The use of 2D‐DIGE to understand the regeneration of somatic embryos in avocado

Abstract

Avocado embryogenic cell cultures can be classified into two groups based on their morphology when cultured on a medium containing auxin: somatic embryo (SE) and proembryonic masses (PEM) type cultures. The calli of SE-type cell lines are able to go through the maturation process, whereas the calli of PEM cell lines rarely mature. We have investigated four independent avocado cell cultures (two SE and two PEM). The aim of this study was to link the differential regeneration capacity of the four cell cultures to a proteomic pattern and to gain insight into the regeneration capacity. A 2D-DIGE analysis followed by a blind multivariate analysis was able to separate the two SE lines from the PEM lines indicating that the protein profiles of SE and PEM calli are different. Based on the variable importance, that is, the differential protein pattern, we hypothesize that the regeneration capacity in avocado is correlated to the ability to overcome the physicochemical stress stimuli associated with the in vitro culture. Our identical culture conditions do not seem to trigger an appropriate response in PEM lines.

Quantitative proteomic analysis of Araucaria angustifolia (Bertol.) Kuntze cell lines with contrasting embryogenic potential

“…In studies of A. angustifolia and Picea glauca, experimental manipulation of the glutathione redox state by medium culture supplementation with reduced glutathione (GSH) or glutathione disulfide (GSSG) in EC cultures was shown to enhance the number of early somatic embryos [39,40], and GST isoforms have also been described in the EC proteomes of Vitis vinifera [41], Persea americana Mill. [42], and Theobroma cacao L [43].…”

The Current Status of Proteomic Studies in Somatic Embryogenesis

“…In this chapter, we compile the most relevant proteomic information by considering prominent proteomic studies in embryogenic and non-embryogenic cultures from different plant species (Varhanikova et al 2014;Correia et al 2012;Marsoni et al 2008;Sharifi et al 2012;Almeida et al 2012;Guzmán-García et al 2013). We group the identified proteins from each plant species by functional category.…”

“…7.1). These pairs of distinct cultures have been analyzed using proteomics tools with the goal of identifying proteins markers associated with embryogenic potency (Varhanikova et al 2014;Correia et al 2012;Marsoni et al 2008;Sharifi et al 2012;Almeida et al 2012;Guzmán-García et al 2013). However, these studies have not used sufficiently robust proteomics pipelines and therefore have not covered a comprehensive proportion of the proteome during initiation of the acquisition of the embryogenic potency (Rode et al 2011(Rode et al , 2012.…”

“…Many include the addition of growth regulators (auxin and/or cytokinin) during the induction stage (Imin et al 2005;Nolan et al 2003;Schmidt et al 1997). Several types of explants have been used for the establishment of SE including leaflets, stems, cambium, immature zygotic embryos, cell suspensions, and callus (Imin et al 2005;Varhanikova et al 2014;Correia et al 2012;Sharifi et al 2012;Guzmán-García et al 2013). SE can be induced directly from the explant (direct embryogenesis) or indirectly through the formation of somatic embryos from callus or suspension cultures (George et al 2008).…”

“…However, these studies have not used sufficiently robust proteomics pipelines and therefore have not covered a comprehensive proportion of the proteome during initiation of the acquisition of the embryogenic potency (Rode et al 2011(Rode et al , 2012. In most of the proteomics studies of somatic embryogenesis, proteins have been analyzed with 2D-SDS-PAGE and mass spectrometry tools (Rode et al 2011(Rode et al , 2012Varhanikova et al 2014;Correia et al 2012;Marsoni et al 2008;Sharifi et al 2012;Almeida et al 2012;Guzmán-García et al 2013;Vale Ede et al 2014).…”