Screening for Lipids from Marine Microalgae Using Nile Red

Johnson, Zackary I.; Bidigare, Robert R.; Blinebry, Sara K.; Brown, Susan L.; Cullen, John J.; Loftus, Sarah E.; Redalje, Donald G.; Swink, Courtney; Mooy, Benjamin A. S. Van

doi:10.1007/978-3-319-50436-0_382

Cited by 6 publications

(4 citation statements)

References 37 publications

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“…First, we performed chemical fixation of cells and their Nile Red staining of TAGs (see Section 2). Nile Red is an efficient TAG marker and its fluorescence was reported to be tightly correlated with TAG concentration in C. reinhardtii (Johnson et al, 2017). Afterwards, we optimized confocal microscopy conditions (see Section 2) to separate Nile Red signal from cellular autofluorescence and acquired Z‐stacks of prepared specimens to capture Nile Red signal in 3D.…”

Section: Resultsmentioning

confidence: 99%

Chlamydomonas reinhardtii exhibits stress memory in the accumulation of triacylglycerols induced by nitrogen deprivation

Mikulski

Santos‐Aberturas

2022

Plant Enviro Interactions

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Stress memory is a phenomenon whereby exposure to initial stress event influences a response to subsequent stress exposures. Studying stress memory is important to understand the cellular behavior in dynamic environment, especially nowadays, in times with growing environmental instability. Stress memory has been characterized in vascular plants but its occurrence in nonvascular plant species has been rarely investigated. We hypothesized that stress memory occurs in nonvascular plants in relation to metabolic stress. We sought to test it using accumulation of lipids (triacylglycerols) in model green alga Chlamydomonas reinhardtii subjected to nitrogen deprivation stress as a model system. Here, we established stress memory protocol on C. reinhardtii cells. Using a blend of microscopy and gas chromatography methods, we showed that the cells exposed to recurrent stress show differential accumulation of triacylglycerols on the quantitative level without qualitative changes in lipid composition, comparing to single stress controls. Overall, our results suggest that metabolic stress memory does occur in nonvascular plant C. reinhardtii and provides a starting point to characterize mechanistic principles of metabolic stress memory. Due to the commercial potential of algae, our findings are relevant for basic science, as well as industrial production of algae‐derived compounds.

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Section: Resultsmentioning

confidence: 99%

Chlamydomonas reinhardtii exhibits stress memory in the accumulation of triacylglycerols induced by nitrogen deprivation

Mikulski

Santos‐Aberturas

2022

Plant Enviro Interactions

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“…Culture aliquots and 0.2-μm filtrate were measured for total lipids and extracellular lipids, respectively, following Johnson et al (2017). Briefly, samples were preserved with 5 μL/mL of 25% glutaraldehyde (final concentration 0.12%) and stored at −80°C.…”

Section: Methodsmentioning

confidence: 99%

Reused Cultivation Water Accumulates Dissolved Organic Carbon and Uniquely Influences Different Marine Microalgae

Loftus

Johnson

2019

Front. Bioeng. Biotechnol.

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Reusing growth medium (water supplemented with nutrients) for microalgae cultivation is required for economical and environmentally sustainable production of algae bioproducts (fuels, feed, and food). However, reused medium often contains microbes and dissolved organic matter that may affect algae growth. While the accumulation of dissolved organic carbon (DOC) in reused medium has been demonstrated, it is unclear whether DOC concentrations affect algae growth or subsequent rates of algal DOC release. To address these questions, lab-scale experiments were conducted with three marine microalgae strains, Navicula sp. SFP, Staurosira sp. C323, and Chlorella sp. D046, grown in medium reused up to four times. Navicula sp. and Chlorella sp. grew similarly in reused medium as in fresh medium, while Staurosira sp. became completely inhibited in reused medium. Across the three algae, there was no broad trend between initial DOC concentration in reused medium and algae growth response. Navicula sp. released less DOC overall in reused medium than in fresh medium, but DOC release rates did not decrease proportionally with increased DOC concentrations. Net DOC accumulation was much lower than gross DOC released by Navicula sp. and Staurosira sp., indicating the majority of released DOC was degraded. Additionally, biodegradation experiments with reused media showed no further net decrease in DOC, suggesting the accumulated DOC was recalcitrant to the associated bacteria. Overall, these results suggest that taxa-specific factors may be responsible for algae growth response in reused medium, and that DOC release and accumulation are insensitive to prior cultivation rounds. Choosing an algae strain that is uninhibited by accumulated DOC is therefore critical to ensure successful water reuse in the algae industry.

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“…The fluorescent stain Nile Red has been used for the quantification of lipids in microalgae as this stain is economically saving and sensitive to use for screening purposes (Johnson et al, 2017). Microalgae strain with high intracellular lipid contents is important for downstream application such as animal feedstock production (Doan et al, 2011).…”

Section: Nile Red Fluorescence Of Microalgaementioning

confidence: 99%

Isolation and Nile Red Screening of Indigenous Microalgae Species From Pahang Lakes as Potential Lipid Source in Aquaculture Feed

Zakaria

Haris

Wahid

et al. 2020

MABJ

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The high nutritional composition of microalgae that includes lipids is considered as one of the promising alternative lipid sources for animal feeds enrichment. Since microalgae have higher photosynthetic efficiencies than terrestrial plants, highly adaptable to environmental changes, and do not compete with conventional agriculture for resources, the inclusion of lipid sources from microalgae in feed could help the sustainability of livestock production systems. This study has reported the indigenous microalgae strains from Pahang lakes and determined lipid-rich strains that have the potential for alternative lipid sources. Pure isolated strains were identified using an 18S rDNA marker. The microalgae strain biomass was determined for 15 days. Meanwhile, the screening for high-rich lipid contents in microalgae strains was carried out using Nile Red fluorescent dye. A total of 11 strains were successfully isolated that consist of 8 different species (Carteria radiosa, Spongiosaccinopsis terrestris, Desmodesmus sp., Desmodesmus abundans, Dendodesmus brasiliensis, Chlamydomonas reinhardtii, Mychonastes timauensis, and Mychonastes ovahimbae). Results showed that PHG C01, PHG C02, and PHG F03 have the highest biomass production among all strains. Meanwhile, for lipid production, PHG B01 and PHG F03 showed the highest results. Thus, PHG B01 and PHG F03 strains were chosen as potential candidates to be used as an alternative lipid source in animal feed feedstock.

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Screening for Lipids from Marine Microalgae Using Nile Red

Cited by 6 publications

References 37 publications

Chlamydomonas reinhardtii exhibits stress memory in the accumulation of triacylglycerols induced by nitrogen deprivation

Chlamydomonas reinhardtii exhibits stress memory in the accumulation of triacylglycerols induced by nitrogen deprivation

Reused Cultivation Water Accumulates Dissolved Organic Carbon and Uniquely Influences Different Marine Microalgae

Isolation and Nile Red Screening of Indigenous Microalgae Species From Pahang Lakes as Potential Lipid Source in Aquaculture Feed

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