Strains

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires nicotinamide

This is an allele at the NIC7 locus (please see CC-85 for more information). It requires nicotinamide, and is killed by 7.5 micrograms/ml 3-acetyl pyridine.

• Locus:
• NIC7 [QSA1]
• Chromosome:
• 6

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires nicotinamide

This is an allele at the NIC7 locus (see CC-85). It requires nicotinamide, and is killed by 7.5 micrograms/ml 3-acetyl pyridine.

• Locus:
• NIC7 [QSA1]
• Chromosome:
• 6

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires thiamine; motility impaired

This strain maps both arms of linkage group VI. The thi10 marker is leaky but scorable on minimal medium; pf14 is easy to score.

• Locus:
• PF14 [RSP3], THI10
• Chromosome:
• 6

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires nicotinamide

This is an allele at the NIC15 locus. Please see CC-14 for more information on this locus.

• Locus:
• NIC15 [ASO1]
• Chromosome:
• 12

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires nicotinamide

This mutation was UV-induced by Eversole. So far as we can determine it was never analyzed genetically.

Eversole RA (1956) Biochemical Mutants of Chlamydomonas reinhardi. Amer J Bot 43:404-407

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires acetate and nicotinamide; antibiotic and inhibitor resistant (cycloheximide, methionine sulfoximine, streptomycin)

This is a multiply marked strain constructed in Ebersold’s lab. There are other strains that mark more linkage groups, but this one is useful in having two markers on linkage group I and two on VI. The ac14 marker is not especially easy to score, however, and other strains may be better for linkage group I.

• Locus:
• AC14, ACT2 [RPL36a], CAN1, MSR1, NIC7 [QSA1], SR1
• Chromosome:
• 1,6,9

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires acetate and arginine; antibiotic resistant (cycloheximide, streptomycin); altered phototaxis

This strain has been retained in the stock collection mainly for the np mutation, after confirmation in 1981 that it showed negative phototaxis. The np mutation has not been analyzed further, however. The other markers – ac14, arg2, can1 and sr1 – appear in other strains that may be more useful for mapping crosses.

• Locus:
• AC14, ARG7 CAN1, SR1
• Chromosome:
• 1,7,9

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires nicotinamide; antibiotic resistant (cycloheximide)

This strain contains three unlinked, easily scored markers in combination.

• Locus:
• ACT2 [RPL36a], NIC1, PF12
• Chromosome:
• 2,6,14

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: antibiotic resistant (cycloheximide, streptomycin); altered phototaxis; impaired motility

We suspect the np mutation in this stock is the same as the one in CC-639, but it’s difficult to understand why it was retained, since it would be difficult if not impossible to score phototaxis in the presence of the pf7 mutation which severely impairs motility. This is however the only stock in which pf7 is coupled with other markers.

• Locus:
• ACT2 [RPL36a], PF7, SR1
• Chromosome:
• 6,9,17

From Maureen Hanson, Bogorad lab, Harvard University, August 1978

Phenotype: requires p-aminobenzoic acid

This mutant will grow on yeast extract medium, or on 50 micrograms/ml p-aminobenzoic acid. It can be scored by failure to grow on minimal medium, or by sensitivity to 1 microgram/ml sulfanilamide.

Eversole RA (1956) Biochemical Mutants of Chlamydomonas reinhardi. Amer J Bot 43:404-407

Ebersold WT, Levine RP, Levine EE, Olmsted MA (1962) Linkage maps in Chlamydomonas reinhardi. Genetics 47:531-543

Lawrence CW and Davies DR (1967) The mechanism of recombination in Chlamydomonas reinhardii. I Mut Res 4:137-146

• Locus:
• PAB2
• Chromosome:
• 1

From Kathryn Wilson, Indiana University at Indianapolis, October 1978

Phenotype: altered lipid metabolism; antibiotic resistant (nystatin)

This mutant has an altered sterol composition and is resistant to nystatin. It has not been mapped.

Bard M, Wilson KJ, Thompson RM (1978) Isolation of sterol mutants inChlamydomonas reinhardi: Chromatographic analyses. Lipids 13:533-539

From Kathryn Wilson, Indiana University at Indianapolis, October 1978

Phenotype: altered lipid metabolism; antibiotic resistant (nystatin)

This mutant has an altered sterol composition and is resistant to nystatin. It has not been mapped.

