Strains

From Emilio Fernandez, University of Cordoba, October 1995, his strain 307d

Mutants at the NIT3 locus are deficient in the molybdenum cofactor for nitrate reductase and xanthine dehydrogenase.

Sosa FM, Ortega T, Barea JL (1978) Mutants from Chlamydomonas reinhardii affected in their nitrate assimilation capabiligy. Plant Sci Lett 11:51-58

Fernández E, Cárdenas J (1982) Regulation of the nitrate-reducing system enzymes in wild-type and mutant strains of Chlamydomonas reinhardii. Mol Gen Genet 186:164-169

Fernández E, Matagne RF (1984) Genetic analysis of nitrate reductase-deficient mutants in Chlamydomonas reinhardii. Curr Genet 8:635-640

From Ursula Rüffer and Jürgen Pfau, Universität Marburg, October 1995, as 622E

Phenotype: abnormal eyespot

This is an eyespot-deficient mutant originally obtained from M.E. Feinleib. We don’t know whether it corresponds to one of the EYE loci defined by Dieckmann and colleagues.

Rüffer U, Nultsch W (1998) Flagellar coordination in Chlamydomonas cells held on micropipettes. Cell Motil Cytoskeleton 41:297-307

From Olivier Vallon, Institut de Biologie Physico-Chimique, November 1995

Phenotype: does not mate

The gag1 mutation is an allele at the SAG2 locus. Mutants at this locus do not agglutinate with cells of either mating type, but can be mated as mt+ by artificial agglutination with antibodies.

Notes from Olivier Vallon, September 1999: “The sterility phenotype is sex-limited (expressed only in mt+ cells) but the gene is unlinked to the mating type locus. The primary defect is in O-glycosylation: at 32 deg C, in both mating types. The major O-glycoproteins show increased electrophoretic mobility and altered antigenicity. Cells form clumps on plates and palmelloids in liquid culture. These defects are not observed when cells are grown at 18 deg C. However, the sex-limited sterility phenotype is expressed regardless of the growth temperature.”

This strain is the one shown in Figure 1, lane 1, of the following paper:

Vallon O, Wollman FA (1995) Mutations Affecting O-Glycosylation in Chlamydomonas reinhardtii Cause Delayed Cell Wall Degradation and Sex-Limited Sterility. Plant Physiol 108:703-712

From Olivier Vallon, Institut de Biologie Physico-Chimique, November 1995

Phenotype: does not mate

The gag1 mutation is an allele at the SAG2 locus. Mutants at this locus do not agglutinate with cells of either mating type, but can be mated as mt+ by artificial agglutination with antibodies.

Notes from Olivier Vallon, September 1999: “The sterility phenotype is sex-limited (expressed only in mt+ cells) but the gene is unlinked to the mating type locus. The primary defect is in O-glycosylation: at 32 deg C, in both mating types. The major O-glycoproteins show increased electrophoretic mobility and altered antigenicity. Cells form clumps on plates and palmelloids in liquid culture. These defects are not observed when cells are grown at 18 deg C. However, the sex-limited sterility phenotype is expressed regardless of the growth temperature.”

This strain is the one shown in Figure 1, lane 2, of the following paper:

Vallon O, Wollman FA (1995) Mutations Affecting O-Glycosylation in Chlamydomonas reinhardtii Cause Delayed Cell Wall Degradation and Sex-Limited Sterility. Plant Physiol 108:703-712

From George Witman, Worcester Foundation for Experimental Biology, January 1996

Phenotype: impaired motility

This mutant was generated by insertional mutagenesis of a nit1 agg1 mt+ parental strain, transformed with the plasmid pGP505 containing the wild-type NIT1 allele (see Pazour et al. 1995). NIT+ strains having a slow-swimming, jerky-swimming phenotype were selected. (This phenotype is typical of mutants lacking the outer dynein arms).

In this strain, outer dynein arms are absent, and the ODA9 gene (encoding protein IC78) is deleted (see Wilkerson et al., Figure 8). The NIT+ phenotype appears to segregate with the deletion. The genome contains pUC119 but it is not known if this segregates with the deletion.

