Growth Stimulation by Conditioned Medium and Spermidine in Low-Density Suspension Cultures of Rice

Suspension cultures of Oryza sativa L. var IR 20 grew in Murashigeand Skoog medium (MS) supplemented with 2,4-D and kinetin ina density-dependant manner with a critical minimum inoculation-densityof 10,000 cells ml^–1. Conditioned medium obtained fromthese cultures and added to MS+2,4-D+kinetin induced the growthof cultures at 1,000 cells ml^–1. Growth stimulation byconditioned medium was mimicked by spermidine but not by otherpolyamines viz. putrescine and spermine. This is the first reportof a polyamine substituting for conditioned medium in cultures. ² Present address: Vice-Chancellor, Pondicherry University,Pondicherry 605 014, India.     

Studies on the Growth in Culture of Plant Cells: IV. THE INITIATION OF DIVISION IN SUSPENSIONS OF STATIONARY-PHASE CELLS OF ACER PSEUDOPLATANUS L.

Techniques are described whereby a culture medium can be ‘conditioned‘by separation from a dense cell suspension either by a sinteror by a dialysis membrane. The enhanced growth-promoting activityof the conditioned, as compared with a new medium, is revealedby using a low density of cells (15 x 10³ or less cells perml) to initiate the test cultures from a stationary-phase suspension.The optimum pH of the conditioned medium is c6.4. To obtaina conditioned medium of high activity it is necessary to usean appropriate volume ratio of culture medium to conditioningcell suspension and to limit the conditioning period. Conditioningof the culture medium reduces by a factor of 10 (i.e. down toc. 1000 cells per ml) the minimum effective cell density neededfor self-sustaining growth. There therefore exists a population-dependentrequirement which is not met by the conditioned medium as nowprepared. The retention of the activity of the conditioned mediumin various situations has been studied as a preliminary to workon the chemical basis of conditioning.     

Density and distribution of bacterioplankton and planktonic ciliates in the Bering Sea and North Pacific

The total number of planktonic bacteria in the upper mixed layerof the Bering Sea during the late spring-early summer periodranged between 1 and {small tilde}4 x 10⁶ ml^–1 (biomass10–40mg C m^–3). In the northern Pacific, along 47–52⁶N,the corresponding characteristics of the bacterioplankton densityin the upper mixed water layer were: total number 1–2x 10⁶ cells ml^–1 and biomass 15–46mg C m^–3Below the thermocline at 50–100 m, the density of bacterioplanktonrapidly decreased. At 300 m depth, it stabilized at 0.1–0.2x 106 cells ml^–1. The integrated biomass of bacterioplanktonin the open Bering Sea ranged between 1.2 and 3.6 g C m^–2(wet biomass 6–18 g m^–2) Its production per dayvaried from 2 to 23 mg C m^–3 days^–1 in the upper0–100 m. The numerical abundance of planktonic ciliatesin this layer was estimated to be from 3 to l0 x 10³ cells l^–1,and in the northern Pacific from 0.4 to 4.5 x 10³ l^–2.Their populations were dominated by naked forms of Strombidium,Strombilidium and Tontonia. In some shelf areas, up to 40% ofthe total ciliate population was represented by the symbioticciliate Mesodinium rubrum. The data on the integrated biomassof basic groups of planktonic microheterotrophs are also presented,and their importance in the trophic relationships in pelagiccommunities of subarctic seas is discussed.     

Bacterial biomass and production in a shallow bay

Bacterial biomass (BB, acridine orange) and bacterial secondaryproduction (BSP, [³H]thymidine incubations) were measured forthe first time in Concepción Bay (37°35'S, 73°01'W).BB ranged from 1.8x10⁶ to 22.5x10⁶ cells ml^–1 (the lattervalue observed near to an industrial effluent), BSP from 0.27x10⁶to 2.5x10⁶ cell ml^–1 day^–1 and heterotrophic turnoverrates between 0.15 and 1.0 doublings day^–1.     

