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 Science, Warsaw, Poland.
PLoS One. 2024 May 31;19(5):e0304586. doi: 10.1371/journal.pone.0304586. eCollection 2024.
The integration of nanoparticles (NPs) holds promising potential to bring substantial advancements to plant cryopreservation, a crucial technique in biodiversity conservation. To date, little attention has been focused on using nanoparticles in cryobiology research. This study aimed to assess the effectiveness of NPs in enhancing the efficiency of plant cryopreservation. In-vitro-derived shoot tips of bleeding heart (Lamprocapnos spectabilis (L.) Fukuhara) 'Gold Heart' and 'Valentine' were used as the plant material. The encapsulation-vitrification cryopreservation protocol included preculture, encapsulation, dehydration, storage in liquid nitrogen, rewarming, and recovery steps. Gold (AuNPs), silver (AgNPs), or zinc oxide (ZnONPs) nanoparticles were added at various concentrations either into the preculture medium or the protective bead matrix during encapsulation. The explant survival and further morphogenic and biochemical events were studied. Results showed that the impact of NPs on cryopreservation outcomes was cultivar-specific. In the 'Valentine' cultivar, incorporating 5 ppm AgNPs within the alginate bead matrix significantly improved cryopreservation efficiency by up to 12%. On the other hand, the 'Gold Heart' cultivar benefited from alginate supplementation with 5 ppm AgNPs and 5-15 ppm ZnONPs, leading to an over 28% increase in the survival rate of shoot tips. Interestingly, adding NPs to the preculture medium was less effective and sometimes counterproductive, despite promoting greater shoot proliferation and elongation in 'Valentine' explants compared to the control. Moreover, nanoparticles often induced oxidative stress (and enhanced the activity of APX, GPOX, and SOD enzymes), which in turn affected the biosynthesis of plant primary and secondary metabolites. It was found that supplementation of preculture medium with higher concentration (15 ppm) of gold, silver and zinc oxide nanoparticles stimulated the production of plant pigments, but in a cultivar-dependent matter. Our study confirmed the beneficial action of nanoparticles during cryopreservation of plant tissues.
纳米粒子(NPs)的整合有望为植物冷冻保存带来重大进展,这是生物多样性保护的关键技术。迄今为止,在冷冻生物学研究中很少关注使用纳米粒子。本研究旨在评估 NPs 在提高植物冷冻保存效率方面的有效性。以离体衍生的 bleeding heart(Lamprocapnos spectabilis(L.)Fukuhara)'Gold Heart'和'Valentine'的茎尖为植物材料。包埋-玻璃化冷冻保存方案包括预培养、包埋、脱水、液氮储存、复温和解冻步骤。在预培养培养基或包埋过程中的保护珠基质中添加不同浓度的金(AuNPs)、银(AgNPs)或氧化锌(ZnONPs)纳米粒子。研究了外植体的存活率以及进一步的形态发生和生化事件。结果表明,纳米粒子对冷冻保存结果的影响因品种而异。在'Valentine'品种中,在藻酸盐珠基质中添加 5 ppm 的 AgNPs 可将冷冻保存效率提高 12%。另一方面,'Gold Heart'品种受益于添加 5 ppm AgNPs 和 5-15 ppm ZnONPs 的藻酸盐,导致茎尖存活率提高了 28%以上。有趣的是,尽管与对照相比,添加 NPs 到预培养培养基中对'Valentine'外植体的增殖和伸长有促进作用,但效果较差,有时甚至适得其反。此外,纳米粒子经常诱导氧化应激(并增强 APX、GPOX 和 SOD 酶的活性),从而影响植物初级和次级代谢物的生物合成。研究发现,在预培养培养基中添加更高浓度(15 ppm)的金、银和氧化锌纳米粒子刺激了植物色素的产生,但这取决于品种。我们的研究证实了纳米粒子在植物组织冷冻保存过程中的有益作用。
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