Physicochemical characterization of mitochondrial DNA from soybean

Mitochondrial DNA (mtDNA) of soybean (Glycine max L.) was isolated and its buoyant density was contrasted with that of nuclear (nDNA) and chloroplast (ctDNA) DNA. Each of the three DNAs banded at a single, characteristic buoyant density when centrifuged to equilibrium in a CsCl gradient. Buoyant densities were 1.694 g/cm³ for nDNA and 1.706 g/cm³ for mtDNA. These values correspond to G-C contents of 34.7 and 46.9%, respectively. Covalently closed, circular mtDNA molecules were isolated from soybean hypocotyls by ethidium bromide-cesium chloride density gradient centrifugation. Considerable variation in mtDNA circle size was observed by electron microscopy. There were seven apparent size classes with mean lengths of 5.9 μm (class 1), 10 μm (class 2), 12.9 μm (class 3), 16.6 μm (class 4), 20.4 μm (class 5), 24.5 μm (class 6), and 29.9 μm (class 7). In addition, minicircles were observed in all preparations. Partially denatured, circular mtDNA molecules with at least one representative from six of the seven observed size classes were mapped. In class 4, there appear to be at least three distinct denaturation patterns, indicating heterogeneity within this class. It is proposed that the mitochondrial genome of soybean is distributed among the different size circular molecules, several copies of the genome are contained within these classes and that the majority of the various size molecules may be a result of recombination events between circular molecules.     

THE EFFECTS OF RED BLOOD CELL EXTRACTS ON THE PROLIFERATION OF ERYTHROCYTE PRECURSOR CELLS, IN VIVO

Saline incubation extracts of mature erythrocytes were assayed in vivo by a variety of techniques in order to study their ability to modify the proliferation of maturing erythroid cells. Using comparable extracts from granulocytes and lymphocytes, the specificity of the effect of the red cell extract for erythroid cells was confirmed by measurement of autoradiographic labelling indices, radio-iron incorporation and spleen colony growth. The erythroid cells were found to be very sensitive to the effects of the extract, as little as 10 μg per mouse producing a maximum effect on iron incorporation. It was found that the extract does not block erythroid cell proliferation completely but simply lengthens the cell cycle, mainly by increasing the G₁ phase of the cycle. There was no effect on the committed erythroid precursor cells. The in vivo activity, specificity and non-toxicity to the cells, together with the cells' sensitivity to red cell extract suggest, therefore, that this inhibitor may play a physiological role in the control of red cell production.     

Biological control ofMythimna separata [Lep.: noctuidae] in New Zealand and its bearing on biological control strategy

A survey of parasites ofAgrotis spp. was carried out in Pakistan to find promising species for trial against noctuids especiallyAgrotis spp. andMythimna separata (Walker) in New Zealand. The parasites recorded wereApanteles ruficrus Haliday,Macrocentrus collaris Spinola,Periscepsia carbonaria Panzer,Turanogonia smirnovi,Rohdain,Ctenichneumon panzeri Wesmael andAnthrax sp. All exceptAnthrax sp. were supplied to New Zealand whereA. ruficrus andM. collaris were released in large numbers.A. ruficrus became established and is giving excellent control ofM. separata resulting in enormous economic gains. Recently it has also been recovered fromAgrotis spp. This example of biological control is significant in thatM. separata has been controlled by a parasite that was previously known from it in New Zealand. The specific status of the “A. ruficrus” already present in New Zealand requires investigation.     

正常大鼠肾脏细胞溶酶体膜的构成蛋白

溶酶体是细胞内对其吞噬之物质溶解及消化之主要场所,同时也是细胞自噬作用的主要细胞器。为了进一步了解此细胞器的功能与结构,我们采用免疫荧光标记法,通过5种针对大鼠肝细胞溶酶体膜蛋白的特异性单克隆抗体,对大鼠正常肾脏细胞溶酶体膜蛋白进行了标记,并通过NH_4Cl溶液对溶酶体作了膜膨胀处理,结果显示:(1)细胞内溶酶体膜蛋白是由多种蛋白所构成,其各种蛋白的含量是不同的;(2)所有溶酶体膜蛋白均表达于该细胞器之表面;(3)NH_4Cl溶液能有效地使溶酶体扩张,这将有和于进一步研究溶酶体的结构。     

