The response ofRhizobium meliloti to L-methionine DL-sulphoximine

A strain ofRhizobium meliloti has been shown to be capable of growth in the presence of methionine sulphoximine concentrations at least two orders of magnitude higher than that required for the complete inhibition of glutamine synthetase activity. Neither the specific growth rate, nor the nutritional requirements of the organism were affected by methionine sulphoximine in the medium.Rhizobium meliloti appeared to assimilate ammoniavia the glutamate dehydrogenase pathway during growth in the presence of methionine sulphoximine. This suggests thatRhizobium meliloti may have some regulatory mechanism controlling ammonia assimilation that is not present in other enterobacteria possessing similar enzymatic machinery     

Size-independent growth in fishes: patterns, models and metrics

A combination of a dynamic energy budget (DEB) model, field data on Atlantic salmon Salmo salar and brown trout Salmo trutta and laboratory data on Atlantic salmon was used to assess the underlying assumptions of three different metrics of growth including specific growth rate (G), standardized mass‐specific growth rate (G_S) and absolute growth rate in length (G_L) in salmonids. Close agreement was found between predictions of the DEB model and the assumptions of linear growth in length and parabolic growth in mass. Field data comparing spring growth rates of age 1+ year and 2+ year Atlantic salmon demonstrated that in all years the larger age 2+ year fish exhibited a significantly lower G, but differences in growth in terms of G_S and G_L depended on the year examined. For brown trout, larger age 2+ year fish also consistently exhibited slower growth rates in terms of G but grew at similar rates as age 1+ year fish in terms of G_S and G_L. Laboratory results revealed that during the age 0+ year (autumn) the divergence in growth between future Atlantic salmon smolts and non‐smolts was similar in terms of all three metrics with smolts displaying higher growth than non‐smolts, however, both G_S and G_L indicated that smolts maintain relatively fast growth into the late autumn where G suggested that both smolts and non‐smolts exhibit a sharp decrease in growth from October to November. During the spring, patterns of growth in length were significantly decoupled from patterns in growth in mass. Smolts maintained relatively fast growth though April in length but not in mass. These results suggest G_S can be a useful alternative to G as a size‐independent measure of growth rate in immature salmonids. In addition, during certain growth stanzas, G_S may be highly correlated with G_L. The decoupling of growth in mass from growth in length over ontogeny, however, may necessitate a combination of metrics to adequately describe variation in growth depending on ontogenetic stage particularly if life histories differ.     

Distribution of cells in different stages of the cell cycle and some age-related physiological characteristics in dependence on the dilution rate in chemostat cultures ofCandida utilis

Position of cells in their cell cycle was determined microscopically in chemostat cultures ofCandida utilis. Proportion of cells in phase G₁ decreased in a linear manner from 86% to 58% with dilution rate. Proportion of cells in phase S increased in the same range ofD from 5.6 to 13.5% and in the (G₂+M) phase from 8.4 to 28.5%, again linearly. Differential centrifugation was used to separate chemostat cultures to mother and daughter cells. Analyses showed that, relative to mother cells, daughter cells contain 2.1–11.9% more protein and 25.5–34.6% more RNA in dry matter. Their mass is 34.4–5.6% lower and volume is 154–19% smaller.     

Long term delayed luminescence: A possible fast and convenient assay for nutrition deficiencies and environmental pollution damages in plants

Long Term Delayed Luminescence (LDL) of green plants ranging from 0.3 s up to several minutes after irradiation has been found to exhibit complex decay kinetics which are highly dependent on nutritional deficiencies and environmental pollutive components. As a model organism we utilized the unicellular green algaScenedesmus obliquus with fairly well understood properties ofLDL probably involving both photosynthetic reaction centers PS I and PS II. It is shown thatLDL is strongly affected both by depletion of the growth medium of various essential elements such as N, Fe, Ca, Mg or K, as well as by growth in the presence of environmental stress factors such as Cu, Hg, Cd or NO ₂ ^– . Therefore,LDL offers itself as a convenient, highly sensitive and specific assay for a number of stress factors in photosynthesizing plantsin vivo and in the field.     

