Diurnal fluctuations in cotton leaf carbon export, carbohydrate content, and sucrose synthesizing enzymes

In fully expanded leaves of greenhouse-grown cotton (Gossypium hirsutum L., cv Coker 100) plants, carbon export, starch accumulation rate, and carbon exchange rate exhibited different behavior during the light period. Starch accumulation rates were relatively constant during the light period, whereas carbon export rate was greater in the afternoon than in the morning even though the carbon exchange rate peaked about noon. Sucrose levels increased throughout the light period and dropped sharply with the onset of darkness; hexose levels were relatively constant except for a slight peak in the early morning. Sucrose synthase, usually thought to be a degradative enzyme, was found in unusually high activities in cotton leaf. Both sucrose synthase and sucrose phosphate synthetase activities were found to fluctuate diurnally in cotton leaves but with different rhythms. Diurnal fluctuations in the rate of sucrose export were generally aligned with sucrose phosphate synthase activity during the light period but not with sucrose synthase activity; neither enzyme activity correlated with carbon export during the dark. Cotton leaf sucrose phosphate synthase activity was sufficient to account for the observed carbon export rates; there is no need to invoke sucrose synthase as a synthetic enzyme in mature cotton leaves. During the dark a significant correlation was found between starch degradation rate and leaf carbon export. These results indicate that carbon partitioning in cotton leaf is somewhat independent of the carbon exchange rate and that leaf carbon export rate may be linked to sucrose formation and content during the light period and to starch breakdown in the dark.     

Isolation of a Bacillus stearothermophilus mutant exhibiting increased thermostability in its restriction endonuclease.

A procedure was developed for the selection of spontaneous mutants of Bacillus stearothermophilus NUB31 that are more efficient than the wild type in the restriction of phage at elevated temperatures. Inactivation studies revealed that two mutants contained a more thermostable restriction enzyme and one mutant contained three times more enzyme than the wild type. The restriction endonucleases from the wild type and one of the mutants were purified to apparent homogeneity. The mutant enzyme was more thermostable than the wild-type enzyme. The subunit molecular weight, amino acid composition, N-terminal and C-terminal amino acid residues, tryptic peptide map, and catalytic properties of the two enzymes were determined. The two enzymes have similar catalytic properties, but the molecular size of the mutant enzyme is approximately 6 to 7 kilodaltons larger than that of the wild-type enzyme. The mutant enzyme contains 54 additional amino acid residues, of which 26 to 28 are aspartate/asparagine, 8 to 15 are glutamate/glutamine, and 8 to 9 are tyrosine residues. The two enzymes contained similar amounts of the other amino acids, identical N-terminal residues, and different C-terminal residues. Tryptic peptide analyses revealed a high degree of homology between the two enzymes. The increased thermostability observed in the mutant enzyme appears to have been achieved by a mutation that resulted in the addition of amino acid residues to the wild-type enzyme. A number of mechanisms are discussed that could account for the observed difference between the mutant and wild-type enzymes.     

Lymphatic endothelial celline (CH3) from a recurrent retroperitoneal lymphangioma

Summary An endothelial cell line derived from a massive recurrent chyle-containing retroperitoneal lymphangioma was isolated in monolayer culture. Scanning and transmission electron microscopy and immunohistochemistry confirmed a close resemblance to blood vascular endothelium with typical cobblestone morphology, positive immunofluorescence staining for endothelial marker Factor VIII-associated antigen and fibronectin, and prominent Weibel-Palade bodies. The endothelial cells also exhibited other ultrastructural features characteristic of lymphatic endothelium, including sparse microvillous surface projections, overlapping intercellular junctions, and abundant intermediate filaments. This endothelial cell line represents a new source of proliferating lymphatic endothelium for future study, including structural and functional comparison to blood vascular endothelium. Supported in part by Arizona Disease Control Research Commission contracts 8277-000000-1-1-AT-6625 and ZB-7492. Presented in part at the 10th International Congress of Lymphology in Adelaide, Australia, August 1985.     

Genethics: project accountability via evaluation of teacher and student growth.

Accountability through demonstrated learning is increasingly being demanded by agencies funding science education projects. For example, the National Science Foundation requires evidence of the educational impact of programs designed to increase the scientific understanding and competencies of teachers and their students. The purpose of this paper is to share our human genetics educational experiences and accountability model with colleagues interested in serving the genetics educational needs of in-service secondary school science teachers and their students. Our accountability model is facilitated through (1) identifying the educational needs of the population of teachers to be served, (2) articulating goals and measurable objectives to meet these needs, and (3) then designing and implementing pretest/posttest questions to measure whether the objectives have been achieved. Comparison of entry and exit levels of performance on a 50-item test showed that teacher-participants learned a statistically significant amount of genetics content in our NSF-funded workshops. Teachers, in turn, administered a 25-item pretest/posttest to their secondary school students, and collective data from 121 classrooms across the United States revealed statistically significant increases in student knowledge of genetics content. Methods describing our attempts to evaluate teachers' use of pedagogical techniques and bioethical decision-making skills are briefly addressed.     

