Abstract
Marine microalga Isochrysis galbana is an important feed species with a high nutritional value. Different from other unicellular algae, its cell contains two chloroplasts which are the major sites for lipid synthesis. Here, we optimized a chloroplast isolation approach suitable for the isolation of I. galbana chloroplasts and determined the purity and integrity of the isolated chloroplasts through microscopic observations and enzyme activity assay. The chloroplast lipids were analyzed with a ultrahigh-performance liquid chromatography-Q Exactive Orbitrap-mass spectrometry. This newly developed isolation approach is simple and reliable to isolate chloroplasts with high integrity and purity. The average yield of intact chloroplasts was 15.3%±0.1%. Glycolipids and acylglycerols were the main chloroplast lipids. Glycolipids accounted for 56.6% of chloroplast lipid. Digalactosyldiacylglycerol (DGDG), monogalactosyldiacylglycerol (MGDG) and sulfoquinovosyldiacylglycerol (SQDG) were the main glyceroglycolipids. The fatty acyl R1/R2 were mostly 18:4/16:1, 18:3/16:1 and 18:4/18:5 in DGDGs, 14:0/18:4, 18:4/18:5, 18:4/18:4 and 18:3/18:4 in MGDGs and 16:0/14:0, 16:0/18:3, and 18:4/18:3 in SQDGs. In addition, diacylglycerol (DAG) was the most abundant acylglycerols; the content of 22:6/18:4-DAG was the highest. There was a little amount of glycosphingolipid (GSL) in chloroplast. Digalactosylmonoglyceride (DGMG), monogalactosylmonoglyceride (MGMG), sulfoquinovosylmonoacylglycerol (SQMG), monoglyceride (MAG), phospholipids (PLs), ceramide (Cer) and betaine lipids were nearly undetectable in chloroplast. The fatty acid proportions of DGDGs, MGDGs, SQDGs, DAGs, triglycerides (TAGs) and GSLs were either higher or lower than or similar to those of whole-cell. Collectively, our isolation approach is applicable to many aspects of chloroplast biology, and may offer a reference for the isolation of chloroplasts from other marine microalgae.
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Acknowledgements
This research was supported by the Ningbo Science and Technology Research Projects (No. 2019B10006), the National Key Research and Development Program of China (No. 2019YFD0900400), the Zhejiang Major Science Project (No. 2019C02057), the China Agriculture Research System of MOF and MARA, the Natural Science Foundation of Ningbo (No. 2019A610416), the Ningbo Science and Technology Research Projects (No. 2019C10023), and the National Natural Science Foundation of China (No. 31801724).
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Zhong, Y., Li, Y., Xu, J. et al. Isolation of Chloroplasts from Marine Microalga Isochrysis galbana Parke for Their Lipid Composition Analysis. J. Ocean Univ. China 21, 225–235 (2022). https://doi.org/10.1007/s11802-022-4834-4
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DOI: https://doi.org/10.1007/s11802-022-4834-4