The structure ofHLA-B35 suggests that it is derived fromHLA-Bw58 by two genetic mechanisms

The primary structure ofHLA-B51 andHLA-Bw52 suggested thatHLA-B51 was derived fromHLA-Bw52 by the combination of a genetic exchange withHLA-B8 and a point mutation. To investigate the evolution of theHLA-B5 cross reactive group, theHLA-B35 gene was cloned and the primary structure was determined.HLA-B35 is identical toHLA-Bw58 except in the α₁ domain. The α₁ domain ofHLA-B35 except Bw4/Bw6-associated amino acids is identical to that ofHLA-B51 ^*, which was suspected to be an intermediate gene betweenHLA-B51 andHLA-Bw52. These data suggest thatHLA-B35 has evolved fromHLA-Bw58 in two steps; an in vivo replacement of the α₁ domain withHLA-B51 and genetic exchange with one of theHLA-Bw6 genes. These three genes andHLA-Bw58 are postulated to share a common ancestor.     

Genetic pathways controlling carpel development inArabidopsis thaliana

Carpel development inArabidopsis is known to be controlled by the organ identity geneAGAMOUS. However, even in the absence of AGAMOUS function, many carpel properties can arise suggesting that other genes are also involved. Two new carpel genes,CRABS CLAW andSPATULA, have been recognised by their specific disruptions to carpel development in mutant plants. These disruptions suggest thatCRABS CLAW normally plays a role in promoting the growth of specific regions of the carpel wall, whereasSPATULA apparently has a primary function in promoting development of the transmitting tract. When the function of these genes is also compromised along with that ofAGAMOUS in multiply mutant plants, carpelloid properties vanish. ThusAGAMOUS, CRABS CLAW andSPATULA act together in specifying carpel development, although none can do this alone. BecauseSPATULA mutants are epistatic to mutants of another carpel development gene,ETTIN, the latter may normally act by suppressing the action ofSPATULA in specific regions of the developing gynoecium. There is indirect evidence thatETTIN, and another morphogenetic gene,PINOID, act through regulating auxin-induced growth in specific regions of the developing flower, but it is not yet known how this could result in the suppression of SPATULA function. The extended abstract of a paper presented at the 13th International Symposium in Conjugation with Award of the International Prize for Biology “Frontier of Plant Biology”     

Note on a combinatorial problem

In the first part of this paper we have assembled some properties of the quantitiesR _m ⁿ , whereR _m ⁿ denotes the number of distributions ofn different objects intom indifferent parcels, with no empty parcels allowed. We then discuss the following problem (N. Rashevsky, 1954, 1955 a,b, 1956): to find the total number,G _n , of graphs that can be obtained from the biotopological transformation (T ⁽¹⁾ X) for a given value of the parametern. This is related to the distribution ofn indifferent objects intom different boxes. A formula forG _n is given which, however, is not very convenient for practical computations because it involves a summation over certain “admissible partitions” of the numbermn (m is a second parameter of the transformation). Some theorems are derived; with their help we can simplify the calculation ofG _n to a small extent. The numbersG _n are calculated forn≤9 and estimated forn=10. It is found thatG ₇≈5.4×10⁴,G ₈≈8.3×10⁵,G ₉≈1.4×10⁷, andG ₁₀≈3×10⁸. These values ofn are those which might be used in connection with N. Rashevsky’s work (cf. Rashevsky, 1956).     

Lack of genetic relatedness betweenYersinia philomiragia (the “philomiragia” bacterium) andYersinia species

Four strains ofYersinia philomiragia were studied biochemically and genetically. They showed a high degree of phenotypic similarity and formed a homogenous DNA relatedness group. We could not demonstrate any significant DNA relatedness between this group and otherYersinia species, other Enterobacteriaceae, orPasteurella multocida. Y. philomiragia is phenotypically quite different from other yersiniae and Enterobacteriaceae. These DNA and phenotypic data indicate thatY. philomiragia is not a member of the genusYersinia or a member of the family Enterobacteriaceae. It is suggested that, pending further study, this organism be referred to as the “Philomiragia” bacterium.     

Characterization ofClostridium bifermentans and its biotransformation of 2,4,6-trinitrotoluene (TNT) and 1,3,5-triaza-1,3,5-trinitrocyclohexane (RDX)

Summary We have determined that an organism able to degrade both RDX and TNT in a pure culture is a strain ofClostridium bifermentans. The consortium from which this organism is derived also degrades these compounds, and we suspect thatC. bifermentans is also the responsible organism within that consortium. The bioconversion of RDX and TNT occurs under anaerobic conditions both in the consortium and in pure culture without the need of an added reductant. The presence of co-metabolites speeded these biotransformations.     

