Andrew Wiseman, Boynton-Gillham laboratory, Duke University, 1977

Phenotype: dies in the dark

Isolated by Andrew Wiseman in the Boynton-Gillham laboratory, nitrosoguanidine-induced in wild type mt-. This is the original mutant.

This strain was originally described as respiratory deficient, unable to grow on acetate in the dark. It showed non-Mendelian segregation, suggestive of a possible mitochondrial mutation, but was not analyzed in detail because Wiseman was unable to obtain a homoplasmic stock with a consistent phenotype. Subsequently the original strain acquired what appears to be a secondary nuclear mutation. At last check, the strain still showed a dark-dier phenotype.


Wiseman A, Gillham NW, Boynton JE (1977) The mitochondrial genome of Chlamydomonas. II. Genetic analysis of non-mendelian obligate photautotrophic mutants. Mol Gen Genet 150:109-118

Andrew Wiseman, Boynton-Gillham laboratory, Duke University, 1977

Phenotype: dies in the dark

Isolated by Andrew Wiseman in the Boynton-Gillham laboratory, nitrosoguanidine-induced in wild type mt+. This is the original mutant.

This strain has reduced levels of cyanide-sensitive respiration and cytochrome oxidase, but essentially normal mitochondrial ultrastructure. At last check, the strain still showed a dark-dier phenotype.


Wiseman A, Gillham NW, Boynton JE (1977) Nuclear mutations affecting mitochondrial structure and function in Chlamydomonas. J Cell Biol 73:56-77

Andrew Wiseman, Boynton-Gillham laboratory, Duke University, 1977

Phenotype: dies in the dark

Isolated by Andrew Wiseman in the Boynton-Gillham laboratory, nitrosoguanidine-induced in wild type mt+. This is the original mutant.

This strain has reduced levels of cyanide-sensitive respiration and cytochrome oxidase, but essentially normal mitochondrial ultrastructure. At last check, the strain showed a leaky dark-dier phenotype.


Wiseman A, Gillham NW, Boynton JE (1977) Nuclear mutations affecting mitochondrial structure and function in Chlamydomonas. J Cell Biol 73:56-77

Andrew Wiseman, Boynton-Gillham laboratory, Duke University, 1977

Phenotype: dies in the dark

Isolated by Andrew Wiseman in the Boynton-Gillham laboratory, nitrosoguanidine-induced in wild type mt-. This is the original mutant.

This strain has reduced levels of cytochrome oxidase, but normal NADH-cytochrome c reductase; cristae appear normal, but mitochondrial matrix material is precipitated along surface of the cristae. At last check, the strain showed a leaky dark-dier phenotype.


Wiseman A, Gillham NW, Boynton JE (1977) Nuclear mutations affecting mitochondrial structure and function in Chlamydomonas. J Cell Biol 73:56-77

Boynton-Gillham laboratory, Duke University

Phenotype: antibiotic resistant (erythromycin)

From cross CC-148 er-u-37 sr-u-2-60 x CC-146 nr1-1 spr-u-1-6-2 mt-. This strain is resistant to erythromycin but sensitive to streptomycin, neamine and spectinomycin.

This is a mt+ stock equivalent to CC-67 er-u-37 mt-. er-u-37 is a good chloroplast marker for erythromycin resistance, and has been used in many crosses. It maps to the center of the 23S rRNA gene, proximal to the intron. Please see CC-67 for more information on this mutation.


  • Locus:
  • rrnL
  • Chromosome:
  • chloroplast

Jean Forster, Boynton-Gillham laboratory, Duke University, 1976; originally isolated by Wei-Yeh Wang

The r-1 mutation enhances the accumulation of protoporphyrin by brs1 mutants (see CC-339). This strain carries the r-1 mutation alone, and has no obvious phenotype. Unlike the double mutant r-1 brs1, it is not light-sensitive.


