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Biomedical subjects

T M Ching

Publications and source records attributed to T M Ching.

At least 19 recordsLinked to original sources
PubMed1988

Induction of barley leaf urease.

Foliar urea application on barley plants increased leaf urease activity for 5 hours with a peak of 20-fold at 2 hours. To discern the mode of urease induction, urea with or without inhibitors and [(35)S]methionine were incubated with leaf sections for different lengths of time. Urease was extracted, partially purified, electrophoresed, and then quantified by fluorogram. Five urease (U) isozymes were separated by PAGE. U(a) and U(b) might be polymers or complexes that occurred only at the peak of induced activity. U(1) and U(2) appeared at 0.5 and 0.75 hour, respectively, after urea induction, peaked at 2 hours, and persisted only in treated leaves for several additional hours indicating that they are transient inducible forms. U(3) was the constitutive form present in control and treated leaves. Induction with cordycepin or cycloheximide completely prevented urea stimulated activity and nullified the existence of isozymes U(a), U(b), U(1), and U(2). (35)S-U(1), which was labeled in the last hour of induction, appeared on fluorogram 1 hour after induction, peaked at 2 hours, and declined at 3 hours. Results indicated that de novo synthesis of urease is activated by the influx of urea.

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PubMed1987

Purification and properties of Acid phosphatase from plump and shriveled seeds of triticale.

A major triticale (X Triticosecale Wittmack) endosperm acid phosphatase (EC 3.1.2.2) (APase) from sib-lines producing plump and shriveled seed was purified 140- and 230-fold to a specific activity of 94 and 153 micromoles per minute per milligram protein respectively, by ammonium sulfate fractionation, ion-exchange chromatography, chromatofocusing, affinity column chromatography, and gel filtration. The purified enzyme from both materials is a monomeric glycoprotein with an apparent molecular weight of 45,700 +/- 500 containing 12% carbohydrate and an apparent isoelectric point of pH 5.9. It hydrolyzes tri- and di-phosphate of nucleosides as well as phosphate esters and exhibits characteristics of ATP-hydrolase and phosphatase. About 2-fold more of the APase was isolated from shriveled seeds, and the purified enzyme exhibited 3- and 5-fold higher V(max) for p-nitrophenyl phosphate and ATP, respectively, than that of plump seed. The I(50) for Pi concentration was 5.5-fold higher in APase of shriveled seed than the plump one. These varied quantitative and kinetic properties substantiate the role of APase in lines with shriveled seeds being reduction of starch accumulation by depleting substrates and energy supply in the cytosol.

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PubMed1984

Acid phosphatases and seed shriveling in triticale.

Seed shriveling in the man-made intergeneric hybrid, triticale (x Triticosecale Wittmack) appears to be related to increased activity of endosperm acid phosphatases including para-nitrophenyl phosphatase, ATPase, ADPase, phosphatidic phosphatase, and glucose-1-phosphatase that occur specifically at later stages of seed development. These hydrolases may reduce endogenous substrates for starch synthesis, deplete energy supply for maintenance and biosynthesis of tissue growth, and deassemble membrane structures resulting in a partially filled endosperm and localized necrosis. Electrophoretic isozyme patterns of endosperm acid phosphatase exhibited distinctive differences between lines producing plump and shriveled seeds indicating a divergent role of the isozymes in these two different seed conformations.

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PubMed1983

Starch synthesis in shriveled and plump triticale seeds.

Seven lines of triticale (X Triticosecale Wittmack) with either shriveled or plump seed characteristics were planted in the field, and seed developmental changes in weight, starch content, the activity of starch biosynthetic and degradative enzymes, and ATP content were studied in three consecutive years in Oregon. Experimental results varied among genotypes and with growing environment, but overall indicated that: (a) amylase activity was higher in shriveled lines, but was not directly synchronized with the occurrence of shriveling at later stages of seed formation; (b) ADP-glucose starch synthase contributed to starch accumulation in triticale, but it appeared not to be associated with shriveledness as no stage-related changes were observed; and (c) ADP-glucose pyrophosphorylase activity was 2- to 3-fold higher in plump lines than that of shriveled lines, indicating that this enzyme may play an important role in the degree of plumpness or stach accumulation of triticale seeds.

