Continuous culture of Humicola lutea 120-5 for acid proteinase production

Acid proteinase production by the fungus Humicola lutea 120-5 in continuous culture was studied. The maximum activity of the culture broth reached 2200 U/ml at a dilution rate (D) of 0.05/h. The continuous process was carried out for 1 month without any bacterial contamination, due to low pH (3.0–3.5) during the cultivation.     

Continuous culture with deep seawater of a benthic food diatom Nitzschia sp.

A continuous culture system for a benthic food diatom Nitzschia sp. wasestablished by using properties of high nutrient and clean of deep seawater(DSW). DSW collected from 320 m depth in Muroto City, Japan, was introducedinto a glass-pipe bioreactor (14 cm length, 3 cm diam.) containing glassbeads of 0.5 cm diam. as substrata for the alga, and it was incubated at18°C · 80Em^–2sec^–1 · L:D=14:10. The chlorophyll a yield of benthicdiatoms in a reactor as a unit of surface area of the substratum was only0.001–0.003 g cm^–2 when the flow rate of DSW was 0(batch culture conditions). However, when DSW was supplied continuously to areactor, the yield increased to 1.4 g-chl.a cm^–2 alongwith the increase in flow rate of DSW. Moreover, amounts of chl.a washed outof the system were negligible, 0.0014 to 0.0045%, even though theflow rate of DSW was as much as 25 times h^–1, suggesting thatsloughing of benthic diatoms from the substratum was minimized. Although theyield of diatoms fluctuated significantly at the time that the DSW wascollected, the variation could be minimized by increasing the flow rate ofDSW. These results indicate that the continuous culturing system with DSWsupports the stable and effective mass culture of benthic food diatom.     

Myelin basic protein purified on an ion-exchange continuous polymer bed in the presence of ethylene glycol and salt possesses activity againstp-nitrophenyl acetate

In this paper we describe a fast and mild method based on the use of a unique cation exchanger and buffers containing ethylene glycol and salt for the purification of the myelin basic protein (MBP; MW 18.5 kDa). MBP thus purified hydrolyses catalytically p-nitrophenyl acetate. This esterase activity facilitates not only the purification of MBP but also indicates that probably it is in its native state, i.e. there is a good chance that the purified molecules are structurally and chemically identical. This is a prerequisite to obtain crystals appropriate for x-ray diffraction and other studies.Abbreviations used MBP myelin basic protein - MW molecular weight - kDa kilo Dalton - octyl-POE n-octylpolydisperse oligooxyethylene - CHAPS 3-3-cholamidopropyl dimethylammonio-1-propane-sulfonate - CTAB cetyltrimethylammonium bromide - SDS sodium dodecyl sulfate - SDS-PAGE polyacrylamide gel electrophoresis in the presence of SDS - G 3707 heptaoxyethylene lauryl ether - TWEEN-20 polyoxyethylenesorbitan-monolaurat - EDTA ethylenediaminetetraacetic acid - HEPES N-(2-hydroxyethyl)-piperazine-N-(2-ethanesulfonic acid)     

Carotenoid content in growing cells of Haematococcus pluvialis during a sunlight cycle

Under certain culture conditions, cells of the chlorophyte Haematococcus pluvialis accumulate significant amounts of astaxanthin. This study describes biomass and carotenoid production during a sunlight cycle in a continuous culture of growing cells of H. pluvialis and shows that these two parameters are under the control of irradiance. The hourly carotenoid production increases with light intensity and, in our culture conditions, carotenoid accumulation occurs in a few hours and without any morphological change in the algae. These carotenoids seem to be efficient in protecting algal cells against photoinhibition damage if their content is greater than 1% dry biomass. Below this concentration, that is to say in the early hours of high light intensity, dry biomass decreases due to cell lysis. The results demonstrate that secondary carotenoid accumulation in H. pluvialis may occur in the active growth phase and is stimulated from the first hours of sunlight illumination.     

