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Transcriptome sequencing and annotation of the halophytic microalga Dunaliella salina

盐生海藻杜氏盐藻的转录组测序以及注释

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Abstract

The unicellular green alga Dunaliella salina is well adapted to salt stress and contains compounds (including β-carotene and vitamins) with potential commercial value. A large transcriptome database of D. salina during the adjustment, exponential and stationary growth phases was generated using a high throughput sequencing platform. We characterized the metabolic processes in D. salina with a focus on valuable metabolites, with the aim of manipulating D. salina to achieve greater economic value in large-scale production through a bioengineering strategy. Gene expression profiles under salt stress verified using quantitative polymerase chain reaction (qPCR) implied that salt can regulate the expression of key genes. This study generated a substantial fraction of D. salina transcriptional sequences for the entire growth cycle, providing a basis for the discovery of novel genes. This first full-scale transcriptome study of D. salina establishes a foundation for further comparative genomic studies.

摘要

目 的

解析杜氏盐藻代谢过程, 主要关注盐胁迫下累积的代谢物 (渗透平衡产物、多胺和类胡萝卜素) 的代谢。

创新点

本研究通过高通量测序产生了大量来自杜氏盐藻整个生长周期的转录组数据, 描述了杜氏盐藻在盐胁迫下累积的渗透平衡产物、 多胺和类胡萝卜素的代谢过程。另外通过该手段也进一步分析了盐胁迫处理下, 抑制精胺合成底物的供应可能会缓解盐藻增殖对胡萝卜素含量的影响。

方 法

以来自3 个不同生长时期的杜氏盐藻为材料, 进行大规模转录组测序。 在转录组功能注释的基础上, 预测了杜氏盐藻盐胁迫下累积的渗透平衡产物 (图3)、 多胺 (图4) 和类胡萝卜素 (图5) 的代谢路径。 利用相对定量聚合酶链反应 (qPCR) 技术构建了相关代谢路径中关键基因的表达谱 (图6)。

结 论

通过杜氏盐藻转录组测序共获取了 39820 条单一序列。 在功能注释和聚类分析的基础上预测了杜氏盐藻盐胁迫下累积的渗透平衡产物 (甘油和脯氨酸)、 多胺以及类胡萝卜素的代谢路径。 相关代谢途径的关键酶的表达谱分析, 说明盐能够调节甘油、脯氨酸以及多胺的代谢过程。 抑制精胺合成底物的供应可能会缓解盐藻增殖对胡萝卜素含量的影响。

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Authors and Affiliations

  1. Department of Genetics and Developmental Biology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China

    Ling Hong, Jun-li Liu & Samira Z. Midoun

  2. Systems Biology Research Group, Bioengineering Department, University of California, 9500 Gilman Dr. San Diego, La Jolla, CA, 92093, USA

    Philip C. Miller

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  1. Ling Hong
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  2. Jun-li Liu
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  3. Samira Z. Midoun
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Correspondence to Philip C. Miller.

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Project supported by the National High-Tech R&D Program (863) of China (No. 2007AA09Z449)

Electronic supplementary material

Transcriptome sequencing and annotation of the halophytic microalga Dunaliella salina

Transcriptome sequencing and annotation of the halophytic microalga Dunaliella salina

Transcriptome sequencing and annotation of the halophytic microalga Dunaliella salina

Transcriptome sequencing and annotation of the halophytic microalga Dunaliella salina

Table S1 Primers of those selective genes involved in the metabolic processes in D. salina

Table S2 Summary of annotation of D. salina transcriptome

Table S3 Top-hit species (viridiplantae) list of D. salina BLAST-annotated uniseqs

11585_2017_260_MOESM8_ESM.xlsx

Table S4 Enzymes identified in metabolism of osmolytes (glycerol and proline), polyamines, and carotenoid through annotation of D. salina transcriptome

Table S5 The best hit of the highlighted enzymes in the metabolic processes of D. salina

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Hong, L., Liu, Jl., Midoun, S.Z. et al. Transcriptome sequencing and annotation of the halophytic microalga Dunaliella salina . J. Zhejiang Univ. Sci. B 18, 833–844 (2017). https://doi.org/10.1631/jzus.B1700088

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  • DOI: https://doi.org/10.1631/jzus.B1700088

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