Activity of β-galactosidase in homogenates and isolated microvilli fraction of jejunal mucosa from suckling rats

1. beta-Galactosidase activity was studied in homogenates and isolated microvilli fraction of jejunal mucosa from 14-day-old suckling rats. o-Nitrophenyl beta-d-galactoside served as the substrate. 2. The microvilli fraction contains about one-third of the total activity of the original homogenate. 3. The pH optimum of the beta-galactosidase was 3.5 in the total homogenate and supernatant fraction, whereas in the microvilli fraction the maximum activity was at pH5.5. 4. This work gives further support to the view that two beta-galactosidases exist in the jejunal mucosa.     

Germination and regeneration mechanisms in Mediterranean degenerate forests

Abstract. Germination and developmental behaviour during juvenile stages are of importance in relation to succession mechanisms acting on plant communities. This is particularly so in the Mediterranean region, where the climatic summer drought causes stress. We investigated and compared germination and development of four tree species in two parts of the French Mediterranean region where the original dominant tree species (Quercus pubescens) has been largely replaced by Quercus ilex in the southern part, and Castanea sativa in the northern part. Observed differences in germination rates and developmental behaviour, especially shootroot ratio and the role of water supply, suggest that other species may eventually replace the current dominant species.     

Physical characterization of the plasmid pON5300

Summary The antibiotic resistance plasmid Rldrd19Km-which has spontaneously lost its kanamycin resistance marker, and its derivative pON5300, were analysed using the restriction endonucleases SalI, BamHI, HindIII and EcoRI. The fragment patterns were compared with that of the Rldrd19 and the fragments responsible for kanamycin resistance were found to be missing in Rldrd19Km ^– and pON5300. In these plasmids a 7 Mg/mol EcoRI fragment was observed instead of the D (6.3 Mg/mol) fragment of Rldrd19. Further a new 6 Mg/mol EcoRI restriction fragment was observed in pON5300. Using double digestions it was shown that the new fragment does not carry restriction sites for HindIII, BamHI and SalI endonucleases. The non-homology of the analysed plasmid was proved electron microscopically by heteroduplex techniques. The possibility of amplification in the regulatory region for the expression of R-determinants in pON5300 is discussed.     

The mechanism of resistance to streptomycin inEscherichia coli. Functional analysis of the permeability barrier of cells harbouring the Rldrd-19Km plasmid

The mechanism of phenotypically altered SM resistance in mutants of Escherichia coli JC5455 (Rldrd-19Km-) lrs and JC5455 (pON5300) was compared with that of the standard strain JC5455 (Rldrd-19Km-). On analyzing the membrane polypeptides in polyacrylamide gel both mutants were found to possess a protein spectrum different from that of ths standard strain. Transport of D-xylose and L-arginine was the same in all strains, transport of L-proline was decreased in JC5455 (pON5300) which may indicate a mutational interference with energy metabolism. The basic uptake of dihydrostreptomycin was the same in all strains but there were differences after preincubation of cells with streptomycin or glucose. The increased resistance of JC5455 (Rldrd-19Km-) lrs may be due to observed quantitative differences in membrane polypeptides that might play a role in the binding and functional expression of aminoglycoside-3'adenylyl transferase which modifies streptomycin. The increased sensitivity toward streptomycin in JC5455 (pON5300) can be explained by a mutation due to N-methyl-N'-nitro-N-nitrosoguanidine in the host cell since this change of sensitivity to streptomycin could not be transferred by transformation into a nonmutagenized strain. The coincidence of inducibility of increased transport of streptomycin by this antibiotic and the altered frequency of reversion to high levels of streptomycin resistance in JC5455 (pON5300) and in the transformant JC5455 (pON5302) may indicate that the altered reversibility toward phenotypically high resistance to streptomycin is a property of pON5300 and is transferred by transformation.     

Mutation affecting normal colony formation inEscherichia coli K-12

The effect of a mutation in a gene located near thepro marker and determining the normal colony formation and the cell density inEscherichia coli has been analyzed. Thencf mutation has no effect on the permeability of the cell to actinomycin D. On the other hand, a higher level of ATP and a defect in the accumulation of the reserve material in the mutant cell were detected.     

