Purification and properties of indole 2,3-dioxygenase from maize leaves

An indole 2,3-dioxygenase was purified ca 38-fold from maize leaves. The enzyme had an MW of about 98000, an optimum pH of 5.0 and the energy of activation was 9.1 kcal/mol. The K_max for indole was 1.4 × 10^?4 M. The enzyme was inhibited by diethyldithiocarbamate, salicylaldoxime and sodium dithionite. The inhibition by diethyldithiocarbamate was specifically reversed by Cu²⁺. The dialysed enzyme was stimulated by Cu²⁺. Four atoms of oxygen were utilized in the disappearance of 1 mole of indole. Inhibition of the enzyme by -SH compounds and -SH group inhibitors, and their partial removal by Cu²⁺ only, suggested the involvement of -SH groups in binding of Cu²⁺ at the catalytic site.     

Adaptation of beta-ray gauge technique for measurement of water deficit of intact rice leaves

A linear relationship was obtained between the logarithm of count rate and absorber’s (Aluminum foil and leaf) effective thickness in the range of 4.25 to 18.00 mg cm^?2. Also relative water content (RWC) and absorber effective thickness was linearly related (Y = ?89.80 + + 15.84 X, and r = 0.99). From these relationships it was possible to determine the RWC of a rice leaf simply by measuring the radiation intensity of fresh, fully turgid and completely dry leaf by beta ray gauge technique without the added necessity of determining the mass of the leaf.     

Factors affecting in vitro clonal propagation of Prosopis cineraria

Genotype, age of tree, nature of explant and size (length and diameter), season of explant collection, explant position on medium, plant growth regulators and certain additives (ascorbic and citric acids, adenine sulphate, L-arginine, glutamine and ammonium citrate), incubation conditions, and subculturing period greatly influenced the in vitro clonal propagation of P. cineraria. The maximum number of 10–12 shoots were induced from the nodal shoot segment from pruned thorny adult trees on Murashige and Skoog's (MS) medium containing 0.1 mgl^-1 indole- 3-acetic acid (IAA)+2.5 mgl^-1 benzylaminopurine (BAP)+additives. Higher temperature (31+-2°C) and mixed (fluorescent and incandescent) light of 50 mol m^-2 s^-1 photon flux density for 12 h per day photoperiod favoured shoot induction and subsequent growth. Explants from thornless trees produced 6–8 shoots per explant on MS medium containing 0.1 mgl^-1 IAA+5.0 mgl^-1 BAP + additives. Nodal shoot segments obtained from root and stump sprouts produced multiple shoots. Root segments differentiated into multiple shoots on MS medium containing 0.5 mgl^-1 indolebutyric acid (IBA)+2.5 mgl^-1 BAP.Differentiated shoots multiplied best on MS medium containing 0.1 mgl^-1 naphthalene acetic acid (NAA)+1.0 mgl^-1 BAP + additives. To yield multiple shoots the original explant was transferred 6 times on fresh medium after harvesting the differentiated shoots. Shoots were rooted by pulsing with 100 mgl^-1 IBA for 4 h and then culturing on hormone-free half strength MS medium. Initial dark incubation for 5 days at high temperature (33±2°C) was found essential for root induction from shoots which was 63% within two weeks. The rooted plantlets contained a consistent number of chromosomes (2n=28). It is suggested that the protocol developed could be useful for cloning of mature and tested trees of P. cineraria.     

Mitochondrial DNA sequence evolution in sharks: rates, patterns, and phylogenetic inferences

Abundant representation of sharks in the fossil record makes this group a superb system in which to investigate rates and patterns of molecular evolution and to explore the strengths and weaknesses of phylogenetic inferences from molecular data. In this report, the molecular evolution of the cytochrome b gene in sharks is described and the information related to results from phylogenetic analysis of the data evaluated in the light of a phylogeny derived independently of the molecular data. Across divergent lineages of sharks there is evidence for significant substitution rate variation, departure from compositional equilibrium, and substantial homoplasy; nevertheless, the signal of evolutionary history is evident in patterns of shared transversions and amino acid replacements.      

Metabolic rate and directional nucleotide substitution in animal mitochondrial DNA

There is marked heterogeneity of nucleotide composition in mitochondrial DNA across divergent animals. Differences in nucleotide composition presumably reflect differences in directional nucleotide substitution for A+T or G+C nucleotides. In mitochondrial DNA, there is A+T directional nucleotide substitution in most (if not all) animals surveyed, and the magnitude of directional A+T nucleotide substitution differs greatly within and among groups. Differences in directional nucleotide substitution among lineages of mammals can be explained by changes in metabolic physiology. This relationship is thought to be mediated by the effect of oxygen radicals because these toxic compounds are by-products of aerobic metabolism and are known mutagens. Association between metabolism and nucleotide composition provides additional evidence in favor of the hypothesis that rates and patterns of nucleotide substitution in mitochondrial DNA can be influenced by factors that impinge on rates of endogenous DNA damage.      

