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1.
Planta ; 185(1): 97-104, 1991 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-24186285

RESUMO

If Norflurazon-treated mustard (Sinapis alba L.) seedlings are grown in low-fluence-rate white light, accumulation of carotenoids is completely inhibited, while levels of chlorophyll (Chl) a and b are comparable to those of control seedlings. Measurements of fluorescence yield and oxygen evolution indicate that carotenoid-free, green cotyledons are unable to perform leephotosynthesis in vivo. When thylakoid membranes were prepared and electron transport was measured in vitro, only PSI but not PSII activity was detected. Solubilization of the photosystems from thylakoid membranes and separation by sucrose-gradient centrifugation confirmed that PSII is absent in carotenoid-free seedlings, while PSI is present. Western blot analysis for representative proteins of the four photosynthetic complexes showed that subunits 1 and 2 of PSI, the Rieske-iron sulfur-protein, the α-subunit of the CF1 moiety of the ATP-synthase complex, cytochrome b 559 and the lumenal 33-kDa protein of the water-splitting apparatus of PSII are present in comparable amounts in Norflurazon-treated and control plants, while the amounts of Chl-binding proteins of PSII (the major light-harvesting Chl-a/b-binding protein of the antenna complex and the 51- and 44-kDa Chl-a-binding proteins) and two components of the PSII reaction center, (the D1 and D2 protein) are substantially reduced. The data indicate that accumulation of PSII polypeptides is either not inhibited or not completely inhibited in carotenoid-free mustard seedlings, but that assembly of a functional PSII complex does not occur. If Norflurazon-treated seedlings are transferred to water, lutein accumulates rapidly and reaches about 80% of the level detectable in control plants, while the level of other carotenoids is still less than 1%. The accumulation kinetics for lutein are similar to the kinetics for the appearance of PSII activity. This indicates that the availibility of lutein rather than that of other carotenoids might be rate-limiting for the appearance of PSII activity.

2.
Plant Physiol ; 96(3): 971-9, 1991 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16668283

RESUMO

In concurrence with earlier results, the following enzymes showed latency in intact spinach (Spinacia oleracea L.) leaf peroxisomes: malate dehydrogenase (89%), hydroxypyruvate reductase (85%), serine glyoxylate aminotransferase (75%), glutamate glyoxylate aminotransferase (41%), and catalase (70%). In contrast, glycolate oxidase was not latent. Aging of peroxisomes for several hours resulted in a reduction in latency accompanied by a partial solubilization of the above mentioned enzymes. The extent of enzyme solubilization was different, being highest with glutamate glyoxylate aminotransferase and lowest with malate dehydrogenase. Osmotic shock resulted in only a partial reduction of enzyme latency. Electron microscopy revealed that the osmotically shocked peroxisomes remained compact, with smaller particle size and pleomorphic morphology but without a continuous boundary membrane. Neither in intact nor in osmotically shocked peroxisomes was a lag phase observed in the formation of glycerate upon the addition of glycolate, serine, malate, and NAD. Apparently, the intermediates, glyoxylate, hydroxypyruvate, and NADH, were confined within the peroxisomal matrix in such a way that they did not readily leak out into the surrounding medium. We conclude that the observed compartmentation of peroxisomal metabolism is not due to the peroxisomal boundary membrane as a permeability barrier, but is a function of the structural arrangement of enzymes in the peroxisomal matrix allowing metabolite channeling.

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