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PubMed1979

Hydrogen-dependent nitrogenase activity and ATP formation in Rhizobium japonicum bacteroids.

Rhizobium japonicum 122 DES bacteroids from soybean nodules possess an active H(2)-oxidizing system that recycles all of the H(2) lost through nitrogenase-dependent H(2) evolution. The addition of 72 muM H(2) to suspensions of bacteroids increased O(2) uptake 300% and the rate of C(2)H(2) reduction 300 to 500%. The optimal partial pressure of O(2) was increased, and the partial pressure of O(2) range for C(2)H(2) reduction was extended by adding H(2). A supply of succinate to bacteroids resulted in effects similar to those obtained by adding H(2). Both H(2) and succinate provided respiratory protection for the N(2)-fixing system in bacteroids. The oxidation of H(2) by bacteroids increased the steady-state pool of ATP by 20 to 40%. In the presence of 50 mM iodoacetate, which caused much greater inhibition of endogenous respiration than of H(2) oxidation, the addition of H(2) increased the steady-state pool of ATP in bacteroids by 500%. Inhibitor evidence and an absolute requirement for O(2) indicated that the H(2)-stimulated ATP synthesis occurred through oxidative phosphorylation. In the presence of 50 mM iodoacetate, H(2)-dependent ATP synthesis occurred at a rate sufficient to support nitrogenase activity. The addition of H(2) to H(2) uptake-negative strains of R. japonicum had no effect on ATP formation or C(2)H(2) reduction. It is concluded that the H(2)-oxidizing system in H(2) uptake-positive bacteroids benefits the N(2)-fixing process by providing respiratory protection of the O(2)-labile nitrogenase proteins and generating ATP to support maximal rates of C(2)H(2) reduction by oxidation of the H(2) produced from the nitrogenase system.

Adenosine Triphosphate↗
PubMed1978

Developmental Differences in Embryos of High and Low Protein Wheat Seeds during Germination.

Developmental patterns of embryos from high and low protein wheat (Triticum aestivum) grain produced under varied fertilizer conditions were compared. High protein grain produced seedlings 25% heavier with 25% more total RNA, 30% more DNA, 40% more amino acids, 60% more ribosomes, and 80% more soluble protein content than that of low protein seed. Consistently higher glutamine synthetase and alpha-amylase and lower acid phosphatase activities were observed in high protein seeds, though the isozyme pattern of alpha-amylase was not different in the two kinds of seeds. The high total ribosomes and particularly, polysome content observed in high protein seeds may be responsible for the rapid growth and high yield of these seeds.

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PubMed1977

Isolation of bacteria, transforming bacteria, and bacteroids from soybean nodules.

Postnuclei supernatant of soybean (Glycine max cv. Chippewa 64) nodule homogenate was fractionated by stepwise sucrose density gradient centrifugation into supernatant, endoplasmic reticulum and mitochondria, and three distinct bands with 1.22, 1.25, and 1.27 g/cm(3) of peak density. Based on their enzymic activities, composition of electron transport components, and ultrastructural characteristics, the lightest band appears to be the mature bacteroids; the intermediate band the transforming bacteria; and the heaviest, the bacteria. The isolation procedure separates nodule symbionts into different functional and developmental fractions, and it may be a valuable tool for studies involving development, regulation, and senescence of bacteroids in the nodule.

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PubMed1975

Energy State and Dinitrogen Fixation in Soybean Nodules of Dark-grown Plants.

Dark treatment of 25-day-old greenhouse-grown plants of inoculated soybean (Glycine max var. Chippewa) for 1 day reduced ATP by 70%, sucrose by 60%, total adenosine phosphates by 60%, ATP/ADP ratio by 55%, nitrogenase activity by 50%, and energy charge by 15% in nodules. The close correlation between nitrogenase activity and these energy parameters indicates that they may play a major role in regulating dinitrogen fixation in the symbiotic system.

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PubMed1975

Temperature regulation of germination in crimson clover seeds.

