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Horizontal Gene Transfer of Phytochelatin Synthases from Bacteria to Extremophilic Green Algae

  • Microbiology of Aquatic Systems
  • Published:
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Abstract

Transcriptomic sequencing together with bioinformatic analyses and an automated annotation process led us to identify novel phytochelatin synthase (PCS) genes from two extremophilic green algae (Chlamydomonas acidophila and Dunaliella acidophila). These genes are of intermediate length compared to known PCS genes from eukaryotes and PCS-like genes from prokaryotes. A detailed phylogenetic analysis gives new insight into the complicated evolutionary history of PCS genes and provides evidence for multiple horizontal gene transfer events from bacteria to eukaryotes within the gene family. A separate subgroup containing PCS-like genes within the PCS gene family is not supported since the PCS genes are monophyletic only when the PCS-like genes are included. The presence and functionality of the novel genes in the organisms were verified by genomic sequencing and qRT-PCR. Furthermore, the novel PCS gene in Chlamydomonas acidophila showed very strong induction by cadmium. Cloning and expression of the gene in Escherichia coli clearly improves its cadmium resistance. The gene in Dunaliella was not induced, most likely due to gene duplication.

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Acknowledgments

This work was supported by the Spanish Ministry of Economy and Competitivity (MINECO) under Grant CGL-2011-22540 to AA. We acknowledge CSC-IT Center for Science Ltd. (Finland) for the allocation of computational resources. Dr. Salvador Mirete (Centro de Astrobiología CSIC-INTA) is acknowledged for assistance with the growth curves for the analysis of cadmium resistance in E. coli. FP-S was supported by a JAE-pre fellowship from the Spanish Consejo Superior de Investigaciones Científicas (CSIC).

Author information

Authors and Affiliations

  1. Department of Agricultural Sciences, University of Helsinki, P.O. Box 27, 00014, Helsinki, Finland

    Sanna Olsson

  2. Centro de Astrobiología (CSIC-INTA), Carretera de Ajalvir Km 4, Torrejón de Ardoz, 28850, Madrid, Spain

    Sanna Olsson, Vanessa Penacho, Fernando Puente-Sánchez, José Eduardo Gonzalez-Pastor & Angeles Aguilera

  3. Bioarray, S.L., Parque Científico y Empresarial de la UMH. Edificio Quorum III, Avenida de la Universidad s/n, 03202, Elche, Alicante, Spain

    Vanessa Penacho

  4. Departamento de Microbiología-III, Facultad de Biología, Universidad Complutense (UCM), Madrid, Spain

    Silvia Díaz

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  1. Sanna Olsson
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  2. Vanessa Penacho
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  3. Fernando Puente-Sánchez
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  4. Silvia Díaz
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  6. Angeles Aguilera
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Corresponding author

Correspondence to Sanna Olsson.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Online Resource 1

ART II medium used for the cultivation of Dunaliella acidophila strain RT5. The volume was brought to 1 l with distilled water and the pH adjusted with 0.1 N H2SO4. (PDF 256 kb)

Online Resource 2

Quantitative reverse transcription PCR standard-curve parameters for target gene (novel phytochelatin synthase in Chlamydomonas acidophila CaPCS2) and the expression control (housekeeping) genes 18S rRNA and actin. E = amplification efficiency, S = slope, R2 = correlation coefficient. (PDF 393 kb)

Online Resource 3

Flow chart on the process from culture to extraction to analyses for the sequences used in the current study. (PDF 359 kb)

Online Resource 4

Results for BLASTx search using the open reading frame of transcripts coding for CaPCS2 (isotig12422) and DaPCS (comp2951_c3_seq1 and comp2951_c3_seq2) as query. (TXT 110 kb)

Online Resource 5

Introns in the predicted PCS genes from C. acidophila and D. acidophila. There are three spliceosomal introns in CaPCS2, four in CaPCS1 and three in DaPCS1. The introns are either the most common GT-AG splice site or variations of it. (PDF 19 kb)

Online Resource 6

Different contigs of component comp17619_c0 coding for the CaPCS1 gene in Chlamydomonas acidophila. (TXT 11 kb)

Online Resource 7

Different contigs of component comp11852_c0 coding for the CaPCS2 gene in Chlamydomonas acidophila. (TXT 18 kb)

Online Resource 8

PCR products amplified in Dunaliella acidophila strain RT5 using gradient annealing run on 1 % agarose gel (1X TBE). The gel electrophoresis shows amplification of products sized 1198 bp and 542 bp and several non-specific amplification products. The intensity of the non-specific bands depends on the used annealing temperature and PCR program. Wells 1: Ladder 1 kb (Promega Corporation), marker sizes from 250 kb to 10,000 kb, well 2: annealing 52-55 °C. Well 3: annealing 55-60 °C. (PDF 51 kb)

Online Resource 9

Phylogram based on the novel phytochelatin synthase genes and the most similar genes obtained with a BLASTx search against NCBI’s databases. The tree represents the majority consensus of trees sampled after stationarity in the Bayesian analysis. PP values from the Bayesian inference > 0.50 are indicated above each branch. The amino acid alignment was 383 aa long. (PDF 871 kb)

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Olsson, S., Penacho, V., Puente-Sánchez, F. et al. Horizontal Gene Transfer of Phytochelatin Synthases from Bacteria to Extremophilic Green Algae. Microb Ecol 73, 50–60 (2017). https://doi.org/10.1007/s00248-016-0848-z

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  • DOI: https://doi.org/10.1007/s00248-016-0848-z

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