Studies on the Inoculation and Competitiveness of a Rhizobium leguminosarum Strain in Soils Containing Indigenous Rhizobia

The competitiveness of a Rhizobium leguminosarum strain was investigated at two separate locations in field inoculation studies on commercially grown peas. The soil at each location (sites I and II) contained an indigenous R. leguminosarum population of ca. 3 × 10⁴ rhizobia per g of soil. At site I it was necessary to use an inoculum concentration as large as 4 × 10⁷ CFU ml⁻¹ (2 × 10⁶ bacteria seed⁻¹) to establish the inoculum strain in the majority of nodules (73%). However, at site II the inoculum strain formed only 33% of nodules when applied at this (10⁷ CFU ml⁻¹) level. Establishment could not be further improved by increasing the inoculum concentration even as high as 10⁹ CFU ml⁻¹ (9.6 × 10⁷ bacteria seed⁻¹). The inoculum strain could be detected at both sites 19 months after inoculation. Analysis by intrinsic antibiotic resistance patterns and plasmid DNA profiles indicated that a dominant strain(s) and plasmid pool existed among the indigenous population at site II. Competition experiments were carried out under laboratory conditions between a dominant indigenous isolate and the inoculum strain. Both strains were shown to be equally competitive.     

Catalysis by heteropentalenes of electron transfer to oxygen from ferredoxin-NADP oxidoreductase reduced by NADPH

A range of heteropentalene and bipyridinium compounds have been tested as catalysts of electron transfer to oxygen from spinach ferredoxin-NADP⁺ oxidoreductase reduced by NADPH. For a particular class of compound, the rate of oxygen reduction increased with increasing midpoint potential of the compound under conditions in which reduction of the compound was rate-limiting. Compounds with similar midpoint potentials from different structural classes showed marked differences in rate, attributed to specificity in the interaction with ferredoxin-NADP⁺ oxidoreductase.     

Applicability of imprints to immuno-ultrastructural studies of lymphoid tissues

In order to evaluate the applicability of imprints to immuno-ultrastructural studies of lymphoid tissues, we compared distribution pattern and morphology of B cells, T cells, T-cell subsets, and follicular dendritic reticulum cells (DRC) at the light and ultrastructural level in imprints and sections of tonsils and lymph nodes. The surface antigenic profile of lymphoid cells was revealed with monoclonal antibodies in an avidin-biotin-peroxidase complex (ABC) method. Distribution of lymphoid cells in coherent areas of imprints recapitulated their disposition in sections of respective lymphoid tissues. Preservation of microanatomical relationships and ultrastructure of lymphoid cells in imprints allowed evaluation of associations and fine structural detail of lymphoid cells. Morphologic configurations of B cells in imprints, confined to round aggregates, were similar to fine structural morphology of B cells in mantle zones (MZ) and germinal centers (GC). Processes of DRCs in imprints formed conformations resembling their meshwork within follicles and mantled lymphoid cells. In imprints and sections, lymphocytes of cytotoxic/suppressor phenotype had a large amount of cytoplasm with many organelles. In contrast, cells of helper/inducer phenotype displayed a high nucleocytoplasmic (N/C) ratio and small numbers of organelles. Thus, imprints represent an easy, fast, and reliable method that lends itself to immunoultrastructural studies of lymphoid tissues.     

A restriction endonuclease map of Streptomyces phage VWB

Summary A physical map of the actinophage VWB has been constructed using the restriction endonucleases BglII, ClaI, EcoRI, EcoRV, HindIII, KpnI and SphI. Phage VWB, genome size 47.3 kb, propagates on Streptomyces venezuelae, and it can also lysogenise this species. The three BglII-generated fragments of VWB DNA were cloned in pBR322, and subsequently mapped. In this manner the restriction map of the VWB phage genome was constructed.Abbreviations dam DNA adenine methylase activity - kb kilobase pairs - :: novel joint     

Effects of temperature on the development,growth, and survival of larvae and pupae of a north-temperate chrysomelid beetle

