RESUMEN

Fluorescence methods are widely used for the study of marine and freshwater phytoplankton communities. However, the identification of different microalgae populations by the analysis of autofluorescence signals remains a challenge. Addressing the issue, we developed a novel approach using the flexibility of spectral flow cytometry analysis (SFC) and generating a matrix of virtual filters (VF) which allowed thorough examination of autofluorescence spectra. Using this matrix, different spectral emission regions of algae species were analyzed, and five major algal taxa were discriminated. These results were further applied for tracing particular microalgae taxa in the complex mixtures of laboratory and environmental algal populations. An integrated analysis of single algal events combined with unique spectral emission fingerprints and light scattering parameters of microalgae can be used to differentiate major microalgal taxa. We propose a protocol for the quantitative assessment of heterogenous phytoplankton communities at the single-cell level and monitoring of phytoplankton bloom detection using a virtual filtering approach on a spectral flow cytometer (SFC-VF).

Asunto(s)

Microalgas , Citometría de Flujo/métodos , Fitoplancton , Agua Dulce , Coloración y Etiquetado

RESUMEN

Regulated cell death (RCD) is central to the development, integrity, and functionality of multicellular organisms. In the last decade, evidence has accumulated that RCD is a universal phenomenon in all life domains. Cyanobacteria are of specific interest due to their importance in aquatic and terrestrial habitats and their role as primary producers in global nutrient cycling. Current knowledge on cyanobacterial RCD is based mainly on biochemical and morphological observations, often by methods directly transferred from vertebrate research and with limited understanding of the molecular genetic basis. However, the metabolism of different cyanobacteria groups relies on photosynthesis and nitrogen fixation, whereas mitochondria are the central executioner of cell death in vertebrates. Moreover, cyanobacteria chosen as biological models in RCD studies are mainly colonial or filamentous multicellular organisms. On the other hand, unicellular cyanobacteria have regulated programs of cellular survival (RCS) such as chlorosis and post-chlorosis resuscitation. The co-existence of different genetically regulated programs in cyanobacterial populations may have been a top engine in life diversification. Development of cyanobacteria-specific methods for identification and characterization of RCD and wider use of single-cell analysis combined with intelligent image-based cell sorting and metagenomics would shed more light on the underlying molecular mechanisms and help us to address the complex colonial interactions during these events. In this review, we focus on the functional implications of RCD in cyanobacterial communities.