Project description:Coffee is one of the most important commodities cultivated worldwide and has great economic impact in producing countries. Although 130 different species belonging to the coffea gender have been described, only two of them are commercially exploited: Coffea arabica and Coffea canephora. C. arabica is responsible for 61% of the world production (Van der Vossen et al., 2015). However, due to the narrow genetic back ground, classical genetic breeding is time consuming and takes around 30 years (Santana-Buzzy et al., 2007; Hendre et al., 2014). Several genetic engineering and biotechnological tools have been successfully applied in coffee breeding. Somatic embryogenesis (SE) is a process in which new viable embryos are produced from somatic tissues. It is one of the most promising production processes (Santana-Buzzy et al, 2007; Marsoni et al., 2008). A better understanding of the molecular basis related to somatic embryogenesis will give insight into the process of embryo formation and totipotency and will allow the development of new in vitro culture strategies for the propagation and genetic manipulation of elite cultivars (Marsoni et al., 2008). High throughput proteomics in coffee is limited so far to 2D gel based proteomics techniques. Although really useful and the most common technique for plants, 2DE is limited in throughput and a gel free technique allow to go a step further (Carpentier & America, 2014; Vanhove et al., 2015). To improve the knowledge about somatic embryogenesis, we present the first high throughput proteome profile (1051 confident protein identifications) of coffee embryogenic cell suspensions developed from leaves of Coffea arabica cultivar Catuaí.
Project description:Coffea arabica is one of the most important crops worldwide. In vitro culture is an alternative for achieving Coffea regeneration, propagation, conservation, genetic improvement, and genome editing. The aim of this work is to identify proteins involved in auxin homeostasis by isobaric tandem mass tag (TMT) and the synchronous precursor selection (SPS)-based MS3 technology on the Orbitrap Fusion™ Tribrid mass spectrometer™ in three types of biological material corre-sponding to C. arabica: plantlet leaves, calli, and suspension culture. Proteins included in the β-oxidation of indole butyric acid, and in the signaling, transport, and conjugation of in-dole-3-acetic acid were identified, such as the indole butyric response (IBR), the auxin binding protein (ABP), the ATP-binding cassette transporters (ABC), the Gretchen-Hagen 3 proteins (GH3), and the indole-3-acetic-leucine resistant proteins (ILR). A more significant accumulation of pro-teins involved in auxin homeostasis was found in the suspension cultures vs the plantlet com-parison, followed by callus vs plantlet and suspension culture vs callus, suggesting greater par-ticipation of these proteins as cell differentiation increases.