Pazour GJ, Sineshchekov OA, Witman GB (1995) Mutational analysis of the phototransduction pathway of Chlamydomonas reinhardtii. J Cell Biol 131:427-440

Wilkerson CG, King SM, Koutoulis A, Pazour GJ, Witman GB (1995) The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly. J Cell Biol 129:169-178

• Locus:
• ODA9 [DIC1]
• Chromosome:
• 12

From George Witman, Worcester Foundation for Experimental Biology, January 1996

Phenotype: impaired motility

This mutant was generated by insertional mutagenesis of a nit1 agg1 mt+ parental strain, transformed with the plasmid pGP505 containing the wild-type NIT1 allele (see Pazour et al. 1995). NIT+ strains having a slow-swimming, jerky-swimming phenotype were selected. (This phenotype is typical of mutants lacking the outer dynein arms).

In this strain, outer dynein arms are absent, and the ODA9 gene is deleted (see Wilkerson et al., Figure 8). The NIT+ phenotype can be segregated away from the mutation, and pUC119 is not present in the genome. Thus, derivatives of this strain would be useful for further transformations using NIT1 as a selectable marker.

Pazour GJ, Sineshchekov OA, Witman GB (1995) Mutational analysis of the phototransduction pathway of Chlamydomonas reinhardtii. J Cell Biol 131:427-440

Wilkerson CG, King SM, Koutoulis A, Pazour GJ, Witman GB (1995) The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly. J Cell Biol 129:169-178

• Locus:
• ODA9 [DIC1]
• Chromosome:
• 12

From George Witman, Worcester Foundation for Experimental Biology, January 1996

Phenotype: impaired motility

This mutant was generated by insertional mutagenesis of a nit1 agg1 mt+ parental strain, transformed with the plasmid pGP505 containing the wild-type NIT1 allele (see Pazour et al. 1995). NIT+ strains having a slow-swimming, jerky-swimming phenotype were selected. (This phenotype is typical of mutants lacking the outer dynein arms).

In this strain, outer dynein arms are absent, and the ODA9 gene is disrupted by a large insertion (see Wilkerson et al., Figure 8). The NIT+ phenotype segregates with the insertion. The genome contains pUC119 but it is not known if this segregates with the insertion. This mutation does not complement the original oda9 in diploids.

Pazour GJ, Sineshchekov OA, Witman GB (1995) Mutational analysis of the phototransduction pathway of Chlamydomonas reinhardtii. J Cell Biol 131:427-440

Wilkerson CG, King SM, Koutoulis A, Pazour GJ, Witman GB (1995) The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly. J Cell Biol 129:169-178

• Locus:
• ODA9 [DIC1]
• Chromosome:
• 12

From George Witman, Worcester Foundation for Experimental Biology, January 1996

Phenotype: impaired motility

This mutant was generated by insertional mutagenesis of a nit1 agg1 mt+ parental strain, transformed with the plasmid pGP505 containing the wild-type NIT1 allele (see Pazour et al. 1995). NIT+ strains having a slow-swimming, jerky-swimming phenotype were selected. (This phenotype is typical of mutants lacking the outer dynein arms).

In this strain, outer dynein arms are absent, and the ODA9 gene is deleted (see Wilkerson et al., Figure 8). The NIT+ phenotype can be segregated away from the mutation Thus, derivatives of this strain would be useful for further transformations using NIT1 as a selectable marker. The genome contains pUC119 but it is not known if the plasmid sequence segregates with the deletion. This mutation does not complement oda9 in diploids.

