Pérez-Pérez Reneé, Pinski Artur, Zaranek Magdalena, Beckmann Manfred, Mur Luis A J, Nowak Katarzyna, Rojek-Jelonek Magdalena, Kostecka-Gugała Anna, Petryszak Przemysław, Grzebelus Ewa, Betekhtin Alexander
Institute of Biology, Biotechnology and Environmental Protection, Faculty of Natural Sciences, University of Silesia in Katowice, Katowice, Poland.
Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Penglais Campus, Aberystwyth, Wales, SY23 2DA, UK.
BMC Plant Biol. 2025 Apr 15;25(1):469. doi: 10.1186/s12870-025-06440-x.
Fagopyrum tataricum (Tartary buckwheat) is known for its high phenolic content, particularly rutin. High concentrations of these compounds secreted in the tissue culture medium can lead to its darkening and the eventual death of explants in in vitro cultures. This study aims to enhance the morphogenesis of F. tataricum callus cultures by utilising phenylalanine ammonia-lyase (PAL) inhibitors and polyvinylpyrrolidone (PVP) to mitigate oxidative browning and improve tissue viability.
We analysed the response of protoplasts isolated from morphogenic callus to media supplemented with varying concentrations of PAL inhibitors (AIP, AOPP, OBHA) and PVP. The flow cytometry results revealed that 10 µM AIP and 1% PVP yielded exclusively diploid plants, whereas higher concentrations (100 µM AIP and 3% PVP) failed to regenerate plants. Moreover, AOPP and OBHA addition resulted in the regeneration of tetraploid plants. Further analysis of proembryogenic cell complexes (PECCs) isolated from Tartary buckwheat morphogenic calli responses to AIP and PVP indicated that 100 µM AIP was most effective for plant regeneration. Metabolomic analysis showed that AIP treatments reduced phenolic compounds, notably rutin, and increased the GSH/GSSG ratio, indicating reduced oxidative stress. Gene expression analysis highlighted elevated expression of somatic embryogenesis-related genes (LEC2, BBM) and WUSCHEL in AIP-treated callus.
This study demonstrates that AIP enhances the regeneration potential of F. tataricum callus cultures, offering valuable insights for optimising tissue culture techniques for industrial crops. Additionally, we have detailed the metabolomic changes in calli treated with PVP and AIP, highlighting their impact on metabolism.
苦荞麦以其高酚含量，尤其是芦丁而闻名。这些化合物在组织培养基中高浓度分泌会导致其变黑，并最终导致体外培养的外植体死亡。本研究旨在通过利用苯丙氨酸解氨酶（PAL）抑制剂和聚乙烯吡咯烷酮（PVP）减轻氧化褐变并提高组织活力，从而增强苦荞麦愈伤组织培养的形态发生。
我们分析了从形态发生愈伤组织中分离的原生质体对添加不同浓度PAL抑制剂（AIP、AOPP、OBHA）和PVP的培养基的反应。流式细胞术结果显示，10 μM AIP和1% PVP仅产生二倍体植株，而较高浓度（100 μM AIP和3% PVP）未能再生植株。此外，添加AOPP和OBHA导致四倍体植株的再生。对从苦荞麦形态发生愈伤组织中分离的原胚性细胞复合体（PECCs）对AIP和PVP的反应的进一步分析表明，100 μM AIP对植株再生最有效。代谢组学分析表明，AIP处理降低了酚类化合物，尤其是芦丁，并提高了GSH/GSSG比值，表明氧化应激降低。基因表达分析突出了体细胞胚胎发生相关基因（LEC2、BBM）和WUSCHEL在AIP处理的愈伤组织中的表达升高。
本研究表明，AIP增强了苦荞麦愈伤组织培养的再生潜力，为优化工业作物的组织培养技术提供了有价值的见解。此外，我们详细阐述了用PVP和AIP处理的愈伤组织中的代谢组学变化，突出了它们对代谢的影响。
