Products of Mitochondrial Protein Synthesis in Yeast

Analysis of membrane proteins of Saccharomyces cerevisiae mitochondria in cycloheximide-inhibited cells shows that they are encoded on mitochondrial DNA.     

A Protein from Immature Raspberry Fruits which Inhibits Endopolygalacturonases from Botrytis cinerea and other Micro-organisms

A polygalacturonase-inhibiting protein (PGIP) was purified fromimmature raspberry fruits using ion exchange chromatography.The protein was composed of a single polypeptide chain withM_r of 38·5 kDa and a pI residing above pH 10. Kineticstudies suggested that the inhibition was of a non-competitivenature. The PGIP inhibited two endopolygalacturonases (endo-PG)purified from Botrytis cinerea and an endo-PG produced by Aspergillusniger to varying degrees but did not inhibit two exo-PGs purifiedfrom B. cinerea, bacterial endopectate lyases and bacterialendo-PGs. The concentration of PGIP at various stages of flowerand fruit development was determined. The inhibitor was notdetected in the flower, but reached a maximum of 69 units g^–1in the immature green fruit decreasing to 9 units g^–1as fruits matured. The N-terminal amino-acid sequence was determined. Key words: Polygalacturonase-inhibiting protein, Rubus idaeus, red raspberry, Botrytis cinerea, pectinases     

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.     

A technique for scoring midge blight of red raspberry, a disease complex caused by Resseliella theobaldi and associated fungi

A scoring technique is described for assessing midge blight using maps representing vascular lesions beneath periderm damaged by second generation larvae of Resseliella theobaldi . Individual canes were scored and some components of yield examined. Yield was not affected until lesions covered more than 20% of the stele surface at the base of canes. Midge blight scores increased with cane girth, probably because splitting of the cortex was most extensive in stout canes.     

Fungi on red raspberry from lesions associated with feeding wounds of cane midge (Resseliella theobaldi)

Two types of vascular lesion are described from the base of canes in plantations infested by raspberry cane midge ( Resseliella theobaldi ); (1) brown lobate lesions ('patches') confined to midge feeding areas, (2) brown lesions spreading proximally and distally from the point of infection ('stripes'). Either or both types of lesion may be presented in individual canes. Isolations from (1) produced principally Fusarium avenaceum ; isolations from tissues where (1) and (2) are contiguous gave Leptosphaeria coniothyrium and F. avenaceum ; isolations from (2) arising from old cane stub wounds in the absence of midge gave L. coniothyrium .
The important secondary role of fungi in the midge blight complex is confirmed, but the involvement of L. coniothyrium in the complex is unclear because it also infects stub wounds on midge-infested canes.     

Localization of Phosphatases in Trypanosoma rhodesiense1

ABSTRACT. Phosphatase activity in Trypanosoma rhodesiense has been examined histochemically by light and electron microscopy and by enzymatic assay in homogenate fractions. Using a method with lead as capture ion, acid phosphatase was found in lysosome-like vesicles and in the flagellar pocket. No alkaline adenosine triphosphatase (ATPase) was detectable by this method. Direct assay of p-nitrophenylphosphatase activity in homogenate fractions showed that acid phosphatase activity was strongly membrane-bound, but that activity at pH 9 was minimal in both soluble and particulate fractions. “Endogenous” ATPase activity was localized specifically and reproducibly in the mitochondrial membranes and under the plasma membrane of the flagellum. This nonenzymic reaction product could not be eradicated by glycerol extraction or glucose depletion. Unlike the membrane staining, which was manifest only after lead treatment, heat-resistant electron-dense material was found in the matrix of lysosomal vesicles in trypanosomes fixed in glutaraldehyde only and not subjected to further treatment with heavy metal reagents. X-ray emission analysis showed the presence of calcium and phosphorus, indicating that the matrix might have a phosphate storage function.     

The effect of oxamyl on the growth of peas attacked by pea cyst nematode

Oxamyl treatment at drilling greatly increased the growth of the plants, the yield and quality of the peas harvested, in soil infested by small or large numbers of Heterodera goettingiana. Treated plants were continuing to set pods when non-treated plants were senescing. Rhizobium nodulation, sap nitrate and tissue nitrogen content were increased in treated plants. Oxamyl applied to growing plants reduced later nematode invasion and enabled root nodulation to proceed with an increase in tissue nitrogen but no increase in yield. Oxamyl applied at drilling in April was inactive against invasion by H. goettingiana in June, which enabled the nematode to multiply equally in treated and non-treated plots. The most economic treatments were at the smallest rates, giving the greatest increase in yield/kg of oxamyl applied.