Summary Developoment, growth, and survival of larvae and pupae of the red turnip beetle, Entomoscelis americana Brown, were studied in 10 constant and four alternating temperature regimes (10 to 32.5° C), in field-cages, and in natural populations in Manitoba. This beetle has a northtemperate distribution in North America. Larval and pupal development occurs in spring and normally is completed before the end of June. Growth and development occurred at all constant temperatures tested, but survival was low at the extreme temperatures. Therefore, the threshold and upper limit were near 10 and 32.5° C. The developmental times of the sexes did not differ and decreased with temperature, except possibly at 32.5° C. The average weight of adult females increased with temperature up to 32.5° C and those of males up to 25° C. Considering developmental rate, survival, adult weight, and incidence of malformed adults, the optimum temperature was estimated to be near 27.5° C.Development was accelerated significantly (6 to 9%) in alternating regimes with temperatures differing by 10° C, but not in regimes differing by 5 and 15° C. All alternating regimes increased adult weight, 5 to 17% for females and 2 to 10% for males. Field cage studies confirmed the increase in adult weight, but not the acceleration in development.A three-parameter normal function described accurately the relationship between developmental rate and constant temperature. A computer simulation model based on this equation estimated developmental times in field cages to within one to five days. For natural populations the model overestimated the developmental times by five to 16 days. The discrepancies between model estimates and observed developmental times in natural populations apparently were due to the elevation of larval and pupal body temperatures above air temperatures by behavioral thermoregulation. The elevation of body temperature was estimated to be equivalent to the addition of 5 to 6° C to the maximum daily air temperature. The adaptations and responses of this beetle to the cool spring temperatures of the north-temperate region are discussed.Contribution No. 1164, Agriculture Canada, Research Station, Winnipeg, Manitoba, Canada     

Distribution and recovery of15N after fertilization of Douglas-fir saplings with different nitrogen sources

Summary Four-year-old Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) saplings planted in pots with a sand and peat mix (11) were fertilized at the rate of 200 kg N/ha of (¹⁵NH₂)₂CO (U-15),¹⁵NH₄NO₃ (A-15) and NH₄ ¹⁵NO₃(An-15). They were placed in a shadehouse and watered regularly to maintain soil moisture at field capacity over periods of one and two years. Quantity of¹⁵N in foliage generally increased from old to current growth, irrespective of the nitrogen source. Utilization of¹⁵N fertilizers by saplings after the first and second growing seasons following fertilization was greatest with nitrate labelled ammonium nitrate AN-15, and nearly equal for urea U-15 and ammonium labelled ammonium nitrate A-15. The soil immobilized more fertilizer nitrogen-15 from U-15 and A-15 than from AN-15. Data from the present study, in which leaching losses of fertilizer were minimized, demonstrated that in terms of nitrogen uptake by the saplings the nitrate fertilizer was superior to ammonium fertilizer.     

Isolation and cryopreservation of O-methylthreonine-resistant Rosa cell lines altered in the feedback sensitivity of l-threonine deaminase

O-Methylthreonine (OMT) inhibits the growth of plated Rosa cells (ID₅₀6·10^-6M). Isoleucine is able to reverse efficiently and specifically this OMT toxicity. From OMT-resistant colonies occurring at a frequency of 1.58·10^-7 variants per cell plated at 10^-4M OMT, the variant strains OMT^R-1 and OMT^R-2 were isolated, cloned via protoplasts and characterized. Both variants were ten times more resistant to OMT than the wildtype and were cross-resistant to another isoleucine analog, dl-4-thiaisoleucine. The resistant variants retained their resistance after storage for three years in liquid nitrogen. Both resistant strains were stable for several months when subcultured in the absence of OMT although it was shown in a reconstitution experiment that wildtype cells overgrow OMT^R-2 variant cells if co-cultivated for many passages in drug-free medium. One case of instability was observed upon long-term subculturing in drug-free medium: the strain OMT^R-1D^* partially lost phenotypic properties. Resistance to OMT was followed qualitatively by a new method based on inhibition-zone formation in cell suspensions plated in agar medium. The OMT-resistant variants showed a reduction in sensitivity of the enzyme l-threonine deaminase to feedback inhibition by isoleucine, a decreased stability of l-threonine deaminase when stored at-18°C or incubated at +55°C and a two- to threefold increase of the free isoleucine pool within the cells. The genetical events and the biochemical mechanisms which might lead to the observed stable and biochemically defined character are discussed with particular reference to the high ploidy level of the Rosa cell line.Abbreviations OMT l-O-methylthreonine - TD l-threonine deaminase     

