Table 5. Summary of the environmental conditions used to culture phytoplankton species and strains in the present study.

		Protocol
Organism	Laboratory	A.	B.	C.	D.	E.	F.	G.	H.
Trichodesmium spp.,Crocosphaera spp.	Hutchins/Fu	Yes	Yes (6 replicates)	Yes	Yes (150)	Yes	Yes	Yes	No15
Thalassiosira pseudonana	Litchman	Yes	Yes (6 replicates)	Yes	Yes (100)	Yes6	No8	No12	No15
Proboscia inermis	Boyd/Strzepek	Yes	Yes (6 replicates)	Yes	Yes (90)	Yes	Yes9	No12	Yes
Prorocentrum donghaiense; Synechococcus	Mulholland	No1	Yes	Yes4	Yes (35;100)	Yes	No10	Yes13	Yes
Akashiwo sanguinea	Kudela	No2	Yes (5 replicates)	Yes	Yes (125)	Yes	Yes	Yes14	Yes
Thalassiosira weissflogii	Passow	No1	Yes (4 replicates)	Yes5	No (35)	Yes7	Yes	Yes14	Yes
Thalassiosira rotula	Rynearson	No3	Yes (3–5 replicates)	Yes	No (112)	Yes	No11	Yes 14	Yes

A. Growth rates were determined at a minimum of six temperature conditions. B. A minimum of three replicate growth rates were determined. C. All other environmental variables were held constant within each individual experiment, other than temperature. These include day length, culture medium, and culture protocols. Saturating nutrients were used to avoid nutrient-induced growth limitation. D. Isolates grown at saturating light intensity (µmol quanta m⁻² s⁻¹). E. Semi-continuous cultures were diluted using media that was previously adjusted to the appropriate temperature. Dilution frequencies were set so that cells were maintained in constant exponential growth phase and growth rates were reported when cultures were fully acclimated to the experimental conditions, after statistically invariant growth rates were recorded for at least 3–5 generations [98] F. Upper and lower thermal limits were tested repeatedly (at least 3 times) G. Multiple biomass parameter/proxies were used to determine daily abundance and included cell counts, extracted chlorophyll a, and in vivo chlorophyll a fluorescence. Each method could be used reliably to determine steady-state acclimation. H. At each temperature, the maximum acclimated specific growth rate (d⁻¹) for each isolate was determined by regressing the change in the log of fluorescence, cell count or chlorophyll a over time and testing the equality of slopes from at least three serial cultures (α = 0.05) [99]. If slopes of serial growth curves were not significantly different, the average regression coefficient was used to estimate the common slope, which represented the average acclimated growth rate and the standard error.

Footnotes:

¹Growth rates were determined at five temperatures.

²Growth rates were determined at 4 temperatures. Cells failed to grow at 35°C and reliable growth estimates could not be obtained at 10°C.

³Growth rates were determined for 4 temperatures, cells exhibited no growth at 35°C.

⁴Isolates also grown in 4 different nitrogen species (nitrate, ammonium, urea, and glutamate).

⁵Carbonate system also held within a specific range at ambient conditions.

⁶Recorded growth for 5 days after acclimation (not necessarily 3–5 generations).

⁷After >8 generations.

⁸This was performed for upper limit only.

⁹Upper limited tested repeatedly. Lower limit was below 0°C, the lowest temperature tested.

¹⁰Upper and lower limits were tested for P. donghaiense, however only lower limit was tested for Synechococcus. Upper and lower limit tests were performed twice, not three times.

¹¹This was performed for upper limit in all isolates, and in one isolate for the lower limit.

¹²Fluorescence alone was used.

¹³ In vivo fluorescence reported, but Chl measured at the first and last culture days, as well as PN and PC.

¹⁴Growth rates were determined using in vivo fluorescence, but were not significantly different from growth rates determined using cell counts.

¹⁵Significant differences between slopes of replicate cultures were not tested. Instead, the mean slope was used. Variation within a temperature treatment was much smaller than variation between temperatures.