Pazour GJ, Sineshchekov OA, Witman GB (1995) Mutational analysis of the phototransduction pathway of Chlamydomonas reinhardtii. J Cell Biol 131:427-440

Wilkerson CG, King SM, Koutoulis A, Pazour GJ, Witman GB (1995) The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly. J Cell Biol 129:169-178

• Locus:
• ODA9 [DIC1]
• Chromosome:
• 12

From George Witman, Worcester Foundation for Experimental Biology, January 1996

Phenotype: impaired motility

This mutant was generated by insertional mutagenesis of a nit1 agg1 mt+ parental strain, transformed with the plasmid pGP505 containing the wild-type NIT1 allele (see Pazour et al. 1995). NIT+ strains having a slow-swimming, jerky-swimming phenotype were selected. (This phenotype is typical of mutants lacking the outer dynein arms).

In this strain, the beta dynein heavy chain (ODA4) is deleted (see Figure 8 of Wilkerson et al. 1995). This deletion includes both the 3′ and 5′ ends and thus is presumed to be complete. The genome contains pUC119 but it is not known if this segregates with the deletion.

Pazour GJ, Sineshchekov OA, Witman GB (1995) Mutational analysis of the phototransduction pathway of Chlamydomonas reinhardtii. J Cell Biol 131:427-440

Wilkerson CG, King SM, Koutoulis A, Pazour GJ, Witman GB (1995) The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly. J Cell Biol 12:169-178

• Locus:
• ODA4 [DHC14]
• Chromosome:
• 9

From George Witman, Worcester Foundation for Experimental Biology, January 1996

Phenotype: impaired motility

This mutant was generated by insertional mutagenesis of a nit1 agg1 mt+ parental strain, transformed with the plasmid pGP505 containing the wild-type NIT1 allele (see Pazour et al. 1995). NIT+ strains having a slow-swimming, jerky-swimming phenotype were selected. (This phenotype is typical of mutants lacking the outer dynein arms).

In this strain, the beta dynein heavy chain (ODA4) is deleted (see Figure 8 of Wilkerson et al. 1995). This deletion includes the 5′ end but not the 3′ end. The genome does not contain pUC119. It is not known if the NIT+ phenotype segregates with the deletion.

Pazour GJ, Sineshchekov OA, Witman GB (1995) Mutational analysis of the phototransduction pathway of Chlamydomonas reinhardtii. J Cell Biol 131:427-440

Wilkerson CG, King SM, Koutoulis A, Pazour GJ, Witman GB (1995) The 78,000 M(r) intermediate chain of Chlamydomonas outer arm dynein isa WD-repeat protein required for arm assembly. J Cell Biol 12:169-178

• Locus:
• ODA4 [DHC14]
• Chromosome:
• 9

From Bruce Kohorn, Duke University, February 1996

Phenotype: impaired motility

From a cross of CC-1387 pf27 mt+ x CC-124 wild type, by Benoit Baillet in Bruce Kohorn’s laboratory at Duke, February 1996.

In pf27 cells, five polypeptides associated with radial spokes are not phosphorylated. This mutant is non-motile and easy to score. See CC-1387 for the same mutant in mt+.

• Locus:
• PF27
• Chromosome:
• 12

From Patrick Ferris, Goodenough lab, Washington University, August 1996

This is a revertant of bs-37 (see CC-2392) that has a 3 bp insertion. It fuses somewhat less well than wild type.

Ferris PJ, Woessner JP, Goodenough UW (1996) A sex recognition glycoprotein is encoded by the plus mating-type gene fus1 of Chlamydomonas reinhardtii. Mol Biol Cell 7:1235-1248

• Locus:
• FUS1
• Chromosome:
• 6

From Patrick Ferris, Goodenough lab, Washington University, August 1996

Phenotype: does not mate

This is a spontaneous fus1 mutation that arose in CC-125.

Ferris PJ, Woessner JP, Goodenough UW (1996) A sex recognition glycoprotein is encoded by the plus mating-type gene fus1 of Chlamydomonas reinhardtii. Mol Biol Cell 7:1235-1248

• Locus:
• FUS1
• Chromosome:
• 6

From Patrick Ferris, Goodenough lab, Washington University, August 1996

This is a revertant of imp1, probably the result of incomplete excision of the transposon that generated this mutation. This strain has the 2.5 kb insert remaining, and fuses less well than wild type.