Bard M, Wilson KJ, Thompson RM (1978) Isolation of sterol mutants in Chlamydomonas reinhardi: Chromatographic analyses. Lipids 13:533-539

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: altered phototaxis

This strain grows on all media, but showed a poor phototaxis response in tests in 1981.

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: altered phototaxis

This strain showed negative phototaxis in 1981. It was not checked for the y1 yellow-in-the-dark phenotype.

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires acetate at high pH

Although labeled np, this strain showed normal phototaxis (like wild type) when tested in 1981. The ac6 mutation must be scored on minimal medium at high pH (make TAP medium, omitting acetic acid and using HCl to bring pH to 8.3).

Please see CC-2 for more information on ac6.

• Locus:
• AC6
• Chromosome:
• 7

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: requires acetate

So far as we know, this corresponds to the original ac7 mutant. Please see CC-520 ac7a, an allelic mutant, for more information on the AC7 locus.

• Locus:
• AC7
• Chromosome:
• 11

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

The markers in this strain are uncertain, and no explanation was received from Ebersold. It seems to have a leaky acetate requirement.

From Kathleen Peterson and W.T. Ebersold, UCLA, June 1978

Phenotype: altered phototaxis

This strain grows on all media, and showed a poor phototaxis response when tested in 1981. The y1 mutation was not checked.

Chlamydomonas Genetics Center, Duke University, 1978

Phenotype: requires acetate (suppressed)

Strains carrying the ac46 mutation have a strong tendency to acquire second-site suppressors, but we have always been able to recover the original mutation by outcrossing to wild type. The y1 mutant is a better genetic marker for this linkage group.

CC-705 is from CC-125 x CC-563, a suppressed ac46 strain no longer in the collection. It was scored as cleanly acetate-requiring in 1978, In a cross in 1982 of CC-705 x CC-124, complete tetrads were obtained with clear 2:2 segregation. CC-705 was subsequently outcrossed to produce CC-1361.

The ac46 mutant appears to map within 1-2 map units of its own centromere but in early crosses did not show linkage to any of 36 markers tested on other linkage groups. Crosses by Dutcher et al. and by the Chlamydomonas Genetics Center subsequently showed that AC46 and Y1 constitute a single linkage group.

Levine and Goodenough listed the ac46 mutant as non-photosynthetic; the cause of the defect was not determined. Subsequent research by Lemaire and Wollmann, and by Majeran et al., demonstrated that the ac46 locus affects expression of the chloroplast atpH and atpI genes encoding subunits of chloroplast ATP synthase.

Levine RP, Volkmann D (1961) Mutants with impaired photosynthesis in Chlamydomonas reinhardi. Biochem Biophys Res Commun 6:264-269

Levine RP, Goodenough UW (1970) The genetics of photosynthesis and of the chloroplast in Chlamydomonas reinhardi. Annu Rev Genet 4:397-408

Smyth RD, Martinek GW, Ebersold WT (1975) Linkage of six genes in Chlamydomonas reinhardtii and the construction of linkage test strains. J Bacteriol 124:1615-1617

Lemaire C, Wollman FA (1989) The chloroplast ATP synthase in Chlamydomonas reinhardtii. II. Biochemical studies on its biogenesis using mutants defective in photophosphorylation. J Biol Chem 264:10235-10242

Dutcher SK, Power J, Galloway RE, Porter ME (1991) Reappraisal of the genetic map of Chlamydomonas reinhardtii. J Hered 82:295-301

Majeran W, Olive J, Drapier D, Vallon O, Wollman FA (2001) The light sensitivity of ATP synthase mutants of Chlamydomonas reinhardtii. Plant Physiol 126:421-433

• Locus:
• AC46
• Chromosome:
• 15

Boynton-Gillham laboratory, Duke University, 1979

Phenotype: antibiotic resistant (spectinomycin)

Obtained from a cross of CC-105 (a derivative of the original chloroplast mutant spr-u-1-27-3 strain; see CC-104) to CC-126, a spectinomycin-sensitive strain no longer in the stock collection. CC-706 was used in crosses in the Boynton-Gillham laboratory at Duke. It is easily scored as resistant to 100 micrograms/ml spectinomycin on agar.