Quantification and correlation of vesicular-arbuscular mycorrhizal propagules with soil properties of some mollisols of northern India

The populations of vesicular-arbuscular mycorrhizae (VAM) propagules by the most probable number method in some mollisols and their correlations with some important soil properties were determined. On average, the six soils, Phoolbagh clay loam, Beni silty clay loam, Haldi loam, Nagla loam, Khamia sandy loam and Patherchatta sandy loam contained 4.9, 4.0, 7.9, 7.9, 3.3 and 13.0 propagules/g soil, respectively, i.e. none of the soils was found to be high in VAM. The size of the VAM population was compared to soil properties such as pH, organic carbon, sand content, available phosphorus and available potassium, cation-exchange capacity, silt and clay contents. A significant positive correlation (r=0.586) was only found with available soil phosphorus (P<0.05) and a significant negative correlation (r=-0.555) with soil clay content (P<0.05).Directorate research paper series No. 7862     

Elemental distribution in striated muscle and the effects of hypertonicity: Electron probe analysis of cryo sections

A method of rapid freezing in supercooled Freon 22 (monochlorodifluoromethane) followed by cryoultramicrotomy is described and shown to yield ultrathin sections in which both the cellular ultrastructure and the distribution of diffusible ions across the cell membrane are preserved and intracellular compartmentalization of diffusabler ions can be quantitated. Quantitative electron probe analysis (Shuman, H., A.V. Somlyo, and A.P. Somlyo. 1976. Ultramicros. 1:317-339.) of freeze-dried ultrathin cryto sections was found to provide a valid measure of the composition of cells and cellular organelles and was used to determine the ionic composition of the in situ terminal cisternae of the sarcoplasmic reticulum (SR), the distribution of CI in skeletal muscle, and the effects of hypertonic solutions on the subcellular composition if striated muscle. There was no evidence of sequestered CI in the terminal cisternae of resting muscles, although calcium (66mmol/kg dry wt +/- 4.6 SE) was detected. The values of [C1](i) determined with small (50-100 nm) diameter probes over cytoplasm excluding organelles over nuclei or terminal cisternae were not significantly different. Mitochondria partially excluded C1, with a cytoplasmic/ mitochondrial Ci ratio of 2.4 +/- 0.88 SD. The elemental concentrations (mmol/kg dry wt +/- SD) of muscle fibers measured with 0.5-9-μm diameter electron probes in normal frog striated muscle were: P, 302 +/- 4.3; S, 189 +/- 2.9;C1, 24 +/- 1.1;K, 404 +/- 4.3, and Mg, 39 +/- 2.1. It is concluded that: (a) in normal muscle the "excess CI" measured with previous bulk chemical analyses and flux studies is not compartmentalized in the SR or in other cellular organelles, and (b) the cytoplasmic C1 in low [K](0) solutions exceeds that predicted by a passive electrochemical distribution. Hypertonic 2.2 X NaCl, 2.5 X sucrose, or 2.2 X Na isethionate produced: (a) swollen vacuoles, frequently paired, adjacent to the Z lines and containing significantly higher than cytoplasmic concentrations of Na and Cl or S (isethionate), but no detectable Ca, and (b) granules of Ca, Mg, and P = approximately (6 Ca + 1 Mg)/6P in the longitudinal SR. It is concluded that hypertonicity produces compartmentalized domains of extracellular solutes within the muscle fibers and translocates Ca into the longitudinal tubules.     

Effect of soil moisture stress on legume-rhizobium symbiosis in soybeans

Summary In a pot culture experiment, the influence of soil moisture stress at different physiological stages of soybean, cv. Hark, on nodulation, symbiosis and nitrogen accumulation was studied. Moisture stress reduced leghemoglobin content of root nodules and nitrogen uptake by plants. It had no effect on number of bacteroids. Stress at mid bloom and rapid pod filling stages reduced yield and seed protein content. However, these parameters were not affected by stress at nodule initiation and early flowering stages, though, flower initiation and maturity of the plant were delayed. Moisture stress at any stage did not alter nitrogen status of roots.     

Effect of dimethyl sulfoxide (DMSO) on zinc availability (L-value), growth and metabolic activities of rice plants

Summary Effects of soil application (0.01, 0.1 and 1%) and foliar sprays (0.001, 0.01 and 0.1%) of dimethyl sulfoxide (DMSO) and soil application of Zn (10 and 20 ppm) on growth, yield, photosynthetic pigments and some enzymatic activities of rice (Oryza sativa L. variety Jaya) were investigated in a Zn-deficient soil under pot culture trials. Control plants showed typical Zn deficiency symptoms, very low dry matter and chlorophyll contents and significantly lower activities of carbonic anhydrase and tryptophan synthetase. Application of 10 or 20 ppm Zn to the soil markedly improved plant growth, chlorophyll content, enzymatic activities, Zn availability (L-value), and grain yield. Application of 1% DMSO to the soil proved to be severly phytotoxic for plant growth and dry matter accumulation. Application of lower doses of DMSO to the soil (0.01 and 0.1%) or as foliar sprays (0.001 and 0.01%), however, slightly increased dry weights of all plant parts at 45 days after transplanting. Grain yield was significantly increased by all DMSO treatments, except 1% soil application which completely suppressed the grain formation.Enzymatic activities and chlorophyll and carotenoid accumulation showed concentration-dependent stimulation or inhibition by DMSO treatments. Carbonic anhydrase activity was significantly increased by most of the DMSO treatments whereas tryptophan synthetase activity was stimulated only by the lowest dose of soil (0.01%) and foliar (0.001%) applications. Chlorophyll content was stimulated by lower doses of foliar application of DMSO (0.001 and 0.01%) but the other DMSO treatments brought about a concentration-dependent decrease in chlorophyll accumulation. Carotenoid accumulation was inversely related to that of chlorophylls. Chlorophyll: carotenoid ratio was increased by all DMSO and Zn treatments except 1% soil application of DMSO. Zn availability (L-value) of the soil was increased by all DMSO treatments. Zn content of leaf blades was positively correlated with chlorophyll content (r=0.579), tryptophan synthetase (r=0.700) and carbonic anhydrase (r=0.537) activities but negatively correlated with that of carotenoid content (r=–0.896). The possible mechanisms of DMSO on plants are discussed in relation to its possible agricultural utility.Publication No. 811 under Journal Series of the Experiment Station, G.B. Pant of Agriculture & Technology, Pantnagar (Nainital), India.