In this study, the effects of limited and excess nitrate on biomass, lipid production, and fatty acid profile in Messastrum gracile SE-MC4 were determined. The expression of fatty acid desaturase genes, namely stearoyl-ACP desaturase (SAD), omega-6 fatty acid desaturase (ω-6 FAD), omega-3 fatty acid desaturase isoform 1 (ω-3 FADi1), and omega-3 fatty acid desaturase isoform 2 (ω-3 FADi2) was also assessed. It was found that nitrate limitation generally increased the total oil, α-linolenic acid (C18:3n3) and total polyunsaturated fatty acid (PUFA) contents in M. gracile. The reduction of nitrate concentration from 1.76 to 0.11 mM increased the total oil content significantly from 32.5 to 41.85% (dry weight). Palmitic (C16:0) and oleic (C18:1) acids as the predominant fatty acids in this microalgae constituted between 82 and 87% of the total oil content and were relatively consistent throughout all nitrate concentrations tested. The expression of SAD, ω-6 FAD, and ω-3 FADi2 genes increased under nitrate limitation, especially at 0.11 mM nitrate. The ω-3 FADi1 demonstrated a binary up-regulation pattern of expression under both nitrate-deficient (0.11 mM) and -excess (3.55 mM) conditions. Thus, findings from this study suggested that limited or excess nitrate could be used as part of a cultivation strategy to increase oil and PUFA content following media optimisation and more efficient culture methodology. Data obtained from the expression of desaturase genes would provide valuable insights into their roles under excess and limited nitrate conditions in M. gracile, potentially paving the way for future genetic modifications.
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Álvarez-Díaz PD, Ruiz J, Arbib Z, Barragán J, Garrido-Pérez C, Perales JA (2014) Lipid production of microalga Ankistrodesmus falcatus increased by nutrient and light starvation in a two-stage cultivation process. Appl Biochem Biotechnol 174(4):1471–1483
Asselborn V, Fernández C, Zalocar Y, Parodi ER (2015) Effects of chlorpyrifos on the growth and ultrastructure of green algae, Messastrum gracile. Ecotoxicol Environ Saf 120:334–341
Bogen C, Klassen V, Wichmann J, Russa ML, Doebbe A, Grundmann M, Uronen P, Kruse O, Mussgnug JH (2013) Identification of Monoraphidium contortum as a promising species for liquid biofuel production. Bioresour Technol 133:622–626
Breuer G, Lamers PP, Martens DE, Braaisma RB, Wijffels RH (2012) The impact of nitrogen starvation on the dynamics of triacylglycerol accumulation in nine microalgae strains. Bioresour Technol 124:217–226
Cha TS, Chen JW, Goh EG, Aziz A, Loh SH (2011) Differential regulation of fatty acid biosynthesis in two Chlorella in response to nitrate treatments and the potential of binary blending of microalgae oils for biodiesel application. Bioresour Technol 102:10633–10640
Choi GG, Kim BH, Ahn CY, Oh HM (2011) Effect of nitrogen limitation on oleic acid biosynthesis in Botryococcus braunii. J Appl Phycol 23:1031–1037
Dong CJ, Cao N, Zhang ZG, Shang QM (2016) Characterization of the fatty acid desaturase genes in cucumber: structure, phylogeny, and expression patterns. PLoS ONE 11(3):e0149917
Griffiths MJ, van Hille RP, Harrison ST (2012) Lipid productivity, settling potential and fatty acid profile of 11 microalgal species grown under nitrogen replete and limited conditions. J Appl Phycol 24(5):989–1001
Guillard RRL, Ryther JH (1962) Studies of marine planktonic diatoms Cyclotella nana, Hustedt and Detonula confervacae (Cleve) Gran. Can J Microbiol 8:229–239
He Q, Yang H, Hu C (2015a) Optimizing light regimes on growth and lipid accumulation in Ankistrodesmus fusiformis H1 for biodiesel production. Bioresour Technol 198:876–893
He Q, Yang H, Wu L, Hu C (2015b) Effect of light intensity on physiological changes, carbon allocation and neutral lipid accumulation in oleaginous microalgae. Bioresour Technol 191:219–228
Hoa LTP, Quang DN, Ha NTH, Tri NH (2011) Isolating and screening mangrove microalgae for anticancer activity. Res J Phytochem 5:156–162
Hu Q, Sommerfeld M, Jarvis E, Grihardi M, Posewitz M, Seibert M, Darzins A (2008) Miroalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant J 54:621–639
Jusoh M, Loh SH, Aziz A, Cha TS (2019) Gibberellin promotes cell growth and induces changes in fatty acid biosynthesis and upregulates fatty acid biosynthesis genes in Chlorella vulgaris UMT-M1. Appl Biochem Biotechnol 188:450–459
Kachroo A, Shanklin J, Whittle E, Lapchyk L, Hildebrand D, Kachroo P (2007) The Arabidopsis stearoyl-acyl carrier protein-desaturase family and the contribution of leaf isoforms to oleic acid synthesis. Plant Mol Biol 63:257–271
