Plant Physiology

Thompson, Robert John; Mosig, Gisela

doi: 10.1104/pp.76.1.1pmid: 16663777

Abstract We have looked for chloroplast genes whose expression is controlled by light by comparing the abundance of specific chloroplast transcripts in light-grown and dark-grown cells of Chlamydomonas reinhardtii. In addition, we have investigated whether genetic components influence expression of such genes. Northern blot analyses of specific separated transcripts showed that in certain strains (e.g. CC-278), several chloroplast transcripts from the Hpa II 5 region of the chloroplast chromosome were more abundant in dark-grown cells than in light-grown cells. The increased abundance of these Hpa II 5-specific transcripts was dependent on the genetic background: we have isolated mutants and we have found distantly related wild type strains which do not overaccumulate these transcripts in the dark. The strains which overaccumulated the Hpa II 5-specific transcripts, and only these strains, died after several cell divisions following transfer to the dark. Overaccumulation appears to be a necessary but not sufficient prerequisite for commitment to cell death in the dark. By analogy to bacterial systems, we speculate that a critical event associated with accumulation of one of these specific transcripts involves control at the level of transcription initiation or termination-antitermination. 1 Supported by National Cancer Institute Training Grant CA09385 and Cellular Molecular Biology Training Grant 2T32 07319 (R. J. T.), by National Institutes of Health Grant GM 13221-18 (G. M.), and by the Vanderbilt University Natural Science Committee Fund. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Beale, Samuel I.; Cornejo, Juan

doi: 10.1104/pp.76.1.7pmid: 16663825

Abstract Cell-free extracts of the unicellular red alga Cyanidium caldarium catalyze the transformation of biliverdin to a product indistinguishable from phycocyanobilin, the free bilin derived from phycocyanin by methanolysis. Crude cell-free extract requires biliverdin as the only substrate, but after removal of low molecular weight components by gel filtration, the reaction shows an additional requirement for a reduced pyridine nucleotide. Boiled extract is enzymically inactive, activity is not sedimented by high-speed centrifugation, and mesobiliverdin cannot serve as a substrate. Incubation of cell extracts with biliverdin yields two products with very similar spectrophotometric properties in acidic methanol, but which are separable by reverse-phase high pressure liquid chromatography. The same two products are formed by methanolysis of protein-bound phycocyanin chromophore, with the late-eluting one predominating. The two products derived from either phycocyanin methanolysis or cell extract incubation with biliverdin are partially interconvertible and they form the same ethylidine-free isomeric derivative, mesobiliverdin. Their absorption spectra correspond to those of the Z- and E-ethylidine isomers of phycocyanobilin. Based on previous work showing that the major methanolysis product has the E-ethylidine configuration, the other product of methanolysis and enzymic biliverdin transformation is therefore the Z-ethylidine isomer. The time course for formation of the two products during incubation suggests that the early-eluting product is the precursor of the late-eluting one. These results suggest that Z-ethylidine phycocyanobilin is the precursor of the E-ethylidine isomer, and that the latter may be a normal cellular precursor to protein-bound phycocyanin chromophore. 1 Supported by National Science Foundation Grant PCM-8213948, United States Department of Agriculture Grant 81-CRCR-1-0679, and National Institutes of Health Biomedical Research Support Grant 2-S07-RR07085-17. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Adeli, Khosrow; Allan-Wojtas, Paula; Altosaar, Illimar