Bacterivory in seawater samples estimated by a dual radioactive-labelling technique

A dual radioactive-labelled bacteria technique using Vibrio(DRLV), developed for laboratory studies on bacterivory, hasbeen refined for use at the concentrations of prey and predatorstypcially found at sea. Experiments with estuarine water collectedin spring and in autumn showed that bacterivorous nanoflagellates(HNF) (concentration 1.38±0.35x10³ HNF ml^–1) ingested2.7±0.96 DRLV flagellate1^–1 h^–1 at concentrationsof 0.8–2.2x10⁶ DRLV ml^–1 in the presence of 2.04±0.68x10⁶natural bacteria ml^–1. The method was also applied tosamples collected in October in the Celtic Sea, when on average1 ml of water from the surface layer contained 1.41±0.16x10⁶natural bacteria, 14.6x10³ cyanobacteria, 530±170 HNF,7.3±3.0x103 phototrophic nanoflagellates (1.5–4µm), 49.0±26.5 phototrophic dinoflagellates, 36.3±12.6heterotrophic dinoflagellates and 21.3±9.5 Leucocryptosmarina. Under these conditions the grazing rate in most samplesdid not exceed the coefficient of variation of the method (2%),although we estimate the grazing rate was -1.6 DRLV HNF^–1h^–1 and on one occasion a rate of 2.45 was recorded. Thegross growth efficiency for protein of -30% displayed by naturalHNF means that they could release about     

Discrimination of female sex pheromone by male Trichoplusia ni (Hubner)

The olfactory discrimination process of male cabbage loopermoths, Trichoplusa ni (Hübner), was assessed by measuringtheir response to one of two emission sources within a windtunnel. The males discriminated between (1) Z7–12:Ac concentrations;(2) Z7–12:Ac alone and the volatile emission from excisedfemale sex pheromone glands; and (3) Z7–12:Ac and theemission from a mixture of six synthetic pheromone componentsthat mimics the volatile emissions of a female gland. Althoughmales could discriminate between a freshly excised female sexpheromone gland and 7.4x 10^–11 M Z7–12:Ac, theycould not discriminate between a gland and 78.5x10^–11M Z7–12:Ac. Males also could not discriminate betweenthe mixture of six volatile compounds and 28.7x10^–11 Mof Z7–12:Ac. The data show that male cabbage looper mothshave difficulty discriminating between Z7–12:Ac aloneand in mixtures with other female-emitted volatile compounds.     

Initiation and Growth of Callus and Cell Suspensions of Theobroma cacao L

Callus and suspension cultures of Theobroma cacao L., initiatedfrom immature cotyledons of beans from pods harvested 120–130days after pollination were established. A modified B-5 or Murashige—Skoogagar medium sustained growth of callus without loss of vigourafter each sub-culture. A 15-fold weight increase occurred duringthe 4 week culture periods at 30 ± 1 °C. Coconutwater improved callus growth substantially. The optimum hormonalconcentrations for growth of suspensions were 0.5 mg 1^–1of 2, 4-dichlorophenoxyacetic acid and 0.1 mg I^–1 of kinetinin a Murashige—Skoog basal medium liquid medium. The optimumtemperature for growth of suspensions was 25–30 °C.The cell number and cell mass of suspensions increased 20-foldin 14 days. No organogenesis or embryogenesis was observed. Theobroma cacao L., acao, cell culture, suspension culture, tissue culture.     

Population structure and swarm formation of the cyclopoid copepod Dioithona oculata near mangrove cays

The cyclopoid copepod Dioithona oculata forms swarms in water>30 on deep among prop roots of red mangroves (Rhizophoramangle) which fringe protected areas of two lagoonal cays, TwinCays, Belize. During 7 of 8 months surveyed by in situ observation,swarms were present but differed in size from small cylindricalswarms (5–10 cm diameter) to bands extending up to 1200m Swarms were never observed at night Swarms formed at dawnwhen light intensities reached an average value of 13.82 (logioquanta cm^–Abstract. s^–1) and dispersed at dusk atsimilar intensities Swarms observed in June formed earlier anddispersed later in the day than swarms observed in January,their swarming behavior followed seasonal changes in light intensityMean dioithonan density in swarms (10 ml^–1) was much higherthan the mean density (0 15 ml^–1) of non-swarming dioithonansaround mangrove prop roots. In open water 3–5 m away fromthe mangroves, mean dioithonan density was 7 9 x 10^–5ml¹ during the day, and 2 68 x10^–3 ml^–1 at nightSwarms were composed predominantly of adults and copepodid stagesIV and V, although younger copepodid stages could be presentNauplii were never present. The ‘average copepodid stage’for all 95 swarms sampled was 5 3, where 6 0 represents a swarmwith only adults In open water 3–5 m away from the mangroves,the youngest copepodids (stage one) dominated the dioithonanpopulation during the day. At night when swarms dispersed toopen waters, average copepodid stage was higher (3 5) comparedwith the day value (1.2) in open waters. Although densitiesin swarms were higher in June than January, average copepodidstage in June was higher (5 6) than that in January (4.9). Ahigher percentage of adults were females during June than January.Therefore higher densities did not result from increases ofsmaller stages in swarms, but perhaps changes in behavior orpopulation structure.     

