Abstract
Despite the decline in the national hop production, a part of hop products, made of Ukrainian aroma hop, is highly estimated in the international market and exported. Therefore, a relevant prerequisite of ensuring a suffi cient level of competitiveness of domestic hop production is the expansion of its assortment. To satisfy this need the breeders face the task of creating varieties with unique characteristics, which requires searching for new forms and strains of hop with increased content of different biologically active compounds in cones. In this regard national research program aimed to investigate genetic stab ility of hop varieties during multiple planting and storing of explants in in vitro conditions while introducing them into the collection, adapting the composition of Murashige and Skoog culture medium for specifi c varieties is discussed. Studies conducted included the analysis of plants by morphological and variety-specifi c traits, the identifi cation of varieties by biochemical criteria, improvement of hop regenerants using ELISA, molecular-genetic analysis based on PCR (polymerase chain reaction) for identifi cation of hop genotypes and determination of genetic stability, and the improvement of method of microclonal reproduction of hop. As a result of the perennial research of cultivating hop planting material using the Murashige and Skoog complex of nutrients, the foundations of creating and maintaining the in vitro collection of hop varieties were fi rst elaborated in Ukraine which allow for the possibility of decreasing the concentration of nutrients in the culture medium by 50 %, replacing expensive gel-forming preparation for the maintenance of plants in the culture medium - agar-agar, the share of whose cost in the medium composition is up to 70 %, with a cheaper substance - modifi ed starch DDKamod or agroperlite, and reducing the expenses for the maintenance of genetic pool in the in vitro collection. It was determined that the spectra of microsatellite loci of the amplifi ed DNA of the explants, cultivated in vitro, during the study period did not differ from the spectra of plant DNA prior to cloning, which demonstrated DNA stability and allowed cultivating hop varieties in the in vitro culture without any changes in the genome. The composition of media for cultivation and long- term storing of hop varieties in in vitro conditions was selected.
Highlights
Unique organic substances – bitter principles, essential oils and polyphenols – are synthesized in hop cones (Humulus lupulus L.), due to which hop is widely used in different branches of industry
As a result of the perennial research of cultivating hop planting material using the Murashige and Skoog complex of nutrients, the foundations of creating and maintaining the in vitro collection of hop varieties were first elaborated in Ukraine which allow for the possibility of decreasing the concentration of nutrients in the culture medium by 50 %, replacing expensive gel-forming preparation for the maintenance of plants in the culture medium – agar-agar, the share of whose cost in the medium composition is up to 70 %, with a cheaper substance – modified starch DDKamod or agroperlite, and reducing the expenses for the maintenance of genetic pool in the in vitro collection
It was determined that the spectra of microsatellite loci of the amplified DNA of the explants, cultivated in vitro, during the study period did not differ from the spectra of plant DNA prior to cloning, which demonstrated DNA stability and allowed cultivating hop varieties in the in vitro culture without any changes in the genome
Summary
Slovianka Aroma female plant (Ukraine)/male form І2 of variety F-108 (Great Britain). Aroma mid-season variety, yield – 1.9–2.8 t/ha, content of alpha-acids 5.4-8.5 %. DNA analysis was conducted to determine genome stability of regenerants during the process of their cultivation in the in vitro conditions. The plants of the abovementioned varieties, cultivated for two and more years, with the presumably possible DNA changes, were selected from the in vitro collection. The remaining plants were subjected to positive selection by quality traits – they were not to have the signs of disease and were noted for high productivity The cones from these plants underwent biochemical analysis for the determination of the main varietal biochemical traits: the ratio between beta-acids and alphaacids, which is a constant for each variety, the amount of cohumulone in the composition of alpha-acids and the amount of colupulone in the composition of beta-acids.
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