Abstract
The popularity of nanoparticles (NPs) is continuously increasing. To date, however, there has been little research on the application of NPs in plant cryopreservation, i.e. storage of tissues in liquid nitrogen (LN). The aim of this study is to analyze the effect and evaluate the usefulness of gold nanoparticles (AuNPs) in regard to cryobiology studies. In vitro-derived shoot tips of Lamprocapnos spectabilis ‘Valentine’ were cryopreserved with the encapsulation-vitrification protocol. Gold nanoparticles (at 10–30 ppm concentration; 13 nm in size) were added either into the preculture medium; to the protective bead matrix during encapsulation; or to the recovery medium after rewarming of samples. The control plants were produced from cryopreserved explants non-treated with nanoparticles or treated with colloid dispersion medium without NPs. A non-LN-treated standard was also considered. The influence of AuNPs on the cryopreservation efficiency was determined by evaluating the recovery rate of explants and their morphogenic response; the membrane stability index (MSI); the concentration of pigments in shoots; and the antioxidant enzymes activity. The genetic stability of the plant material was evaluated using Start Codon Targeted Polymorphism (SCoT) markers. It was found that 10 ppm of AuNPs added into the alginate bead matrix improved the recovery level of LN-derived shoot tips (70.0%) compared to the non-NPs-treated cryopreserved control (50.5%). On the other hand, the presence of nanoparticles in the recovery medium had a deleterious effect on the survival of explants. AuNPs usually had no impact on the MSI (73.9–85.9%), except for those added into the recovery medium at the concentration of 30 ppm (decline to 55.8%). All LN-derived shoots were shorter and contained less chlorophyll and carotenoids than the untreated standard. Moreover, the application of AuNPs affected the enzymatic activity in L. spectabilis. Minor genetic variation was found in 8.6% of plants if AuNPs were added either into the preculture medium (at 10 and 20 ppm) or to the alginate matrix (at 30 ppm). In conclusion, AuNPs added at a lower concentration (10 ppm) into the protective bead matrix can significantly improve the cryopreservation efficiency in L. spectabilis with no alternation in the DNA sequence.
Highlights
The protection of biodiversity is important for systematics research, species evolution understanding, and in breeding programs (Deliège and Neuteleers 2015)
As for the cryopreservation-derived shoot tips, the highest recovery level (70.0%) was reported after adding 10 ppm of AuNPs into the protective bead matrix
A high 61.7–62.4% recovery was found after supplementing the preculture medium with 10 ppm AuNPs and the alginate matrix with 20 ppm AuNPs, it was not significantly different from the NPs-free control (50.5%)
Summary
The protection of biodiversity is important for systematics research, species evolution understanding, and in breeding programs (Deliège and Neuteleers 2015). Cryopreservation, i.e. maintenance of biological material at the cryogenic temperature of liquid nitrogen (LN; − 196 °C), is currently considered the most effective long-term storage method. This approach has been applied to numerous plant species. After storage in LN, the explants are again exposed to osmotic shock related to their rehydration. All of those steps may induce oxidative stress through the formation of free radicals, which may (micro) fracture the cell membrane system or affect the stability of plants (Zevallos et al 2013b). Nanoparticles (NPs) may significantly affect the effectiveness of long-term storage protocols and ought to be scrupulously tested
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