Open access
Announcement
4 June 2020

Draft Genome Sequence of the Astaxanthin-Producing Microalga Haematococcus lacustris Strain NIES-144

ABSTRACT

Haematococcus lacustris is an industrially important eukaryotic microalga that is thought to be a great source of natural astaxanthin with strong antioxidant activity. Here, we report the draft assembly and annotation results of the genome of H. lacustris NIES-144. These data will expand our knowledge of the molecular biological features of this microalga.

ANNOUNCEMENT

Haematococcus is a genus of eukaryotic Chlorophyceae microalgae that can form a red immotile cyst and accumulate the highest content of natural astaxanthin reported to date under stress conditions (1). Although these microalgae have been studied as a natural resource for astaxanthin, which is a high-value carotenoid with strong antioxidant activity (2), the genomic information is limited to H. lacustris strain SAG192.80 (3).
To expand our molecular biological knowledge of these industrially important microalgae, we determined the draft genome sequence of H. lacustris NIES-144, which was obtained from the Natural Institute for Environmental Studies (NIES, Japan). H. lacustris NIES-144 was cultured in C medium (4) under 14/10-h light/dark photocycles at 25°C. Extraction of genomic DNA from Haematococcus cells was performed using a FastDNA Spin kit for soil (MP Biomedical, USA). Paired-end and mate pair libraries (3 kb and 10 kb, respectively) were prepared using a combination of the Covaris (USA) sonicator and the TruSeq DNA LT sample prep kit or the Nextera mate pair sample preparation kit (Illumina), respectively. The paired-end library was sequenced using the TruSeq rapid sequencing by synthesis (SBS) kit on the Illumina HiSeq 2500, while the mate pair library was sequenced using the TruSeq SBS kit v3 on the Illumina HiSeq 2000 platform.
The mate pair reads (average, 154,807,864 reads) were processed with cutadapt 1.2.1 (5) to remove adapter sequences. The paired-end reads (215,289,986 reads) and trimmed mate pair reads (average, 105,701,143 reads) were assembled into 9,693 scaffolds with a total length of 172 Mb (genome coverage, 186×; GC content, 58.4%; N50 scaffold length, 38,941 bp) using ALLPATHS-LG R45226 (6) with the following parameters: GENOME_SIZE: 125,000,000; FRAG_COVERAGE: 100; JUMP_COVERAGE: 100; and HAPLOIDFY: True. The completeness of the draft genome was 57.7% based on the Benchmarking Universal Single-Copy Orthologs (BUSCO) software v3.1.0 (eukaryota_odb9 database) (7). Prior to gene structure prediction, the repeat sequences of the H. lacustris NIES-144 genome were identified and masked by RepeatMasker v4.0.9 (8) with default parameters. The gene structure of the masked Haematococcus genome was predicted by using MAKER v2.31.10 (9) in collaboration with AUGUSTUS 3.3.2 (10), SNAP v2006-07-28 (11), and GeneMark-ES 4.3.0 (12) (model parameters, Chlamydomonas, Arabidopsis thaliana, and Chlamydomonas reinhardtii, respectively). For RNA and protein homology evidence in the MAKER prediction, we also recruited the transcriptome data of H. lacustris NIES-144 (SRA accession number SRX3729494) (13) and the protein sequences of representative eukaryotic species, including H. lacustris strain SAG192.80 (3). A total of 13,309 genes were functionally annotated for H. lacustris NIES-144 by BLASTp analysis against the UniProtKB SWISS-PROT and TrEMBL databases (14) with E value thresholds of <1.0 × 10−5 and InterProScan v5.36-75.0 (15) analysis against the Pfam database (16). Also, 277 tRNAs were predicted using tRNAscan-SE v2.0 (17). This genome will provide the prerequisite information for genetic engineering and spur the further development of efficient astaxanthin production by this microalga.

Data availability.

This whole-genome shotgun project has been deposited at DDBJ/ENA/GenBank under BioProject number PRJDB8952 (BioSample number SAMD00192397). This version of the project has the accession number BLLF00000000 and consists of sequences deposited under the accession numbers BLLF01000001 to BLLF01009693. The raw reads can be accessed under the SRA accession number DRP005830.

ACKNOWLEDGMENTS

We thank Ayano Nakatani for technical assistance. Genome sequencing and assembly were conducted at Dragon Genomics Center, TaKaRa Bio, Inc. (Otsu, Japan). The computational analysis in this study was performed using the Super Computer System at the Institute for Chemical Research, Kyoto University.
We declare that this research was conducted in the absence of any commercial or financial relationships.

