The problem statement. Hazelnut is a valuable nut culture, which is quite profitable in economic way. A deterrent to an extensive cultivation of hazelnut in Ukraine is a low ratio of breeding in a conventional methods. The alternative to solving this problem may be the method of microclonal propagation, which is actively implemented in commercial purposes. The difficulties of hazelnut microclonal propagation exist on every stage of this technology: 1) introduction to aseptic conditions; 2) multiplication in vitro; 3) rhizogenesis induction; 4) postaseptic adaptation. The aim of the research. The article deals with problem aspects of hazelnut microclonal propagation and analyzes the ways of solving these problems based on the own research results. In particular, the influence of phenol emergence, culture medium, type, concentration and method of phytohormones application on root formation and proliferation are examined. Materials and methods. The research was held in a standart laboratory conditions. The object of research are hazelnut plants variaties such as Córylus Trapezund, Corylus avellana Syrena, Corylus colurna. It is established that rhizogenesis and proliferation processes are induced by trophic and hormone determinants. Results and discussion. Using the DKW culture medium is recommended to optimize the hazelnut micriclonal propagation process. I was found out that the use of activated carbon and explants transplantation on the early stages neutralizes phenol emergence. In order to resolve the difficulties of the phenol emergence the effectiveness of such points as cultivation of mother plans in the presence of diffused light in depositary condition, introduction of plant though by meristemas separation, buds awakening, the addition of PPM Plant Preservative Mixture biocide and polyvinylpyrolidone into the culture medium were established. At the multiplication stage 1.5 mg/l of benzylaminopurine is added into the culture medium. The influence of different concnetrations of activated carbon on rhizogenesis on the background of 3 mg/l of auxin indolebutyric acid was stidued. The activated carbon obscures the culture medium, adsorbes toxines, therefore it has an effective impact on root formation. Among the comparative concentration the optimal one is 2.5g/l of the medium. The possibility of using the greenhouse for postaceptic regenerants adaptation is shown. Conclusions. Processing plants and substrate with Previcur Energi improves their establishment and stimulates the growth. Key words: microclonal propagation, decontamination, phenol self-poisoning, phytohormones, rhizogenesis induction, postaseptic adaptation.
Hazelnuts and almonds are promising industrial crops that can differentiate the risks posed by climate change. Domestic almond varieties adapted to local conditions have been created in Ukraine, which require rapid propagation. Creating highly productive and disease-resistant varieties of hazelnuts also requires the reproduction of this crop in signifcant quantities. Of all the methods of vegetative propagation microclonal is the most promising one. Prior to introduction into aseptic culture, mother plants are grown in conditions that minimize endogenous accumulation in tissues of contaminants and phenolic substances. Sodium hypochlorite and Blanidas 300 are used for decontamination of explants. The addition of PPM biocide to the nutrient medium is promising to improve the sterilization of hazelnut explants. The decrease in contamination of almond explants decreases with increasing concentration of Cu, Ag ions in the environment. Mineral nutrients and phytohormones are hte main determinants of in vitro ontogeny. Along with the classic media (DKW, QL, MS, WPM) NRM environment is promising for hazelnuts, and NAM environment – for almonds. Excess in the environment of some elements reduces the assimilation of others, which is manifested visually. Thus, an excess of nitrogen causes symptoms of insufcient absorption of calcium, copper. This process is intensifed as the temperature increases. In addition, regenerating shoots are shortened, thickened and have excessive tissue hydration. Turkish scientists have proposed an original method of creating recipes for almonds and hazelnuts on the quantitative content of elements in the seeds of these plants. Cytokine are the most common phytohormones at the multiplication stage. For both cultures, most researchers prefer benzylaminopurine. Indolylbutyric acid is most often used to induce rhizogenesis. A perlite-based substrate is effective for the regenerants adaptation. It is suggested to inoculate regenerants on bitter almond seedlings as a type of adaptation. A method of photoautrophic microclonal propagation has been developed in Ukraine. These technological methods, which, along with vegetative propagation, allow the adaptation of regenerants in a short time. The efciency is achieved by activating the processes of photosynthesis due to increased carbon dioxide content and light intensity. Key words: hazelnuts, almonds, climate change, microclonal reproduction, determinants, nutrient media, phytohormones.
The purpose of the research was to identify the physiological–biochemical and anatomical–morphological peculiarities found during in vitro cultivation of Corylus avellana L. and Prunus dulcis (Mill.) D.A.Webb, which occur as a result of the action of trophic and phytohormonal determinants. The research was conducted on three hazelnut varieties (Trapezund, Jefferson, Tonda Gentile Romana) and four almond varieties (E5 Borozan, M41 Alex, Georgia, Louise). A theoretical and experimental rationale for their use in the microclonal propagation of these cultures has been developed. The effectiveness of the preparatory stage before introduction into aseptic conditions for growing mother plants in closed soil conditions with scattered artificial lighting and microbiological protection has been proven. This reduced self-intoxication by oxidation products with phenol-like substances and microflora contamination of primary explants. Among the nutrient media compared, the best was Nas and Read (NRM) for hazelnuts and Nas Almond Medium (NAM) for almonds. Passaging on the same media leads to a decrease in regeneration indicators. In regenerants of almonds, the rosette of the shoots was noted, and in hazelnuts, the death of apical buds was also found. To prevent this, alternating NRM and driver and kuniyuki walnut (DKW) media for hazelnuts and NAM and Quirin and Lepoivre (QL) for almonds was effective. Long-term cultivation on media with a high content of synthetic analogues of phytohormones leads to the accumulation of phytotoxic effects with each subsequent passage. The phytotoxic effect of cytokinins was manifested in hyperhydration of shoots and of auxins in callus fertilisation. On comparing the ontogenesis of regenerants from explants isolated from mother plants aged 30–180 days, it was found that the optimal age for hazelnut and almond was 90 days. To reset the trophic and hormonal determinants, the introduction of explant donors into a state of dormancy has been successfully used. At the final fourth stage of microclonal propagation, in order to adapt plants simultaneously with an increase in the number of regenerants, it is effective to use the photoautotrophic method of microclonal propagation with intensive lighting and air enriched with carbon dioxide. For the transition of plants from conventional heterotrophic propagation to autotrophic cultivation, an intermediate stage of pre-adaptation of regenerants in wet chamber conditions is effective.
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