Seeds of Dixie crimson clover (Trifolium incarnatum cv. Dixie) completed germination in 36 hours at 20 C. At 10 C germination was delayed by 24 hours. At 30 C only 20% germinated and the rest remained viable for a long time but not germinable. Different patterns of adenylate energy state and zymograms of acid phosphatase and esterase were observed from seeds grown under the three temperatures for 24 hours. Varied specific activities of protease, alpha-amylase, ATPase, RNase, acid phosphatase, glutamine synthetase, and fumarase were also found. Protein-synthesizing ability was proportional to temperature. These data indicate that temperature regulates seed germination at multiple sites.

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PubMed1974

Adenylate energy pool and energy charge in maturing rape seeds.

A study of energy state and chemical composition of pod walls and seeds of maturing rape (Brassica napus L.) was conducted on two varieties, Victor and Gorczanski. Total adenosine phosphates, ATP, and adenylate energy charge increased with increasing cell number and cellular synthesis during the early stages, remained high at maximum dry weight accumulation and maximum substrate influx time, and decreased with ripening. A temporal control of energy supply and ATP concentration is evident in developing tissues with determined functions; whereas the association of a high energy charge and active cellular biosynthesis occurs only in tissues with a stabilized cell number.

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PubMed1973

Compartmental utilization of carboxyl-C-tripalmitin by tissue homogenate of pine seeds.

A tissue homogenate of megagemetophyte of germinating seeds of Jeffrey pine (Pinus Jefferii Grev. and Balf.) was incubated with sonication-dispersed and albumin-carried (14)C-tripalmitin in order to elucidate the sequential and quantitative role of cellular organelles in utilizing lipid reserve in seeds. After 5 minutes at 30 C, 25% of the tracer was localized in the fat body fraction, 9% in the pellet containing mitochondria and glyoxysomes, 14% in the supernatant, and 2% was found as CO(2). Radioactivity increased with time of incubation in the latter three fractions indicating the forward direction of utilization. Fat bodies contained mainly lipases and hydrolyzed the tracer to palmitate with diglyceride and monoglyceride as intermediates. About two-thirds of the palmitate had left the fat bodies in 5 minutes and entered the pellet fraction within which the tracer was distributed 1:2 in mitochondria and glyoxysomes, respectively. Longer incubation reduced the ratio to 1:3 while both organelles acquired more radioactive intermediates. Labeled acetyl-CoA and intermediate of beta-oxidation were found in both organelle-containing fractions. The supernatant fraction contained radioactive diglycerides, monoglycerides, palmitate, sterol esters, and phospholipids, indicating lipase activity and direct utilization of fatty acid for the synthesis of sterol esters and polar lipids.

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PubMed1973

Adenosine triphosphate content and seed vigor.

ATP content in imbibed seeds is significantly correlated with seedling size in fatty, starchy and proteinaceous seeds, and it indicates viability in seed lots. ATP content thus appears to be a useful biochemical index of seed vigor.

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PubMed1972

Content of adenosine phosphates and adenylate energy charge in germinating ponderosa pine seeds.

An average of 540 picomoles of total adenosine phosphates was found in the embryo of mature seeds of ponderosa pine (Pinus ponderosa Laws.) and 1140 picomoles in the gametophyte. Adenylate energy charges were 0.44 and 0.26, respectively. After stratification, total adenosine phosphates increased 7-fold and 6-fold in embryo and gametophyte, respectively, and energy charges rose to 0.85 and 0.75. During germination, total adenosine phosphates increased to a 20-fold peak on the 9th day in gametophytic tissue, parallel with the peak of reserve regradation and organellar synthesis, and then decreased. In embryo and seedling, total adenosine phosphates elevated 80-fold with two distinct oscillating increases of AMP and ADP. The oscillating increases occurred before the emergence of radicle and cotyledons during which the highest mitotic index prevailed in all tissues. Energy charges fluctuated between 0.65 at the rapid cell dividing stage to 0.85 at the fully differentiated stage of the seedling, while energy charges remained around 0.75 in the gametophyte. These data indicated that the content of adenosine phosphates of germinating seeds reflects growth, organogenesis, and morphogenesis, and that a compartmentalized energy metabolism must exist in dividing and growing plant cells.

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