Population structure and mycelial phenotypic variability of the ectomycorrhizal basidiomycete Laccaria bicolor (Maire) Orton

Mating type allele distribution and phenotypic variability were investigated in field populations of Laccaria bicolor. Sporophores associated with Norway spruce (Picea abies), colonized by natural sources of inoculum and growing in a seed orchard, were sampled to obtain dikaryotic strains and assay their phenotypic variability for traits important to the symbiosis. Basid-iospores were also collected for mating type analysis of different mycelia. Four sporophore mating types were identified containing seven A and five B factors. Out-breeding efficiency was estimated at 73.8% and the population was slightly inbred. Crosses with previously characterized L. bicolor strains from two nearby populations identified in total six sporophore mating types and ten A and nine B factors, for an estimated outbreeding efficiency (85.7%) similar to previous studies of more spatially disparate Laccaria spp. populations. Dikaryotic strains were tested for mycelial growth rate, as a measure of their competitive ability, on agar media containing a soluble (NaH₂PO₄), or an insoluble (CaHPO₄) phosphate source. Their ability to solubilize the latter was also tested to assess their relative capacity to access insoluble, inorganic phosphate. In most cases, significant variation was detected among strains from the same site for all variables. On three sites (VC4, VC5 and VC7), each determined previously to possess a uniform mycelial genotype, phenotypic variability was likely due to epigenetic variation among different strains of the same genotype. Possible evidence for dikaryon-monokaryon crosses was observed in vivo on one sample site (VC2) where adjacent mycelia shared two mating factors. The phenotypic variability of different mycelial genotypes reflected their genetic variability observed as mating type allele diversity and underlined the importance of basidiospore dispersal in introducing new genotypes into the population. The reproductive strategies of L. bicolor are discussed and compared to those of other basidiomycete species.     

Differentiation of vegetation zones and species strategies in the subalpine region of Mt. Fuji

The floristic and structural differentiation of vegetation along the altitudinal gradient in four subalpine forests of different developmental stages on Mt. Fuji has been studied. Near the forest limit a micropattern of vegetation corresponding to the altitudinal zonation has been observed which elucidated the mechanisms of development of the vegetation zonation. As to early stages of vegetation development only two types can be distinguished: the volcanic desert above 1500 m and the pioneer forests below. As to later stages a differentiation of subzones includes from higher to lower altitudes: the Alnus maximowiczii, Betula ermanii, Abies veitchii and Tsuga diversifolia forests. Larix leptolepis and Sorbus americana ssp. japonica, appear as co-dominants in ecotonal communities between the principal subzones and are also important pioneers in early stages. Similarity analyses reveal that the upper subalpine Alnus-Betula forests can be regarded as early successional phases of the climax Abies-Tsuga forests of the lower subalpine zone. The regular arrangement of A. maximowiczii-B. ermanii-A. veitchii is studied along the gradient from the margin to the interior of the forest growing near the forest limit where locally favourable conditions prevail. Growth form, height growth, photosynthetic activity, seed supply, and seedling distribution of the three principal species have been compared, as well as biomass and production relations in contiguous forests of these species. The marginal Alnus type community is productive and disturbance-tolerant, and has a wide ecological and sociological amplitude along the gradient, while the central Abies community is accumulative and disturbance-intolerant, and has a narrower tolerance range, but is superior in competition under stable habitat conditions. A vegetation organization, ‘temporal multi-storeyed structure’, is suggested which means that a zonal pattern of vegetation within a climax region develops by successive replacement of successional species along an environmental gradient.     

Evolutionary stable strategies and game dynamics for n-person games

This note contains a generalization of the definition of an evolutionary stable strategy and of the corresponding game dynamics from 2-person to n-person games. This broader framework also allows modelling of several interacting populations or of populations containing different types of individuals, for example males and females.     