Ferris PJ, Woessner JP, Goodenough UW (1996) A sex recognition glycoprotein is encoded by the plus mating-type gene fus1 of Chlamydomonas reinhardtii. Mol Biol Cell 7:1235-1248

• Locus:
• FUS1
• Chromosome:
• 6

From Patrick Ferris, Goodenough lab, Washington University, August 1996

This is a revertant of imp1, probably the result of incomplete excision of the transposon that generated this mutation. This strain has the 1.5 kb insert remaining, and fuses less well than wild type.

Ferris PJ, Woessner JP, Goodenough UW (1996) A sex recognition glycoprotein is encoded by the plus mating-type gene fus1 of Chlamydomonas reinhardtii. Mol Biol Cell 7:1235-1248

• Locus:
• FUS1
• Chromosome:
• 6

Sammlung von Algenkulturen (SAG), October 1996

This is the SAG isolate of Tsubo’s strain C8 (see CC-407). It can grow on nitrate.

Sammlung von Algenkulturen (SAG), October 1996

SAG isolates 81.72 and 7.73, originally designated as C. globosa and C. incerta respectively, appear to be identical with respect to chloroplast DNA restriction fragments and the ITS sequences that separate the nuclear ribosomal RNA genes, which have proved to be excellent diagnostic markers for phylogenetic analysis in Chlamydomonas. Since C. globosa was recorded as an isolate from the Netherlands, and C. incerta from the zoological garden in Havana, Cuba, one of these strains may have replaced the other in the SAG collection at some time in the past. Alternatively, the Cuban zoological garden might have inadvertently acquired a European Chlamydomonas.

SAG now lists both these strains as C. reinhardtii, although neither mates with laboratory C. reinhardtii strains.

Harris EH, Boynton JE, Gillham NW, Burkhart BD, Newman SM (1991) Chloroplast genome organization in Chlamydomonas. Arc. Protistenk 139:183-192

Pröschold T, Harris EH, Coleman AW (2005) Portrait of a species: Chlamydomonas reinhardtii. Genetics 170:1601-1610

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3351 is the product of a cross of the original far1-1 (an insertional mutant obtained by transformation of strain 21gr with the CRY1 [RPS14] gene) to a mt- strain carrying the sr1 mutation.

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1
• Chromosome:
• 1

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3352 is the far1-2 mutant, which was isolated in a pf14 background after transformation with the wild-type RPS3 (PF14) gene as a reporter for expression from the NIT1 promoter).

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1, PF14 [RSP3]
• Chromosome:
• 1,6

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3353 is the far1-3 mutant, which was isolated in a pf14 background after transformation with the wild-type RPS3 (PF14) gene as a reporter for expression from the NIT1 promoter).

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1, PF14 [RSP3]
• Chromosome:
• 1,6

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3354 is the far1-4 mutant, which was isolated in a pf14 background after transformation with the wild-type RPS3 (PF14) gene as a reporter for expression from the NIT1 promoter).

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1, PF14 [RSP3]
• Chromosome:
• 1,6

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3355 is the far1-5 mutant, which was isolated in a pf14 background after transformation with the wild-type RPS3 (PF14) gene as a reporter for expression from the NIT1 promoter). This strain is the product of a cross of the original far1-5 to a strain carrying the supcs1-2 mutation, conferring cold sensitivity.

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1
• Chromosome:
• 1

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3356 is the far1-6 mutant, which was isolated in a pf14 background after transformation with the wild-type RPS3 (PF14) gene as a reporter for expression from the NIT1 promoter). This strain is the product of a cross of the original far1-6 to a strain carrying the supcs1-2 mutation, conferring cold sensitivity.

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1
• Chromosome:
• 1

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3357 is the far1-7 mutant, which was isolated in a pf14 background after transformation with the wild-type RPS3 (PF14) gene as a reporter for expression from the NIT1 promoter).