Effect of abscisic acid on elongation and kinetin-induced tuberization of isolated stolons of Solarium tuberosum L.

The response of isolated stolons cultured in vitro, to abscisicacid (ABA) has been studied in the presence and absence of kinetin(6-furfurylaminopurine). ABA alone in concentrations from 7.5x 10^–4 mM to 7.5 x 10^–2 mM, inhibited stolon elongationbut failed to promote tuber initiation. In the presence of kinetin,ABA at concentrations of 3.0 x 10^–2 and 7.5 x 10^–2mM markedly inhibited kinetin-induced tuber initiation and stolonelongation, but at 7.5 x 10^–4 and 7.5 x 10^–3 mMABA did not prevent tuber initiation. When stolons were incubated on a medium containing kinetin andlater transferred to one containing ABA with or without kinetin,the inhibitory effect of ABA decreased appreciably as the timeof incubation on kinetin is increased. The results are discussed in relation to the role of ABA inthe inhibition of nucleic acid and protein synthesis and theinteraction with cytokinins and the possible effect of ABA onkinetin uptake, transport and accumulation at the locus of action. (Received February 26, 1969; )     

GROWTH OF JUVENILE BITHYNIA TENTACULATA (PROSOBRANCHIA, BITHYNIIDAE) UNDER DIFFERENT FOOD REGIMES: A LONG-TERM LABORATORY STUDY

The growth of juvenile Bithynia tentaculata (Proso-branchia,Bithyniidae) was measured under controlled laboratory conditionsover 18 months. The animals were fed with different concentrationsof suspended food (Chlamydomonas reinhardii at 2, 000 cells.ml^–1and at 10, 000 cells.ml^–1), solid food (lettuce) and combinationsof both (lettuce with 2, 000 cells.ml^–1 and with 10, 000cells.ml^–1 Chlamydomonas). The growth of all animals wasmeasured weekly, and after 1 years final shell sizes, shelldry weights and ash-free dry weights, as well as soft body dryweights and ash-free dry weights were determined. Survival rateof the animals increased from 20% when fed with 2, 000 algalcells.ml^–1 to 75% with lettuce and 10, 000 cells.ml^–1as food. Final shell sizes and shell weights were not influencedby food, but soft body dry weights were significantly reducedwhen animals were fed on suspended food only. Reproduction (i.e.egg laying) was observed at the end of the second summer whenlettuce (with or without algal addition) was given as food.The ecological consequences of these results are discussed. (Received 7 April 1993; accepted 9 August 1994)     

Contribution of Picocyanobacteria to total primary production and community respiratory losses in a backwater system

The contribution of autotrophic picoplankton (APP) to phytoplanktonicprimary production, investigated during the phytoplankton growingseason (March–September) in a macrophyte-dominated backwatersystem near Vienna, showed that APP mainly consisted of rod-shapedand coccoid cyanobacteria. Two stations were examined, exhibitingsimilar seasonal patterns in the development of picocyanobacteria,although the two sites differed in picocyanobacterial cell numbersand biomass by a factor of 1.5. Cell numbers determined by epifluorescencemicroscopy varied between 0.29 x 10⁴ and 34.5 x 10⁴ cells ml^–1at Station 1, and between 0.23 x 10⁴ and 19.1 x 10⁴ cells ml^–1at Station 2. At both sites, the mean cell volume of picocyanobacteriawas 0.5 µm³. Carbon fixation in the planktonic communityof the Kühwörter Wasser was dominated primarily bylarger phytoplankton, although the picoplankton community sometimessupplied up to 74% (mean: 35%) of total primary production.Distinct differences in chlorophyll a concentrations and primaryproduction between the two sites refer to a greater competitionbetween phytoplankton and macrophytes at Station 2. Communityrespiration deviated greatly in time and in level at the twostations, showing a higher dynamic in community metabolism atStation 1. At this site, community respiration losses rangedbetween 12 and 100% of gross production. Hence, community metabolismcomprised net autotrophic, balanced, and net heterotrophic situationsover the investigation period, whereas at Station 2, only netautotrophic situations could be determined.     