doi: 10.1104/pp.76.1.16pmid: 16663789

Abstract The synthesis, transport, and posttranslational processing of reserve globulin in Avena sativa L. seeds were studied by pulse-chase labeling. Developing oat seeds were labeled with radioactive sulfate and tissue homogenates were used for globulin extraction. Two globulin precursors (58-62 kilodaltons) were labeled after 1 hour pulse. The α and β globulin subunits appeared between 2 and 10 hours later, while simultaneously the 58 to 62 kilodaltons polypeptides gradually disappeared. This confirmed a precursor-product relationship. In a second pulse-chase experiment, the tissue extracts were fractionated on a sucrose gradient. The major portion of radioactivity was initially (1 hour pulse) associated with the endoplasmic reticulum. However, a significant amount of radioactivity shifted from the endoplasmic reticulum to protein bodies after 20 hours chase, suggesting the transport of the newly synthesized proteins. Protein bodies isolated from pulse-chased seeds were analyzed for the arrival of the newly synthesized globulin. Labeled precursors were detected after 2 hours chase and gradually disappeared. The α and β subunits appeared during the same chase period and assembled into a 12S oligomer. The data indicated that oat globulin was synthesized as two large precursors which were transported from endoplasmic reticulum into protein bodies where they were processed to the α and β subunits forming a 12S oligomer. 1 Supported in part by Natural Science and Engineering Research Council grant A6711 and Agriculture Canada Research Contract No. 0SU81-00411. Contribution 583 from the Food Research Institute, Agriculture Canada, Ottawa. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Tobin, Alyson K.; Givan, Curtis V.

doi: 10.1104/pp.76.1.21pmid: 16663800

Abstract We have studied the effects of ATP and ADP on the oxidation of malate by coupled and uncoupled mitochondria prepared from etiolated hypocotyls of mung bean (Vigna radiata L.). In coupled mitochondria, ATP (1 millimolar) increased pyruvate production and decreased oxaloacetate formation without altering the rate of oxygen consumption. ATP also significantly decreased oxaloacetate production and increased pyruvate production in mitochondria that were uncoupled by carbonyl cyanide p-trifluoromethoxyphenyl hydrazone plus oligomycin. In coupled mitochondria, ADP (1 millimolar) increased the production of both pyruvate and oxaloacetate concomitantly with the acceleration of oxygen uptake to the state 3 rate. The effects of ADP were largely eliminated in uncoupled mitochondria. These results indicate that, whereas the ADP stimulation of oxaloacetate and pyruvate production in the coupled mitochondria is brought about primarily as the result of the accelerated rates of electron transport and NADH oxidation by the respiratory chain in state 3, ATP has significant regulatory effects independent of those that might be exerted by control of electron transport. 2 Present address: Biochemistry Department, University of Sussex, Falmer, Brighton, Sussex BN1 9QG England. 1 Supported by a postgraduate research studentship from the Science and Engineering Research Council (U.K.) to A. K. T. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Briskin, Donald P.; Poole, Ronald J.

doi: 10.1104/pp.76.1.26pmid: 16663811

Abstract The plasma membrane ATP-phosphohydrolase (ATPase) from red beet (Beta vulgaris L.) storage tissue was solubilized with the zwitterionic detergent Zwittergent 3-14 from a plasma membrane-enriched fraction which was extracted with the anionic detergent, sodium deoxycholate. For both the extraction of extraneous proteins by deoxycholate and the solubilization of active plasma membrane ATPase by Zwittergent 3-14, the optimal concentration of detergent was 0.1% (weight per volume) with a detergent to protein ratio of 1.0 (milligram per milligram). The properties of the solubilized ATPase were found to be similar to the membrane-bound enzyme with respect to pH optimum, substrate specificity, inhibitor sensitivity, and kinetics of K+ stimulation. The solubilized ATPase preparation formed a rapidly turning over phosphoenzyme, the breakdown velocity of which was increased in the presence of 50 millimolar KCl. Solubilization with 0.1% Zwittergent 3-14 following extraction with 0.1% deoxycholate resulted in an increase in both ATPase activity and steady state phosphoenzyme level; however, a direct correspondence between the increase in ATPase activity and phosphorylation level did not exist. It is proposed that this discrepancy may be the result of a detergent-mediated modification of kinetic rate constants in the mechanism of the enzyme. 2 Present Address: Plant Biochemistry and Bioregulation Laboratory, USDA-ARS, Utah State University, UMC 63, Logan, UT 84322. 1 Supported by the Natural Sciences and Engineering Research Council of Canada and the Department of Education of Quebec. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Tripepi, Robert R.; Mitchell, Cary A.