Wang W, Boynton JE, Gillham NW (1975) Genetic control of chlorophyll biosynthesis in chlamydomonas: analysis of a mutant affecting synthesis of delta-aminolevulinic acid. Cell 6:75-84

Wang W, Boynton JE, Gillham NW (1977) Genetic control of chlorophyll biosynthesis: effect of increased δ-aminolevulinic acid synthesis on the phenotype of the y-1 mutant of Chlamydomonas. Mol Gen Genet 152:7-12

Jean Forster, Boynton-Gillham laboratory, Duke University, 1976; originally isolated by Wei-Yeh Wang. Original stock lost, replaced by Wang in December 1982 and again in February 1984

Phenotype: chlorophyll deficient, accumulates protoporphyrin

The brs1 mutant, which was UV-induced by Wei-Yeh Wang in strain CC-125, is chlorophyll-deficient and accumulates protoporphyrin IX. It is very light sensitive and must be grown in the dark. Chekounova et al. determined that the brs1 mutation is a frameshift in exon 10 of the CHLH gene.

Cultures of the original brs1 mutant were brown. The r1 mutation enhances the accumulation of protoporphyrin. This strain appears dark brown and is very light-sensitive.


Wang WY, Wang WL, Boynton JE, Gillham NW (1974) Genetic control of chlorophyll biosynthesis in Chlamydomonas. Analysis of mutants at two loci mediating the conversion of protoporphyrin-IX to magnesium protoporphyrin. J Cell Biol 63:806-823

Wang WY (1978) Genetic control of chlorophyll biosynthesis in Chlamydomonas reinhardtii. Int. Rev Cytol suppl 8:335-364

Johanningmeier U and Howell SH (1984) Regulation of light-harvesting chlorophyll-binding protein mRNA accumulation in Chlamydomonas reinhardi. Possible involvement of chlorophyll synthesis precursors. J Biol Chem 259:13541-13549

Chekounova E, Voronetskaya V, Papenbrock J, Grimm B, Beck CF (2001) Characterization of Chlamydomonas mutants defective in the H subunit of Mg-chelatase. Mol Gen Genet 266:363-373

Von Gromoff ED, Alawady A, Meinecke L, Grimm B, Beck CF (2008) Heme, a plastid-derived regulator of nuclear gene expression in Chlamydomonas. Plant Cell 20:552-567


  • Locus:
  • CHLH
  • Chromosome:
  • 7

Obtained by Barbara Sears in Boynton-Gillham Laboratory, Duke University, 1976, from Robert Smyth, University of Colorado

Phenotype: requires acetate and nicotinamide

This strain is a rare nic7/ac29a recombinant isolated by Robert Smyth.


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


  • Locus:
  • AC29 [ALB3], NIC7 [QSA1]
  • Chromosome:
  • 6

Hurley Shepherd in Boynton-Gillham laboratory, Duke University, 1979

Phenotype: requires acetate

The ac-u-b-1-5 mutant is deficient in ATP synthase activity, but the mutational lesion has not been identified. Chloroplast DNA appears normal when digested with BamHI or EcoRI.


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

Hurley Shepherd in Boynton-Gillham laboratory, Duke University, 1979

Phenotype: requires acetate

This is a frameshift mutation near the 5′ end of the atpE gene. CC-367 is easy to score as a clean acetate-requiring strain.


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

Woessner JP, Masson A, Harris EH, Bennoun P, Gillham NW, Boynton JE (1984) Molecular and genetic analysis of the chloroplast ATPase of chlamydomonas. Plant Mol Biol 3:177-190

Robertson D, Boynton JE, Gillham NW (1990) Cotranscription of the wild-type chloroplast atpE gene encoding the CF1/CF0 epsilon subunit with the 3' half of the rps7 gene in Chlamydomonas reinhardtii and characterization of frameshift mutations in atpE. Mol Gen Genet 221:155-163


  • Locus:
  • atpE
  • Chromosome:
  • chloroplast

Hurley Shepherd in Boynton-Gillham laboratory, Duke University, 1979

Phenotype: requires acetate

This is a deletion mutation in the atpB gene. CC-368 is easy to score as a clean acetate-requiring strain. Another deletion mutant, CC-373 ac-u-c-2-21, is more frequently used as a recipient for atpB transformation of the chloroplast, however.


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

Woessner JP, Masson A, Harris EH, Bennoun P, Gillham NW, Boynton JE (1984) Molecular and genetic analysis of the chloroplast ATPase of chlamydomonas. Plant Mol Biol 3:177-190


  • Locus:
  • atpB
  • Chromosome:
  • chloroplast

Hurley Shepherd in Boynton-Gillham laboratory, Duke University, 1979

Phenotype: requires acetate

This is a deletion mutation in the atpB gene.