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1, PF14 [RSP3]
• Chromosome:
• 1,6

From Donghong Zhang, Washington University, March 1997

The far1 (free from ammonia repression) mutants were isolated by insertional mutagenesis or by selection of spontaneous mutants with constitutive expression of the NIT1 (nitrate reductase) gene in the presence of ammonia. CC-3358 is the product of a cross of the original far1-1 (an insertional mutant obtained by transformation of strain 21gr with the CRY1 [RPS14] gene) to a mt- strain carrying the sr1 mutation.

Zhang D, Lefebvre PA (1997) FAR1, a negative regulatory locus required for the repression of the nitrate reductase gene in Chlamydomonas reinhardtii. Genetics 146:121-133

• Locus:
• FAR1
• Chromosome:
• 1

From Karen Kindle, Cornell University, November 1996

Phenotype: requires arginine

This is arg7 in a strain that can grow on nitrate. CC-3373 is the same combination in mt-.

For more information on the ARG7 locus see CC-48.

• Locus:
• ARG7
• Chromosome:
• 1

From Karen Kindle, Cornell University, November 1996

Phenotype: requires arginine; can grow on nitrate

This is arg7 in a strain that can grow on nitrate. CC-3372 is the same combination in mt+.

For more information on the ARG7 locus see CC-48.

• Locus:
• ARG7
• Chromosome:
• 1

From Roel Funke and Don Weeks, December 1996

Phenotype: requires arginine

This strain has the wild type allele at the NIT2 locus, and can be transformed with either ARG7 or NIT1.

• Locus:
• ARG7, NIT1
• Chromosome:
• 1,9

Anita Lardans, Boynton-Gillham laboratory, Duke University

Phenotype: requires acetate; herbicide resistant (atrazine, bromacil, metribuzin)

This strain contains a site-directed mutation A251V in the psbA gene, and a silent mutation (indicated by * in the strain name) creating an SspI site by substituting a T for a C at position 247. CC-125 wild-type cells were co-transformed with the psbA plasmid and with plasmid P-228, which confers spectinomycin resistance by a mutation in the gene encoding the chloroplast 16S ribosomal RNA.

The A251V mutation confers resistance to atrazine, bromacil and metribuzin, but not to DCMU, and results in poor photoautotrophic growth, although it is not as severely impaired as the Leu substitution at the same residue (see CC-3389).

Förster B, Heifetz PB, Lardans A, Boynton JE, Gillham NW (1997) Herbicide resistance and growth of D1 Ala251mutants in Chlamydomonas. Z Naturforsch 52:654-664

Lardans A, Gillham NW, Boynton JE (1997) Site-directed mutations at residue 251 of the photosystem II D1 protein of Chlamydomonas that result in a nonphotosynthetic phenotype and impair D1 synthesis and accumulation. J Biol Chem 272:210-216

• Locus:
• psbA
• Chromosome:
• chloroplast

From Anita Lardans, Boynton-Gillham laboratory, Duke University

This strain contains a site-directed mutation A251I in the psbA gene, and a silent mutation (indicated by * in the strain name) creating an SspI site by substituting a T for a C at position 247. CC-125 wild-type cells were co-transformed with the psbA plasmid and with plasmid P-228, which confers spectinomycin resistance by a mutation in the gene encoding the chloroplast 16S ribosomal RNA.

The A251I mutation confers resistance to atrazine, bromacil and metribuzin, but not to DCMU, and results in poor photoautotrophic growth, although it is not as severely impaired as the Leu substitution at the same residue (see CC-3389).

Förster B, Heifetz PB, Lardans A, Boynton JE, Gillham NW (1997) Herbicide resistance and growth of D1 Ala251mutants in Chlamydomonas. Z Naturforsch 52:654-664

Lardans A, Gillham NW, Boynton JE (1997) Site-directed mutations at residue 251 of the photosystem II D1 protein of Chlamydomonas that result in a nonphotosynthetic phenotype and impair D1 synthesis and accumulation. J Biol Chem 272:210-216

• Locus:
• psbA
• Chromosome:
• chloroplast