Strains

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Using plasmid p699 (GuhaMajumdar et al. 2008), Sriram Satagopan in Spreitzer’s group performed rbcL direct mutagenesis, chloroplast transformation of wild-type 2137 mt+ (cloned), selection for spectinomycin resistance in the dark, and screening for a homoplasmic acetate-requiring phenotype to create a K175R substitution (AAA-CGT) in the Rubisco large subunit. The K175R substitution causes a 50% decrease in Rubisco holoenzyme and a 99% decrease in Rubisco carboxylase activity (Lim and Spreitzer, unpublished). The x-ray crystal structure of this K175R Rubisco enzyme has been solved (Andersson and Spreitzer, unpublished). This mutant was created to investigate the role of active-site Lys-175 in catalysis (Harpel et al. 2002). It has been maintained with acetate medium in darkness since its creation.

GuhaMajumdar M, Dawson-Baglien E, Sears BB (2008) Creation of a chloroplast microsatellite reporter for detection of replication slippage in Chlamydomonas reinhardtii. Eukaryot Cell 7:639-646

Harpel MR, Larimer FW, Hartman FC (2002) Multifaceted roles of Lys166 of ribulose-bisphosphate carboxylase/oxygenase as discerned by product analysis and chemical rescue of site-directed mutants. Biochemistry 41:1390-1397

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Using plasmid p699 (GuhaMajumdar et al. 2008), Sriram Satagopan in Spreitzer’s group performed rbcL direct mutagenesis, chloroplast transformation of wild-type 2137 mt+ (cloned), selection for spectinomycin resistance in the dark, and screening for a homoplasmic acetate-requiring phenotype to create a K175G substitution (AAA-TCA) in the Rubisco large subunit. The K175S substitution causes a 90% decrease in Rubisco holoenzyme and a 99% decrease in Rubisco carboxylase activity (Lim and Spreitzer, unpublished). This mutant was created to investigate the role of active-site Lys-175 in catalysis (Harpel et al. 2002). It has been maintained with acetate medium in darkness since its creation.

GuhaMajumdar M, Dawson-Baglien E, Sears BB (2008) Creation of a chloroplast microsatellite reporter for detection of replication slippage in Chlamydomonas reinhardtii. Eukaryot Cell 7:639-646

Harpel MR, Larimer FW, Hartman FC (2002) Multifaceted roles of Lys166 of ribulose-bisphosphate carboxylase/oxygenase as discerned by product analysis and chemical rescue of site-directed mutants. Biochemistry 41:1390-1397

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Using plasmid p699 (GuhaMajumdar et al. 2008), Boon Hoe Lim in Spreitzer’s group performed rbcL direct mutagenesis, chloroplast transformation of wild-type 2137 mt+ (cloned), selection for spectinomycin resistance in the dark, and screening for a homoplasmic acetate-requiring phenotype to create a K201E substitution in the Rubisco large subunit. This active-site mutant was created to see whether suppressor substitutions could be selected that would complement a deficiency in Rubisco activation (Spreitzer and Salvucci 2002). It has been maintained with acetate medium in darkness since its creation.

GuhaMajumdar M, Dawson-Baglien E, Sears BB (2008) Creation of a chloroplast microsatellite reporter for detection of replication slippage in Chlamydomonas reinhardtii. Eukaryot Cell 7:639-646

Harpel MR, Larimer FW, Hartman FC (2002) Multifaceted roles of Lys166 of ribulose-bisphosphate carboxylase/oxygenase as discerned by product analysis and chemical rescue of site-directed mutants. Biochemistry 41:1390-1397

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Using plasmid p699 (GuhaMajumdar et al. 2008), Boon Hoe Lim in Spreitzer’s group performed rbcL direct mutagenesis, chloroplast transformation of wild-type 2137 mt+ (cloned), selection for spectinomycin resistance in the dark, and screening for a homoplasmic acetate-requiring phenotype to create a K334R substitution in the Rubisco large subunit. This active-site mutant was created to see whether suppressor substitutions could be selected that would complement a deficiency in Rubisco carboxylation (Spreitzer and Salvucci 2002). It has been maintained with acetate medium in darkness since its creation.

