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Biomedical subjects

T C Fox

Publications and source records attributed to T C Fox.

18 recordsLinked to original sources

Mollusc/algal chloroplast symbiosis: how can isolated chloroplasts continue to function for months in the cytosol of a sea slug in the absence of an algal nucleus?

A marine sea slug, Elysia chlorotica, has acquired the ability to carry out photosynthesis as a result of forming an intracellular symbiotic association with chloroplasts of the chromophytic alga, Vaucheria litorea. The symbiont chloroplasts (kleptoplasts) are functional, i.e. they evolve oxygen and fix CO(2) and actively transcribe and translate proteins for several months in the sea slug cytosol. Considering the dependency of plastid function on nuclear genes, the level of kleptoplast activity observed in the animal cell is quite remarkable. Possible factors contributing to this long-lasting functional association that are considered here include: the presence of an algal nuclear genome in the sea slug, autonomous chloroplasts, unusual chloroplast/protein stability, re-directing of animal proteins to the kleptoplast, and lateral gene transfer. Based on our current understanding, the acquisition and incorporation of intact algal plastids by E. chlorotica is aided by the robustness of the plastids and the long-term functional activity of the kleptoplasts appears to be supported by both plastid and protein stability and contributions from the sea slug.

Journal Article↗

Mollusc-algal chloroplast endosymbiosis. Photosynthesis, thylakoid protein maintenance, and chloroplast gene expression continue for many months in the absence of the algal nucleus.

Early in its life cycle, the marine mollusc Elysia chlorotica Gould forms an intracellular endosymbiotic association with chloroplasts of the chromophytic alga Vaucheria litorea C. Agardh. As a result, the dark green sea slug can be sustained in culture solely by photoautotrophic CO(2) fixation for at least 9 months if provided with only light and a source of CO(2). Here we demonstrate that the sea slug symbiont chloroplasts maintain photosynthetic oxygen evolution and electron transport activity through photosystems I and II for several months in the absence of any external algal food supply. This activity is correlated to the maintenance of functional levels of chloroplast-encoded photosystem proteins, due in part at least to de novo protein synthesis of chloroplast proteins in the sea slug. Levels of at least one putative algal nuclear encoded protein, a light-harvesting complex protein homolog, were also maintained throughout the 9-month culture period. The chloroplast genome of V. litorea was found to be 119.1 kb, similar to that of other chromophytic algae. Southern analysis and polymerase chain reaction did not detect an algal nuclear genome in the slug, in agreement with earlier microscopic observations. Therefore, the maintenance of photosynthetic activity in the captured chloroplasts is regulated solely by the algal chloroplast and animal nuclear genomes.

Algal Proteins↗

Hydrogeologic modeling for permeable reactive barriers.

The permeable reactive barrier technology for in situ treatment of chlorinated solvents and other groundwater contaminants is becoming increasingly popular. Field scale implementation of this and other in situ technologies requires careful design based on the site-specific hydrogeology and contaminant plume characteristics. Groundwater flow modeling is an important tool in understanding the hydraulic behavior of the site and optimizing the reactive barrier design. A combination of groundwater flow modeling and particle tracking techniques was used to illustrate the effect of hydraulic conductivity of the aquifer and reactive media on key permeable barrier design parameters, such as the capture zone width, residence time, flow velocity, and discharge. Similar techniques were used to illustrate the modeling approach for design of different configurations of reactive barriers in homogeneous and heterogeneous settings.

Computer Simulation↗

Identification and gene expression of anaerobically induced enolase in Echinochloa phyllopogon and Echinochloa crus-pavonis.

Enolase (2-phospho-D-glycerate hydrolase, EC 4.2.1.11) has been identified as an anaerobic stress protein in Echinochloa oryzoides based on the homology of its internal amino acid sequence with those of enolases from other organisms, by immunological reactivity, and induction of catalytic activity during anaerobic stress. Enolase activity was induced 5-fold in anoxically treated seedlings of three flood-tolerant species (E. oryzoides, Echinochloa phyllopogon, and rice [Oryza sativa L.]) but not in the flood-intolerant species (Echinochloa crus-pavonis). A 540-bp fragment of the enolase gene was amplified by polymerase chain reaction from cDNAs of E. phyllopogon and maize (Zea mays L.) and used to estimate the number of enolase genes and to study the expression of enolase transcripts in E. phyllopogon, E. crus-pavonis, and maize. Southern blot analysis indicated that only one enolase gene is present in either E. phyllopogon or E. crus-pavonis. Three patterns of enolase gene expression were observed in the three species studied. In E. phyllopogon, enolase induction at both the mRNA and enzyme activity levels was sustained at all times with a further induction after 48 h of anoxia. In contrast, enolase was induced in hypoxically treated maize root tips only at the mRNA level. In E. crus-pavonis, enolase mRNA and enzyme activity were induced during hypoxia, but activity was only transiently elevated. These results suggest that enolase expression in maize and E. crus-pavonis during anoxia are similarly regulated at the transcriptional level but differ in posttranslational regulation, whereas enolase is fully induced in E. phyllopogon during anaerobiosis.

