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J E Varner

Publications and source records attributed to J E Varner.

At least 73 records · Page 4Linked to original sources
PubMed1971

Hormone-controlled synthesis of endoplasmic reticulum in barley aleurone cells.

The rate of synthesis of the endoplasmic reticulum in barley aleurone cells after treatment with gibberellic acid was determined by measurement of [(14)C]-choline incorporation into acid-insoluble material in a semipurified fraction containing the endoplasmic reticulum. 94% of the (14)C incorporated into this fraction is extractable by lipid solvents and only 9% is removed by procedures for nucleic acid extraction. Gibberellic acid increases the rate of synthesis of the endoplasmic reticulum 4- to 8-fold, starting about 4 hr after addition of the hormone (at about the same time as polysome formation). Abscisic acid inhibits this gibberellic acid-enhanced increase in the rate of synthesis of endoplasmic reticulum.

Carbon Isotopes↗
PubMed1971

Intact tissue assay for nitrite reductase in barley aleurone layers.

A method has been devised for the detection and measurement of nitrite reductase activity in intact barley (Hordeum vulgare L. cv. Himalaya) aleurone layers. The technique involves feeding aleurone layers nitrite and measuring nitrite disappearance after a given time period. The method also allows simultaneous determination of nitrite uptake by the tissue. Quantitative recovery of nitrite is obtained by rapid heating of tissue in the presence of dimethyl sulfoxide.Using the procedure described, nitrite reductase activity in intact barley aleurone layers was determined. Enzyme activity was increased by prior incubation of the tissue with nitrate, but considerable activity was present in tissue incubated without nitrate. Nitrate-induced activity was inhibited by cycloheximide but not by actinomycin D. Enzyme activity in induced layers was inhibited by 2,4-dinitrophenol, and partially by antimycin A and 2-n-heptyl-4-hydroxyquinoline N-oxide. Activity in noninduced tissue appeared to be less sensitive to these respiratory inhibitors. In contrast, both activities were inhibited more than 90% by anaerobiosis; but nitrate-induced and noninduced aleurone layers were able to reduce nitrite anaerobically when the concentration of substrate in the assay medium was reduced from 250 mum to 25 mum. Nitrite uptake was relatively insensitive to anaerobiosis and to the inhibitors tested.Nitrite depletion from the medium by aleurone layers was rapid at pH 4.5 and negligible at pH 7.5. Nitrite accumulated at pH 4.5 under anaerobic conditions was rapidly released when the tissue was transferred to medium at pH 7.5. Nitrite release at pH 7.5 occurred whether the tissue was maintained under anaerobic or aerobic conditions.

Journal Article↗
PubMed1970

Control of nitrate reductase activity in barley aleurone layers.

Nitrate reductase activity in barley (Hordeum vulgare L. cv. Himalaya) aleurone layers has been determined in the intact tissue, using two different methods. The first method measures the rate of appearance of H(2) (18)O produced during the reduction of KN(18)O(3). The second assay measures excreted nitrite resulting from nitrate reduction under anaerobic conditions. Nitrite production in this anaerobic, intact-tissue assay was dependent upon the presence of phosphate (pH 7.5) and was increased by ethanol and bisulfite.After ten hours of nitrate induction, nitrate reductase activities measured by the KN(18)O(3) assay are one-sixth, and those measured by the anaerobic intact-tissue assay are one-third, of those observed in cell-free extracts of aleurone layers. Addition of ethanol to the anaerobic intact-tissue medium increased the rate of nitrate reduction to a level greater than that found in the cell-free assay.Oxygen inhibited nitrite release in the anaerobic intact-tissue assay. However, under aerobic conditions and in the presence of 2-heptyl-4-hydroxyquinoline-N-oxide or antimycin A, nitrate reduction increased to rates comparable to those observed under anaerobiosis. Neither of these electron transport inhibitors affected anaerobic nitrate reduction, though they were effective in inhibiting oxygen uptake in separate experiments.

Journal Article↗
PubMed1970

Deuterium oxide as a density label of peroxidases in germinating barley embryos.

Density labeling with deuterium oxide, gel electrophoresis, and isopycnic equilibrium sedimentation were used to study the appearance and disappearance of individual peroxidases in the embryos of germinating barley. No detectable label was incorporated into those peroxidases which are present in the embryo of the dry seed and disappear during germination. Deuterium was incorporated into the additional peroxidases which appeared in the embryo during germination. This incorporation is not due to deuterium-hydrogen exchange into preformed proteins. The results indicate that the newly appearing peroxidases arise by synthesis during germination.

Journal Article↗
PubMed1970

Characterization of rapidly labeled ribonucleic acid from dwarf peas.

The ribonucleic acid synthesized by excised shoots of dwarf pea (Pisum sativum L. cv. Progress No. 9) during short labeling periods has been characterized. Thirty percent of the total (32)P(i) incorporated in 1 hour is found in the ribosomal fraction. This labeled RNA was polydisperse (6-18 Svedberg units) and after chromatography on a methylated albumin-kieselguhr column about 80% of the radioactivity appeared in two peaks. One of these appeared on the shoulder of heavy ribosomal RNA ("mRNA") while the other was tenaciously bound to the column (TB-RNA). In the presence of high NaCl concentration, about half of the polydisperse RNA interacted with ribosomal RNA and eluted as "mRNA" while the remainder eluted as TB-RNA. This interaction in the presence of salt seems to result in the alteration of secondary structure because the "mRNA" fraction had a high sedimentation coefficient (45-50 Svedberg units). The polydisperse RNA approaches DNA in low cytidylate and guanylate content. After short periods of labeling TB-RNA showed higher adenylate content than "mRNA." The radioactivity from the "mRNA" peak can be chased, and these counts may represent a class of shortlived messenger RNA molecules with an average half-life of 10 to 15 minutes. The other component, TB-RNA, could not be chased and accumulated radioactivity during the chase period.