GuhaMajumdar M, Dawson-Baglien E, Sears BB (2008) Creation of a chloroplast microsatellite reporter for detection of replication slippage in Chlamydomonas reinhardtii. Eukaryot Cell 7:639-646

Harpel MR, Larimer FW, Hartman FC (2002) Multifaceted roles of Lys166 of ribulose-bisphosphate carboxylase/oxygenase as discerned by product analysis and chemical rescue of site-directed mutants. Biochemistry 41:1390-1397

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), a Q401T substitution (CAG-ACT) was created in the Rubisco large subunit. This mutant was created to investigate phylogenetic differences in the Rubisco active site (Spreitzer et al. 1988; Chene et al. 1997). See also rbcL-T173I (CC-4819). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

William Zerges reported CC-4874 (Q401T) has no Rbcl gene. The pcr band is the same size as the deletion mutant from three trials/independent preps. (personal communication, 12/8/20)

Chene P, Day AG, Fersht AR (1997) Role of isoleucine-164 at the active site of Rubisco from Rhodospirillum rubrum. Biochem Biophys Res Com 232:482-486

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Spreitzer RJ, Brown T, Chen Z, Zhang D, Al-Abed SR (1988) Missense mutation in the Chlamydomonas chloroplast gene that encodes the Rubisco large subunit. Plant Physiol 86:987-989

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), an R32K substitution was created in the Rubisco large subunit. This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S. The R32K substitution does not affect Rubisco catalysis or pyrenoid formation (Lim and Meyer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), an A222S substitution was created in the Rubisco large subunit. This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S. The A222S substitution does not affect Rubisco catalysis or pyrenoid formation (Lim and Meyer, unpublished). An A222T substitution suppresses the low CO2/O2 specificity and thermal instability of rbcL-L290F Rubisco (Du and Spreitzer 2000). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Du YC, Spreitzer RJ (2000) Suppressor mutations in the chloroplast-encoded large subunit improve the thermal stability of wild-type ribulose-1,5-bisphosphate carboxylase/oxygenase. J Biol Chem 275:19844-19847

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), a C247S substitution was created in the Rubisco large subunit. This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S. The C247S substitution does not affect Rubisco catalysis or pyrenoid formation (Lim and Meyer, unpublished). Cys-247 forms a disulfide bond with Cys-247 in a neighboring large subunit, which may be the only disulfide bond in the Rubisco holoenzyme (Knight et al. 1990). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Knight S, Andersson I, Branden CI (1990) Crystallographic analysis of ribulose 1,5-bisphosphate carboxylase from spinach at 2.4 A resolution. Subunit interactions and active site. J Mol Biol 215:113-160

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), three substitutions (R32K, A222S, and C247S) were created together in the Rubisco large subunit. This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K, rbcL-A222S, and rbcL-C247S. The three combined substitutions do not affect Rubisco catalysis or pyrenoid formation (Lim and Meyer, unpublished). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), a V145C substitution was created in the Rubisco large subunit (Lim and Meyer, unpublished). This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S and rbcL-R32K/V145C/P210S/A222S/I225L/C247S/I309M. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), a P210S substitution was created in the Rubisco large subunit (Lim and Meyer, unpublished). This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S and rbcL-R32K/V145C/P210S/A222S/I225L/C247S/I309M. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), an I225L substitution was created in the Rubisco large subunit (Lim and Meyer, unpublished). This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S and rbcL-R32K/V145C/P210S/A222S/I225L/C247S/I309M. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), an I309M substitution was created in the Rubisco large subunit (Lim and Meyer, unpublished). This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). See also rbcL-R32K/A222S/C247S and rbcL-R32K/V145C/P210S/A222S/I225L/C247S/I309M. An I309M substitution in plant Rubisco alters carboxylation catalytic efficiency (Whitney et al. 2011). This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

Whitney S, Sharwood RE, Orr D, White SJ, Alonso H, Galmes J (2011) Isoleucine 309 acts as a C4 catalytic switch that increases ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) carboxylation rate in Flaveria. Proc Natl Acad Sci USA 108:14688–14693

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Using standard methods of directed mutagenesis and chloroplast transformation of rbcL∆-MX3312 mt+ (CC-4696) (Satagopan and Spreitzer 2004), seven substitutions were created together in the Rubisco large subunit. This mutant was created to investigate phylogenetic differences in the Rubisco large subunit that may be responsible for pyrenoid formation in algae (Nozaki et al. 2002). The seven combined substitutions do not block pyrenoid formation, but the strain grows slower than wild type under photosynthetic growth conditions (Lim and Meyer, unpublished). See also rbcL-R32K/A222S/C247S. This strain has been maintained with acetate medium in darkness to prevent selection for secondary mutations that may improve Rubisco function.

