Zaranek Magdalena, Pinski Artur, Skupien-Rabian Bozena, Jankowska Urszula, Godel-Jedrychowska Kamila, Sala-Cholewa Katarzyna, Nowak Katarzyna, Kurczyńska Ewa, Grzebelus Ewa, Betekhtin Alexander
Institute of Biology, Biotechnology and Environmental Protection, Faculty of Natural Sciences, University of Silesia in Katowice, St. Jagiellonska 28, Katowice, 40-032, Poland.
Proteomics and Mass Spectrometry Core Facility, Malopolska Centre of Biotechnology, Jagiellonian University, St. Gronostajowa 7A, Krakow, 30-387, Poland.
BMC Plant Biol. 2025 Jan 24;25(1):102. doi: 10.1186/s12870-025-06119-3.
Due to the totipotency of plant cells, which allows them to reprogram from a differentiated to a dedifferentiated state, plants exhibit a remarkable regenerative capacity, including under in vitro culture conditions. When exposed to plant hormones, primarily auxins and cytokinins, explant cells cultured in vitro can undergo differentiation through callus formation. Protoplast culture serves as a valuable research model for studying these processes in detail. This knowledge is particularly relevant for improving common and Tartary buckwheat species. To gain deeper insights into the stages of cell development from protoplasts-such as cell division, cell colony formation, and microcalli development-we focused on analyzing proteomes, cell wall composition, and changes in the expression profiles of selected genes in Fagopyrum protoplast cultures.
The results demonstrate a significant accumulation of somatic embryogenesis-related proteins like late embryogenesis abundant proteins (embryogenic protein-DC-8-like, seed biotin-containing protein) and endochitinases during the developmental path of protoplast-derived cultures. Additionally, we noted an extensive increase in seed storage proteins like vicilin, oleosins, and seed biotin-containing proteins during the culture. Investigation of somatic embryogenesis-associated transcription factors revealed massive up-regulation of LEAFY COTYLEDON1 for the 50th day of F. tataricum protoplast-derived cultures. However, for BABY BOOM, the transcription factor was noted to be down-regulated during the development of cell colonies. Furthermore, we demonstrated the variable distribution of cell wall components like pectin side chains, arabinogalactan proteins (AGPs) and extensins (EXTs), indicating the reorganisation of cell wall composition during the culture period.
This study revealed changes correlating with regaining embryogenic competence during the development of Fagopyrum protoplast-derived cell colonies. Our findings revealed variable expression levels of genes and proteins associated with somatic embryogenesis. This analysis identified an increase in seed storage proteins that play a significant role in the somatic somatic embryogenesis pathway of regeneration. Furthermore, the relationship between transcription factors and these processes seems to be connected with regaining somatic cells' totipotency and promoting embryogenic competence of protoplast-derived cell colonies. Additionally, we observed dynamic changes in cell wall composition during the development of the protoplast-derived cultures.
Not applicable.
由于植物细胞的全能性，使其能够从分化状态重新编程为去分化状态，植物表现出显著的再生能力，包括在体外培养条件下。当暴露于主要是生长素和细胞分裂素的植物激素时，体外培养的外植体细胞可通过愈伤组织形成进行分化。原生质体培养是详细研究这些过程的有价值的研究模型。这一知识对于改良普通荞麦和苦荞麦品种尤为重要。为了更深入了解原生质体的细胞发育阶段，如细胞分裂、细胞集落形成和微愈伤组织发育，我们专注于分析荞麦原生质体培养物中的蛋白质组、细胞壁组成以及所选基因表达谱的变化。
结果表明，在原生质体衍生培养物的发育过程中，体细胞胚胎发生相关蛋白如晚期胚胎发生丰富蛋白（胚胎发生蛋白-DC-8样、含种子生物素蛋白）和内切几丁质酶大量积累。此外，我们注意到培养过程中种子贮藏蛋白如豌豆球蛋白、油质蛋白和含种子生物素蛋白大量增加。对体细胞胚胎发生相关转录因子的研究表明，在苦荞原生质体衍生培养物的第50天，LEAFY COTYLEDON1大量上调。然而，对于BABY BOOM，该转录因子在细胞集落发育过程中被发现下调。此外，我们证明了细胞壁成分如果胶侧链、阿拉伯半乳聚糖蛋白（AGPs）和伸展蛋白（EXTs）的可变分布，表明培养期间细胞壁组成的重组。
本研究揭示了与荞麦原生质体衍生细胞集落发育过程中恢复胚胎发生能力相关的变化。我们的研究结果揭示了与体细胞胚胎发生相关的基因和蛋白质的可变表达水平。该分析确定了种子贮藏蛋白的增加，这些蛋白在体细胞胚胎发生再生途径中起重要作用。此外，转录因子与这些过程之间的关系似乎与恢复体细胞全能性和促进原生质体衍生细胞集落的胚胎发生能力有关。此外，我们观察到原生质体衍生培养物发育过程中细胞壁组成的动态变化。
不适用。