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

Woessner JP, Masson A, Harris EH, Bennoun P, Gillham NW, Boynton JE (1984) Molecular and genetic analysis of the chloroplast ATPase of chlamydomonas. Plant Mol Biol 3:177-190


  • Locus:
  • atpB
  • Chromosome:
  • chloroplast

Hurley Shepherd in Boynton-Gillham laboratory, Duke University, 1979

Phenotype: requires acetate

ac-u-c-2-21 is a deletion mutation in the chloroplast atpB gene. CC-373 is a clean, non-reverting acetate-requiring mutant, and has been widely used as a recipient for chloroplast transformation experiments. For the same mutation in mt-, see CC-728.


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

Woessner JP, Masson A, Harris EH, Bennoun P, Gillham NW, Boynton JE (1984) Molecular and genetic analysis of the chloroplast ATPase of chlamydomonas. Plant Mol Biol 3:177-190

Boynton JE, Gillham NW, Harris EH, Hosler JP, Johnson AM, Jones AR, Randolph-Anderson BL, Robertson D, Klein TM, Shark KB (1988) Chloroplast transformation in Chlamydomonas with high velocity microprojectiles. Science 240:1534-1538


  • Locus:
  • atpB
  • Chromosome:
  • chloroplast

From John Cross, Cal Tech, spring 1976

Phenotype: temperature sensitive, cannot grow at 33 degrees

This is a temperature-sensitive lethal, introduced in an arg2 strain by nitrosoguanidine mutagenesis followed by canavainine selection.


McMahon D (1971) The isolation of mutants conditionally defective in protein synthesis in Chlamydomonas reinhardi. Mol Gen Genet 112:80-86

Cross J and McMahon D (1976) Temperature-sensitive breakdown in vivo of polysomes with mutants of Chlamydomonas reinhardii. Mol Gen Genet 147:169-177

From John Cross, CalTech, spring 1976

Phenotype: temperature sensitive, cannot grow at 33 degrees

This is a temperature-sensitive lethal, introduced in an arg2 strain by nitrosoguanidine mutagenesis followed by canavainine selection.


McMahon D (1971) The isolation of mutants conditionally defective in protein synthesis in Chlamydomonas reinhardi. Mol Gen Genet 112:80-86

Cross J and McMahon D (1976) Temperature-sensitive breakdown in vivo of polysomes with mutants of Chlamydomonas reinhardii. Mol Gen Genet 147:169-177

From Ruth Sager, Sidney Farber Cancer Institute, December 1976

Phenotype: antibiotic resistant (clindamycin, erythromycin)

This mutant was isolated by Sager in her strain 21 gr background, as resistant to 50-100 micrograms/ml clindamycin (Cleocin), and designated by her as cle1 or cle. This mutant is cross-resistant to erythromycin, and was found by Bartlett et al. to be genetically allelic with the er-u-37 mutant. Harris et al. determined that this mutation is an A -> G change in the peptidyl transferase region of the 23S rRNA gene, at the base equivalent to E. coli 2058. The base pair change in this mutant is identical to that in the independently isolated erythromycin-resistant mutant er-u-AW-17.


Bartlett SG, Harris EH, Grabowy CT, Gillham NW, Boynton JE (1979) Ribosomal subunits affected by antibiotic resistance mutations at seven chloroplast loci in Chlamydomonas reinhardtii. Mol Gen Genet 176:199-208

Harris EH, Burkhart BD, Gillham NW, Boynton JE (1989) Antibiotic resistance mutations in the chloroplast 16S and 23S rRNA genes of Chlamydomonas reinhardtii: correlation of genetic and physical maps of the chloroplast genome. Genetics 123:281-292


  • Locus:
  • rrnL
  • Chromosome:
  • chloroplast

From Mike Adams in David Luck’s laboratory, Rockfeller University, May 1977

Phenotype: antibiotic resistant (anisomycin)

This is a spontaneous mutant isolated in David Luck’s laboratory, in strain 137c. James and Lefebvre showed that ani1 is an allele at the PDR1 locus. It is easily scored as resistant to 10 micrograms/ml anisomycin on plates.

This strain is a good marker for linkage group XVIII.