Nozaki H, Onishi K, Morita E (2002) Differences in pyrenoid morphology are correlated with differences in the rbcL genes of members of the Chloromonas lineage (volvocales, chlorophyceae). J Mol Evol 55:414-430

Satagopan S, Spreitzer RJ (2004) Substitutions at the Asp-473 latch residue of Chlamydomonas ribulosebisphosphate carboxylase/oxygenase cause decreases in carboxylation efficiency and CO2/O2 specificity. J Biol Chem 279:14240-14244

• Locus:
• rbcL
• Chromosome:
• chloroplast

From Jason Petersen, Sioux Falls VA Healthcare System, August 2014

Phenotype: uvs9 is UV sensitive and is blocked in excision repair

The UVS9 gene is on Chromosome 10 (Cre10.g429050) and encodes an XPG homolog. The uvs9 allele has a nonsense mutation in exon 9.

The original uvs9 was generated by chemical mutagenesis (N-methyl-N’-nitro-N-nitrosoguanidine) of “137c mt(+)” by Gary Small (personal communication) and shared with Daniel Vlcek. This strain is a descendant of uvs9 from the Vlcek lab.

• Locus:
• UVS9
• Chromosome:
• 10

From Jason Petersen, Sioux Falls VA Healthcare System, August 2014

Phenotype: uvs9 is UV sensitive and is blocked in excision repair

The UVS9 gene is on Chromosome 10 (Cre10.g429050) and encodes an XPG homolog. The uvs9 allele has a nonsense mutation in exon 9.

The original uvs9 was generated by chemical mutagenesis (N-methyl-N’-nitro-N-nitrosoguanidine) of “137c mt(+)” by Gary Small (personal communication) and shared with Daniel Vlcek. This strain is a descendant of uvs9 from the Vlcek lab.

• Locus:
• UVS9
• Chromosome:
• 10

From Robert J. Spreitzer, University of Nebraska, August 2014

Wild-type 2137 mt+ was recovered from a cross between 21gr mt+ (from Sager) and 137c mt- (from Levine) (Spreitzer 1980; Spreitzer and Mets 1981). Wild-type 21gr mt+ grew in clumps in minimal medium, was green in the dark, and was positively phototactic. Wild-type 137c mt- was yellow in the dark (y1) and negatively phototactic. Wild-type 2137 mt+ was chosen as having good-mating, non-clumping, green-in-the-dark, and negatively-phototactic phenotypes (Spreitzer 1980). This is the original 2137 strain. It has been maintained with acetate medium in darkness since it was recovered.

Spreitzer RJ (1980) A uniparental mutant of Chlamydomonas renhardii with altered ribulose-1,5-bisphosphate carboxylase. Ph. D. thesis, Case Western Reserve University

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas reinhardtii with associated light-sensitive phenotypes. Plant Physiol 67:565-569

From Robert J. Spreitzer, University of Nebraska, August 2014

Wild-type 2137 mt+ was recovered from a cross between 21gr mt+ (from Sager) and 137c mt- (from Levine) (Spreitzer 1980; Spreitzer and Mets 1981). Wild-type 21gr mt+ grew in clumps in minimal medium, was green in the dark, and was positively phototactic. Wild-type 137c mt- was yellow in the dark (y1) and negatively phototactic. Wild-type 2137 mt+ was chosen as having good-mating, non-clumping, green-in-the-dark, and negatively-phototactic phenotypes (Spreitzer 1980). The original 2137 mt+ strain has been cloned four times since 1980 to retain its original phenotype. This is the strain used as wild type in Spreitzer’s group (Spreitzer 1998). It has been continuously maintained with acetate medium in darkness.

