Abstract
Embryogenic suspension cultures of Araucaria angustifolia (Bertol.) Kuntze (Araucariaceae) can be used as a model to test the effects of compounds added to the culture medium on the cellular growth and morphology of Pro-Embryogenic Masses (PEMs). PEMs are formed by embryogenic and suspensor-type cells. To measure changes in the cellular growth of embryogenic cultures, we performed sedimented cell volume (SCV) quantification, which is a non-destructive method. Morphological analysis by microscopy allowed for the observation of growth and development of PEMs and the alterations in embryogenic and suspensor-type cells. The methods used here provide an efficient means for monitoring the cellular growth of PEMs and identifying morphological changes during the development of embryogenic cultures. These studies can also be combined with biochemical and molecular analyses, such as proteomics, to further investigate embryo growth and morphology.
Keywords: Somatic embryogenesis, Size, Sedimented cell volume
Background
Silveira et al. (2006) used SCV measurements to analyze the effects of exogenous polyamines on the morphological changes of A. angustifolia PEMs and Osti et al. (2010) tested the effect of different nitric oxide donors on cellular growth and PEM morphology. Recently, Douétts-Peres et al. (2016) studied the effect of a cellular growth inhibitor on cellular growth and PEM morphology using SCV, fresh and dry weight, PEM area, and individual diameters of embryogenic-type cells, including the length and width of the suspensor-type cells. In addition, alterations to cellular growth and morphology in response to endogenous compounds, such as polyamines, nitric oxide and specific proteins have been evaluated using this method (Silveira et al., 2006; Osti et al., 2010; Douétts-Peres et al., 2016).
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Acknowledgments
This work was supported by FAPERJ (Foundation for Research Support of the State of Rio de Janeiro) and CNPq (National Counsel of Technological and Scientific Development). The VCS method presented was derived from Silveira et al. (2006).
References
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