Strains

Boynton-Gillham laboratory, Duke University

Phenotype: antibiotic resistant (kanamycin)

The kr-u-24-2 mutant was solated by N.W. Gillham at Duke, using nitrosoguanidine as mutagen.

This strain is resistant to 50-100 micrograms/ml kanamycin or neamine in agar. The mutation is a change from C to U at position 1341 (equivalent to E. coli 1409) in the chloroplast 16S rRNA.

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:
• rrnS
• Chromosome:
• chloroplast

Boynton-Gillham laboratory, Duke University

Phenotype: antibiotic resistant (erythromycin, streptomycin)

From a cross of nr-u-2-1 sr-u-2-60 spr-u-1-H-4 mt+ to CC-67 er-u-37 mt-, in the Boynton-Gillham lab at Duke; derived from progeny of a spontaneous biparental zygote.

This recombinant product does not carry either the nr-u-2-1 or spr-u-1-H-4 markers. It has been used extensively in genetic analysis at Duke and is a good choice when a strain with two easily distinguished chloroplast antibiotic resistance markers is desired.

• Locus:
• rrnL, rrnS
• Chromosome:
• chloroplast

From Ruth Sager, Hunter College, April 1974

Phenotype: antibiotic resistant (cycloheximide, streptomycin)

This is one of the strains used by Sager and Ramanis in their series of papers on mapping chloroplast genes by pedigree and cosegregation analysis. It combines the chloroplast marker sr-u-sm2 (in the rps12 gene; see Liu et al. 1989) with the nuclear marker act1, which permitted identification of second-division segregation events in meiosis.

Phenotypic tests at Duke showed that CC-215 carries a nuclear neamine resistance marker, presumably nr1 although allelism tests have not been done to confirm this. The cycloheximide resistance marker was shown to be act1 in a cross to ac12.

Like most of Sager’s strains, CC-215 grows on nitrate as sole N source. This property is in contrast to most strains from the Ebersold/Levine background, which carry the nit1 and nit2 mutations.

Liu XQ, Gillham NW, Boynton JE (1989) Chloroplast ribosomal protein gene rps12 of Chlamydomonas reinhardtii. Wild-type sequence, mutation to streptomycin resistance and dependence, and function in Escherichia coli. J Biol Chem 264:16100-16108

• Locus:
• ACT1, rps12
• Chromosome:
• 2,chloroplast

From Ruth Sager, Hunter College, April 1974

Phenotype: antibiotic resistant (erythromycin)

Like most of Sager’s strains, CC-216 grows on nitrate as sole N source . This property is in contrast to most strains from the Ebersold/Levine background, which carry the nit1 and nit2 mutations.

er-u-11 is a C-> G base change in the 23S rRNA at the position equivalent to E. coli 2611. This is the same nucleotide that is mutated to T in er-u-1a.

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 Ruth Sager, Hunter College, April 1974

Phenotype: antibiotic resistant (spectinomycin)

The spr-u-sp-23 mutant (originally sp23) was isolated by Sager and later shown by Harris et al. to have an A to C change at position 1123 in the chloroplast 16S rRNA. This mutation confers high level spectinomycin resistance under all growth conditions.

Like most of Sager’s strains, CC-217 grows on nitrate as sole N source (indicated by the NIT allele designation). This property is in contrast to most strains from the Ebersold/Levine background, which carry the nit1 and nit2 mutations.

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:
• rrnS
• Chromosome:
• chloroplast

Andrew Wiseman, Boynton-Gillham laboratory, Duke University

Phenotype: antibiotic resistant (erythromycin)

This is a mutant isolated by Andrew Wiseman in the Boynton-Gillham laboratory at Duke. The base pair change is an A>G in the chloroplast 23S rRNA at the position equivalent to E. coli 2058. In contrast to er-u-37, which is a mutation in the adjacent nucleotide in the 23S rDNA, er-u-AW-17 is cross-resistant to lincomycin and clindamycin.

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

Elizabeth Harris, Boynton-Gillham laboratory, Duke University

Phenotype: cold sensitive on minimal medium

The cr5 mutant is deficient in chloroplast ribosomes and accumulates 54S subunit particles. It is not allelic with the other cr mutants with similar phenotypes.

Elizabeth Harris, Boynton-Gillham laboratory, Duke University

Phenotype: cold sensitive on minimal medium

The cr5 mutant is deficient in chloroplast ribosomes and accumulates 54S subunit particles. It is not allelic with the other cr mutants with similar phenotypes.

Elizabeth Harris, Boynton-Gillham laboratory, Duke University

Phenotype: cold sensitive on minimal medium

The cr6 mutant is deficient in 70S chloroplast ribosomes and accumulates some 41S and some 54S subunit particles and/or poorly defined mixed subunit material. Initial characterization suggested that this mutant was distinguishable from another mutant, cr7, based on the amount of ribosomal subunit material accumulated when analyzed on sucrose gradients. In subsequent analysis by Myers et al. these two mutants proved to be indistinguishable, and were shown to be alleles at the same locus. Myers et al. showed that cr6 and cr7 fail to assemble to proteins of the large subunit of the chloroplast ribosome, one normally synthesized in the chloroplast and one in the cytoplasm.

