Pernis Miroslav, Salaj Terézia, Bellová Jana, Danchenko Maksym, Baráth Peter, Klubicová Katarína
Institute of Plant Genetics and Biotechnology, Plant Science and Biodiversity Center, Slovak Academy of Sciences, Nitra, Slovakia.
Institute of Chemistry, Slovak Academy of Sciences, Bratislava, Slovakia.
Front Plant Sci. 2023 Aug 4;14:1225424. doi: 10.3389/fpls.2023.1225424. eCollection 2023.
Somatic embryogenesis is an efficient mean for rapid micropropagation and preservation of the germplasm of valuable coniferous trees. Little is known about how the composition of secretome tracks down the level of embryogenic capacity. Unlike embryogenic tissue on solid medium, suspension cell cultures enable the study of extracellular proteins secreted into a liquid cultivation medium, avoiding contamination from destructured cells. Here, we present proteomic data of the secretome of cell lines with contrasting embryogenic capacity, accounting for variability between genotypes. Our results showed that cell wall-related and carbohydrate-acting proteins were the most differentially accumulated. Peroxidases, extensin, α-amylase, plant basic secretory family protein (BSP), and basic secretory protease (S) were more abundant in the medium from the lines with high embryogenic capacity. In contrast, the medium from the low embryogenic capacity cell lines contained a higher amount of polygalacturonases, hothead protein, and expansin, which are generally associated with cell wall loosening or softening. These results corroborated the microscopic findings in cell lines with low embryogenic capacity-long suspensor cells without proper assembly. Furthermore, proteomic data were subsequently validated by peroxidase and α-amylase activity assays, and hence, we conclude that both tested enzyme activities can be considered potential markers of high embryogenic capacity.
体细胞胚胎发生是针叶树优良品种快速微繁殖和种质保存的有效手段。目前对于分泌蛋白质组的组成如何追踪胚胎发生能力的水平了解甚少。与固体培养基上的胚性组织不同,悬浮细胞培养能够研究分泌到液体培养基中的细胞外蛋白质,避免了来自破碎细胞的污染。在此,我们展示了具有不同胚胎发生能力的细胞系分泌蛋白质组的蛋白质组学数据,解释了基因型之间的变异性。我们的结果表明,与细胞壁相关和具有碳水化合物作用的蛋白质积累差异最大。过氧化物酶、伸展蛋白、α-淀粉酶、植物碱性分泌家族蛋白(BSP)和碱性分泌蛋白酶(S)在胚胎发生能力高的细胞系培养基中含量更高。相比之下,胚胎发生能力低的细胞系培养基中含有更多的多聚半乳糖醛酸酶、热头蛋白和扩张蛋白,这些通常与细胞壁松弛或软化有关。这些结果证实了胚胎发生能力低的细胞系中的显微镜观察结果——长而无适当组装的胚柄细胞。此外,蛋白质组学数据随后通过过氧化物酶和α-淀粉酶活性测定得到验证,因此,我们得出结论,这两种测试的酶活性都可被视为高胚胎发生能力的潜在标志物。