II.A.1.g. Statistical Analysis of Multivariate Effects on Microalgal Growth and Lipid Content

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National Renewable Energy Laboratory

Publications:

Berglund, D.; Cooksey, B.; Cooksey, K.E.; Priscu, L.R. (1987) ACollection and screening of

microalgae for lipid production: possible use of a flow cytometer for lipid analysis.@ FY 1986

Aquatic Species Program Annual Report, Solar Energy Research Institute, Golden, Colorado,

SERI/SP-231-3071, pp. 41-52.

Cooksey, K.E.; Guckert, J.B.; Williams, S.A.; Collis, P.R. (1987) AFluorometric determination

of the neutral lipid content of microalgal cells using Nile Red.@ J. Microbiol. Methods 6:333-345.

Johansen, J.; Lemke, P; Barclay, W.; Nagle, N. (1987) ACollection, screening, and

characterization of lipid producing microalgae: Progress during Fiscal Year 1987.@ FY 1987

Aquatic Species Program Annual Report, Solar Energy Research Institute, Golden, Colorado,

SERI/SP-231-3206, pp. 27-42.

Lien, S. (1981a) In Proc. Brighton Int. Conf. On Energy from Biomass (Palz, W.; Chartier, P.; and

Hall, D.O., eds.) Applied Science Publishers, London, pp. 697-702.

Sommerfeld, M.; Ellingson, S.; Tyler, P. (1987) AScreening microalgae isolated from the

southwest for growth potential and lipid yield.@ FY 1987 Aquatic Species Program Annual

Report, Solar Energy Research Institute, Golden, Colorado, SERI/SP-231-3206, pp. 43-57.

Additional references:

Greenspan. P.; Mayer, E.P.; Fowler, S.D. (1985) ANile Red: A selective fluorescent stain for

intracellular lipid droplets.@ J. Cell Biol: 100:965-973.

Greenspan. P.; Fowler, S.D. (1985) J. Lipid Res. 26:787

II.A.1.g. Statistical Analysis of Multivariate Effects on Microalgal Growth and Lipid Content

As discussed earlier, environmental variables (particularly nutrient status) can have great effects

on growth and the quantity and quality of lipids in microalgae. To determine the effects of several

environmental variables alone and in combination on the growth and lipid contents of microalgae,

a multivariate, fractional factorial design experiment was carried out with two promising diatoms,

N. saprophila (NAVIC1) and C. muelleri (CHAET9). For these experiments, cells were grown

in modified SERI Type II/25 medium in which the alkalinity was adjusted by adding sodium

carbonate and sodium bicarbonate and the conductivity was adjusted by adding sodium chloride.

Cultures were grown on the temperature gradient table described previously at 200 µE$m-2$s-1.

The following variables were tested in the multivariate analysis: conductivity (20 to 80

mmho$cm-1), temperature (17° to 321C), N (urea) concentration (0 to 144 mg$L-1), sodium

silicate concentration (0 to 500 mg$L-1), and alkalinity (8.8 to 88 meq$L-1). In these experiments,

growth was measured by changes in AFDW and lipid was measured by the use of the Nile Red

fluorometric assay.

A Look Back at the Aquatic Species Program--Technical Review