Strains

From Takako Kato-Minoura, Chuo University-Japan, October 2021

Kato-Minoura T, Ogiwara Y, Yamano T, Fukuzawa H, Kamiya R. Chlamydomonas reinhardtii tubulin-gene disruptants for efficient isolation of strains bearing tubulin mutations. PLoS One. 2020 Nov 23;15(11):e0242694. doi: 10.1371/journal.pone.0242694. PMID: 33227038; PMCID: PMC7682851.

From Takako Kato-Minoura, Chuo University-Japan, October 2021

Kato-Minoura T, Ogiwara Y, Yamano T, Fukuzawa H, Kamiya R. Chlamydomonas reinhardtii tubulin-gene disruptants for efficient isolation of strains bearing tubulin mutations. PLoS One. 2020 Nov 23;15(11):e0242694. doi: 10.1371/journal.pone.0242694. PMID: 33227038; PMCID: PMC7682851.

From Takako Kato-Minoura, Chuo University-Japan, October 2021

Kato-Minoura T, Ogiwara Y, Yamano T, Fukuzawa H, Kamiya R. Chlamydomonas reinhardtii tubulin-gene disruptants for efficient isolation of strains bearing tubulin mutations. PLoS One. 2020 Nov 23;15(11):e0242694. doi: 10.1371/journal.pone.0242694. PMID: 33227038; PMCID: PMC7682851.

From Takako Kato-Minoura, Chuo University-Japan, October 2021

Kato-Minoura T, Ogiwara Y, Yamano T, Fukuzawa H, Kamiya R. Chlamydomonas reinhardtii tubulin-gene disruptants for efficient isolation of strains bearing tubulin mutations. PLoS One. 2020 Nov 23;15(11):e0242694. doi: 10.1371/journal.pone.0242694. PMID: 33227038; PMCID: PMC7682851.

From Takako Kato-Minoura, Chuo University-Japan, October 2021

Kato-Minoura T, Ogiwara Y, Yamano T, Fukuzawa H, Kamiya R. Chlamydomonas reinhardtii tubulin-gene disruptants for efficient isolation of strains bearing tubulin mutations. PLoS One. 2020 Nov 23;15(11):e0242694. doi: 10.1371/journal.pone.0242694. PMID: 33227038; PMCID: PMC7682851.

From Takako Kato-Minoura, Chuo University-Japan, October 2021

Kato-Minoura T, Ogiwara Y, Yamano T, Fukuzawa H, Kamiya R. Chlamydomonas reinhardtii tubulin-gene disruptants for efficient isolation of strains bearing tubulin mutations. PLoS One. 2020 Nov 23;15(11):e0242694. doi: 10.1371/journal.pone.0242694. PMID: 33227038; PMCID: PMC7682851.

From Antonia Schad, University of Leipzig, July 2021

This strain was obtained from a conventional crossing using two mutants: LMJ.SG0182.017965 (deficient in the gene for glycolate dehydrogenase) and CC-2702 cia5 (deficient in carbon concentrating mechanisms). Prior to this, both mutant strains were crossed with wild type strains to improve fitness: LMJ.SG0182.017965 x SAG 11-32b and CC-2702 cia5 x CC-410.

The strain requires elevated CO2 for growth on minimal medium due to its photorespiratory phenotype. It grows well on acetate medium. Since it contains a paromomycin resistance cassette, it can also be cultured on medium with antibiotic (Zhang et al., 2014).

CMD6cia5/GYD is a double mutant deficient in the genes for cia5/CCM1 and GYD1. Cia5 has previously been identified as master regulator of the carbon concentrating mechanism in Chlamydomonas (Fukuzawa et al, 2001; Xiang et al, 2001). GYD1 is the glycolate dehydrogenase that converts glycolate from photorespiration into glyoxylate.

Schad A, Rössler S, Nagel R, Wagner H, Wilhelm C. Crossing and selection of Chlamydomonas reinhardtii strains for biotechnological glycolate production. Appl Microbiol Biotechnol. 2022 May 5. doi: 10.1007/s00253-022-11933-y. Epub ahead of print. PMID: 35511277.

  • Locus:
  • CIA5 [CCM1], GYD1
  • Chromosome:
  • 2,6

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain fails to accumulate mRNAs of psbB, psbT and psbH.

Monod C, Goldschmidt-Clermont M, Rochaix JD. Accumulation of chloroplast psbB RNA requires a nuclear factor in Chlamydomonas reinhardtii. Mol Gen Genet. 1992 Feb;231(3):449-59. doi: 10.1007/BF00292715. PMID: 1371579.