• Locus:
• rrnS
• Chromosome:
• chloroplast

Boynton-Gillham laboratory, Duke University, 1979

Phenotype: requires acetate

ac-u-c-2-21 is a deletion mutation in the chloroplast atpB gene, which results in a clean, non-reverting acetate-requiring phenotype. CC-373, which is the same mutation in mt+, has been widely used as a recipient for chloroplast transformation experiments. Please see CC-373 for additional information and reference citations.

• Locus:
• atpB
• Chromosome:
• chloroplast

Boynton-Gillham laboratory, Duke University, 1979

Phenotype: requires acetate

This is a chloroplast mutation, not yet identified with a specific gene. Shepherd et al. found no obvious defects in any photosynthetic components. Chloroplast DNA appears normal on digestion with BamHI or Eco RI.

Shepherd HS, Boynton JE, Gillham NW (1979) Mutations in nine chloroplast loci of Chlamydomonas affecting different photosynthetic functions. Proc Natl Acad Sci USA 76:1353-1357

Myers AM, Grant DM, Rabert DK, Harris EH, Boynton JE, Gillham NW (1982) Mutants of Chlamydomonas reinhardtii with physical alterations in their chloroplast DNA. Plasmid 7:133-151

• Locus:
• Chromosome:
• chloroplast

From Itzhak Ohad, Hebrew University of Jerusalem, July 1979

Phenotype: yellow in the dark

This is Sager’s original y1 strain, as used by Ohad and others in their extensive series of experiments on Chlamydomonas greening. As y1 strains go, it seems to be reasonably stable, but users are advised to subclone for a good yellow-in-the-dark phenotype before using, since we maintain the strain in continuous light without selection for the yellow character. This strain can grow on nitrate.

CC-3992 is an equivalent strain, obtained by Hoober from Ohad. CC-1168 is a more stable allele at the same locus, and CC-1179 and CC-1180 contain a stable but temperature-conditional allele.

Sager R, Palade GE (1954) Chloroplast structure in green and yellow strains of Chlamydomonas. Exp Cell Res 7:584-588

Ohad I, Siekevitz P, Palade GE (1967a) Biogenesis of chloroplast membranes. II. Plastid differentiation during greening of a dark-grown algal mutant (Chlamydomonas reinhardi). J Cell Biol 35:553-584

Ohad I, Siekevitz P, Palade GE (1967b) Biogenesis of chloroplast membranes. II. Plastid differentiation during greening of a dark-grown algal mutant (Chlamydomonas reinhardi). J Cell Biol 35:553-584

Ford C, Wang W (1982) Instability at the y-1 locus of Chlamydomonas reinhardtii. Mol Gen Genet 187:286-290

Bednarik DP, Hoober JK ( 1985) Synthesis of Chlorophyllide b from Protochlorophyllide in Chlamydomonas reinhardtii y-1. Science 230:450-453

Dutcher SK, Power J, Galloway RE, Porter ME (1991) Reappraisal of the genetic map of Chlamydomonas reinhardtii. J Hered 82:295-301

White RA, Hoober JK (1994) Biogenesis of Thylakoid Membranes in Chlamydomonas reinhardtii y1 (A Kinetic Study of Initial Greening). Plant Physiol 106:583-590

• Locus:
• Y1
• Chromosome:
• 15

From Pierre Bennoun, Institut de Biologie Physico-Chimique, September 1979

Phenotype: requires acetate

This is a chloroplast mutant deficient in the cytochrome b6/f complex. It was obtained by Bennoun after treatment with 5-fluorodeoxyuridine.

Finazzi et al. showed that the lesion in FUD2 is a 36 bp duplication in petB, producing a 12 amino acid duplication in the cd loop of cytochrome b6. The mutant protein can assemble into the cytochrome b6/f complex, but these complexes show slower electron transfer and increased degradation in aging cells.

