Results of propagation of Fagus silvatica L. plants in vitro
A number of experimental studies have been conducted regarding the modification of the main stages of microcloning of Fagus silvatica L., namely: features of decontamination of plant material, initiation of growth and rhizogenesis of explants and adaptation of clones to in vivo conditions. The influ...
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| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Ukrainian National Forestry University
2024-09-01
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| Series: | Наукові праці Лісівничої академії наук України |
| Subjects: | |
| Online Access: | http://fasu.nltu.edu.ua/index.php/nplanu/article/view/800 |
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| Summary: | A number of experimental studies have been conducted regarding the modification of the main stages of microcloning of Fagus silvatica L., namely: features of decontamination of plant material, initiation of growth and rhizogenesis of explants and adaptation of clones to in vivo conditions. The influence of the concentration and duration of treatment with chemical sterilant agents on the yield of aseptic explants of the species under study is characterized. The methodology for conducting experimental studies on the reproduction of Fagus silvatica L. in vitro culture is presented. It was found that it is most effective to use apical buds, harvested in the second half of February, as the starting plant material.
A stepwise scheme of chemotherapy for the initial explants was used, which ensured maximum decontamination: a 15-second treatment with 96% C2H5OH (ethanol); a 24-hour treatment with running water with detergent; a 15-minute treatment with 30% NaClO solution (chlorine); a 10-second treatment with 70% C2H5OH; a 15-second treatment with 2% solution of H2O2 (hydrogen peroxide); a 10-minute treatment with 0.2% AgNO3 (silver nitrate) with washing out with sterile distilled water after each reagent. Immediately before the passage, the explants were treated for 10-15 seconds with a 10% solution of the antibiotic "Cefazolin-KMR".
Studies were conducted on the effectiveness of three types of nutrient media, namely P-24, WPM and MS, which were modified using different ratios and concentrations of auxins: NAA (α-naphthylacetic acid), IAA (indomyl-3-acetic acid), ІВА (β-indolylbutyric acid) and cytokinins: BAP (6-benzylaminopurine) and K (kinetin) in various concentrations and combinations for the initiation of explants. Rhizogenesis of the initiated explants was carried out only on the MS medium, which proved to be the most effective at the initiation stage, modified as follows: the concentration of the main mineral salts was reduced by 2-4 times, the amount of sucrose was reduced to 0.5-1.0%, and in some variants cytokinin was completely extracted. Adaptation of regenerant plants in four types of substrate was carried out: a mixture of soddy soil with coarse-grained sand and peat in a ratio of 1:1:1; coarse-grained sand; soddy soil with coarse-grained sand (1:1); peat tablets (“Jiffy-7 Forestry”). All the substrates were sterilized at a temperature of + 90 °C for 1.5 hours and pre-treated with a 0.5% KMnO₄ solution.
It was found that the MS nutrient medium modified with stimulants 2.0 mg/l NAA + 0.5 mg/l ІВА + 0.5 mg/l K + 0.5 mg/l BAP provides the highest number of initiated explants of Fagus silvatica L. (96%). It was revealed that with an increased content of auxin or cytokinin or their absence in the nutrient medium, the death of the plant material of the species under study occurs. The study showed that the use of a 0.5 MS nutrient medium modified with 1.0 mg/l NAA + 1.0 mg/ ІВА provides a high percentage of adventitious root formation in the plants under study (96%). A 4-fold decrease in the concentration of mineral salts in the nutrient medium makes the ability of Fagus silvatica L. clones to rhizogenesis virtually impossible. It was found that the adaptation of regenerant plants of the studied species is best carried out with the use of sterilized peat tablets, which ensures a success rate of 94% in 30 days of cultivation. |
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| ISSN: | 1991-606X 2616-5015 |