Spectrophotometric Measurements of Phytochrome in vivo and Their Correlation with Photomorphogenic Responses of Phaseolus

Direct in vivo measurements of phytochrome have been made in Phaseolus vulgaris by 2-filter difference spectrophotometry (Ratiospect). All measurements were made at 730 versus 800 nm and it is assumed that the Δ (ΔOD) is directly proportional to the PFR concentration of phytochrome present. Dose response curves were determined for both physiological and spectrophotometric responses for red induction and far-red photoinactivation. For induction, saturation occurs at 100 mj/cm² and for inactivation at 30 mj/cm². The rate of hook opening and the physiological response measured 20 hours after induction are both shown to be directly proportional to the initial amount of PFR present spectrophotometrically. The sensitivity of the tissue correlates well with the absolute amount of phytochrome present, the inner portion of the hook having the maximum concentration of 0.042 Δ (ΔOD)/g fresh weight. If the total reversible phytochrome concentration is reduced by exposure to red light and allowing PFR to decay out of the system the remaining sensitivity of the tissue is shown to be directly correlated with the amount of PR remaining in the tissue. PFR disappears rapidly in the dark at 25°, and is not detectable after 6 hours. There is no indication that PFR reverts in the system to PR. At 4°, PFR does not disappear measurably up to 1 hour and is nearly totally reversible to PR.     

Fine Structure of Thiobacillus thiooxidans

Mahoney, Robert P. (Skidmore College, Saratoga Springs, N.Y.), and Mercedes R. Edwards. Fine structure of Thiobacillus thiooxidans. J. Bacteriol. 92: 487-495. 1966.-Thin section analysis of the chemosynthetic autotroph Thiobacillus thiooxidans revealed structures comparable to gram-negative heterotrophic bacteria. Although this species is unique in that it oxidizes elemental sulfur for energy, uses carbon dioxide as its sole source of carbon, and can withstand a pH of less than 1, thin sections revealed a profile of the cell envelope (cell wall and plasmalemma) similar to other gram-negative species which have more common physiological traits. The cell wall is composed of five layers with an overall width of approximately 200 A, and the plasmalemma appears as a conventional "unit membrane" with a width of about 85 A. Volutin granules and less-dense bodies of similar shape and size were frequently observed in close association with the nucleoplasm. The nature and function of these bodies are unknown at this time.     

A quantitative comparison of dual control of a hormone response element by progestins and glucocorticoids in the same cell line

Progesterone receptor-containing T47D human breast cancer cells are responsive to progestins but fail to respond to other steroid hormones, in particular dexamethasone, because they have no measurable levels of receptors for estrogens, androgens, or glucocorticoids. To quantitatively study dual responsiveness of the mouse mammary tumor virus (MMTV) promoter to progestins and glucocorticoids, we have stably transfected T47D cells with a glucocorticoid receptor (GR) expression vector. A cloned derivative (A1-2) was isolated that expresses a normal, full length GR, as assessed by steroid binding and Western immunoblot with a monoclonal anti-GR antibody. Moreover, GR is expressed at levels (80,000-100,000 molecules per cell) comparable to the high levels of endogenous progesterone receptor (200,000 molecules per cell). In A1-2 cells transiently transfected with an MMTV-chloramphenicol acetyl transferase reporter gene, induction by glucocorticoid was substantially greater (5-fold) than induction mediated by progestins. These results suggest that glucocorticoids may be the primary regulator of MMTV.     

Coordinate regulation of 3-hydroxy-3-methylglutaryl-coenzyme A synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase, and prenyltransferase synthesis but not degradation in HepG2 cells

Human hepatoma HepG2 cells were used to demonstrate coordinate regulation of three enzymes of cholesterol synthesis under a variety of conditions. Addition of either delipidized serum and mevinolin or low density lipoprotein, 25-hydroxycholesterol, or mevalonic acid to HepG2 cells resulted in rapid changes both in the levels of the mRNAs and in the rates of synthesis of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) synthase, HMG-CoA reductase, and farnesyl pyrophosphate synthetase (prenyltranferase). In all cases, the changes in mRNA levels were paralleled by changes in the rates of specific protein synthesis. Pulse-chase techniques were used to determine the half-lives of all three proteins. Addition of low density lipoprotein to the media during the chase increased the rate of degradation of HMG-CoA reductase 4.6-fold but had no affect on the half-lives of HMG-CoA synthase or prenyltransferase. Therefore, we conclude that the coordinate regulation of these three enzymes under a variety of conditions occurs at the level of enzyme synthesis and not at the level of protein stability.     

Spatial Analysis of the Canopy of Kiwifruit Vines as it Relates to the Physical, Chemical and Postharvest Attributes of the Fruit

The position of individual fruit on kiwifruit vines (Actinidiadeliciosa var. deliciosa) grown on a horizontal trellis (pergola)and on a T-bar trellis was determined using a theodolite. Thephysical, chemical, and postharvest attributes of the fruitwere related to their position on the vine during development. Fruit from the pergola vines were more numerous, of lesser weight,with lower concentrations of most mineral nutrients, but greaterconcentrations of soluble solids, and similar flesh firmnessafter 12 weeks of storage at 0 °C, than fruit from the T-barvines. The position on the vine accounted for most of the variationin the attributes of the fruit. Differences between fruit ona single lateral accounted for 43-56% of the variation. Variationbetween vines was relatively small (< 4% of the total variance). The heavier fruit were located at the apical ends of the laterals,while greater concentrations of soluble solids were associatedwith fruit located closer to the cordon. The larger fruit fromthe pergola vines developed from the early opening flowers.A similar relationship existed initially for the T-bar vines,but a reduction in growth of fruit from the early opening flowers8 weeks after anthesis resulted in a more even distributionof fruit size at harvest. The strongest relationship between mineral composition and postharvestattributes of the fruit was with soluble solids concentration(29-46% of the variance). The relationship with flesh firmnesswas weak (r = -0·14 to -0·32). Individual elementscould not be considered in isolation but rather in groups ofelements. Nitrogen was grouped strongly with phosphorus, sulphur,potassium, and copper, while calcium was linked with a secondgroup which included manganese and zinc. These two groups werenegatively related to one another. The greatest proportion of fruit with superior characteristicswas located in the denser parts of the canopy. Fruit with lessdesirable attributes were from the extremities of the canopywhere the leaf area index was low.Copyright 1994, 1999 AcademicPress Actinidia deliciosa, kiwifruit, fruit position, fruit quality, within-vine variation     

Role of proteinaceous amino acids released in root exudates in nutrient acquisition from the rhizosphere

The role of proteinaceous amino acids in rhizosphere nutrient mobilization was assessed both experimentally and theoretically. The degree of adsorption onto the soil's solid phase was dependent on both the amino acid species and on soil properties. On addition of amino acids to both soil and freshly precipitated Fe(OH)₃, no detectable mobilization of nutrients (K, Na, Ca, Mg, Cu, Mn, Zn, Fe, S, P, Si and Al) was observed, indicating a very low complexation ability of the acidic, neutral and basic amino acids. This was supported by results from a solution equilibria computer model which also predicted low levels of amino acid complexation with solutes present in the soil solution. On comparison with the Fe(OH)₃ and equilibria data obtained for the organic acid, citrate, it was concluded that amino acids released into the rhizosphere have a limited role in the direct acquisition of nutrients by plants. The effectiveness of root exudates such as amino acids, phytosiderophores and organic acids in nutrient mobilization from the rhizosphere is discussed with reference to rhizosphere diffusion distances, microbial degradation, rate of complexation and the root's capacity to recapture exudate-metal complexes from the soil.