Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3-8-1, Meguro-ku, Tokyo, 153-8902 Japan.
Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Komaba 3-8-1, Meguro-ku, Tokyo, 153-8902 Japan.
Plant Cell Physiol. 2020 Jan 1;61(1):158-168. doi: 10.1093/pcp/pcz188.
Microalgae such as Chlamydomonas reinhardtii accumulate triacylglycerol (TAG), which is a potential source of biofuels, under stress conditions such as nitrogen deprivation, whereas Chlamydomonas debaryana NIES-2212 has previously been identified and characterized as one of the rare species of Chlamydomonas, which massively accumulates TAG in the stationary phase without external stress. As the high density of the cells in the stationary phase was supposed to act as a trigger for the accumulation of TAG in C. debaryana, in this study, C. debaryana was encapsulated in a Ca2+-alginate gel for the culture with high cell density. We discovered that the growth of the encapsulated cells resulted in the formation of spherical palmelloid colonies with high cell density, where daughter cells with truncated flagella remained wrapped within the mother cell walls. Interestingly, gel encapsulation markedly promoted proliferation of C. debaryana cells, and the encapsulated cells reached the stationary phase earlier than that of the free-living cells. Gel encapsulation also enhanced TAG accumulation. Gene expression analysis revealed that two genes of acyltransferases, DGAT1 and DGTT3, were upregulated in the stationary phase of free-living C. debaryana. In addition, the gene expression of these acyltransferases increased earlier in the encapsulated cells than that in the free-living cells. The enhanced production of TAG by alginate gel encapsulation was not found in C. reinhardtii which is known to use a different repertoire of acyltransferases in lipid accumulation.
微藻如莱茵衣藻在氮饥饿等胁迫条件下积累三酰基甘油(TAG),这是生物燃料的潜在来源,而衣藻巴氏变种 NIES-2212 先前被鉴定为衣藻的稀有物种之一,它在静止期大量积累 TAG 而无需外部胁迫。由于静止期细胞密度高被认为是衣藻积累 TAG 的触发因素,因此在本研究中,衣藻被包封在 Ca2+-藻酸盐凝胶中进行高密度培养。我们发现,包封细胞的生长导致形成具有高密度的球形棕榈状集落,其中带有截断鞭毛的子细胞仍被包裹在母细胞壁内。有趣的是,凝胶包封明显促进了衣藻细胞的增殖,并且包封细胞比自由生活细胞更早进入静止期。凝胶包封还促进了 TAG 的积累。基因表达分析表明,两种酰基转移酶基因,DGAT1 和 DGTT3,在自由生活的衣藻巴氏变种的静止期上调。此外,这些酰基转移酶的基因表达在包封细胞中比在自由生活细胞中更早增加。在已知使用不同酰基转移酶库进行脂质积累的莱茵衣藻中未发现藻酸盐凝胶包封增强 TAG 的产生。