A multicopy suppressor ofnin1-1 of the yeastSaccharomyces cerevisiae is a counterpart of theDrosophila melanogaster diphenol oxidase A2 gene,Dox-A2

NIN1 is an essential gene for growth of the yeastSaccharomyces cerevisiae and was recently found to encode a component of the regulatory subunit of the 26S proteasome. Thenin1-1 mutant is temperature sensitive and its main defect is in G₁/S progression and G₂/M progression at non-permissive temperatures. One of the two multicopy suppressors ofnin1-1, SUN2 (SUppressor of Nin1-1), was found to encode a protein of 523 amino acids whose sequence is similar to those ofDrosophila melanogaster diphenol oxidase A2 and the mouse mast-cell Tum^– transplantation antigen, P91A. The C-terminal half of Sun2p was found to be functional as Sun2p at 25° C, 30° C, and 34° C but not at 37° C. The open reading frame (ORF) of theDrosophila diphenol oxidase A2 gene (Dox-A2) was obtained from a lambda phage cDNA library using the polymerase chain reaction technique. TheDox-A2 ORF driven by theTDH3 promoter complemented the phenotype of a strain deleted forsun2. ThisDox-A2-dependent strain was temperature sensitive and accumulated dumb-bell-shaped cells, with an undivided nucleus at the isthmus, after temperature upshift. This morphology is similar to that ofnin1-1 cells kept at a restrictive temperature. These results suggest thatSUN2 is a functional counterpart ofDox-A2 and that these genes play a pivotal role in the cell cycle in each organism.     

Prototheca filamenta: A fungus not an alga

The genusPrototheca is composed of colorless algae. Plastids, a characteristic feature of colorless algae, are absent fromP. filamenta cells. The growth pattern seen with light and scanning electron microscopes, and the ultrastructure observed with the transmission electron microscope suggest thatP. filamenta is a fungus. Consequently we recommend the removal ofP. filamenta from the genusPrototheca and subsequent investigation of this organism to determine its correct taxonomic placement among the fungi.     

Interaction of alleles of therelA, relC andspoT genes inEscherichia coli: Analysis of the interconversion of GTP, ppGpp and pppGpp

Summary Mutants in thespoT gene have been isolated as stringent second site revertants of therelC mutation. These show varying degrees of the characteristics associated with thespoT1 gene,viz relative amount and absolute levels of both pppGpp and ppGpp and the decay rate of the latter. The entry of³H-guanosine into GTP and ppGpp pools inspoT ⁺ andspoT1 cells either growing exponentially or during amino acid starvation was determined, and the rate of ppGpp synthesis and its decay constant calculated. During exponential growth the ppGpp pool is 2-fold higher, its decay constant 10-fold lower, and its synthesis rate 5-fold lower inspoT ^- than inspoT ⁺ cells; during amino acid starvation the ppGpp pool is 2-fold higher, its decay constant 20-fold lower, and its synthesis rate 10-fold lower inspoT than inspoT ⁺ cells. In one of the “intermediate”spoT mutants the rate of entry of³H-guanosine into GTP, ppGpp and pppGpp was measured during amino acid starvation. The data form the basis of a model for the interconversion of the guanosine nucleotides in which the flow is:GDP→GTP→pppGpp→ppGpp→Y. Calculations of the rates of synthesis and conversion of pppGpp and ppGpp under various conditions in variousspoT ⁺ andspoT ^- strains indicate that the ppGpp concentration indirectly controls the rate of pppGpp synthesis. ThespoT1 allele was introduced into various relaxed mutants. It was shown that many phenomena associated with the relaxed response ofrelC and “intermediate”relA mutants were phenotypically suppressed when thespoT1 allele was introduced into these mutants. These double mutants exhibit ppGpp accumulation, rate of RNA accumulation, rate of β-galactosidase synthesis, and heat lability of β-galactosidase synthesized during amino acid starvation similar to the stringent wild-type. It is concluded that the relaxed response is due directly to the lack of ppGpp and that the stringest response is due directly to ppGpp.     