Autotrophic picoplankton in southern Lake Baikal: abundance, growth and grazing mortality during summer

Autotrophic picoplankton were highly abundant during the thermalstratification period in late July in the pelagic area (waterdepth 500–1300 m) of southern Lake Baikal; maximum numberswere 2 x 10⁶ cells ml^–1 in the euphotic zone ({small tilde}15m). Unicellular cyanobacteria generally dominated the picoplanktoncommunity, although unidentified picoplankton that fluorescedred under blue excitation were also abundant (maximum numbers4 x 10⁵ cells ml^–1) and contributed up to {small tilde}40%of the total autotrophic picoplankton on occasions. Carbon andnitrogen biomasses of autotrophic picoplankton estimated byconversion from biovolumes were 14–84 µg C l^–1and 3.6–21 µg N l^–1. These were comparableto or exceeded the biomass of heterotrophic bacteria. Autotropicpicoplankton and bacteria accounted for as much as 33% of paniculateorganic carbon and 81% of nitrogen in the euphotic zone. Measurementsof the photosynthetic uptake of [^l4C]bicarbonate and the growthof picoplankton in diluted or size-fractionated waters revealedthat 80% of total primary production was due to picoplankton,and that much of this production was consumed by grazers inthe <20 µ.m cell-size category. These results suggestthat picoplankton-protozoan trophic coupling is important inthe pelagic food web and biogeochemical cycling of Lake Baikalduring summer.     

Effect of temperature and food concentration on post-embryonic development, egg production and adult body size of calanoid copepod Eurytemora affinis

Post-embryonic development time, egg production rate and adultbody size of calanoid copepod Eurytemora affinis from Lake Ohnuma,Japan, were determined under six temperature-food conditions(10³,5 x 10³, 10⁴ and 5 x 10⁴ cells ml^–1 at 15°C,5 x 10⁴ cells ml^–1 at 10and20°C) in the laboratory.The measured parameters varied with both temperature and foodconcentration. Development time from hatching to adult femalewas 9.2, 11.4 and 22.8 days at 20, 15 and 10°C respectively,at the highest food concentration. The males developed to adultat one to two days earlier than the females. An effect of foodshortage on development time occurred at the lowest food concentration.This development time was 24.8 days even at 15°C, beingtwice as long as that at the highest food concentration. Prosomelength of these food-limited females was approximately 75% ofwell-fed ones, which reduced by only 10% with increasing temperaturefrom 10 to 20°C. Clutch size (C, eggs clutch^–1) ofwell-fed individuals depended on prosome length of the adultfemale (L, mm), and was expressed as an equation: C = 65.2 L³⁸³. Clutch size of individuals reared at less than 10⁴ cellsml^–1, however, mostly laid below the estimated curve,especially at the lowest food concentration being only 10% ofthat at the highest food concentration. These results suggestthat food availability is a more important factor affectingpopulation growth of E.affinis in Lake Ohnuma than variationof temperature.     

Abundance and productivity of heterotrophic nanoplankton in Georgia coastal waters

The population abundances and rates of biomass production ofheterotrophic nanoplankton (HNAN) in Georgia coastal waterswere evaluated by epifluorescence microscopy. HNAN populations(mostly non-pigmented microflagellates <10 µm in diameter)ranged from 0.3 x 10³ cells ml^–1 in shelf waters 15 kmoffshore to 6.3 x 10³ cells ml^–1 in waters 0.25 km fromthe coast. There was a strong correlation (r = 0.83) betweenHNAN and free bacterioplankton population abundances, but noapparent relation (r = 0.38) between HNAN and phototrophic nanopLankton(PNAN) abundances. HNAN biomass production in estuarine andnearshore shelf waters, as estimated from increases in HNANpopulations during laboratory incubations of natural water samples,ranged from 0.10 to 0.79 mg C m^–3 h^–3, with populationgeneration times of 9.7 to 26.5 h. There was a significant linearrelation (r = 0.95) between HNAN biomass and HNAN productivity.We calculated that HNAN may graze at least 30% to 50% of dailybacterioplankton production in Georgia coastal waters.     

