Laboratório de Biotecnologia, Centro de Biociências e Biotecnologia (CBB) , Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF) , Avenue Alberto Lamego 2000 , Campos dos Goytacazes , Rio de Janeiro 28013-602 , Brazil.
Unidade de Biologia Integrativa, Setor de Genômica e Proteômica, UENF , Avenue Alberto Lamego 2000 , Campos dos Goytacazes , Rio de Janeiro 28013-602 , Brazil.
J Proteome Res. 2018 Aug 3;17(8):2767-2779. doi: 10.1021/acs.jproteome.8b00213. Epub 2018 Jul 5.
Somatic embryogenesis is an important biological process in several plant species, including sugar cane. Proteomics approaches have shown that H pumps are differentially regulated during somatic embryogenesis; however, the relationship between H flux and embryogenic competence is still unclear. This work aimed to elucidate the association between extracellular H flux and somatic embryo maturation in sugar cane. We performed a microsomal proteomics analysis and analyzed changes in extracellular H-flux and H-pump (P-H-ATPase, V-H-ATPase, and H-PPase) activity in embryogenic and non-embryogenic callus. A total of 657 proteins were identified, 16 of which were H pumps. We observed that P-H-ATPase and H-PPase were more abundant in embryogenic callus. Compared to non-embryogenic callus, embryogenic callus showed higher H influx, especially on maturation day 14, as well as higher H-pump activity (mainly, P-H-ATPase and H-PPase activity). H-PPase appears to be the major H pump in embryogenic callus during somatic embryo formation, functioning in both vacuole acidification and PPi homeostasis. These results provide evidence for an association between higher H-pump protein abundance and, consequently, higher H flux and embryogenic competence acquisition in the callus of sugar cane, allowing for the optimization of the somatic embryo conversion process by modulating the activities of these H pumps.
体细胞胚胎发生是包括甘蔗在内的几种植物物种中的一个重要生物学过程。蛋白质组学方法表明，H 泵在体细胞胚胎发生过程中存在差异调节；然而，H 流与胚胎发生能力之间的关系尚不清楚。本研究旨在阐明甘蔗细胞外 H 流与体细胞胚胎成熟之间的关系。我们进行了微粒体蛋白质组学分析，并分析了胚胎发生和非胚胎发生愈伤组织中外源 H 流和 H 泵（P-H-ATPase、V-H-ATPase 和 H-PPase）活性的变化。共鉴定出 657 种蛋白质，其中 16 种是 H 泵。我们观察到 P-H-ATPase 和 H-PPase 在胚胎发生愈伤组织中更为丰富。与非胚胎发生愈伤组织相比，胚胎发生愈伤组织表现出更高的 H 流入，特别是在成熟第 14 天，以及更高的 H 泵活性（主要是 P-H-ATPase 和 H-PPase 活性）。H-PPase 似乎是体细胞胚胎形成过程中胚胎发生愈伤组织中的主要 H 泵，在液泡酸化和 PPi 平衡中起作用。这些结果为 H 泵蛋白丰度较高与甘蔗愈伤组织中较高的 H 流和胚胎发生能力获得之间存在相关性提供了证据，通过调节这些 H 泵的活性，可以优化体细胞胚胎转化过程。