The original cr6 mutant formed diploids with ac20, cr1, cr2, cr3, and cr4; cr5 was not tested.

Myers AM, Harris EH, Gillham NW, Boynton JE (1984) Mutations in a nuclear gene of Chlamydomonas cause the loss of two chloroplast ribosomal proteins, one synthesized in the chloroplast and the other in the cytoplasm. Curr Genet 8:369-378

Elizabeth Harris, Boynton-Gillham laboratory, Duke University

Phenotype: cold sensitive on minimal medium

The cr6 mutant is deficient in 70S chloroplast ribosomes and accumulates some 41S and some 54S subunit particles and/or poorly defined mixed subunit material. Initial characterization suggested that this mutant was distinguishable from another mutant, cr7, based on the amount of ribosomal subunit material accumulated when analyzed on sucrose gradients. In subsequent analysis by Myers et al. these two mutants proved to be indistinguishable, and were shown to be alleles at the same locus. Myers et al. showed that cr6 and cr7 fail to assemble to proteins of the large subunit of the chloroplast ribosome, one normally synthesized in the chloroplast and one in the cytoplasm.

The original cr6 mutant formed diploids with ac20, cr1, cr2, cr3, and cr4; cr5 was not tested.

Myers AM, Harris EH, Gillham NW, Boynton JE (1984) Mutations in a nuclear gene of Chlamydomonas cause the loss of two chloroplast ribosomal proteins, one synthesized in the chloroplast and the other in the cytoplasm. Curr Genet 8:369-378

Elizabeth Harris, Boynton-Gillham laboratory, Duke University

Phenotype: cold sensitive on minimal medium

This mutant accumulates some 41S and some 54S subunits of chloroplast ribosomes, or poorly defined mixed subunit material, and is very deficient in 70S ribosomes.

Myers et al. showed that cr7 and its allele cr6 fail to assemble to proteins of the large subunit of the chloroplast ribosome, one normally synthesized in the chloroplast and one in the cytoplasm.

Myers AM, Harris EH, Gillham NW, Boynton JE (1984) Mutations in a nuclear gene of Chlamydomonas cause the loss of two chloroplast ribosomal proteins, one synthesized in the chloroplast and the other in the cytoplasm. Curr Genet 8:369-378

• Locus:
• CR7
• Chromosome:
• 1

Elizabeth Harris, Boynton-Gillham laboratory, Duke University

Phenotype: cold sensitive on minimal medium

This mutant accumulates some 41S and some 54S subunits of chloroplast ribosomes, or poorly defined mixed subunit material, and is very deficient in 70S ribosomes.

Myers et al. showed that cr7 and its allele cr6 fail to assemble to proteins of the large subunit of the chloroplast ribosome, one normally synthesized in the chloroplast and one in the cytoplasm.

Myers AM, Harris EH, Gillham NW, Boynton JE (1984) Mutations in a nuclear gene of Chlamydomonas cause the loss of two chloroplast ribosomal proteins, one synthesized in the chloroplast and the other in the cytoplasm. Curr Genet 8:369-378

• Locus:
• CR7
• Chromosome:
• 1

Chlamydomonas Genetics Center, Duke University

Phenotype: requires nicotinamide; antibiotic and inhibitor resistant (canavanine, cycloheximide, methionine sulfoximine, pyrithiamine)

From a cross of CC-28 x CC-36, 1974 (Ebersold multiply marked strain x cr3). This isolate does not carry the pf2 marker from CC-28, or the cr3 marker.

This strain was used in mapping crosses by the Chlamydomonas Genetics Center in 1980 to confirm the linkage groups of some pf mutations.

• Locus:
• ACT2 [RPL36a], CAN1, MSR1, NIC13 [NSN1], PYR1
• Chromosome:
• 1,4,6,7,10

Chlamydomonas Genetics Center, Duke University

Phenotype: requires thiamine; impaired motility

From a cross of CC-21 pf15 x CC-24 thi2.

This strain provides two linked markers on group III. The thi2 marker is difficult to score in the presence of acetate-requiring mutations.

• Locus:
• PF15 [KAT2], THI2
• Chromosome:
• 3

Chlamydomonas Genetics Center, Duke University

Phenotype: requires thiamine; impaired motility

From a cross of CC-15 pf1 x CC-25 thi8.

This strain provides two markers on linkage group V.

• Locus:
• PF1 [RSP4], THI8
• Chromosome:
• 5

Chlamydomonas Genetics Center, Duke University

Phenotype: requires nicotinamide; impaired motility

From a cross of CC-13 nic11 x CC-22 pf20.

This strain provides two markers for linkage group IV.

• Locus:
• NIC11, PF20
• Chromosome:
• 4

From Jeff Davidson in Lawrence Bogorad’s laboratory, Harvard University, 1974

Phenotype: impaired motility

Cells with the pf17 mutation lack flagellar spokeheads and are missing five polypeptides of the radial spoke complex. Yang et al. showed that this is a mutation in the gene encoding radial spoke protein 9.