Comparison of growth and cellulolytic enzyme production inAspergillus chevalieri andPenicillium steckii from mouldy cacao beans

Aspergillus chevalieri andPenicillium steckii grew best at 30°C and at pH of 6.5–7.5. Among the carbon sources employed, sucrose supported maximum growth ofA. chevalieri while glucose was best forP. steckii. Growth of both organisms was optimal on ammonium tartrate as the sole source of nitrogen.A. chevalieri andP. steckii grew in synthetic media containing, respectively, soluble or insoluble cellulose as the sole carbon source, releasing a cellulolytic enzyme into the medium. The enzymes from each organism were separated and partially purified by molecular exclusion and ion-exchange chromatography into two components. There was synergism between the components of enzymes from each organism in that they together released more glucose units from insoluble cellulose than could be predicted from their activities alone. The molar mass of the enzymes estimated from the elution volume on Sephadex was approximately 110 kg/mol forA. chevalieri and 94 kg/mol forP. steckii.     

Optimal strategy of plant antiherbivore defense: Implications for apparency and resource-availability theories

Plants produce chemicals as methods against animal herbivory. Such chemical defenses are classified into two major categories: (i) quantitative defenses with massive production of indigestible substances; and (ii) qualitative defenses with production of poisonous substances. A mathematical model was developed that identified factors that favored the evolution of quantitative defenses. Selecting an annual plant for simplicity, the allocation of photosynthetic production between growth substances and defense substances was considered. If the plant invests more in defense substances, it can protect itself more efficiently from herbivory but with a reduced growth rate. If it invests more in growth substances, the contrary holds. Using Pontoryagin's maximum principle, the following results were obtained: (i) the plant should conduct quantitative defenses when the growth rate (G), reflecting resource-availability, is low and the growth period (T) is long as well; (ii) if the plant invests in quantitative defenses, the optimal proportion of defense substances (χ^*) should be higher asG is smaller, but it is independent ofT; and (iii) the value of χ^* is not monotone for the effectiveness of defense substance (A), but has a maximum at an intermediate value ofA. Predictions of the model partly supported both Feeny's apparency theory, claiming that apparent plants or their parts for herbivores should quantitatively defend themselves, and Coley's resource-availability theory, claiming that plants with rich resources should invest in growth rather than defense.     

Optimization of culture conditions and continuous production of chitosan by the fungi,Absidia coerulea

The production of chitosan from the mycelia ofAbsidia coerulea was studied to improve cell growth and chitosan productivity. Culture conditions were optimized in batch cultivation (pH 4.5 agitator speed of 250 rpm, and aeration rate of, 2 vvm) and the maximum chitosan concentration achieved was 2.3 g/L under optimized conditions. Continuous culture was carried out successfully by the formation of new growth spots under optimized conditions, with a chitosan productivity of 0.052 gL⁻¹ h⁻¹, which is the highest value to date, and was obtained at a dilution rate of 0.05 h⁻¹. Cell chitosan concentrations reached about 14% in the steady state, which is similar to that achieved in batch culture. This study shows that for the continuous culture ofAbsidia coerulea it is vital to control the medium composition.     

Infection of procaryotic and eucaryotic microorganisms withMetallogenium

To elucidate the biological characteristics ofMetallogenium, a study was undertaken into the effect of infection of such organisms unable to oxidize manganese as fungi, yeasts, and bacteria with an ultrafiltrate of this organism on the subsequent behavior of the resulting binary cultures. The infection of microorganism cultures is accompanied by (a) acquisition by the binary cultures of a persistent capacity to oxidize manganese, (b) evolution of the characteristic structures ofMetallogenium (c) inhibition of the growth of the inoculated cultures; the inhibition manifests itself by fungi losing their capacity for spore formation and pigmentation, the upsetting of cell division processes, and lysis of the cells of the infected cultures all the way to death. When a heated ultrafiltrate ofMetallogenium was used to infect microorganism cultures, no signs of infection were detected.Metallogenium may be hosted by an extremely broad range of microorganisms that are in no way allied to one another. The experimental results suggest thatMetallogenium is an organism capable of parasitizing lower eucaryotic and procaryotic microorganisms. Analysis of 134 strains of fungi isolated from freshwater bodies is indicative of possible parasitism ofMetallogenium on microorganisms in their natural habitats.     