doi: 10.1104/pp.76.1.31pmid: 16663817

Abstract Flood tolerance of woody plants has been attributed to internal oxygen diffusion from shoot to root, metabolic adaptation within the root, or both. The purpose of this study was to compare several biochemical and physiological responses of birch roots to hypoxia in order to determine the nature of root metabolic adaptation to low oxygen tension. One-year-old seedlings of flood-tolerant river birch (Betula nigra L.) and flood-intolerant European birch (Betula pendula Roth) were transferred to solution culture, and the solutions were bubbled with air or nitrogen. After 18 days of hypoxia, total adenosine phosphate and ATP contents of river birch roots were 35% and 23% of controls, respectively, whereas those of European birch roots were 13% and 8%. Adenylate energy charge of river birch roots decreased between 6 and 12 days of hypoxia. In contrast, energy charge of European birch roots decreased after only 1 day of hypoxia. In vitro activity of cytochrome c oxidase and oxygen consumption capacity of excised roots from both birch species decreased under hypoxia. In vitro activity of alcohol dehydrogenase from roots of both species increased after 1 day of hypoxia. However, alcohol dehydrogenase activity from river birch roots increased 25-fold after 6 days of hypoxia, whereas that from European birch decreased back to control levels. Hypoxia decreased malate content of roots from both species. Metabolic adaptation within the root, rather than internal oxygen diffusion, appears to be responsible for the relative tolerance of river birch to hypoxia. 2 Present address: Department of Plant, Soil, and Entomological Sciences, University of Idaho, Moscow, ID 83843. 1 Supported in part by a grant to R. R. T. from the International Society of Arborists. Purdue Agricultural Experiment Station Journal paper number 9755. Project conducted by the senior author in partial fulfillment of the Ph.D. degree. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Cline, Gary R.; Reid, C. P. Patrick; Powell, Paul E.; Szaniszlo, Paul J.

doi: 10.1104/pp.76.1.36pmid: 16663818

Abstract When Fe was supplied at 100 micromolar in nutrient solution of pH 7.5, 10 and 1 micromolar levels of the siderophore desferrioxamine B (DFOB), a microbial iron transport compound, significantly (α = 0.05) enhanced growth and reduced chlorosis of an Fe-inefficient variety of sorghum (Sorghum bicolor L.). Although significantly adverse effects resulted when both Fe and desferrioxamine B (DFOB) were added at 100 micromolar as FeDFOB, the plants were relatively healthy when grown with 100 micromolar DFOB plus 200 micromolar Fe. It was concluded that sorghum absorbed Fe from the pool of nonchelated, solubilized Fe, and utilized DFOB as a shuttle agent, in equilibrium with this pool, to transport Fe from finely suspended solid phase Fe particles to the membrane of absorbing root cells. In contrast to sorghum, absorption of Fe by the Fe-efficient species sunflower (Helianthus annuus L.) was related to the level of FeDFOB and independent of the level of solubilized, nonchelated Fe. The latter was decreased whenever the concentration of DFOB was equal to or greater than the concentration of total Fe. For an Fe concentration of 10 micromolar, significantly larger and greener plants were obtained when DFOB was present at 1, 10, or 100 micromolar than in the absence of DFOB. When grown with 100 micromolar FeDFOB, sunflower plants appeared larger and less chlorotic than those supplied with 100 micromolar Fe and no DFOB. Sunflower apparently was able to utilize FeDFOB more directly than was sorghum. It is suggested that sunflower acquires Fe after binding FeDFOB at membrane sites and/or by producing sufficient reductants in the rhizosphere to reduce biologically significant levels of Fe(III)DFOB to the less stable Fe(II)DFOB. 2 Present address: Caesar Kleberg Wildlife Research Institute, College of Agriculture, Texas A & I University, Kingsville, TX 78363. 1 Supported by National Science Foundation grant DEB-79-11276. From a dissertation by the senior author submitted to the Academic Faculty of Colorado State University in partial fulfillment of the requirements for the Ph.D. degree. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Gumber, Subhash C.; Loewus, Mary W.; Loewus, Frank A.

