Evaluation of the regenerative potential of callus tissue in Actinidia chinensis var. deliciosa (A.Chev.) C.F. Liang & A.R. Ferguson cultivars in in vitro culture

Background. The use of modern agrobiotechnological methods allows for obtaining qualitatively new highly profitable Actinidia cultivars with an increased content of bioactive substances. Achieving such results is possible through a comprehensive study and use of genetic resources of plants from the VIR collection, including in vitro aseptic conditions. In this regard, it is relevant to study the issues of callusogenesis induction with subsequent proliferation of promising cultivars of Actinidia chinensis var. deliciosa (A. Chev.) C.F. Liang & A.R. Ferguson in vitro with the aim of preserving them for further selection. Materials and methods. The objects of the study were the cultivars: ‘Hayward’, ‘Bruno’, ‘Monty’, and ‘Allison’ of A. chinensis var. deliciosa from the VIR collection. Young shoots of plantlets from seeds germinated in culture vessels in a climatic chamber were used as explants for introduction into artificial culture conditions. At this stage, three variants of sterilizing agents were tested for identifying the main substance for multi-stage sterilization of explants. Shoots were placed in test tubes with a nutrient medium according to the Murashige and Skoog (MS) prescription. On day 10, the efficiency of introduction applying different types of sterilizing substances was assessed. To induce callusogenesis, leaf segments from Actinidia microplants were planted on four variants of the MS nutrient medium with different combinations of growth regulators. In order to determine the efficiency of 3-Hydroxytetrahydrofuran (Riftal) plant growth regulator, a steadily growing callus was transplanted onto four variants of the MS nutrient medium with the addition of 1 ml/l of the growth regulator at different concentrations. Calluses of Actinidia cultivars maintained in vitro for three years were used directly for the induction of organogenesis. Results and discussion. The best result in terms of sterilization efficiency and explant viability was shown by the variant of shoot sterilization with a 10% solution of “Belizna”, a universal detergent, disinfectant and bleaching agent. Primary callus formation was noted on all variants of nutrient media for callusogenesis induction. However, the subsequent cultivation of callus tissue on MS nutrient media containing 2,4-D, resulted in formation of loose, hydrated, chlorophyll-free calli. In the experiment focused on the effect of Riftal, steady callus growth was noted on all variants of nutrient media and root formation was recorded in cultivars ‘Hayward’, ‘Allison’, and ‘Bruno’. Organogenesis was noted in callus tissue of all studied cultivars. Conclusions. Evaluation of the regenerative potential of A. chinensis var. deliciosa cultivars ‘Hayward’, ‘Allison’, ‘Bruno’, and ‘Monty’ has shown that ‘Hayward’ and ‘Allison’ have the highest number of regenerants per organogenic callus: 2.2 and 3.7, respectively. The obtained results will serve for intensifying the breeding within the genus Actinidia using modern molecular genetics and biotechnological methods, including CRISPR/Cas genome editing.

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