Calluna and its associated species: Some aspects of co-existence in communities

Summary Using heath communities (dominated byCalluna vulgaris) as an example, an attempt is made to identify those characteristics of other species wiich enable them to co-exist with the dominant. In the course of its life-span,Calluna passes through a series of growth phases (pioneer, building, mature, degenerate) during which there are marked changes in the condition affecting associated species. These display a variety of ‘complementary strategies’ which enable them to co-exist withCalluna during a part, or the whole, of the sequence. Changes with time in the flora of even-aged stands ofCalluna, produced as a result of management by burning, are accounted for in similar terms. It is also shown that invasion of heath communities by tree species (e.g.Betula spp.,Pinus sylvestris), which may eventually eliminateCalluna, is related to the sequence of growth-phases. Contribution to the Symposium on plant species and plant communities, held at Nijmegen, 11–12 November 1976, on the occasion of the 60th birthday of Professor Victor Westhoff-to whom the author conveys the good wishes of Scottish botanists and conservationists, and particularly those of the University of Aberdeen. Nomenclature follows Clapham, Tutin & Warburg (1962) for vascular plants, Watson (1968) for bryophytes, and Duncan (1970) for lichens. Following common practiceCalluna vulgaris is referred to simply asCalluna. Grateful acknowledgement is made to several present and former research students who have kindly permitted the use of their data: particularly P. Barclay-Estrup, Edith M. French, Gong Wooi Khoon, C.J. Legg, Evelyn W. Paterson, S.D. Ward. I am especially grateful to C.J. Legg for valuable discussions on the subject of this paper.     

Aspects of population dynamics in Halimione portulacoides communities

Summary Studies on sample plots inHalimione portulacoides communities show that environmental disturbances, either natural or induced by man, start a sequence of partly overlapping density maxima inSuaeda maritima, Aster tripolium andPuccinellia maritima successively, before the originalHalimione community totally recovers. When succession time before recovering is long enough, there are tendencies in redundancy of this sequence stressing the unilinear character of the succession. Minor environmental impacts induce a longer time-lag period of theSuaeda density maximum, suggesting threshold values of these impacts for the species to maintain minimal population densities or to become locally extinct. This sequence of interim species starting after an environmental disturbance, suggests also a gradient character in various biological attributes, for instance in life-time, propagation, nutrient and genetic plasticity strategies. The mechanism described can therefore be interpreted as a complex of mostly well-adapted and well-integrated inherent species strategies capable of absorbing environmental shocks. It is suggested that in the salt-marsh ccosystem the pattern of spatial variation in densities and that of temporal variation in fluctuations of the three species populations under natural conditions reflect corresponding patterns of environmental disturbances in the vegetation taking into account a timelag associated with the magnitude of the impact concerned.Contribution to the Symposium on Plant species and plant communities held at Nijmegen, 11–12 November 1976, on the occasion of the 60th birthday of Professor Victor Westhoff.Nomenclature follows Heukels-van Ooststroom. Flora van Nederland, 18e druk, 1975. Wolters-Noodhoff, Groningen.The authors are greatly indebted to Dr K.F. Vaas (Yerseke) for reviewing the English text.Communication Nr. 160.     

Estimating the maintenance energy and biomass concentration of Saccharomyces cerevisiae by continuous calorimetry

Continuous calorimetry has been applied to monitoring the heat evolution of Saccharomyces cerevisiae grown on d-glucose. The heat evolution, together with the energy and carbon balances, was used to evaluate the energetic efficiency of biomass, by-product biosynthesis, fermentative heat evolution as well as the maintenance energy of S. cerevisiae in ‘aerobic fermentation’ and ‘aerobic respiration’. In aerobic fermentation, under catabolite repression, the fraction of substrate energy converted to heat evolution, maintenance requirement, and biomass decreased with the increase of d-glucose concentration. The fraction of substrate energy converted to ethanol is the highest value and it could contribute up to 70% of the total substrate energy. In aerobic respiration, 43% of the total substrate energy was evolved as heat. While 50% of the total substrate energy was converted into biomass, only 7% of the total substrate energy was used for maintenance functions. The maintenance energy coefficient of S. cerevisiae was determined to be 0.427 MJ kg^?1 cell h^?1 (0.102 kcal g^?1 cell h^?1). For the first time, heat evolution together with yield-maintenance energy was used to predict biomass concentration during the fed-batch cultivation of S. cerevisiae.