Plant growth promoting bacteria in Brachiaria brizantha

Brachiaria brizantha is considered one of the preferred fodders among farmers for having high forage yield and large production of root mass. The association of beneficial bacteria with these grasses can be very valuable in the recovery of the pasture areas with nutritional deficiency. With the aim of studying this possibility, we carried out the sampling of soil and roots of B. brizantha in three areas (Nova Odessa-SP, S?o Carlos-SP and Campo Verde-MT, Brazil). Seventy-two bacterial strains were isolated and used in tests to evaluate their biotechnological potential. Almost all isolates presented at least one positive feature. Sixty-eight isolates produced analogues of indole-3-acetic acid, ten showed nitrogenase activity when subjected to the method of increasing the concentration of total nitrogen (total N) in the culture medium and sixty-five isolates showed nitrogenase activity when subjected to acetylene reduction technique. The partial sequencing of 16S rRNA of these isolates allowed the identification of seven main groups, with the prevalence of those affiliated to the genus Stenotrophomonas (69?%). At the end, this work elected the strains C4 (Pseudomonadaceae) and C7 (Rhodospirillaceae) as promising organisms for the development of inoculants due to their higher nitrogenase activity.     

High-Andean vegetation and environmental gradients in northwestern Patagonia,Argentina

Abstract. We analysed the heterogeneity of high-elevation vegetation on three mountains along a west-east transect at 41 °S lat. in the Andes of northwestern Patagonia, Argentina. In this area, high-Andean vegetation occurs as islands on mountain tops above Nothofagus pumilio forests ? with the timberline at ca. 1700 m a.s.l. We recorded floristic, topographic and substrate data in 166 sites stratified according to longitude, altitude, slope and aspect. Vegetation data were classified with TWINSPAN and ordinated with Detrended Correspondence Analysis. The relationship between environmental and floristic variation was analysed using Canonical Correspondence Analysis. In order of importance, geographical longitude, altitude and aspect were the major determinants of vegetation variation, whereas substrate texture and slope appeared less important. The combination of these factors resulted in two main vegetation gradients. The first gradient is related to a moisture availability gradient, primarily determined by longitude and secondarily to variation in wind exposure (east vs. west aspect). The second vegetation gradient is related to variation in temperature, primarily determined by altitude, and secondarily by variation in insolation related to the contrast between north and south aspects. The four communities obtained with TWINSPAN are therefore associated with four characteristic habitat types: moist-cold, moist-warm, dry-cold and dry-warm. The community of warm and dry environments is the richest and has elements in common with dry steppe communities situated at lower elevations to the east, while the vegetation of the cold-moist habitat type has unique elements that are typical of the southern Andes. Although current climatic factors appear to be the major determinants of high-Andean vegetation gradients, historical events of Pleistocene times probably also affected the vegetation patterns we see today.     

The promoter of the wheat puroindoline-a gene (<Emphasis Type="Italic">PinA</Emphasis>) exhibits a more complex pattern of activity than that of the <Emphasis Type="Italic">PinB</Emphasis> gene and is induced by wounding and pathogen attack in rice

Puroindolines form the molecular basis of wheat grain hardness. However, little is known about puroindoline gene regulation. We previously reported that the Triticum aestivum puroindoline-b gene (PinB) promoter directs β-glucuronidase gene (uidA) seed-specific expression in transgenic rice. In this study, we isolated a puroindoline-a gene (PinA), analyzed PinA promoter activity by 5′ deletions and compared PinA and PinB promoters in transgenic rice. Seeds of PinA-1214 and PinB-1063 transgenic plants strongly expressed uidA in endosperm, in the aleurone layer and in epidermis cells in a developmentally regulated manner. The GUS activity was also observed in PinA-1214 embryos. Whereas the PinB promoter is seed specific, the PinA promoter also directed, but to a lower level, uidA expression in roots of seedlings and in the vascular tissues of palea and pollen grains of dehiscent anthers during flower development. In addition, the PinA promoter was induced by wounding and by Magnaporthe grisea. By deletion analysis, we showed that the “390-bp” PinA promoter drives the same expression pattern as the “1214-bp” promoter. Moreover, the “214-bp” PinA promoter drives uidA expression solely in pollen grains of dehiscent anthers. The presence of putative cis-regulatory elements that may be related to PinA expression is discussed from an evolutionary point of view. By electrophoretic mobility shift assay, we showed that putative cis-elements (WUN-box, TCA motifs and as-1-like binding sites) whose presence in the PinA promoter may be related to wounding and/or the pathogen response form complexes with nuclear extracts isolated from wounded wheat leaves.