James SW and Lefebvre PA (1989) Isolation and characterization of dominant, pleiotropic drug-resistance mutants in Chlamydomonas reinhardtii. Curr Genet 15:443-452


  • Locus:
  • PDR1
  • Chromosome:
  • 16

From K.-S. Chiang, University of Chicago, originally from S.J. Surzycki; brought to Duke by David Grant ca. 1978

Phenotype: wall deficient

This is the most widely used wall-deficient mutant, and this mutation is present in many strains in the Chlamydomonas Resource Center collection. Davies characterized it as a mutant in his class C, defined as mutants in which cell walls are absent or produced in greatly reduced quantity compared to wild type.

Wall-deficient mutants form flat, liquid colonies on agar, and the cells lyse in a 1% solution of the nonionic detergent NP-40.

This is an isolate derived by David Grant from a cw15 stock obtained from S.J. Surzycki, while David was in Chiang’s lab.

The original stock required 0.2% sodium acetate for growth in liquid and was quite sensitive to pH and/or CO2; this single clone isolate does not require acetate and is not sensitive.


Davies DR and Plaskitt A (1971) Genetical and structural analyses of cell-wall formation in Chlamydomonas reinhardi. Genet Res 17:33-43

Hyams J and Davies DR (1972) The induction and characterization of cell wall mutants of Chlamydomonas reinhardi. Mut Res 14:381-389

Davies DR and Lyall V (1973) The assembly of a highly ordered component of the cell wall: the role of heritable factors and of physical structure. Mol Gen Genet 124:21-34

From K.-S. Chiang, University of Chicago, originally from
R.P. Levine; brought to Duke by David Grant ca. 1978

Phenotype: wall deficient

This stock is the cw15 strain used at Harvard, obtained from Levine by Chiang’s lab, brought to Duke by David Grant. Please see CC-400 for more information on cw15.

From Eiji Hase, Institute of Applied Microbiology, University of Tokyo, Japan, ca. 1978

This is C. reinhardtii strain C8 from Eiji Hase, Institute of Applied Microbiology, University of Tokyo, Japan, and was brought to Duke from K.S. Chiang’s laboratory by David Grant.

According to information from Matsuda 9/84, strain C8 and strain C9 were acquired by Y. Tsubo from Ruth Sager in 1954 and were donated to the Tokyo stock collection in 1959. Thus these strains are probably equivalent to Sager 21 gr and y1. Unlike 21 gr, this strain appears to be yellow in the dark. This strain can grow on nitrate.


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

From Eiji Hase, Institute of Applied Microbiology, University of Tokyo, Japan, ca. 1978

This is C. reinhardtii strain C9 from Eiji Hase, Institute of Applied Microbiology, University of Tokyo, Japan, and was brought to Duke from K.S. Chiang’s laboratory by David Grant.

According to information from Matsuda 9/84, strain C8 and strain C9 were acquired by Y. Tsubo from Ruth Sager in 1954 and were donated to the Tokyo stock collection in 1959. Thus these strains are probably equivalent to Sager 21 gr and y1. This strain can grow on nitrate.

For more information on the origin of the standard laboratory strains of C. reinhardtii, please see the following paper:


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

This strain was obtained by David Grant from Uwe G. Schlösser, Sammlung von Algenkulturen (SAG), while David was at the University of Chicago, and was brought to Duke University by him. It is equivalent to CC-1010 UTEX 90, and can grow on nitrate.

For more information on the origin of the standard laboratory strains of C. reinhardtii, please see the following paper:


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

This strain was obtained by David Grant from Uwe G. Schlösser, Sammlung von Algenkulturen (SAG), while David was at the University of Chicago, and was brought to Duke University by him. It was originally accessioned by SAG as “Lewin Caroline Islands strain” but its chloroplast DNA is indistinguishable from that of the standard laboratory strain of C. reinhardtii. Ferris reported that its distribution of the Gulliver transposon is also consistent with its being an isolate of the standard strain.

Christoph Beck reported this strain did not mate with his 137c or CC-1373 11/86, whereas our tests showed good mating with CC-125.