Liverwort genomes display extensive structural variations

PIKE, L. M., HU, A., RENZAGLIA, K. S. & MUSICH, P. R., 1992. Liverwort genomes display extensive structural variations. Analyses of the total genomic DNA of eight species of liverworts and two species of green algae by thermal denaturation and CsCl buoyant density gradient centrifugation reveal a high degree of structural complexity and interspecific heterogeneity. The hepatic taxa exhibit two or more DNA components of varying base composition. Average G4-C contents of total cellular DNA calculated from melting profiles are similarly variable, ranging from 38% to 53% G + C. The green alga Chara , a member of the ancestral line to land plants, shows similarities with liverworts in possessing multiple DNA components of comparable complexity, whereas Hydrodiciyon DNA displays a single component. Detailed hybridization analyses of individual density gradient fractions using α-tubulin, rRNA and ribulose 1,5-bisphosphate carboxylase/oxygenase large subunit (rbcL) gene probes were performed to locate the low-copy number and moderately repetitive nuclear genes, and the chloroplast chromosome, respectively. The location of each gene within the density gradient is highly variable among the organisms examined; a-tubulin occurs in fractions ranging from 44–64% G + C, rDNA in 50–64% G + C fractions, and the RbcL gene is located in fractions from 30–59% G + C. For a given species, the two nuclear genes normally overlap in their distributions within the gradient. In most instances, neither gene occurs in the major DNA components, indicating that these components may contain repetitive DNAs. The observed variation in the density of the rbcL gene implies substantial reorganization of the chloroplast genome. The overall differences in the genomic components within and between taxa provide insight into the dynamics of DNA structure that have occurred during the extended evolutionary history of these organisms.     

Oxidation of hydrogen sulfide by flocculated Thiobaccillus denitrificans in a continuous culture

In a continuous fermentation, significant advantages may be gained by immobilization of microbial cells. Immobilization allows cells to be retained in the fermenter or to be readily recovered and recycled. Therefore, the hydraulic retention time and the biomass retention time are decoupled. A novel cell immobilization has been developed for the immobilization of autotrophic bacteria by coculture with floc-forming heterotrophic bacteria with growth of the latter limited by the availability of organic carbon. The result is an immobilization matrix which grows along with the immobilized autotroph. We have previously demonstrated the utility of this approach by immobilizing the chemoautotroph Thiobacillus denitrificans in macroscopic floc by coculture with floc-forming heterotrophs from an activated sludge treatment facility. Floc with excellent settling characteristics were produced. These floc have now been used to remove H(2)S from a gas stream bubbled through continuous cultures. The stoichiometry and kinetics of H(2)S oxidation by immobilized T. denitrificans were comparable to that reported previously for free-cell cultures. Oxygen uptake measurements indicated the growth of both T. denitrificans and the heterotrophs although the medium contained no added organic carbon. Continuous cultures with total biomass recycle were maintained for up to four months indicating the long-term stability of the commensal relationship between the immobilized autotroph and the heterotrophs which composed the immobilization matrix. It was observed that at any given H(2)S loading the biomass concentration reached a maximum and leveled out. The ultimate biomass concentration was dependent upon the H(2)S feed rate.     

Action of some retinol derivatives and their provitamins on microsome-catalyzed formation of benzo[a]pyrene-DNA adduct

Several vitamin A compounds have been tested for their ability to suppress formation of DNA adduct by the carcinogen benzo[a]pyrene (B[a]P) in an in vitro reaction catalyzed by rat liver microsomes. Retinol, retinal, 3-dehydroretinol and 3-hydroxyretinol were found to be effective inhibitors of adduct formation. Certain carotenoids that are precursors of these retinoids also displayed considerable inhibitory capacity. Carotenoids and the 3-substituted retinoids appeared to modulate the DNA adduct formation exclusively through their action on microsomal enzymes, since an effective inhibition in each case was observed on the formation of B[a]P-7,8-diol, a proximate carcinogenic metabolite of B[a]P. Unsubstituted retinoids, on the other hand, had marginal effect on enzymes but were found effective in accelerating inactivation of B[a]P-7,8-diol-9,10-epoxide, the ultimate carcinogenic metabolite that binds to DNA.