Centrifugation, Density Gradient↗
PubMed1970

Multiple forms of amylase induced by gibberellic acid in isolated barley aleurone layers.

The addition of gibberellic acid to isolated aleurone layers of barley (Hordeum vulgare L.) causes the production and secretion of four alpha-amylases. Two of these are stable at pH 3.7 and are not inactivated by ethylenediaminetetraacetate. The other two represent the classical barley alpha-amylases; i.e., they are inactivated at pH 3.7 and by reagents which from complexes with divalent metal ions. All four forms are synthesized de novo in response to the addition of gibberellic acid.

Amylases↗
PubMed1970

Respiration and Protein Synthesis in Dormant and After-ripened Seeds of Avena fatua.

Dormant seeds of Avena fatua, which do not germinate when allowed to imbibe water, have a respiration rate only about 20% less than that of imbibed nondormant (after-ripened) seeds in the period before actual germination and are capable of synthesizing protein at a rate comparable to that of the nondormant seeds. An increase of protein synthesis is observed in nondormant seeds at the beginning of root protrusion. Autoradiography of seeds administered (3)H-leucine shows that protein synthesis occurs in the axis part of the embryo, the scutellum, the coleorhiza, and the aleurone layer. Dormancy in seeds is not a state of general inactivity; rather, it must be due to some specific metabolic block.

Journal Article↗
PubMed1970

Synthesis of Cellulase during Abscission of Phaseolus vulgaris Leaf Explants.

When abscission in leaf explants from Phaseolus vulgaris, cultivar Red Kidney, was allowed to proceed while the explants were in (2)H(2)O, a 1.25% increase in the buoyant density of cellulase in a cesium chloride gradient was observed. These data indicate that the increase in cellulase activity during abscission is a result of the synthesis of new protein. Two differentially soluble forms of cellulase are present in the abscission zone. The form which is soluble only in a high salt buffer seems more closely related to the abscission process than the form which is soluble in dilute buffer. The correlation between changes in pull force and increase in cellulase activity and the effects of several hormones on cellulase activity are discussed.

Journal Article↗
PubMed1969

Substrate induction of nitrate reductase in barley aleurone layers.

Nitrate induces the formation of nitrate reductase activity in barley (Hordeum vulgare L. cv. Himalaya) aleurone layers. Previous work has demonstrated de novo synthesis of alpha-amylase by gibberellic acid in the same tissue. The increase in nitrate reductase activity is inhibited by cycloheximide and 6-methylpurine, but not by actinomycin D. Nitrate does not induce alpha-amylase synthesis, and it has no effect on the gibberellic acid-induced synthesis of alpha-amylase. Also, there is little or no direct effect of gibberellic acid (during the first 6 hr of induction) or of abscisic acid on the nitrate-induced formation of nitrate reductase. Gibberellic acid does interfere with nitrate reductase activity during long-term experiments (greater than 6 hr). However, the time course of this inhibition suggests that the inhibition may be a secondary one. Barley aleurone layers therefore provide a convenient tissue for the study of both substrate- and hormone-induced enzyme formation.

Journal Article↗
PubMed1969

Metabolism of C-Maltose in Avena fatua Seeds During Germination.

Non-dormant and dormant seeds of Avena fatua metabolize (14)C-maltose in different ways: in non-dormant seeds, (14)C-maltose administered to the endosperm is readily converted to sucrose in the scutellum and translocated to the embryo; in dormant seeds, little sucrose is synthesized from (14)C-maltose, and maltose and glucose tend to accumulate in the endosperm. It is suggested that biosynthesis of sucrose is essential for effective transport of the endosperm reserve to the embryonic axis in germinating seeds.

Journal Article↗
PubMed1969

Enzymic degradiation of starch granules in the cotyledons of germinating peas.

Starch, total alpha- and beta-amylase, and phosphorylase levels and the zymogram patterns of these 3 starch-degrading enzymes were determined in the cotyledons of smooth pea (Pisum sativum L.) during the first 15 days of germination. Starch is degraded slowly in the first 6 days; during this time, alpha-amylase is very low, beta-amylase is present at a constant level while phosphorylase gradually increases and reaches a peak on the fifth day. Beginning on the sixth day there is a more rapid degradation of starch which coincides with alpha-amylase production. One phosphorylase band and 2 beta-amylase bands are present in the zymogram of the imbibed cotyledon. An additional phosphorylase band and 1 alpha-amylase band appear during germination. Seeds imbibed in benzyladenine, chloramphenicol, and in cycloheximide show retarded growth and slower starch degradation and enzyme production than the controls. We conclude that alpha-amylase is the major enzyme involved in the initial degradation of starch into more soluble forms while phosphorylase and beta-amylase assist in the further conversion to free sugars.

Amylases↗