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Appl. Sci. 2020, 10, 2531 9 of 18 showed better growth for our strain, and it has been reported that fusiform cells have a greater lipid percentage as compared to other forms [43,51]. Appl. Sci. 2020, 10, x FOR PEER REVIEW 9 of 22 Figure3. AnalysFisguoref3P..AtnrailcysoirsnofuPt.utrmicorbnuiotumbaiosmsa(sAs()Aa)andlilpipidiadccuamccuulatmionu(lBa)tiinoanuto(tBro)phiniccaoundtiotitornosphicconditions under continuous white (W) light, red (R) light, and red with light shift (RS), which is red light during under continuous white (W) light, red (R) light, and red with light shift (RS), which is red light during the exponential phase and then a shift to white light. Experiments were done in triplicate in 50 mL the exponential phase and then a shift to white light. Experiments were done in triplicate in 50 mL liquid culture, and cells were collected at day 10. Results are expressed as mean value ± standard deviation. An asterisk (*) indicates significant differences compared to the white light (W) used as liquid culture, and cells were collected at day 10. Results are expressed as mean value ± standard control. Statistical analysis was done using one-way ANOVA. deviation. An asterisk (*) indicates significant differences compared to the white light (W) used as Total lipids were also analyzed in the same conditions, using dry weight estimation. Nile red control. StatiAsptpil.cSacil. 2a02n0,a1l0y, xsFiOsRwPEaEsR RdEoVnIEeW using one-way ANOVA. 11 of 22 assay was used for rapid screening of lipid and confocal microscopy for visual analysis, as described previouslyalindustrial applicationound321.89 mg/g dry w in W in our study,whichtinRSwas significantlure3B).Itis interesting ng from red light to full It is known thattheusuhtintensity, nutrient de An increase inC/Nratioeenreported [11]. The sa e light shift, whereas the In addition, lipidaccumellasp.[29], whichcanbmbersinred [20]. P. tricornutum produces lipids that are potentially significant for sever s. It has been reported that the crude lipids extracted from P. tricornutum are ar eight [42] or 34% of dry weight [43] and are comparable to the lipid extracted h is 0.4 g/g dry weight in control white light. Furthermore, the lipid dry weig y increased to 2.3-fold and 1.8-fold as compared to W and R, respectively (Fig to note the 1.8 fold increase in lipid from R to RS, which suggests that shifti spectrum during the stationary phase has an impact on lipid accumulation. al change in the lipid composition is influenced by the cellular C/N ratio, lig pletion, and other possible inducible biotic/abiotic stress conditions [28–30]. , lipid, and carbohydrate in red light acclimated P. tricornutum culture has b me study reported an increase in the lipid content from red light to blu reverse shift, from blue to red light, promoted an increase in carbohydrates. ulation under red light has been observed in Botryococcus sp. [7] and Chlor e linked with effective nitrogen consumption rate and a doubling of cell nu aids in increasing the total lipid content. In addition, high light intensity has id biosynthesis in Nannocloropsis gaditana, which has been correlated with th fatty acid synthase of type 1 (FAS1) [44] and polyketide synthase (PKS) an light, which to affect lip of cytosolic been shown e regulation d the down- Figure 4. Microscopic fluorescent images (top) and transmitted with fluorescent overlay (bottom). Figure 4. Microscopic fluorescent images (top) and transmitted with fluorescent overlay (bottom). Visualization of lipid droplets in Phaeodactylum tricornutum using Nile red staining. Each panel Visualization ofrelpirpesiedntsdNriolepreldesttsainiingPofhliapeidodraocptleytsluinmwhtitreilcigohrtncountduitmionsu(as,icn)agndNinirleedlirgehdt(bs,dt)a.Sicnalieng. Each panel represents 100 nm. represents Nile red staining of lipid droplets in white light conditions (a,c) and in red light (b,d). Scale represents 1030.5.nFmatt.y Acid Methyl Esters Analysis (FAMEs) Polyunsaturated fatty acids (PUFA) were present in greatest abundance, followed by monounsaturated fatty acids (MUFA) and lower amounts of saturated fatty acids (SFA), which constituted 41.86%, 31.80%, and 26.34%, respectively (Table 3). MUFA levels were lower in RS (28.91%) compared to W (31.80%) or R (31.85%) (Table 3). In addition, the P. tricornutum PUFA ratio of omega-3/omega-6 was lower in R and RS compared to control W (Table 3). The evidence suggests that a high ratio of dietary omega-3/omega-6 PUFAs reduces the risk of several diseases [52]. Therefore, P. tricornutum cultured in W is a potentially more valuable source of functional food and animal feed components than P. tricornutum cultivated under R or RS conditions. These results suggest that the RS condition regulated the fatty acid content in P. tricornutum, specifically lowering MUFAs and increasing PUFAs. It has been reported that stress conditions such as nitrogen depletion,PDF Image | Red Light Variation Lipid Profiles in Phaeodactylum tricornutum
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