doi: 10.1104/pp.76.1.40pmid: 16663819

Abstract myo-Inositol-1-phosphatase has been purified to homogeneity from Lilium longiflorum pollen using an alternative procedure which includes pH change and phenyl Sepharose column chromatography. Sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis shows that the enzyme is a dimer (subunit molecular weight, 29,000 daltons). The enzyme is stable at low pH values and is inactivated only below pH 3.0. In addition to 1l-and 1d-myo-inositol-1-phosphate, it shows high specificity for 1l-chiro-inositol-3-phosphate. As observed earlier with other primary phosphate esters, d-glucitol-6-phosphate and d-mannitol-6-phosphate are hydrolyzed very slowly. No activity is observed with inorganic pyrophosphate or myo-inositol pentaphosphate as substrate. The enzyme is inhibited by fluoride, sulfate, molybdate, and thiol-directed reagents. Partial protection against N-ethylmaleimide inhibition by substrate and Mg2+ together suggests sulfhydryl involvement at the active site. 1 Supported in part by Grant GM-22427 from the National Institute of Health, United States Public Health Service. Scientific Paper No. 6763, Project 0266, College of Agriculture Research Center, Washington State University, Pullman, WA 99164. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Turgeon, Robert

doi: 10.1104/pp.76.1.45pmid: 16663820

Abstract The sink-source conversion in developing leaves of tobacco (Nicotiana tabacum L.) was studied to determine whether import termination is caused by the onset of export or is related to achievement of positive carbon balance. Albino shoots were grown in vitro and grafted to detopped stems of green tobacco plants. Termination of import was studied by providing mature leaves of the stock plant with 14CO2 and detecting the presence of labeled nutrient in developing albino leaves by whole-leaf autoradiography. In albino leaves, import terminated progressively in the basipetal direction at the same stage of development as in leaves of green shoots. Starch was not present in the plastids of mesophyll cells of mature albino leaves but starch was synthesized when discs were cut from these leaves and incubated on 3 millimolar sucrose. Import ceased progressively in developing green leaves even when photosynthesis was prevented by darkening. It was concluded that cessation of import does not require achievement of positive carbon balance and is not the direct result of export initiation. To determine whether vein loading capacity develops in albino leaves, discs were cut from mature leaves and floated on [14C]sucrose solution. Uptake of label into the veins was detected by autoradiography and this uptake was sensitive to the phloem loading inhibitor p-chloromercuribenzenesulfonic acid. However, the amount of label taken up by veins in albino leaves was less than that taken up by veins of mature green leaves. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)

Kruger, Nicholas J.; Beevers, Harry

doi: 10.1104/pp.76.1.49pmid: 16663821

Abstract The cytoplasmic form of fructose 1,6-bisphosphatase (FBPase) was purified over 60-fold from germinating castor bean endosperm (Ricinus communis). The kinetic properties of the purified enzyme were studied. The preparation was specific for fructose 1,6-bisphosphate and exhibited optimum activity at pH 7.5. The affinity of the enzyme for fructose 1,6-bisphosphate was reduced by AMP, which was a mixed linear inhibitor. Fructose 2,6-bisphosphate also inhibited FBPase and induced a sigmoid response to fructose 1,6-bisphosphate. The effects of fructose 2,6-bisphosphate were enhanced by low levels of AMP. The latter two compounds interacted synergistically in inhibiting FBPase, and their interaction was enhanced by phosphate which, by itself, had little effect. The enzyme was also inhibited by ADP, ATP, UDP and, to a lesser extent, phosphoenolpyruvate. There was no apparent synergism between UDP, a mixed inhibitor, and fructose 2,6-bisphosphate. Similarly ADP, a predominantly competitive inhibitor, did not interact with fructose 2,6-bisphosphate. Possible roles for fructose 2,6-bisphosphate and the other effectors in regulating FBPase are discussed. 2 Present address: Department of Biology, Queen's University, Kingston, Ontario K7L 3N6 Canada. 1 Supported by Grant PCM-78-19575 from the United States National Science Foundation. This content is only available as a PDF. © 1984 American Society of Plant Biologists This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model)