Spreitzer RJ (1980) A uniparental mutant of Chlamydomonas renhardii with altered ribulose-1,5-bisphosphate carboxylase. Ph. D. thesis, Case Western Reserve University

Spreitzer RJ (1998) The Molecular Biology of Chloroplasts and Mitochondria in Chlamydomonas. Genetic engineering of Rubisco 515-527

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas reinhardtii with associated light-sensitive phenotypes. Plant Physiol 67:565-569

From Robert J. Spreitzer, University of Nebraska, August 2014

This centromere-marker strain was obtained from crosses between wild-type 2137 mt+ (original) and pf2 mt-, which was received by Laurens Mets from Paul Levine in the 1970s. This is the strain most often used for genetic analysis in Spreitzer’s group (Spreitzer and Mets 1981; Spreitzer and Ogren 1983; Spreitzer et al. 1988; Gotor et al. 1994; Hong and Spreitzer 1994). The pf2 phenotype is best observed by placing a loop of water on a colony, and watching for swimming with a dissecting microscope. This strain has been maintained with acetate medium in darkness since its recovery.

Gotor C, Hong S, Spreitzer RJ (1994) Temperature-conditional nuclear mutation of Chlamydomonas reinhardtii decreases the CO2/O2 specificity of chloroplast ribulose-bisphosphate carboxylase/oxygenase. Planta 193:313-319

Hong S, Spreitzer RJ (1994) Nuclear mutation inhibits expression of the chloroplast gene that encodes the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. Plant Physiol 106:673-678

Spreitzer RJ, Al-Abed SR, Huether MJ (1988) Temperature-sensitive, photosynthesis-deficient mutants of Chlamydomonas reinhardtii. Plant Physiol 86:773-777

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas reinhardtii with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• PF2
• Chromosome:
• 11

From Robert J. Spreitzer, University of Nebraska, August 2014

This wild-type strain was received by Laurens Mets from Ruth Sager in the 1970s, and was used as the mt+ parent for production of wild-type strain 2137 mt+ (CC-3269) (Spreitzer and Mets 1981). The 21gr mt+ strain (CC-1690) was originally selected for good growth in the dark from Smith’s 137 mt+ strain (Sager 1955). See also Smith and Regnery (1950). This strain has been maintained with acetate medium in darkness since 1980.

Sager R (1955) Inheritance in the green alga Chlamydomonas reinhardi. Genetics 40:476-489

Smith GM, Regnery DC (1950) Inheritance of sexuality in Chlamydomonas reinhardi. Proc Natl Acad Sci USA 36:246-248

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas reinhardtii with associated light-sensitive phenotypes. Plant Physiol 67:565-569

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, leaky

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 2137 mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 11-6F grows slowly on minimal medium in the light and displayed a mendelian pattern of inheritance. It has PSI-deficient fluorescence-induction kinetics, but only a 50% decrease in CO2 fixation (Spreitzer and Mets 1981). This is the original isolate (see also CC-1229 11-6F mt-). It has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants or suppressors.

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 21gr mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 8-31 displayed uniparental inheritance, and has normal fluorescence-induction kinetics (Spreitzer and Mets 1981). Preliminary studies indicated that it lacks photosynthetic phosphorylation (Selman-Reimer et al. 1981), and may be an atp-gene mutant. Recombination tests showed that 8-31 is at the same locus (psp-u-1) as 8-35, 10-2E, 10-7C, and 12-10B (Spreitzer and Ogren 1983a). A different photosynthetic phosphorylation locus (psp-u-2) was defined by mutant 12-5A (CC-2260) (Spreitzer and Ogren 1983a). This strain was recovered from a cross between the original 8-31 mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Selman-Reimer S, Merchant S, Selman BR (1981) Isolation and characterization of the chloroplast coupling factor 1 from wild-type Chlamydomonas reinhardi. In BR Selman, S Selman-Reimer, eds, Energy Coupling in Photosynthesis, pp 341-352

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 21gr mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 8-31 displayed uniparental inheritance, and has normal fluorescence-induction kinetics (Spreitzer and Mets 1981). Preliminary studies indicated that it lacks photosynthetic phosphorylation (Selman-Reimer et al. 1981), and may be an atp-gene mutant. Recombination tests showed that 8-31 is at the same locus (psp-u-1) as 8-35, 10-2E, 10-7C, and 12-10B (Spreitzer and Ogren 1983a). A different photosynthetic phosphorylation locus (psp-u-2) was defined by mutant 12-5A (CC-2260) (Spreitzer and Ogren 1983a). This strain was recovered from a cross between the original 8-31 mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Selman-Reimer S, Merchant S, Selman BR (1981) Isolation and characterization of the chloroplast coupling factor 1 from wild-type Chlamydomonas reinhardi. In BR Selman, S Selman-Reimer, eds, Energy Coupling in Photosynthesis, pp 341-352