Amino Acid Sequence↗

Characterization of the association of nitrate reductase with barley (Hordeum vulgare L.) root membranes.

The nature of the association between nitrate reductase (NR) and membranes was examined. Nitrate reductase activity (NRA) associated with the microsomal fraction of barley (Hordeum vulgare L.) roots amounted to 0.6 to 0.8% of soluble NRA following sonication in the presence of 250 mM KI and repeated osmotic shock. This treatment removed all contaminating soluble NRA from microsomes of uninduced barley roots that had been homogenized in a soluble extract from roots of NO3(-)-induced plants. On continuous sucrose gradients, NRA co-migrated specifically with VO4(-)-sensitive ATPase activity, a plasma membrane (PM) marker; activity of glucose-6-phosphate dehydrogenase, assayed as cytosolic marker, co-migrated with NRA. Microsomal NRA was absent in barley deficient in soluble NR. Perturbation and trypsinolysis experiments with PM vesicles isolated by aqueous two-phase partitioning indicated that NR is associated with the periphery of the cytoplasmic face of the bilayer. These results demonstrate that PM and soluble NRs are essentially the same protein but that the membrane-associated form is tightly bound. Although it is possible that PM-associated NR exists in vivo, unequivocal evidence for this has yet to be shown. However, PM NR is definitely present in vitro.

Adenosine Triphosphatases↗

Anaerobic metabolism in plants.

Exposure to oxygen deficits is more widespread in biological systems than is commonly believed. Until recently, the general perception of anaerobic metabolism was often limited to the induction of alcoholic or lactic acid fermentation as the sole biochemical response to hypoxia/anoxia. Developments in the physiology, biochemistry, and molecular biology of anaerobic responses in invertebrates, lower plants, and higher plants have demonstrated that, depending upon the species, anaerobic metabolism may encompass much more than simple glycolytic metabolism. Here, recent progress in elucidating the mechanism(s) determining tolerance versus intolerance to anaerobic environments in higher plants is discussed, drawing most heavily on experimental systems using seeds or seedlings.

Journal Article↗

Survival of bovine embryos transferred to progesterone-treated asynchronous recipients.

Treatment of recipient cows with 100 mg of progesterone daily from Days 1 to 5 of the oestrous cycle increased plasma progesterone compared with vehicle-treated recipients. Embryo transfer to progesterone-treated recipients which showed oestrus 72 h after the donor cows resulted in pregnancy rates at Day 35 similar to those of synchronous (+/- 12 h) recipients (42 vs. 50%). Only 1 of 22 (4.8%) asynchronous (-72 h) vehicle-treated recipients established pregnancy. Similar treatments of cyclic cows with progesterone shortened (P less than 0.01) the interoestrous interval by 3.2 days. When assessed on Day 7 of pregnancy, administration of progesterone to superovulated donor cows on Days 1-4 of pregnancy did not affect early embryo development compared with superovulated cows treated with vehicle alone. Plasma progesterone increased rapidly in superovulated cows compared with cows during the oestrous cycle. The results indicate that administration of progesterone early in the oestrous cycle of the recipient can effectively advance uterine receptivity for the transfer of older asynchronous embryos.

Animals↗

Activities of isolated mitochondria and mitochondrial enzymes from aerobically and anaerobically germinated barnyard grass (echinochloa) seedlings.