Khozin-Goldberg I (2016) Lipid metabolism in microalgae. In: Borowitzka M, Beardall J, Raven J (eds) The physiology of microalgae. Developments in applied phycology. Springer, Cham, pp 413–484
Kim S, Kim MJ, Jung MG, Lee S, Baek YS, Kang SH, Choi HG (2013) De novo transcriptome analysis of an arctic microalga, Chlamydomonas sp. Genes Genom 35:215–223
Krienitz L, Bock C (2011) SSU rRNA gene phylogeny of morphospecies affiliated to the bioassay alga “Selenastrum capricornutum” recovered the polyphyletic origin of crescent-shaped Chlorophyta. J Phycol 47:880–893
Liu J, Huang J, Fan KW, Jiang Y, Zhong Y, Sun Z, Chen F (2010) Production potential of Chlorella zofingienesis as a feedstock for biodiesel. Bioresour Technol 101:8658–8663
Liu J, Sun Z, Zhong Y, Huang J, Hu Q, Chen F (2012) Stearoyl-acyl carrier protein desaturase gene from the oleaginous microalga Chlorella zofingiensis: cloning, characterization and transcriptional analysis. Planta 236:1665–1676
Meng X, Yang J, Xu X, Zhang L, Nie Q, Xian M (2009) Biodiesel production from oleaginous organism. Renew Energy 34:1–5
Muto M, Kubota C, Tanaka M, Satoh A, Matsumoto M, Yoshino T, Tanaka T (2013) Identification and functional analysis of delta-9 desaturase, a key enzyme in PUFA synthesis, isolated from the oleaginous diatom Fistulifera. PLoS ONE 8:e73507
Nguyen HM, Cuiné S, Beyly-Adriano A, Légeret B, Billon E, Auroy P, Beisson F, Peltier G, Li-Beisson Y (2013) The green microalga Chlamydomonas reinhardtii has a single ω-3 fatty acid desaturase that localizes to the chloroplast and impacts both plastidic and extraplastidic membrane lipids. Plant Physiol 163:914–928
Pancha I, Chokshi K, George B, Ghosh T, Paliwal C, Maurya R, Mishra S (2014) Nitrogen stress triggered biochemical and morphological changes in the microalgae Scenedesmus sp. CCNM 1077. Bioresour Technol 56:146–154
Patidar SK, Mitra M, George B, Soundarya R, Mishra S (2014) Potential of Monoraphidium minutum for carbon sequestration and lipid production in response to varying growth mode. Bioresour Technol 172:32–40
Rocha GS, Parrish CC, Lombardi AT, Melão MDGG (2018) Biochemical and physiological responses of Selenastrum gracile (Chlorophyceae) acclimated to different phosphorus concentrations. J Appl Phycol 30:2167
Schmittgen TD, Livak KJ (2008) Analyzing real-time PCR data by the comparative CT method. Nat Protoc 3:1101–1108
Shah FH, Rashid O, San CT (2000) Temporal regulation of two isoforms of cDNA clones encoding delta 9-stearoyl-ACP desaturase from oil palm (Elaies guineensis). Plant Sci 152:27–33
Sipaúba-Tavares LH, Millan RN, Berchielli FA, Braga FMS (2011) Use of alternative media and different types of recipients in a laboratory culture of Ankistrodesmus gracilis (Reinsch) Korshikov (Chlorophyta). Acta Sci Biol Sci 33:247–253
Teh KY, Wan Afifudeen CL, Aziz A, Wong LL, Loh SH, Cha TS (2019) De novo whole genome sequencing data of two mangrove-isolated microalgae from Terengganu costal waters. Data Brief 27:104680
Wen Z, Chen F (2003) Heterotrophic production of eicosapentaenoic acid by microalgae. Biotechnol Adv 21:273–294
Xue Y, Chen B, Wang R, Win AN, Li J, Chai Y (2018a) Genome-wide survey and characterization of fatty acid desaturase gene family in Brassica napus and its parental species. Appl Biochem Biotechnol 184:582–598
Xue Y, Chen B, Win AN, Fu C, Lian J, Liu X, Wang R, Zhang X, Chai Y (2018b) Omega-3 fatty acid desaturase gene family from two ω-3 sources, Salvia hispanica and Perilla frutescens: cloning, characterization and expression. PLoS ONE 13(1):e0191432
Yee W (2016) Microalgae from the Selenastraceae as emerging candidates for biodiesel production: a mini review. World J Microbiol Biotechnol 32(4):64
You FM, Li P, Kumar S, Ragupathy R, Li Z, Fu YB, Cloutier S (2014) Genome-wide identification and characterization of the gene families controlling fatty acid biosynthesis in Flax (Linum usitatissimum L.). J Proteomic Bioinform 7:10
Zhang S, Liu PH, Yang X, Hao ZD, Zhang L, Luo N, Shi J (2014) Isolation and identification by 18S rDNA sequence of high lipid potential microalgal species for fuel production in Hainan Dao. Biomass Bioenergy 66:197–203
This research project was funded under Science Fund (Project No: 05-01-12-SF1007) from the Ministry of Agriculture (MOA) Malaysia.
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Anne-Marie, K., Yee, W., Loh, S.H. et al. Influence of nitrogen availability on biomass, lipid production, fatty acid profile, and the expression of fatty acid desaturase genes in Messastrum gracile SE-MC4. World J Microbiol Biotechnol 36, 17 (2020). https://doi.org/10.1007/s11274-019-2790-y
- Fatty acid desaturase