Examination of carbohydrate and lipid metabolic changes during Haematococcus pluvialis non-motile cell germination using transcriptome analysis
During the complex Haematococcus pluvialis life cycle, germination from non-motile cells to motile cells is the key stage for cell recovery from resting spores. Three phases can be recognized: first, repeated mitotic events; next, cytokinesis to form the zoospore; and finally, a fast release of motile cells. After HiSeq 2000 sequencing of RNA collected at four time points during non-motile cell germination, a total of 2202 differentially expressed genes (DEGs) were identified. In the expression profiles, there is a consistent increase in the expression level of α-amylase and ACAA1 in the entire process, indicating starch and lipid mobilization. In phase 1, two down-regulated genes (fructose-1, 6-bisphosphate aldolase (FBA) and pyruvate kinase (PK)) of glycolysis limited the provision of acetyl-CoA for the TCA cycle, which could be compensated by fatty acid degradation (up-regulation of ACAA1) in the glyoxysome. In phase 2, nine and eight up-regulated enzymes of carbohydrate (glycolysis, TCA cycle, and pentose phosphate pathway) and lipid (fatty acid synthesis and degradation) metabolism, respectively, would increase the metabolic rate and come into a balance between production and consumption of starch and lipid. Till phase 3, the expression of the vast majority of carbohydrate metabolism-related DEGs remained high, while lipid metabolism did not. This suggested that the carbon flux centered on carbohydrate metabolism in this phase. In addition, several isozymes of FBA, GAPDH, PK, and so on were separated by SMART analysis and are postulated to serve different actions during H. pluvialis non-motile cell germination.
KeywordsHaematococcus pluvialis Non-motile cell Motile cell Germination Metabolic change
This research was supported by the National Natural Science Foundation of China (31702366 and 31572639). We thank Shanghai Personal Biotechnology Co., Ltd. (Shanghai, China) for Illumina transcriptome sequencing and initial data analysis. We thank Ling Li, Fang Su, and Chunhui Zhang for their work in cultivation and collection of alga. Special thanks to Dr. John van der Meer (Pan-American Marine Biotechnology Association) for his assistance with proofreading.
This study was designed by QL and JL. QL performed the experiments. QL and LZ analyzed the data. QL and JL wrote the paper. All authors read and approved the final manuscript.
Compliance with ethical standards
The authors state this research is free of conflicts of interests.
- Droop MR (1953) On the ecology of flagellates from some brackish and fresh water rockpools of Finland. Acta Bot Fenn 51:3–52Google Scholar
- Elliot A (1934) Morphology and life history of Haematococcus pluvialis. Arch Protistenkd 82:250–272Google Scholar
- Grabherr MG, Haas BJ, Yassour M, Levin JZ, Thompson DA, Amit I, Adiconis X, Fan L, Raychowdhury R, Zeng QD, Chen ZH, Mauceli E, Hacohen N, Gnirke A, Rhind N, di Palma F, Birren BW, Nusbaum C, Lindblad-Toh K, Friedman N, Regev A (2011) Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat Biotechnol 29:644–U130CrossRefGoogle Scholar
- Haas BJ, Papanicolaou A, Yassour M, Grabherr M, Blood PD, Bowden J, Couger MB, Eccles D, Li B, Lieber M, MacManes MD, Ott M, Orvis J, Pochet N, Strozzi F, Weeks N, Westerman R, William T, Dewey CN, Henschel R, Leduc RD, Friedman N, Regev A (2013) De novo transcript sequence reconstruction from RNA-seq using the Trinity platform for reference generation and analysis. Nat Protoc 8:1494–1512CrossRefGoogle Scholar
- Liu J, Zhang J (2000) Photosynthetic and respiration rate of Haematococcus pluvialis. Oceanol Limnol Sinica 31(5):490–495Google Scholar
- Lund JWG, Fryxell GAE (1984) Survival strategies of the algae. Cambridge University Press, CambridgeGoogle Scholar
- Miltiadou D, Hager-Theodorides AC, Symeou S, Constantinou C, Psifidi A, Banos G, Tzamaloukas O (2017) Variants in the 3′ untranslated region of the ovine acetyl-coenzyme A acyltransferase 2 gene are associated with dairy traits and exhibit differential allelic expression. J Dairy Sci 100:6285–6297CrossRefGoogle Scholar
- Recht L, Topfer N, Batushansky A, Sikron N, Gibon Y, Fait A, Nikoloski Z, Boussiba S, Zarka A (2014) Metabolite profiling and integrative modeling reveal metabolic constraints for carbon partitioning under nitrogen starvation in the green algae Haematococcus pluvialis. J Biol Chem 289:30387–30403CrossRefGoogle Scholar
- Stincone A, Prigione A, Cramer T, Wamelink MM, Campbell K, Cheung E, Olin-Sandoval V, Gruning NM, Kruger A, Tauqeer Alam M, Keller MA, Breitenbach M, Brindle KM, Rabinowitz JD, Ralser M (2015) The return of metabolism: biochemistry and physiology of the pentose phosphate pathway. Biol Rev Camb Philos Soc 90:927–963CrossRefGoogle Scholar