Integrated proteomics and histochemical analysis of Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae) in embryogenic suspension culture

Abstract

t Cell suspension culture is a useful in vitro model-system for both scaling up and conserving the Brazilian conifer Araucaria angustifolia. In the present work, cell suspension of Araucaria was subjected to proteomics, biochemical and histochemical analyses. The results revealed new insights underlying the molecular mechanism of proembryogenic masses transition in cell suspension. Embryogenic cell cultures were cultivated in a basal liquid medium modified in a Steward apparatus (orbital agitator). Cell growth dynamics was evaluated using cell volume after sedimentation, fresh weight, mitotic index, conductivity, pH, and the number of proembryogenic masses (PEMs: I, II, III). Histochemical parameters, cell viability, and cell death analyses were performed to pinpoint growth rates. Proteomics analysis was performed using two-dimensional electrophoresis, and protein identification was carried out by MALDI-TOF-TOF tandem mass spectrometry. Cell growth dynamics showed a predominance of PEM III. Maximum slope of the exponential phase growth in fresh weight occurred at exponential phase after 15 days (optimal cultivation time), after which cell viability and pH decreased, thereby allowing the identification of stressrelated proteins. Several metabolism and growth proteins were abundant, such as: cytoskeletal, WOX1, cytokinin-related, and auxin-related proteins acting on cell wall modification, suspensor cell formation, and PEM I to PEM III transition.

Nanomaterial-modulated cellular sodium extrusion and vacuolar sequestration for salt tolerance

“…S4a†), referring to a recovery of cell growth. 36 Correspondingly, 0.1 mg L −1 nCeO 2 exposure enhanced the salt tolerance indices (STI) even after only 2 h exposure ( p < 0.05) and the STI had been elevated more significantly by 11.2% after 4 h (Fig. 1a); while significant improvement (by 14.5%) in this salt tolerance was observed until 12 h of 0.5 mg L −1 N-CDs treatment (Fig.…”

Morphological and endogenous phytohormone changes during long-term embryogenic cultures in Korean pine

“…Findings from the integrated cell biology studies may contribute to overcome the bottlenecks encountered with the SE. For example, morphological changes observed during EC proliferation resulted from the subsequent differential competence of the somatic embryos (Guerra et al 2016;Pereira-Dias et al 2020). Based on the observation of histological morphology, in conifers trees, the proliferations in cell cultures were generally driven by proembryogenic masses (PEMs), which divided cyclically into PEM I, II, and III, and development of PEMs signi cantly affected the ability of differentiation of the somatic embryo (Filonova et al 2000;Stasolla and Yeung 2003;Steiner et al 2015).…”