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 21gr mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 8-35 displayed uniparental inheritance, and has normal fluorescence-induction kinetics (Spreitzer and Mets 1981). Preliminary studies indicated that it lacks photosynthetic phosphorylation (Selman-Reimer et al. 1981), and may be an atp-gene mutant. Recombination tests showed that 8-35 is at the same locus (psp-u-1) as 8-31, 10-2E, 10-7C, and 12-10B (Spreitzer and Ogren 1983a). A different photosynthetic phosphorylation locus (psp-u-2) was defined by mutant 12-5A (CC-2260) (Spreitzer and Ogren 1983a). This strain was recovered from a cross between the original 8-35 mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Selman-Reimer S, Merchant S, Selman BR (1981) Isolation and characterization of the chloroplast coupling factor 1 from wild-type Chlamydomonas reinhardi. In BR Selman, S Selman-Reimer, eds, Energy Coupling in Photosynthesis, pp 341-352

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 2137 mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 10-2E displayed uniparental inheritance, and has normal fluorescence-induction kinetics (Spreitzer and Mets 1981). Recombination tests showed that 10-2E is at the same locus (psp-u-1) as 8-31, 8-35, 10-7C, and 12-10B (Spreitzer and Ogren 1983a) indicating that it may lack photosynthetic phosphorylation due to an atp-gene mutation (Selman-Reimer et al. 1981). A different photosynthetic phosphorylation locus (psp-u-2) was defined by mutant 12-5A (CC-2260) (Spreitzer and Ogren 1983a). This strain was recovered from a cross between the original 10-2E mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Selman-Reimer S, Merchant S, Selman BR (1981) Isolation and characterization of the chloroplast coupling factor 1 from wild-type Chlamydomonas reinhardi. In BR Selman, S Selman-Reimer, eds, Energy Coupling in Photosynthesis, pp 341-352

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 2137 mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 10-2E displayed uniparental inheritance, and has normal fluorescence-induction kinetics (Spreitzer and Mets 1981). Recombination tests showed that 10-2E is at the same locus (psp-u-1) as 8-31, 8-35, 10-7C, and 12-10B (Spreitzer and Ogren 1983a) indicating that it may lack photosynthetic phosphorylation due to an atp-gene mutation (Selman-Reimer et al. 1981). A different photosynthetic phosphorylation locus (psp-u-2) was defined by mutant 12-5A (CC-2260) (Spreitzer and Ogren 1983a). This strain was recovered from a cross between the original 10-2E mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Selman-Reimer S, Merchant S, Selman BR (1981) Isolation and characterization of the chloroplast coupling factor 1 from wild-type Chlamydomonas reinhardi. In BR Selman, S Selman-Reimer, eds, Energy Coupling in Photosynthesis, pp 341-352

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 2137 mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 10-7C displayed uniparental inheritance, and has normal fluorescence-induction kinetics (Spreitzer and Mets 1981). Recombination tests showed that 10-7C is at the same locus (psp-u-1) as 8-31, 8-35, 10-2E, and 12-10B (Spreitzer and Ogren 1983a) indicating that it may lack photosynthetic phosphorylation due to an atp-gene mutation (Selman-Reimer et al. 1981). A different photosynthetic phosphorylation locus (psp-u-2) was defined by mutant 12-5A (CC-2260) (Spreitzer and Ogren 1983a). This strain was recovered from a cross between the original 10-7C mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Selman-Reimer S, Merchant S, Selman BR (1981) Isolation and characterization of the chloroplast coupling factor 1 from wild-type Chlamydomonas reinhardi. In BR Selman, S Selman-Reimer, eds, Energy Coupling in Photosynthesis, pp 341-352