REFERENCES

1.
Liu J, Sun Z, Gerken H, Liu Z, Jiang Y, Chen F. 2014. Chlorella zofingiensis as an alternative microalgal producer of astaxanthin: biology and industrial potential. Mar Drugs 12:3487–3515.
2.
Ambati RR, Phang S-M, Ravi S, Aswathanarayana RG. 2014. Astaxanthin: sources, extraction, stability, biological activities and its commercial applications—a review. Mar Drugs 12:128–152.
3.
Luo Q, Bian C, Tao M, Huang Y, Zheng Y, Lv Y, Li J, Wang C, You X, Jia B, Xu J, Li J, Li Z, Shi Q, Hu Z. 2019. Genome and transcriptome sequencing of the astaxanthin-producing green microalga, Haematococcus pluvialis. Genome Biol Evol 11:166–173.
4.
Ichimura T. 1971. Sexual cell division and conjugation-papilla formation in sexual reproduction of Closterium strigosum. Proc 7th Int Symp Seaweed Res 1971:208–214.
5.
Martin M. 2011. Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J 17:10–12.
6.
Gnerre S, MacCallum I, Przybylski D, Ribeiro FJ, Burton JN, Walker BJ, Sharpe T, Hall G, Shea TP, Sykes S, Berlin AM, Aird D, Costello M, Daza R, Williams L, Nicol R, Gnirke A, Nusbaum C, Lander ES, Jaffe DB. 2011. High-quality draft assemblies of mammalian genomes from massively parallel sequence data. Proc Natl Acad Sci U S A 108:1513–1518.
7.
Simão FA, Waterhouse RM, Ioannidis P, Kriventseva EV, Zdobnov EM. 2015. BUSCO: assessing genome assembly and annotation completeness with single-copy orthologs. Bioinformatics 31:3210–3212.
8.
Tarailo-Graovac M, Chen N. 2009. Using RepeatMasker to identify repetitive elements in genomic sequences. Curr Protoc Bioinformatics 25:4.10.1–4.10.14.
9.
Cantarel BL, Korf I, Robb SMC, Parra G, Ross E, Moore B, Holt C, Sánchez Alvarado A, Yandell M. 2008. MAKER: an easy-to-use annotation pipeline designed for emerging model organism genomes. Genome Res 18:188–196.
10.
Stanke M, Schöffmann O, Morgenstern B, Waack S. 2006. Gene prediction in eukaryotes with a generalized hidden Markov model that uses hints from external sources. BMC Bioinformatics 7:62.
11.
Johnson AD, Handsaker RE, Pulit SL, Nizzari MM, O'Donnell CJ, de Bakker PIW. 2008. SNAP: a Web-based tool for identification and annotation of proxy SNPs using HapMap. Bioinformatics 24:2938–2939.
12.
Ter-Hovhannisyan V, Lomsadze A, Chernoff YO, Borodovsky M. 2008. Gene prediction in novel fungal genomes using an ab initio algorithm with unsupervised training. Genome Res 18:1979–1990.
13.
Lee C, Ahn J-W, Kim J-B, Kim JY, Choi Y-E. 2018. Comparative transcriptome analysis of Haematococcus pluvialis on astaxanthin biosynthesis in response to irradiation with red or blue LED wavelength. World J Microbiol Biotechnol 34:96.
14.
Boeckmann B, Bairoch A, Apweiler R, Blatter M-C, Estreicher A, Gasteiger E, Martin MJ, Michoud K, O'Donovan C, Phan I, Pilbout S, Schneider M. 2003. The SWISS-PROT protein knowledgebase and its supplement TrEMBL in 2003. Nucleic Acids Res 31:365–370.
15.
Jones P, Binns D, Chang H-Y, Fraser M, Li W, McAnulla C, McWilliam H, Maslen J, Mitchell A, Nuka G, Pesseat S, Quinn AF, Sangrador-Vegas A, Scheremetjew M, Yong S-Y, Lopez R, Hunter S. 2014. InterProScan 5: genome-scale protein function classification. Bioinformatics 30:1236–1240.
16.
El-Gebali S, Mistry J, Bateman A, Eddy SR, Luciani A, Potter SC, Qureshi M, Richardson LJ, Salazar GA, Smart A, Sonnhammer ELL, Hirsh L, Paladin L, Piovesan D, Tosatto SCE, Finn RD. 2019. The Pfam protein families database in 2019. Nucleic Acids Res 47:D427–D432.
17.
Lowe TM, Eddy SR. 1997. tRNAscan-SE: a program for improved detection of transfer RNA genes in genomic sequence. Nucleic Acids Res 25:955–964.

Information & Contributors

Information

Published In

cover image Microbiology Resource Announcements
Microbiology Resource Announcements
Volume 9Number 234 June 2020
eLocator: 10.1128/mra.00128-20
Editor: Antonis Rokas, Vanderbilt University

History

Received: 1 April 2020
Accepted: 14 May 2020
Published online: 4 June 2020

Contributors

Authors

Daichi Morimoto
Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, Japan
Takashi Yoshida
Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, Japan
Shigeki Sawayama
Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, Japan

Editor

Antonis Rokas
Editor
Vanderbilt University

Notes

Address correspondence to Daichi Morimoto, [email protected], or Shigeki Sawayama, [email protected].

Metrics & Citations

Metrics

Note:

  • For recently published articles, the TOTAL download count will appear as zero until a new month starts.
  • There is a 3- to 4-day delay in article usage, so article usage will not appear immediately after publication.
  • Citation counts come from the Crossref Cited by service.

Citations

If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. For an editable text file, please select Medlars format which will download as a .txt file. Simply select your manager software from the list below and click Download.

View Options

Figures

Tables

Media

Share

Share

Share the article link

Share with email

Email a colleague

Share on social media

American Society for Microbiology ("ASM") is committed to maintaining your confidence and trust with respect to the information we collect from you on websites owned and operated by ASM ("ASM Web Sites") and other sources. This Privacy Policy sets forth the information we collect about you, how we use this information and the choices you have about how we use such information.
FIND OUT MORE about the privacy policy