The integration of nanoparticles into plant cryopreservation protocols holds great promise for improving the survival rates and recovery potential of explants. This study aimed to verify the effect of nanoparticles on the ex-vitro performance of cryopreservation-derived plants. Lamprocapnos spectabilis (L.) Fukuhara (bleeding heart) 'Gold Heart' and 'Valentine' cultivars were used as the plant material. The encapsulation-vitrification cryopreservation protocol of shoot tips included the preculture, encapsulation, dehydration, storage in liquid nitrogen, rewarming, and recovery steps. Gold (AuNPs), silver (AgNPs), or zinc oxide (ZnONPs) nanoparticles were added at varying concentrations, either into the preculture medium or the protective bead matrix during encapsulation. After the in vitro recovery, the plants were transferred to the glasshouse and subjected to detailed biometrical, biochemical and cytogenetic analyses. Nanoparticles had no evident effect on the acclimatization efficiency (80-100% survival) and leaf number in L. spectabilis 'Gold Heart'. Nonetheless, shoots developed from alginate beads supplemented with 5 ppm AuNPs were twice as long as the control, while the leaves of plants grown on the preculture medium with ZnONPs contained significantly more chlorophyll and had higher Leaf Soil-Plant Analysis Development (SPAD) values. Moreover, several NPs treatments stimulated the development of leaves, including their surface area, length, and perimeter. Higher ZnONPs levels enhanced also the replication process, resulting in higher nuclear DNA content. As for L. spectabilis 'Valentine', alginate augmentation with 5 ppm AgNPs or 5 ppm ZnONPs stimulated the elongation of shoots. There was also a tendency suggesting a positive influence of 5 ppm AgNPs in the alginate bead matrix on foliar growth. The effect of nanoparticles on the content of flavonoids, anthocyanins, and stress markers in the plants varied depending on the treatment and cultivar, but also on the organ studied (leaf or stem). Overall, L. spectabilis 'Gold Heart' was more stress-tolerant and genetically stable than 'Valentine' judging by the activity of Photosystem II (PSII) and flow cytometric analyses, respectively. The complex effects of nanoparticles on survival, biometric parameters, physiological responses, and cytogenetic events underscore the intricate interplay between nanoparticles and plant systems. Nonetheless, our research confirmed the positive effect of nanoparticles on the ex-vitro growth and development of L. spectabilis plants after cryostorage.

Department of Crop Sciences and Agroforestry Faculty of Tropical AgriSciences Czech University of Life Sciences Prague Prague Czech Republic

Laboratory of Horticulture Faculty of Agriculture and Biotechnology Bydgoszcz University of Science and Technology Bydgoszcz Poland

Laboratory of Nanostructures Institute of High Pressure Physics Polish Academy of Sciences Warsaw Poland

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