This mutation is a good marker for linkage group VII. Cells wiggle but don’t swim, and are easily scored in crosses.

Yang P, Diener DR, Yang C, Kohno T, Pazour GJ, Dienes JM, Agrin NS, King SM, Sale WS, Kamiya R, Rosenbaum JL, Witman GB (2006) Radial spoke proteins of Chlamydomonas flagella. J Cell Sci. 119(Pt 6):1165-1174

• Locus:
• PF17 [RSP9]
• Chromosome:
• 7

From Ruth Sager, Hunter College, March 1975

Phenotype: antibiotic and inhibitor resistant (methionine sulfoximine, streptomycin)

This is the primary stock of sr-u-sm3 in the Chlamydomonas Resource Center collection. Other streptomycin resistance mutations, notably sr-u-2-60, have been more frequently used in crosses for chloroplast genetics, however. sr-u-sm3 is unique among chloroplast sr mutants examined in having an alteration in the extreme 5′ end of the 16S rRNA molecule (U -> G at nt 14, corresponding to E. coli 13). This is not the same as sr-u-sm3a, which is a mutation in a different region of the chloroplast 16S rRNA.

Like most of Sager’s strains, CC-266 grows on nitrate as sole N source (indicated by the nit+ or NIT allele designation). This property is in contrast to most strains from the Ebersold/Levine background, which carry the nit1 and nit2 mutations.

Sager R and Ramanis Z (1965) Recombination of nonchromosomal genes in Chlamydomonas. Proc Natl Acad Sci USA 53:1053-1061

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:
• MSR1, rrnS
• Chromosome:
• 1,chloroplast

From Ruth Sager, Hunter College, March 1975

Phenotype: antibiotic resistant (streptomycin)

This is the primary stock of sr-u-sm5 in the Chlamydomonas Resource Center collection. Other streptomycin resistance mutations, notably sr-u-2-60, have been more frequently used in crosses for chloroplast genetics, however. This mutant has an alteration in the ’912′ region of the 16S rRNA molecule (A -> C at nt 858, corresponding to E. coli 914).

Like most of Sager’s strains, CC-267 grows on nitrate as sole N source (indicated by the nit+ or NIT allele designation). This property is in contrast to most strains from the Ebersold/Levine background, which carry the nit1 and nit2 mutations.

Sager R and Ramanis Z (1976) Chloroplast genetics of chlamydomonas. I. Allelic segregation ratios. Genetics 83:303-321

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:
• rrnS
• Chromosome:
• chloroplast

From Ruth Sager, Hunter College, March 1975

Phenotype: antibiotic and inhibitor resistant (cycloheximide, methionine sulfoximine, streptomycin)

This is one of the strains reportedly used by Sager in her series of papers on mapping chloroplast genes by pedigree and cosegregation analysis. Although labeled as sensitive to actidione (cycloheximide) and methionine sulfoximine when received, phenotypic tests at Duke showed that it is resistant to both compounds. The cycloheximide resistance marker was shown to be act1 in a cross to ac12 and pf12.

Like most of Sager’s strains, CC-270 grows on nitrate as sole N source (indicated by the nit+ or NIT allele designation). This property is in contrast to most strains from the Ebersold/Levine background, which carry the nit1 and nit2 mutations.

Please see CC-215 for more information on the sr-u-sm2 mutation.

• Locus:
• ACT1, MSR1, rps12
• Chromosome:
• 1,2,chloroplast

Boynton-Gillham laboratory, Duke University

Phenotype: wall deficient

Isolated by Nancy Alexander in Boynton/Gillham laboratory from a contaminated stock of cw15 sent by D.R. Davies in 1972; designated cw15-2. CC-400 is another cw15 mt+ isolate that grows better than this one, and is therefore recommended. Please see CC-400 for more information on the cw15 mutation.

Boynton-Gillham laboratory, Duke University

Phenotype: wall deficient

Isolated by Nancy Alexander from a cross of CC-277 x wild type mt-; isolate cw15-2-4. We recommend using CC-406 or CC-3491 instead, since they are more vigorous cw15 mt- strains.

Boynton-Gillham laboratory, Duke University

Phenotype: requires acetate

Derived as follows: CC-125 x GB-265 (a clean acetate-requiring mutant in a diploid background) produced a haploid progeny strain ac 8.1, which was then crossed to CC-124 to yield CC-297. At last check this strain was still clearly acetate-requiring, but it has never been characterized further.

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

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, but appears to have become suppressed.

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

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, but appears to have become suppressed.

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

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, but appears to have become suppressed.

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 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 and cytochrome oxidase, but essentially normal mitochondrial ultrastructure. At last check, the strain still showed a dark-dier phenotype, with very slow growth at most in the dark.

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 and abnormal mitochondrial ultrastructure. At last check, the strain still showed a dark-dier phenotype, with some very slow growth of light green colonies in the dark.

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 no detectable cytochrome oxidase activity, but essentially normal mitochondrial ultrastructure. At last check, the strain still showed a very clean 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