Microplankton and primary production in the Sea of Okhotsk in summer 1994

Phytoplankton composition, density, vertical distribution andprimary production were investigated in the Sea of Okhotsk andin the adjacent northern north Pacific in July–August1994, together with measurements of density and distributionof planktonic microheterotrophs: bacteria, nanoheterotrophsand ciliates. Different phases of phytoplankton seasonal successionwere encountered during the period of investigation in variousregions of this sea. Primary production measured at 144 stationswas found to be greatest (1.5–4 g C m^-2day^-1) in areasof spring-phase succession along the Sakhalin shelf and theKashevarov bank. Periodic relapses of the spring blooms of ‘heavy’diatoms during the whole growth season were recorded over thisbank. The summer phase of the phytoplankton minimum prevailedin the central and eastern parts of the sea, manifested by thedominance of nanoflagellates in terms of phytoplankton biomass.Primary production was 0.5–1 g C m^-2 day^-1. The earlyautumn phase of succession was typical of the Kurile straitarea and the adjacent north Pacific. Primary production therevaried from 0.7 to 2 g C m^-2 day^-1. The integrated phytoplanktonbiomass in the water column varied from 9–12 g m^-2 inzones supporting the summer minimum assemblage to 15–20g m^-2 in zones of early autumn recovery of phytoplankton growth,and up to 40–70 g m^-2 in areas of remnant or relapseddiatom blooms. The numerical density of bacterioplankton wasbetween 1 x 10⁶ and 3 x 10⁶ cells ml^-1 and its wet biomass wasbetween 100 and 370 mg m^-3. In deep waters it was 8–15mg m^-3. The integrated bacterioplankton biomass in the upperwater column varied from 6 to 29 g m^-2. The numerical densityof zooflagellates varied in the upper layer between 0.8 x 10⁶and 4 x 10⁶ l^-1 and their biomass was between 20 and 50 mg m^-3.In deep waters they were still present at a density of 0.05x 10⁶ to 0.2 x 10⁶ cells l^-1. The biomass of planktonic ciliatesvaried between stations from 20 to 100 mg m^-3. The joint biomassof planktonic protozoa in the water column was between 3 and12 g m^-3 at most of the stations.     

Direct Somatic Embryogenesis in Roots of Cichorium: Is Callose an Early Marker?

Direct somatic embryogenesis can be obtained from epidermaland cortical cells in roots from in vitro Cichorium plantlets.The first embryogenic cells are seen after six days of culturein darkness, at 35 °C, in a liquid medium supplemented withNAA (1 x 10^–7 M), 6-dimethylallyl-amino-purine (2·5x 10^–6 M), sucrose (0.03 M) and glutamine (1·7x 10^–3 M). Embryogenic cells undergo first a linear andthen a globular segmentation, with increasing cytoplasmic density.These cells and young embryoids show aniline blue fluorescence.SEM allows the same microglobular pattern to be seen on thesurface of young embryoids and on young microspores of Cichoriumused as controls. In this root system, callose deposition seemsto be an early marker in somatic embryogenesis. Somatic embryogenesis, callose, Cichorium     

House Production by Oikopleura dioica (Tunicata, Appendicularia) Under Laboratory Conditions

We examined the generation time and the house renewal rate ofOikopleura dioica under various conditions. Animals were fedtwo flagellates, Isochrysis galbana and Tetraselmis sp., withconcurrent determination of the carbon contents of body andhouse to estimate house production. The generation time was6 days at 15°C, 4 days at 20°C and 3 days at 25°Cat both 25 and 30 p.s.u. with a food concentration of 4 x 10⁴cells ml^–1. The carbon content of newly secreted housesranged from 0.5 to 0.8 µg, corresponding to 15.3 ±4.8% of body carbon. The house renewal rates increased withincreasing temperature and decreasing salinity. Food concentrationsranging from 100 to 16 x 10⁴ cells ml^–1, body size andlight condition had no effect on house renewal rate. Cloggingof the inlet filter by adding the large diatom Ditylum sol causedan increase in house renewal rates. The total number and carboncontent of houses during an animal's lifetime ranged from 46to 53 houses and from 6.5 to 10.4 µg, respectively. Sincedaily house production calculated for the O. dioica populationcorresponded to 130–290% of its biomass and daily discardedhouse materials corresponded to 490–1100% of the biomass,this organism must play an important role as a producer of macroscopicaggregates.     