Analysis of the sheep MHC using HLA class I,II, and C4 cDNA probes

Four cDNA probes for the human major histocompatibility complex (MHC) were used to investigate the sheep MHC, in conjunction with serological typing for ovine lymphocyte antigen (OLA). Lymphocytes from a family (two parents and five offspring) of Romanov sheep were subjected to genomic DNA digestion by the restriction endonuclease Eco RI, followed by gel electrophoresis. A single Southern blot representing all seven individuals was then consecutively hybridized with the class I, alpha-DC, beta-DR, and C4 probes, which were originally designed to identify HLA class I, class II (DC and DR), and C4 products, respectively. Using each of the three class I/class II probes, several bands showing DNA polymorphism were detected. The segregation of these bands in the five offspring exactly paralleled the OLA haplotype segregation established by serological typing. A further eight individuals carrying haplotypes which were phenotypically identical to those in the above-mentioned family showed bands in the corresponding positions when tested with the same three probes. Using the C4 probe, no polymorphism was detected in these fifteen individuals.Abbreviations used in this paper MHC major histocompatibility complex - OLA ovine lymphocyte antigen - kbp kilobase pair(s) - MLR mixed lymphocyte reaction - RFLP restriction fragment length polymorphism     

Risk of predation,hydrodynamic efficiency and their influence on school structure

Synopsis Laboratory studies were conducted on 15 schools of blackchin shiners, Notropis heterodon, to determine if they altered their structure in response to changing environmental demands. The hypothesis tested was that fish schools should sacrifice a flat, hydrodynamically efficient structure in favour of an unobstructed visual field in the presence of a predator by staggering in the vertical plane. Ten schools were exposed for two weeks to a simple environment with only a current. For the next two-week period a predator was added. Five control schools were exposed to the simple environment for both two-week periods. Six of the ten treated schools increased their staggering in the vertical dimension as predicted while none of the control schools changed. This result was suggestive that hydrodynamic advantages were sacrificed. Respirometer experiments indicated these fish were capable of achieving some hydrodynamic benefits from schooling but these benefits may be a function of fish size.     

Histamine-induced pulmonary edema distal to pulmonary arterial occlusion

Histological studies provide evidence that the bronchial veins are a site of leakage in histamine-induced pulmonary edema, but the physiological importance of this finding is not known. To determine if a lung perfused by only the bronchial arteries could develop pulmonary edema, we infused histamine for 2 h in anesthetized sheep with no pulmonary arterial blood flow to the right lung. In control sheep the postmortem extravascular lung water volume (EVLW) in both the right (occluded) and left (perfused) lung was 3.7 +/- 0.4 ml X g dry lung wt-1. Following histamine infusion, EVLW increased to 4.4 +/- 0.7 ml X g dry lung wt-1 in the right (occluded) lung (P less than 0.01) and to 5.3 +/- 1.0 ml X g dry wt-1 in the left (perfused) lung (P less than 0.01). Biopsies from the right (occluded) lungs scored for the presence of edema showed a significantly higher score in the lungs that received histamine (P less than 0.02). Some leakage from the pulmonary circulation of the right lung, perfused via anastomoses from the bronchial circulation, cannot be excluded but should be modest considering the low pressures in the pulmonary circulation following occlusion of the right pulmonary artery. These data show that perfusion via the pulmonary arteries is not a requirement for the production of histamine-induced pulmonary edema.