Antioxidants in sun and shade leaves of sour orange trees (Citrus aurantium) after long-term acclimation to elevated CO2

Antioxidative systems and the contents of pigments, malondialdehyde,soluble protein, and carbohydrate were investigated in sun-and shade-acclimated leaves of sour orange (Citrus aurantium)trees that had been grown for 7.5 years under ambient and elevated(+300 µmol mol^–1) atmospheric CO₂ concentrations.Sunacclimated leaves contained higher ascorbate, glutathioneand soluble carbohydrate contents and higher catalase activitiesthan shade-acclimated leaves. The activities of superoxide dismutases,which belonged to the family of Cu/Zn-isozymes, were similarin sunand shade-acclimated leaves and decreased in responseto enhanced CO₂. In shade-acclimated leaves, none of the otherparameters studied was affected by elevated CO₂. In sun-acclimatedleaves elevated CO₂ caused increases in carbohydrate and ascorbatecontents. There was no evidence for enhanced lipid peroxidationas assessed from the determination of the malondialdehyde contentsunder either conditions. Key words: Ascorbate, catalase, CO₂ enrichment, global change, glutathione, superoxide dismutase     

Plant growth analysis: an evaluation of experimental design and computational methods

Journal of Experimental Botany, Vol. 47, No. 302, pp. 1343–1351,September 1996 On page 1345, equation (2) should have read:      

Cell growth patterns and lens geometry: a quantitative study from three-dimensional reconstructions

The growth of the lens of the sea lamprey, Petromyzon marinus, was studied over the 5 years of larval development. Whole lenses (25) and Golgi-impregnated cells (393) were reconstructed with computer-assisted microscopy. Several cellular geometric parameters (length, width, curvature, surface, volume, shape) were correlated with the position of the cell's base on the lens capsular perimeter. Based on these correlations, the cells formed four groups that correspond to the central anterior, germinative, transitional and cortical fiber zones. A fifth zone, containing nuclear fiber cells, never stained. Lens growth is exponential during the 5 years. The anterior epithelium increases in size and in cell number by cell growth and division. The posterior mass increases in cell number by recruitment and increases in size by cell growth. A model is proposed to account for the size and shape of the lens based upon the coupling of anterior and posterior growth patterns. Four zonal boundaries are defined by changes in cell growth patterns. With growth, cells are subsumed into adjacent zones and zonal boundaries move away from the lens center. We find no support for the suggestion that cells migrate centrally.     

Polarity of meiotic gene conversion is 5′ to 3′ within the niaD gene of Aspergillus nidulans

We have examined polarity of meiotic gene conversion in the niiA-niaD gene cluster of Aspergillus nidulans in two-point crosses. The type and position of the mutations represented by the niaD alleles and the correlation between the relative frequency of gene conversion and the physical position of these mutations were determined. We show that polarity of meiotic gene conversion is 5 to 3 (transcribed strand) within the niaD gene. Additional crosses involving a niiA allele and a niaD allele show little polarity of gene conversion, which suggests that the recombination events leading to restoration of the niaD gene are initiated upstream of the coding region of the niaD gene but within the niiA-niaD gene cluster, possibly within the intergenic promoter region.     

Collagen in the egg shell membranes of the hen

Collagen-like proteins have been found in the egg shell membranes of the hen. Materials similar to types I and V collagens were detected in each of the two layers of this membrane, the thick outer membrane and the thin inner membrane. Collagen was extracted by acid-pepsin digestion and isolated by differential salt precipitation. Identification of type-specific collagen-like material was established by coelectrophoresis on SDS-polyacrylamide gels using known collagen standards. These bands were susceptible to digestion by bacterial collagenase. From differential staining of the gels it was estimated that the ratio of collagen types I:V was approximately 100:1. Further confirmation of these biochemical results was obtained with immunofluorescence microscopy using type-specific antisera against chicken types I and V collagen with the indirect sandwich technique. Both the inner and outer shell membranes contained the two types of collagen. Within each membrane, the large, coarse 2.5-micron fibers contained predominantly type I collagen-like material, while type V collagen was mainly associated with the delicate narrower fibers of approximately 0.6-micron diameter. These tended to be concentrated in the inner membrane. At the electron microscopic level, both types of fibers were coated with glycoproteins that stained positively with ruthenium red. The deposition of these collagen-like substances by the hen oviduct on to the surface of the developing egg is an additional example of interstitial-type collagen synthesis and secretion by epithelial rather than by mesenchymal cells.