Activity of mitochondria isolated from whole seedlings of Echinochloa crus-galli (L.) Beauv. var oryzicola germinated under aerobic and anaerobic conditions for 5 to 7 days was investigated. Mitochondria from both treatments exhibited good respiratory control and ADP/O ratios. Although O(2) uptake was low in anaerobic mitochondria, activity rapidly increased when the seedlings were transferred to air. Mitochondria from both aerobically and anaerobically grown seedlings of E. crus-galli var oryzicola maintained up to 66% of their initial respiration rate in the presence of both cyanide and salicylhydroxamic acid, and the inhibitory effects of cyanide and azide were additive. In addition, antimycin A was not an effective inhibitor of respiration. Reduced-minus-oxidized absorption spectra revealed that cytochromes a, a(3), and b were reduced to a greater extent and cytochrome c was reduced to a lesser extent in anaerobically germinated seedlings relative to that in aerobically germinated seedlings. An absorption maximum in the cytochrome d region of the spectrum was reduced to the same extent under both germination conditions and an absorption maximum at 577 nm was present only in anaerobically germinated seedlings. Anaerobically germinated seedlings contained 70% of the cytochrome c oxidase activity found in air grown seedlings. Upon exposure to air, the developmental pattern of this enzyme in anaerobically germinated seedlings was similar to air controls. Succinate dehydrogenase activity in anaerobic seedlings was only 45% of the activity found in aerobically germinated seeds, but within 1 hour of exposure to air, the activity had increased to control levels. The results suggest that mitochondria isolated from E. crus-galli var oryzicola differ from other plants studied and that the potential for mitochondrial function during anaerobiosis exists.

Journal Article↗

Osmotic Response of Sugar Beet Source Leaves at CO(2) Compensation Point.

As sugar beet source leaves lowered the CO(2) concentration to compensation point in a closed atmosphere, leaf thickness and relative water content decreased. Leaf water potential declined rapidly from -0.5 to -1.4 megapascals. At 340 microliters CO(2) per liter, water potential and sucrose, glucose, and fructose contents were steady in photosynthesizing source leaves. Within 90 minutes after leaves were exposed to a CO(2) concentration at the compensation point, leaf sucrose content declined to 60% of the preteatment level, rapidly in the first 30 minutes and then more slowly. During the subsequent 200 minutes, sucrose content increased to 180% of pretreatment level. Glucose and fructose remained unchanged during the treatment. Degradation of starch was sufficient to account for the additional sucrose that accumulated. Labeled carbon lost from starch appeared in sucrose and several other compounds that likely contributed to the recovery in leaf water content.

Journal Article↗

Developmental Changes in Photosynthetic Gas Exchange in the Polyol-Synthesizing Species, Apium graveolens L. (Celery).

Developmental changes in photosynthetic gas exchange were investigated in the mannitol synthesizing plant celery (Apium graveolens L. ;Giant Pascal'). Greenhouse-grown plants had unusually high photosynthetic rates for a C(3) plant, but consistent with field productivity data reported elsewhere for this plant. In most respects, celery exhibited typical C(3) photosynthetic characteristics; light saturation occurred at 600 micromoles photons per square meter per second, with a broad temperature optimum, peaking at 26 degrees C. At 2% O(2), photosynthesis was enhanced 15 to 25% compared to rates at 21% O(2). However, celery had low CO(2) compensation points, averaging 7 to 20 microliters per liter throughout the canopy. Conventional mechanisms for concentrating CO(2) were not detectable.

Journal Article↗

Effects of decreased net carbon exchange on carbohydrate metabolism in sugar beet source leaves.

The relationship between CO(2) concentration and starch synthesis and degradation was studied by measuring leaf starch content and disappearance of (14)C-starch. At a concentration of 340 microliters CO(2) per liter, starch accumulated without degradation of previously synthesized starch. Degradation of starch began when CO(2) concentration was lowered, but its synthesis continued. At 120 microliters CO(2) per liter rates of synthesis and degradation were equal. Even at the CO(2) compensation point, synthesis of starch continued. Concomitant starch synthesis and mobilization supported export from the leaf. Changes in starch metabolism that occur when photosynthesis is CO(2)-limited provide a means to study regulation of starch metabolism and carbon allocation in translocating leaves.

Journal Article↗

Sources of sucrose translocated from illuminated sugar beet source leaves.

A search for source leaf sucrose pools that differed in their relation to export was carried out in photosynthesizing leaves of Beta vulgaris L. The time course of depletion of [(14)C]sucrose in a leaf in unlabeled CO(2) following steady state labeling provided evidence for two distinct sucrose pools. After the start of the light period, leaf blade sucrose remained constant although it exchanged between the two pools. Newly synthesized sucrose destined for export passed through one pool more rapidly than through the other. All of the leaf blade sucrose appeared to exchange with export sucrose. Modeling and regression analysis of [(14)C]sucrose data provided a means for estimating the size of the two pools. From 20 to 40% of the sucrose was calculated to be present in the pool that provided the less direct path to export; this was likely vacuolar sucrose. The remainder of the sucrose in the blade was probably in the cytoplasm and veins. Added amounts of leaf blade sucrose, produced in response to elevated CO(2), appeared to be stored mainly in the vacuolar compartment.

Journal Article↗