New insights into phenotypic heterogeneity for the distinct lipid accumulation of Schizochytrium sp. H016.
New insights into phenotypic heterogeneity for the distinct lipid accumulation of Schizochytrium sp. H016.
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DOI：
10.1186/s13068-022-02126-w
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发表时间：
2022-03-25
期刊：
影响因子：
--
通讯作者：
Yu L
中科院分区：
文献类型：
--
作者：
Bao Z;Zhu Y;Zhang K;Feng Y;Zhang M;Li R;Yu L
Schizochytrium sp.是一种海洋异养原生生物，也是一种重要的高附加值二十二碳六烯酸可持续资源。裂殖壶菌（Schizochytrium sp.）在连续传代过程中产生不同的表型，导致油脂产量严重降低，并使该菌株在科学研究和工业生产中的应用复杂化。因此，获得一个更好的理解的表型差异和分子机制的细胞间异质性的裂殖壶菌属是必要的。在连续传代培养后，Schizochytrium sp. H 016分化为两个不同形态的亚群，并表现出产脂能力下降。具有降解脂滴的细胞亚群的存在导致总体脂质产量的大幅降低。在这里，提出了一种基于荧光激活细胞分选的快速筛选策略，根据其脂质产生能力快速分类和分离亚群。具有高细胞脂质含量的亚群的最终生物量和脂质产量（即，H 016-H）分别为38.83和17.22 g/L，分别是低脂亚群（即H 016-H）的2.07和5.38倍，H016-L）。随后，进行时间分辨转录组分析以阐明不同亚群中表型异质性的机制。结果表明，H 016-L中细胞周期和脂质降解相关基因的表达显著上调，而H 016-H中脂肪酸合成和甘油酯积累相关代谢途径的表达显著上调。本研究创新性地利用流式细胞术结合转录组技术，对裂殖壶菌不同细胞亚群的表型异质性提供了新的认识，为指导高脂产油微生物的选育奠定了基础。在线版本包含补充材料，可通过10.1186/s13068-022-02126-w获得。
Schizochytrium sp. is a marine heterotrophic protist and an important sustainable resource for high value-added docosahexaenoic acid in the future. The production of different phenotypes during the continuous subculture of Schizochytrium sp. results in a serious reduction in lipid yield and complicates the used of this strain in scientific research and industrial production. Hence, obtaining an improved understanding of the phenotypic differences and molecular mechanisms underlying the cell-to-cell heterogeneity of Schizochytrium sp. is necessary. After continuous culture passage, Schizochytrium sp. H016 differentiated into two subpopulations with different morphologies and showed decreased capacity for lipid production. The presence of cell subpopulations with degraded lipid droplets led to a substantial decrease in overall lipid yield. Here, a rapid screening strategy based on fluorescence-activated cell sorting was proposed to classify and isolate subpopulations quickly in accordance with their lipid-producing capability. The final biomass and lipid yield of the subpopulation with high cell lipid content (i.e., H016-H) were 38.83 and 17.22 g/L, respectively, which were 2.07- and 5.38-fold higher than those of the subpopulation with low lipid content (i.e., H016-L), respectively. Subsequently, time‑resolved transcriptome analysis was performed to elucidate the mechanism of phenotypic heterogeneity in different subpopulations. Results showed that the expression of genes related to the cell cycle and lipid degradation was significantly upregulated in H016-L, whereas the metabolic pathways related to fatty acid synthesis and glyceride accumulation were remarkably upregulated in H016-H. This study innovatively used flow cytometry combined with transcriptome technology to provide new insights into the phenotypic heterogeneity of different cell subpopulations of Schizochytrium sp. Furthermore, these results lay a strong foundation for guiding the breeding of oleaginous microorganisms with high lipid contents. The online version contains supplementary material available at 10.1186/s13068-022-02126-w.
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