Ferris PJ (1989) Characterization of a Chlamydomonas transposon, Gulliver, resembling those in higher plants. Genetics 122:363-377

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

Jean Forster in Boynton-Gillham laboratory, Duke University

Phenotype: antibiotic resistant (cycloheximide, erythromycin, spectinomycin, streptomycin); requires acetate and nicotinamide

Isolated as follows:

CC-147 nr1-8 spr-u-1-6-2 mt+ x CC-29 ac17 can1 nic13 pf2 y1 pyr1 msr1 act2 sr1 mt- –> product 21-5-8, ac spr nic pyr nr act mt- (sensitive to methionine sulfoximine and streptomycin)
CC-227 spr-u-1-6-2 er-u-37 sr-u-2-60 mt+ x 21-5-8 –> product #20, = CC-412


Forster JL, Grabowy CT, Harris EH, Boynton JE, Gillham NW (1980) Behavior of chloroplast genes during the early zygotic divisions of Chlamydomonas reinhardtii. Curr Genet 1, 137-153


  • Locus:
  • AC17, ACT2 [RPL36a], NIC13 [NSN1], rrnL, rrnS
  • Chromosome:
  • 3,6,10,chloroplast

From Barbara Sears in Boynton-Gillham laboratory, Duke University

Phenotype: requires acetate and nicotinamide; antibiotic resistant (spectinomycin)

From a cross by Barbara Sears in the Boynton-Gillham lab, CC-104 spr-u-1-27-3 mt+ x CC-350 nic7 ac29a mt-

Patrick Ferris reports that this strain has a duplication in the mating type region that creates a distinctive RFLP compared to other strains.


  • Locus:
  • AC29 [ALB3], NIC7 [QSA1], rrnS
  • Chromosome:
  • 6,chloroplast

From Barbara Sears in Boynton-Gillham laboratory, Duke University

Phenotype: requires acetate and nicotinamine; antibiotic resistant (kanamycin or neamine)

From a cross by Barbara Sears in 1977, CC-86 nr-u-2-1 mt+ x CC-350 nic7 ac29a mt-


  • Locus:
  • AC29 [ALB3], NIC7 [QSA1], rrnS
  • Chromosome:
  • 6,chloroplast

From Robert Lee, Dalhousie University, October 1977

Phenotype: wall deficient; requires arginine; antibiotic resistant (streptomycin)

Presumably derived from Lee’s cw15 isolate, which was originally from Ledoux.

This strain was used in transformation experiments in Boynton/Gillham and Kohorn laboratories at Duke, 1993. Transformation rates were good, but subsequent crosses of transformants gave many incomplete tetrads.

Please see CC-48 for more information on arg2, CC-118 for the chloroplast streptomycin resistance mutation sr-u-2-60, and CC-400 for more on cw15.


  • Locus:
  • ARG7, rrnS
  • Chromosome:
  • 1,chloroplast

From Robert Lee, Dalhousie University, October 1977

Phenotype: wall deficient; requires arginine; antibiotic resistant (streptomycin)

Presumably derived from Lee’s cw15 isolate, which was originally from Ledoux.

This strain was used in transformation experiments in Boynton/Gillham and Kohorn laboratories at Duke, 1993. Transformation rates were good, but subsequent crosses of transformants gave many incomplete tetrads.

Please see CC-48 for more information on arg2, CC-118 for the chloroplast streptomycin resistance mutation sr-u-2-60, and CC-400 for more on cw15.


  • Locus:
  • ARG7, rrnS
  • Chromosome:
  • 1, chloroplast

From Robert Lee, Dalhousie University, October 1977

Phenotype: wall deficient; requires nicotinamide; antibiotic resistant (spectinomycin)

Presumably derived from Lee’s cw15 isolate, originally from Ledoux.

Please see CC-400 for more information on cw15, and CC-599 for nic1.


  • Locus:
  • NIC1, rrnS
  • Chromosome:
  • 14, chloroplast

Hurley Shepherd in Boynton-Gillham laboratory, Duke University

Phenotype: requires acetate

Woessner et al. found this mutant to be deficient in chloroplast ATP synthase activity, but Bennoun (pers. comm.) had difficulties in scoring it, and Harris found wild type colonies were produced in all matrix crosses; thus this mutant was not included in the complementation/recombination matrix in Woessner et al. (1984). The genetic lesion has not been identified.

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

Woessner JP, Masson A, Harris EH, Bennoun P, Gillham NW, Boynton JE (1984) Molecular and genetic analysis of the chloroplast ATPase of chlamydomonas. Plant Mol Biol 3:177-190

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