A specific increase in chloroplast gene mutations following growth of Chlamydomonas in 5-fluorodeoxyuridine

Wurtz, Edwin A.; Sears, Barbara B.; Rabert, D K; Shepherd, Hurley S.; Gillham, Nicholas W.; Boynton, John E.

doi:10.1007/bf00267056

Cited by 57 publications

(22 citation statements)

References 45 publications

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“…Cultures were shaken under continuous cool white fluorescent light (1.5 X 104 lux) at 250C. Aliquots of these cultures containing 1 X 106 cells were plated on TAP plates (1.5% agar) containing 2 mM sodium arsenate for positive selection of nonphotosynthetic mutants (14,15). Part The publication costs of this article were defrayed in part by page charge payment.…”

Section: Methodsmentioning

confidence: 99%

“…2D) (27), arsenate selection (15) has proved unsuccessful in our hands. However, FdUrd, which decreases the amount of chloroplast DNA (28), increases the recovery of chloroplast mutations resistant to various antibiotics as well as nonphotosynthetic chloroplast mutations requiring acetate (14).…”

Section: Methodsmentioning

confidence: 99%

“…One of these causes the loss of chlorophyll-protein complex 1 (CP1) from the thylakoid membranes (10), the second alters the electrophoretic mobility of a different thylakoid membrane polypeptide (11), and the third appears to affect photophosphorylation (12,13). This paper describes the isolation and characterization of 16 nonphotosynthetic chloroplast mutations by using 5-fluorodeoxyuridine (FdUrd) which we have found increases specifically the frequency of all types of chloroplast gene mutations (14). These nonphotosynthetic mutants fall into nine distinct loci in the chloroplast genome on the basis of recombination analysis.…”

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confidence: 99%

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Mutations in nine chloroplast loci of Chlamydomonas affecting different photosynthetic functions.

Shepherd¹,

Boynton²,

Gillham³

1979

Proc. Natl. Acad. Sci. U.S.A.

100

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Mutations in nine chloroplast loci of Chlamydomonas affecting different photosynthetic functions.

Shepherd¹,

Boynton²,

Gillham³

1979

Proc. Natl. Acad. Sci. U.S.A.

100

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“…Independent cultures of wild-type mt+ cells, within each separate experiment, were grown in the dark and treated with 1 mm 5-fluorodeoxyuridine at 250C as described previously (10). This treatment reduces the ploidy of the chloroplast genome to about one copy and thus allows chloroplast mutants to be recovered (18). The cells from each culture were then concentrated to 2 x 107 cells/ml in 68 mm potassium phosphate buffer (pH 6.9), and mutagenized with 27 mm methvl methanesulfonate (Aldrich) for 1.5 h in the dark at 25°C.…”

Section: Methodsmentioning

confidence: 99%

Temperature-Sensitive, Photosynthesis-Deficient Mutants of Chlamydomonas reinhardtii

Spreitzer¹,

Al-Abed²,

Huether³

1988

Plant Physiol.

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Mutants of the unicellular, green alga Chlamydomonas reinhardtii were recovered by screening for the absence of photoautotrophic growth at 35°C. Whereas nonconditional mutants required acetate for growth at both 25 and 35°C, the conditional mutants have normal photoautotrophic growth at 25°C. The conditional mutants consisted of two classes: (a) Temperature-sensitive mutants died under all growth conditions at 35°C, but (b) temperature-sensitive, acetate-requiring mutants were capable of heterotrophic growth at 35°C when supplied with acetate in the dark. The majority of mutants within the latter of these two classes had defects in photosynthetic functions. These defects included altered pigmentation, reduced whole-chain electron-transport activity, reduced ribulosebisphosphate carboxylase activity, or pleiotropic alterations in a number of these photosynthetic components. Both nuclear and chloroplast mutants were identified, and a correlation between light-sensitive and photosynthesis-deficient phenotypes was observed.The unicellular green alga Chlanmydomtonas reinhardtii has proven to be a useful organism for the genetic dissection and analysis of photosynthesis in eukaryotes (16). Since C. reinhardtii maintains a functional photosynthetic apparatus when grown in the dark with acetate, photosynthesis-deficient mutants have been identified as light-sensitive, acetate-requiring strains (10). Genetic analysis can also be performed, and mendelian or uniparental (maternal) inheritance defines nuclear or chloroplast mutations, respectively (5).We have been particularly interested in using chloroplast mutants and their revertants to investigate the structure/function relationships of Rubisco& (13). As in higher plants, Rubisco consists of eight nuclear-encoded small subunits and eight chloroplast-encoded large subunits, and the large subunit appears to contain the active-site domain (7). However, of three Rubisco mutants defined at the level of the large-subunit gene sequence (3, 15), two were found to result from nonsense mutations. Calculating from the large-subunit gene sequence (2), one would expect only about 5% nonsense mutations out of the total number of point mutations that could alter the large-subunit protein sequence. It thus appeared that many mutants with amino-acid substitutions in the large-subunit protein were not being re-

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“…The frequency of recovery of uniparental mutants is increased by 5-fluorodeoxyuridine treatment (23). When the cultures reached stationary phase ( mating, zygote maturation, and germination were performed as described previously (16).…”

Section: Methodsmentioning

confidence: 99%

Nuclear Suppressors of the Photosensitivity Associated with Defective Photosynthesis in Chlamydomonas reinhardii

Spreitzer

Ogren²

1983

Plant Physiol.

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A specific increase in chloroplast gene mutations following growth of Chlamydomonas in 5-fluorodeoxyuridine

Cited by 57 publications

References 45 publications

Mutations in nine chloroplast loci of Chlamydomonas affecting different photosynthetic functions.

Mutations in nine chloroplast loci of Chlamydomonas affecting different photosynthetic functions.

Temperature-Sensitive, Photosynthesis-Deficient Mutants of Chlamydomonas reinhardtii

Nuclear Suppressors of the Photosensitivity Associated with Defective Photosynthesis in Chlamydomonas reinhardii

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