Vaistij FE, Goldschmidt-Clermont M, Wostrikoff K, Rochaix JD. Stability determinants in the chloroplast psbB/T/H mRNAs of Chlamydomonas reinhardtii. Plant J. 2000 Mar;21(5):469-82. doi: 10.1046/j.1365-313x.2000.00700.x. PMID: 10758498.

Vaistij FE, Boudreau E, Lemaire SD, Goldschmidt-Clermont M, Rochaix JD. Characterization of Mbb1, a nucleus-encoded tetratricopeptide-like repeat protein required for expression of the chloroplast psbB/psbT/psbH gene cluster in Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 2000 Dec 19;97(26):14813-8. doi: 10.1073/pnas.97.26.14813. PMID: 11121080; PMCID: PMC19001.

  • Locus:
  • MBB1
  • Chromosome:
  • 9

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is defective for trans-splicing of psaA intron 2.

Merendino L, Perron K, Rahire M, Howald I, Rochaix JD, Goldschmidt-Clermont M. A novel multifunctional factor involved in trans-splicing of chloroplast introns in Chlamydomonas. Nucleic Acids Res. 2006 Jan 10;34(1):262-74. doi: 10.1093/nar/gkj429. PMID: 16407333; PMCID: PMC1326246.

  • Locus:
  • RAA1
  • Chromosome:
  • 9

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is defective for trans-splicing of both introns of psaA.

Merendino L, Perron K, Rahire M, Howald I, Rochaix JD, Goldschmidt-Clermont M. A novel multifunctional factor involved in trans-splicing of chloroplast introns in Chlamydomonas. Nucleic Acids Res. 2006 Jan 10;34(1):262-74. doi: 10.1093/nar/gkj429. PMID: 16407333; PMCID: PMC1326246.

  • Locus:
  • RAA1
  • Chromosome:
  • 9

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is defective for trans-splicing of both introns of psaA.

Merendino L, Perron K, Rahire M, Howald I, Rochaix JD, Goldschmidt-Clermont M. A novel multifunctional factor involved in trans-splicing of chloroplast introns in Chlamydomonas. Nucleic Acids Res. 2006 Jan 10;34(1):262-74. doi: 10.1093/nar/gkj429. PMID: 16407333; PMCID: PMC1326246.

  • Locus:
  • RAA1
  • Chromosome:
  • 9

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is defective for trans-splicing of psaA intron 2.

Goldschmidt-Clermont M, Girard-Bascou J, Choquet Y, Rochaix JD. Trans-splicing mutants of Chlamydomonas reinhardtii. Mol Gen Genet. 1990 Sep;223(3):417-25. doi: 10.1007/BF00264448. PMID: 2270082.

  • Locus:
  • RAA2
  • Chromosome:
  • 16

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is defective for trans-splicing of psaA intron 2.

Perron K, Goldschmidt-Clermont M, Rochaix JD. A factor related to pseudouridine synthases is required for chloroplast group II intron trans-splicing in Chlamydomonas reinhardtii. EMBO J. 1999 Nov 15;18(22):6481-90. doi: 10.1093/emboj/18.22.6481. PMID: 10562560; PMCID: PMC1171711.

  • Locus:
  • RAA2
  • Chromosome:
  • 16

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is defective for trans-splicing of psaA intron 2.

Lefebvre-Legendre L, Reifschneider O, Kollipara L, Sickmann A, Wolters D, Kück U, Goldschmidt-Clermont M. A pioneer protein is part of a large complex involved in trans-splicing of a group II intron in the chloroplast of Chlamydomonas reinhardtii. Plant J. 2016 Jan;85(1):57-69. doi: 10.1111/tpj.13089. PMID: 26611495.

  • Locus:
  • RAA7
  • Chromosome:
  • 3

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is defective for trans-splicing of psaA intron 2.

Lefebvre-Legendre L, Reifschneider O, Kollipara L, Sickmann A, Wolters D, Kück U, Goldschmidt-Clermont M. A pioneer protein is part of a large complex involved in trans-splicing of a group II intron in the chloroplast of Chlamydomonas reinhardtii. Plant J. 2016 Jan;85(1):57-69. doi: 10.1111/tpj.13089. PMID: 26611495.

  • Locus:
  • RAA7
  • Chromosome:
  • 3

From Michel Goldschmidt-Clermont, University of Geneva-Switzerland, November 2021

This strain is affected in translation and stability of psaA mRNA.