The growth kinetics ofB. subtilis

There has been considerable discussion by Kubitschek and Cooper concerning the growth rate of cells ofE. coli throughout the cell cycle. Consequently, it is relevant to test Kubitschek's linear model against the exponential model espoused by Cooper (and many others) with another organism and another technique.Burdett et al. measured, by electron microscopy and computer analysis of the microphotographs, the distribution of lengths of a population of cells ofBacillus subtilis grown in 0.4% succinate in a minimal medium. The data were fitted to the extended Collins-Richmond method of Kirkwood & Burdett which subdivided the cell cycle into several phases. I have taken their results and compared them with the linear and exponential growth models for the entire cell cycle after applying correction to the data for the shape of completed and forming poles; i.e., to put the data on a cell-volume basis instead of a cell-length basis. Most of the correction involves no arbitrary assumptions. The conclusion is that global volume growth rate is nearly proportional to cell volume; i.e. growth ofBacillus subtilis is nearly exponential for almost every cell in the growing culture.     

A new model estimating stonefly species production in the West Carpathian torrents

Samples of stoneflies were collected from 15 streams of Slovakia during 1976–2000. The model of growth rate is based on 219 data of 50 stonefly species. The non-linear relationship among growth rate (G) of stoneflies and monthly mean water temperature (T) and body mass (W) is described by equation: G = 0.014W ^−0.19 T ^0.25. The model estimates the species production of the families Taeniopterygidae, Nemouridae, Capniidae, Leuctridae and Perlodidae. This model is combined with Morin’s & Dumont’s (1994) model for Perlidae and Chloroperlidae. There is positive evidence that the total stonefly production of mountain and submountain streams increases with discharge. On the other hand, increasing altitude has a negative influence on production.     

Diazotrophic mixed culture ofAzotobacter vinelandii andRhodobacter capsulatus

The question, whetherAzotobacter vinelandii can provide fixed N for the growth of other organisms, was studied with mixed cultures ofA. vinelandii andRhodobacter capsulatus, grown with aeration in the light. N₂-fixation byR. capsulatus was prevented by growing the cultures on either mannitol, glycerol or ethanol, which cannot be used by this organism. In the course of growth with mannitol, cell numbers of both organisms increased largely in parallel and attained a maximal ratio of about oneA. vinelandii per tenR. capsulatus. Prolonged growth of mixed cultures with mannitol did not lead to an adaptation ofR. capsulatus to this compound. After growth on either one of the three alcohols, mixed cultures exhibited almost twice as high protein levels as pure cultures ofA. vinelandii. Up to 80% of the protein of mixed cultures was incorporated intoR. capsulatus. The results suggest thatA. vinelandii provided an organic N-source for the growth ofR. capsulatus.     

Transfer ofHaemophilus equigenitalis Taylor et al. 1978 to the genusTaylorella gen. nov. asTaylorella equigenitalis comb. nov.

The taxonomic position ofHaemophilus equigenitalis Taylor et al. 1978, the causative agent of contagious equine metritis, was studied in comparison with phenotypically similar organisms such asMoraxella, Legionella, and others.Haemophilus equigenitalis is a microaerophilic, Gram-negative, nonmotile, short rod; the mean base composition of deoxyribonucleic acid of this organism is 36.5±0.5 mol% G+C. It shows best growth on chocolate agar, but very poor or no growth on plain nutrient agar and blood agar, although it requires no X- or V-factors for the growth. It is positive in catalase, oxidase, phosphatase, and phosphoamidase tests, but very unreactive in other biochemical tests for routine use. It produces no acid from any carbohydrates nor glycosidase. Arylamidase activities of the organism to -naphtylamide derivatives of various amino acids, and di- and tripeptides were also compared with those of other taxa. The group of this organism was different from other known taxa in the numerical analysis of its phenotypic characteristics, DNA base composition, and DNA-DNA hybridization. These data indicate thatH. equigenitalis does not belong in the genusHaemophilus nor other known genera, but rather in a new genus. Therefore, we propose thatHaemophilus equigenitalis be reclassified in a new genusTaylorella asTaylorella equigenitalis.