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 21gr mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 8-36C displayed uniparental inheritance, and lacks photosystem II activity (Spreitzer and Mets 1981). Recombination tests showed that 8-36C is at the same locus (pst-u-1) as 10-4, 11-1A, 11-3D, 11-4D, and 12-3C (Spreitzer and Ogren 1983a). Because the 8-36C mutation was shown to be linked with a mutation conferring herbicide resistance (Galloway and Mets 1984), subsequent studies confirmed that 8-36C results from a deletion in the psbA gene similar to that of FUD7 (CC-4147) (Bennoun et al 1986). This strain was recovered from a cross between the original 8-36C mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Locus: psbA

Bennoun P, Spierer-Herz M, Erickson J, Girard-Bascou J, Pierre Y, Delosme M, Rochaix JD (1986) Characterization of photosystem II mutants of Chlamydomonas reinhardii lacking the psbA gene. Plant Mol Biol 6:151-160

Galloway RE, Mets L (1984) Atrazine, bromacil, and diuron resistance in Chlamydomonas: A single non-mendelian genetic locus controls the structure of the thylakoid binding site. Plant Physiol 74:469-474

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• psbA
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 21gr mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 8-36C displayed uniparental inheritance, and lacks photosystem II activity (Spreitzer and Mets 1981). Recombination tests showed that 8-36C is at the same locus (pst-u-1) as 10-4, 11-1A, 11-3D, 11-4D, and 12-3C (Spreitzer and Ogren 1983a). Because the 8-36C mutation was shown to be linked with a mutation conferring herbicide resistance (Galloway and Mets 1984), subsequent studies confirmed that 8-36C results from a deletion in the psbA gene similar to that of FUD7 (CC-4147) (Bennoun et al 1986). This strain was recovered from a cross between the original 8-36C mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Locus: psbA

Bennoun P, Spierer-Herz M, Erickson J, Girard-Bascou J, Pierre Y, Delosme M, Rochaix JD (1986) Characterization of photosystem II mutants of Chlamydomonas reinhardii lacking the psbA gene. Plant Mol Biol 6:151-160

Galloway RE, Mets L (1984) Atrazine, bromacil, and diuron resistance in Chlamydomonas: A single non-mendelian genetic locus controls the structure of the thylakoid binding site. Plant Physiol 74:469-474

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• psbA
• Chromosome:
• chloroplast

From Robert J. Spreitzer, University of Nebraska, August 2014

Phenotype: requires acetate, sensitive to light

Following 5-fluorodeoxyuridine treatment and ethyl-methanesulfonate mutagenesis of wild-type 2137 mt+ cells, colonies were screened for light-sensitive, acetate-requiring phenotypes (Spreitzer and Mets 1981). Mutant 10-4 displayed uniparental inheritance, and lacks photosystem II activity (Spreitzer and Mets 1981). Recombination tests showed that 10-4 is at the same locus (pst-u-1) as 8-36C, 11-1A, 11-3D, 11-4D, and 12-3C (Spreitzer and Ogren 1983a). Because 8-36C, 11-1A, and 11-4D result from deletions in psbA (Bennoun et al 1986), 10-4 may also result from a psbA mutation. This strain was recovered from a cross between the original 10-4 mt+ and pf2 mt- (from Levine), and has been maintained with acetate medium in darkness since 1980 to prevent selection for revertants and non-light-sensitive suppressors (Spreitzer and Ogren 1983b).

Chromosome: chloroplast

Bennoun P, Spierer-Herz M, Erickson J, Girard-Bascou J, Pierre Y, Delosme M, Rochaix JD (1986) Characterization of photosystem II mutants of Chlamydomonas reinhardii lacking the psbA gene. Plant Mol Biol 6:151-160

Spreitzer RJ, Mets L (1981) Photosynthesis-deficient mutants of Chlamydomonas with associated light-sensitive phenotypes. Plant Physiol 67:565-569

Spreitzer RJ, Ogren WL (1983a) Rapid recovery of chloroplast mutations affecting ribulosebisphosphate carboxylase/oxygenase in Chlamydomonas reinhardtii. Proc Natl Acad Sci USA 80:6293-6297

Spreitzer RJ, Ogren WL (1983b) Nuclear suppressors of the photosensitivity associated with defective photosynthesis in Chlamydomonas reinhardtii. Plant Physiol 71:35-39

• Locus:
• Chromosome:
• chloroplast