Functional Analysis of ACP1and RCA2in Chlamy Domonas Reinhardtii Lipid Accumulation
|Keywords||ACP1 RCA2 Chlamydomonas reinhardtii amiRNA lipid accumulation|
Biodiesel, as a renewable energy, is a quality substitute of petroleum and diesel. Although many species of Microalgae, which are great sources of biodiesel, have significant production of neutral fat (triacylglycerol), human do not know much about it. For the perspective of cost, high quality algae that is suitable for large-scale culture is not available and cultivation technology is still immature at present, so it hasn’t been commercialized to take advantage of microalgae for production of biodiesel.It has been reported that ACP1and RCA2are associated with lipid metabolism and photosynthesis in plants respectively, but there is no relevant reports in chlamydomonas reinhardtii. Chlamydomonas reinhardtii is an unicellular eukaryotic green algae. Its genetic transformation of nucleus, chloroplast and mitochondrion had been implemented successfully. Chlamydomonas reinhardtii, which is a great model organism, has a important significance of reference for us to understand regulatory mechanism of lipid metabolism of microalgae in depth. We study the ACP1and RCA2gene of Chlamydomonas reinhardtii by a cell-wall-deficient and arginine-requiring strain. We construct two kinds of amiRNA donor vector by PCR for ACP1and RCA2respectively, then we construct their miRNA receptor vector by mating-assisted genetically integrated clone (MAGIC), finally we put these two interference expression vector into cells respectively.The expression of amiRNA-ACP1and amiRNA-RCA2was high and the silencing effect was remarkable from the results of real-time quantitative PCR. Silence of these two gene had effect on accumulation of lipid content of Chlamydomonas reinhardtii, particularly ACP1. The results had demonstrated that ACP1and RCA2were closely related to the lipid metabolic pathway of Chlamydomonas reinhardtii, and there was great space to explore.