Lefebvre-Legendre L, Choquet Y, Kuras R, Loubéry S, Douchi D, Goldschmidt-Clermont M. A nucleus-encoded chloroplast protein regulated by iron availability governs expression of the photosystem I subunit PsaA in Chlamydomonas reinhardtii. Plant Physiol. 2015 Apr;167(4):1527-40. doi: 10.1104/pp.114.253906. Epub 2015 Feb 11. PMID: 25673777; PMCID: PMC4378161.

  • Locus:
  • OPR22; TAA1
  • Chromosome:
  • 6

From Anthony Berndt, Stephen Mayfield lab, University of California-San Diego, November 2021

Chloroplast directed SARS-CoV-2 Spike RBD fused to mClover fluorescent protein.

Plasmid insertion makes strain Zeocin and Hygromycin resistant. RBD::mClover fusion protein is directed to the Chloroplast using the psaE Chloroplast Transit Sequence. Expression may mildly attenuate over several months on non-selective media but can be restored by re-selection on 30 µg/mL Hygromycin B on regular tap plates.

Berndt AJ, Smalley TN, Ren B, Simkovsky R, Badary A, Sproles AE, Fields FJ, Torres-Tiji Y, Heredia V, Mayfield SP. Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii. PLoS One. 2021 Nov 18;16(11):e0257089. doi: 10.1371/journal.pone.0257089. PMID: 34793485; PMCID: PMC8601568.

From Anthony Berndt, Stephen Mayfield lab, University of California-San Diego, November 2021

Secreted SARS-CoV-2 Spike RBD fused to Clover fluorescent protein.

Plasmid insertion makes strain Zeocin and Hygromycin resistant. RBD::mClover fusion protein is directed for secretion using the PHC2 secretion signal peptide. Expression may mildly attenuate over several months on non-selective media but can be restored by re-selection on 30 µg/mL Hygromycin B on regular tap plates.

Berndt AJ, Smalley TN, Ren B, Simkovsky R, Badary A, Sproles AE, Fields FJ, Torres-Tiji Y, Heredia V, Mayfield SP. Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii. PLoS One. 2021 Nov 18;16(11):e0257089. doi: 10.1371/journal.pone.0257089. PMID: 34793485; PMCID: PMC8601568.

From Anthony Berndt, Stephen Mayfield lab, University of California-San Diego, November 2021

ER-Golgi retained SARS-CoV-2 Spike RBD fused to mClover fluorescent protein.

Plasmid insertion makes strain Zeocin and Hygromycin resistant. RBD::mClover fusion protein is directed for secretion using the PHC2 secretion signal peptide and retained in the ER-Golgi network with a C-terminal KDEL motif. Expression may mildly attenuate over several months on non-selective media but can be restored by re-selection on 30 µg/mL Hygromycin B on regular tap plates.

Berndt AJ, Smalley TN, Ren B, Simkovsky R, Badary A, Sproles AE, Fields FJ, Torres-Tiji Y, Heredia V, Mayfield SP. Recombinant production of a functional SARS-CoV-2 spike receptor binding domain in the green algae Chlamydomonas reinhardtii. PLoS One. 2021 Nov 18;16(11):e0257089. doi: 10.1371/journal.pone.0257089. PMID: 34793485; PMCID: PMC8601568.

From Anthony Berndt, Stephen Mayfield lab, University of California-San Diego, November 2021

From Anthony Berndt, Stephen Mayfield lab, University of California-San Diego, November 2021

From Anthony Berndt, Stephen Mayfield lab, University of California-San Diego, November 2021

From Sunjoo Joo and Jae-Hyeok Lee, University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.

From Sunjoo Joo and Jae-Hyeok Lee, University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.

From Sunjoo Joo and Jae-Hyeok Lee, University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.

From Sunjoo Joo and Jae-Hyeok Lee, University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.

From Minjae Kim, Eonseon Jin, Thamali Kariyawasam and Jae-Hyeok Lee, Hanyang University/University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.

From Minjae Kim, Eonseon Jin, Thamali Kariyawasam and Jae-Hyeok Lee, Hanyang University/University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.

From Sunjoo Joo, Thamali Kariyawasam, and Jae-Hyeok Lee, University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.

From Sunjoo Joo, Thamali Kariyawasam, and Jae-Hyeok Lee, University of British Columbia, December 2021

Joo, S., Kariyawasam, T., Kim, M., Jin, ES., Goodenough, U.W., and Lee, J.-H. (2022) Sex-linked deubiquitinase establishes uniparental transmission of chloroplast DNA.