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C D Miles

Publications and source records attributed to C D Miles.

10 recordsLinked to original sources

Nuclear gene-regulated expression of chloroplast genes for coupling factor one in maize.

In order to gain a better understanding of the interaction between the chloroplast and nuclear genomes in controlling the expression of plastid genes and the biosynthesis of chloroplast proteins, maize (Zea mays) nuclear gene mutant hcf(*)-38, in which alpha and beta subunits of coupling factor one (CF(1)) are almost completely missing was studied. The mutant possesses all the other subunits of CF(1) but several peptides of photosystem II are present in reduced amounts. A competitive hybridization experiment showed the presence of the same plastid mRNA species in mutant and wild-type plants except for slightly lower levels of some transcripts in the mutant. Northern hybridization and dot blot hybridization experiments showed the features of transcripts for alpha and beta subunits of CF(1) in the mutant to be similar to those in the wild-type maize although their levels are somewhat lower in the mutant. In vivo and in organello protein labeling experiments with L-[(35)S]Met have shown that alpha and beta subunits of CF(1) are synthesized, assembled into CF(1), and probably associated with thylakoid membranes in mutant plants. It is concluded that they are subsequently degraded.

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Effect of nuclear mutation in maize on photosynthetic activity and content of chlorophyll-protein complexes.

A number of new nuclear mutants have been isolated from maize by selection for high chlorophyll (Chl) fluorescence. These mutants show reduced rates of photosynthesis and/or are deficient in Chl. Electrophoretic examination of wild type thylakoid membranes revealed five Chl-protein complexes, two containing only Chl a and three containing Chl a and Chl b. A class of nonviable, photosystem I-deficient mutants was found to be lacking one (A-1) of the two Chl a-protein complexes. A second class of nonviable, photosystem I-lacking mutants was found to be missing not only this A-1 complex but also one or more of the three Chl a and b-containing, light-harvesting Chl-protein complexes. Viable mutants were obtained which appeared to have lost just one of the Chl b-containing complexes, whereas a second class of viable mutants was missing all three of the Chl b-complexes. The results confirm that the A-1 band is associated with the P700-Chl a-protein complex characterized previously. The data also indicate the existence of structurally different forms of the light-harvesting Chl a- and b-containing complexes. The results also show a lower molecular weight band (A-2) containing primarily Chl a and which appears to be required for viability.

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Selection of diquat resistance photosynthesis mutants from maize.

Resistance of a seedling to the herbicide 1,1'-ethylene-2,2'-dipyridylium bromide (diquat) can be used as a selective technique for photosynthesis mutants in Zea mays L. Diquat requires reduction by the light reaction in order to kill leaf cells and, therefore, nonphotosynthetic mutants survive. This technique was tested using known mutants and is applicable to larger samples of plants than previous techniques. Resistance to diquat should allow selection of mutants on the oxidizing side of photosystem II which are not previously available in higher plants.

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Chloroplast Reactions of Photosynthetic Mutants in Zea mays.

Three seedling lethal mutants of Zea mays with impaired photosynthesis are described. These recessive mutants were selected on the basis of high chlorophyll fluorescence. They have normal chlorophyll pigmentation but are unable to fix CO(2) fully. Evidence is presented from fluorescence characteristics of isolated chloroplasts that both photosystem I and II mutants were isolated. Using conventional measures of photosynthetic electron transport, we suggest that the photosystem I mutant has limited ability to reduce NADP. The other two mutants are clearly blocked in photosystem II, one possibly lacking the primary electron acceptor.

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Inhibition of Photosystem II in Isolated Chloroplasts by Lead.

Inhibition of photosynthetic electron transport in isolated chloroplasts by lead salts has been demonstrated. Photosystem I activity, as measured by electron transfer from dichlorophenol indophenol to methylviologen, was not reduced by such treatment. However, photosystem II was inhibited by lead salts when electron flow was measured from water to methylviologen and Hill reaction or by chlorophyll fluorescence. Fluorescence induction curves indicated the primary site of inhibition was on the oxidizing side of photosystem II. That this site was between the primary electron donor of photosystem II and the site of water oxidation could be demonstrated by hydroxylamine restoration of normal fluorescence following lead inhibition.

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Acid-bath Phosphorylation in Heptane-extracted Chloroplasts.

When spinach (Spinacia oleracea) chloroplast fragments were lyophilized and extracted with n-heptane, acid-bath phosphorylation was significantly reduced. Phosphorylation could be restored to these chloroplasts, if the extracted material (containing quinones, carotenoids, and other lipids) was added back to dry preparation before rehydration. The results reported suggest it was not quinones which were required for restoration of acid-bath phosphorylation but lipids. It is suggested that the lipids which were extracted by n-heptane serves as structural role in the thylakoid membrane.

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The Differentiation of Pigmentation in Flower Parts, IV. Flavonoid Elaborating Enzymes From Petals of Impatiens balsamina s.

Extracts of the flower petals of Impatiens balsamina L. contain enzymes which catalyze the glycosylation of phenolic compounds. Enzymes have been extracted which glycosylate hydroquinone to arbutin and at least 3 different flavonols to the 3-monoglucoside. The hydroquinone glucosylating enzyme is similar to enzymes previously described except that it requires an unidentified low molecular weight cofactor. The glucosylation of flavonols follows normal enzyme kinetics; it requires a nucleotide diphosphate glucose donor for activity, and is made more evident by the presence of glucono-1:5-lactone, an inhibitor of endogenous glucosidases. It is suggested that the flavonol glucosylating enzyme acts naturally to glucosylate a precursor of both flavonols and anthocyanins to the 3-monoglucoside. The only elaboration of an anthocyanin observed with petal extracts was an acylation of pelargonidin-3-monoglucoside.

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