SYNCHRONIZATION OF BACTERIAL CULTURE BY PREINCUBATION OF CELLS AT HIGH CELL DENSITY

The growth of Escherichia coli strain B in a liquid medium wasfound to cease at a cell density of 5x10⁹ cells per ml. (Thiscritical concentration is designated as the maximum or M-concentration.)Even cells harvested from the logarithmic growth phase couldnot divide at this or higher cell densities. Investigationson the metabolic activities of such cultures, however, showedthat the synthesis of cellular protein and nucleic acid wastaking place under such circumstances, showing that only someprocess (or processes) particularly related to cell divisionwas suppressed at the critical cell concentration in question. This finding led us to devise a new method of synchronizationof E. coli: cells harvested from a logarithmic phase were preincubatedat the critical concentration of 5x10⁹ cells per ml for 45 minutes,and then diluted 100 times with fresh medium. This led to synchronizationof cell division, as shown by a stepwise multiplication in cellnumber. (Received June 20, 1961; )     

Zooplankton feeding ecology: does a diet of Phaeocystis support good copepod grazing, survival, egg production and egg hatching success?

The bloom-forming alga Phaeocystis is ingested by a varietyof zooplankton grazers, but is thought to be a poor source offood. We examined copepod grazing on solitary Phaeocystis cellsby adult females of Temora stylifera, and survival, fecal pelletproduction, egg production and egg hatching success in Calanushelgolandicus and T. stylifera over periods of 15 consecutivedays. Phaeocystis cell concentrations were high (1.2–3.6x 10⁴ cells ml^–1 for C. helgolandicus and 2.5–7.9x 10⁴ cells ml^–1 for T. stylifera), but within the rangeof maxima recorded for natural blooms. Both copepods survivedwell and continuously produced fecal pellets (indicating continuousgrazing) on a diet of Phaeocystis. However, egg production ratesfor both copepods were low, even though hatching success ofthe few eggs produced was high. Clearance rates for T. styliferawere higher than for most previous measurements of other copepodsfeeding on Phaeocystis solitary cells at lower cell concentrations.We conclude that even though copepods feed well upon Phaeocystis,resulting poor fecundity on this diet may inhibit copepod populationincreases during blooms, thereby contributing to the perpetuationof blooms. However, the high egg hatching success on this dietargues against Phaeocystis containing chemical compounds thatact as mitotic inhibitors reducing copepod egg viability, suchas those found in some other phytoplankters.     

Seasonal dynamics in the abundance of Micromonas pusilla (Prasinophyceae) and its viruses in the Gulf of Naples (Mediterranean Sea)

To assess the role of viruses in the bloom dynamics of Micromonaspusilla in the Gulf of Naples (Mediterranean Sea), variationsof host and virus abundance were followed over one annual cycleand in late winter–spring of three consecutive years.Micromonas pusilla was recorded from autumn to spring, withpeak values up to 6.6 x 10³ cells ml^–1, but was undetectablein summer. Free M.pusilla viruses were detectable in all seasons,with concentrations from 0.02 viruses ml^–1 to 1.9 x 10³viruses ml^–1, exceeding host abundances only in one case.We found a great intraspecific variability in host susceptibilityto viruses present in natural samples, with viral titres rangingover one or two orders of magnitude for the same samples incubatedon different M.pusilla strains. Over the winter–springperiods, a highly dynamic situation was evident, with wide fluctuationsfor both host and virus abundances from one week to another.In some cases, peak host concentrations were accompanied byan increase in viral numbers, whereas in other cases the respectivefluctuations were uncoupled. Although fluctuations of M.pusillaabundance could be influenced by viral infection, there wasno evidence that viruses were able to terminate host blooms.The summer decline of M.pusilla populations did not appear tobe related to the impact of viral infection.