Finazzi G, Büschlen S, de Vitry C, Rappaport F, Joliot P, Wollman FA (1997) Function-directed mutagenesis of the cytochrome b6f complex in Chlamydomonas reinhardtii: involvement of the cd loop of cytochrome b6 in quinol binding to the Q(o) site. Biochemistry 36:2867-2874

• Locus:
• petB
• Chromosome:
• chloroplast

Doug Rabert in Boynton-Gillham laboratory, Duke University, November 1979

Phenotype: requires acetate

This mutant was isolated in an arg2/arg7 diploid strain by Doug Rabert in 1979. Cells were pregrown in 0.5 mM 5-fluorodeoxyuridine in liquid cultures, concentrated, and irradiated with 2 minutes X-ray @ 6 krad/min. The irradiated cells were plated on 2 mM arsenate and arsenate-tolerant (non-photosynthesizing) clones were selected.

This strain is a mt+ derivative, obtained by successive crosses to CC-125 wild type of a mutant isolated in an arg2/arg7 diploid strain. This strain is not arginine-requiring. The mutant shows symmetrical 9 kb deletions in both copies of the psbA gene.

This strain can be used as a recipient for transformation with the psbA gene.

Myers AM, Grant DM, Rabert DK, Harris EH, Boynton JE, Gillham NW (1982) Mutants of Chlamydomonas reinhardtii with physical alterations in their chloroplast DNA. Plasmid 7:133-151

• Locus:
• psbA
• Chromosome:
• chloroplast

From Robert Lee, Dalhousie University, October 1979

Phenotype: antibiotic resistant (spectinomycin)

This is a nuclear spectinomycin resistant mutant, induced by mutagenesis with methyl methanesulfonate and easily scored on 100-200 micrograms/ml spectinomycin on agar. Please see CC-770 for additional information on the SPR1 locus.

• Locus:
• SPR1
• Chromosome:
• 16

From Robert Lee, Dalhousie University, October 1979

Phenotype: antibiotic resistant (spectinomycin)

This is a nuclear spectinomycin resistant mutant, induced by mutagenesis with methyl methanesulfonate and easily scored on 100-200 micrograms/ml spectinomycin on agar. Based on its phenotype and map position, we suspect that it is a mutation in the PRPS5 gene encoding chloroplast ribosomal protein S5.

• Locus:
• SPR1
• Chromosome:
• 16

Chlamydomonas Genetics Center, Duke University, deriviative of CC-798 from Jacqueline Hermann-Smith

From a cross CC-1039 wild type mt+ x CC-798 nitA mt-, 1980.
This replaced the original CC-797, which was received from Jacqueline Hermann-Smith 1979 and was supposed to be nitA mt- but turned out to be mt+. Please see CC-798 for more information on this allele.

• Locus:
• NIT1
• Chromosome:
• 9

From Jacqueline Hermann-Smith, University of Swansea, November 1979

The nitA or nit-A mutation was isolated in Sager’s 21 gr strain by Syrett and colleagues. It is specifically deficient in the diaphorase activity of the nitrate reductase complex.

Nichols GL, Syrett PJ (1978) Nitrate reductase deficient mutants of Chlamydomonas reinhardii. Isolation and genetics. J Gen Microbiol 108:71–77

Nichols GL, Shehata SAM, Syrett PJ (1978) Nitrate reductase deficient mutants of Chlamydomonas reinhardii. Biochemical characteristics. J Gen Microbiol 108:79–88

• Locus:
• NIT1
• Chromosome:
• 9

From Jacqueline Hermann-Smith, University of Swansea, November 1979

The nitB or nit-B mutation was isolated in Sager’s 21 gr strain by Syrett and colleagues. It lacks both the diaphorase and terminal reductase activities of the nitrate reductase complex. Fernandez and Matagne showed that this is a nit2 allele.

Nichols GL, Syrett PJ (1978) Nitrate reductase deficient mutants of Chlamydomonas reinhardii. Isolation and genetics. J Gen Microbiol 108:71–77

Nichols GL, Shehata SAM, Syrett PJ (1978) Nitrate reductase deficient mutants of Chlamydomonas reinhardii. Biochemical characteristics. J Gen Microbiol 108:79–88

Fernández E, Matagne RF (1986) In vivo complementation analysis of nitrate reductase-deficient mutants in Chlamydomonas reinhardtii. Curr Genet 10:397-403

• Locus:
• NIT2
• Chromosome:
• 3