing cuticle thickness, epicuticular wax depositions, and plastids density, improving the stomatal ability to close and decreasing the stomatal aperture length while reducing the substomatal chambers and intercellular spaces in the mesophyll. As a result, a significant reduction in water loss % was observed in both in vitro and ex vitro PEG treated leaves as compared to untreated ones, which exhibited rapid wilting when exposed to low humidity for 4 h. PEG application significantly increased Chlorophylls a, b and carotenoids concentrations, especially 10, 20 g L−1 treatments, which were sequentially reflected in increasing the reducing sugar concentration. However, leaves of plantlets treated with PEG at 30 g L−1 became yellow and had necrosis ends with death. In vitro hardening by 20 g L−1 PEG increased the survival rate of plantlets to 90% after ex vitro transfer compared to 63% recorded for the untreated plantlets. Therefore, this application provides normal date palm plantlets developed faster and enhances survival after ex vitro transfer.

scholarly journals Droplet vitrification technique for cryopreservation of a large diversity of blackcurrant (Ribes nigrum L.) cultivars

2020 ◽  
Author(s):  
Saija Rantala ◽  
Janne Kaseva ◽  
Anna Nukari ◽  
Jaana Laamanen ◽  
Saara Tuohimetsä ◽  
...  
Keyword(s):  
Cultural Values ◽  
Core Collection ◽  
Gene Bank ◽  
Ribes Nigrum ◽  
Ex Vitro ◽  
Droplet Vitrification ◽  
Successful Transfer ◽  
Future Utilization

AbstractThe aim of plant gene banks is to preserve genetic resources selected based on their phenotypic, agronomic, historical or other cultural values for future utilization. In the present study the modified PVS2 droplet vitrification technique was tested and optimized for cryopreservation of a large diversity of blackcurrant (R. nigrum L.) accessions propagated in vitro and selected into a national gene bank core collection. Out of four accessions tested to optimize the method, three recovered and regenerated by 89–97% on average, but one recalcitrant in vitro line only by 25%. The tested post-cryopreservation recovery media with different macronutrient and growth regulator levels showed no generalized effect on regenerated shoots, but the effect of recovery media was different between cultivars. When the whole regeneration chain from cryopreservation via micropropagation to greenhouse conditions was tested, shoots at least 1 cm in length were found necessary for successful transfer ex vitro. The long-term cryopreservation of 22 blackcurrant accessions was finally conducted, with practices slightly modified from the tested protocol. The estimated recovery of shoot tips after 9 weeks in vitro was 17–94% with at least 75% recovery in seven accessions and at least 40% recovery in 19 out of 22 accessions. Only one accession had no cryopreservation success. The results demonstrated that the modified droplet vitrification technique is appropriate for a large diversity of blackcurrant accessions. However, cultivar-related differences and recovery procedures are to be considered for success in regeneration and ex vitro adaptation.

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