Egg size and clutch size in two Daphnia species grown at different food levels

The effect of three different Scenedesmus food concentrations(0.04, 0.2 and 1 mg C l^–1) on maternal investment wasstudied in two cladoceran species of similar size, Daphnia pulicariaand D.hyalina. It was observed that as food concentration decreased(between 1 and 0.2 mg C l^–1), there was an increase insize, protein content, lipid content, carbon and mass of theegg, while, at the same time, the clutch size of the femalesbecame smaller. Such an increase in ‘per offspring investment’was reflected in an increase in body length, body carbon andbody mass of neonates as the food available for females decreased.However, in D.pulicaria this tendency was not maintained downat 0.04 mg C l^–1 in which there was a decrease of theegg characteristics mentioned above. Although, there are notavailable all the egg and neonate parameters of D.hyalina at0.04 mg C l^–1, the body length of the neonates was largerthan at 0.2 and 1 mg C l^–1. These results show that, asfood diminishes, these two cladoceran species are able to respondby decreasing clutch size, but increasing the size of egg, therebyincreasing the probability of neonate survival. This tendencyis probably maintained until the food concentration is too lowand the females have to reduce the energy allocated for reproduction.     

Effect of External Supply of Growth Substances on Axillary Proliferation and Development in Dioscorea bulbifera

Bulbil development in cultured nodes of D. bulbifera proceededin the absence of growth substances from the medium. When IAAwas incorporated into the medium at the concentrations of 5mg l^–1 and 10 mg l^–1 the cultured nodes producedlarger bulbils than in its absences. When the concentrationof IAA was increased to 15 mg l^–1, however, the culturednodes produced a callus instead of a properly organized bulbil.The dry weight of bulbils increased when kinetin was added tothe medium at the concentrations of 0.05, 0.5, and 2.5 mg l^–1.The greatest increase was with 0.5 mg l^–1 kinetin. Onincreasing the concentration of kinetin in the medium to 5.0mg l^–1 the tissue produced had smaller dry weight thanthose produced in the absence of growth substances. Additionof different combinations of IAA and kinetin to the basal mediumresulted in the production of normal bulbils, roots, and shootsin some instances (suitable combinations) and in the productionof callus and abnormal shoots in others (non suitable combinations).     

In Vitro Micropropagation of Delphinium malabaricum (Huth) Munz.--a Rare Species

A micropropagation technique was developed for Delphinium malabaricumusing nodes from inflorescence stalks Maximum shoot proliferationwas obtained on Murashige and Skoog's medium supplemented with2-1P (10 mg l^–1) and inositol (100 mg l^–1) Fromthe sixth passage onwards, shoots could be multiplied by omissionof inositol and reduction of 2-1P (0.5 mg l^–1) concentrationBest rooting response was obtained with a 24-h pulse treatmentof shoots with 0.5 mg I^–1 IBA in the dark, transfer oftreated shoots to hormone-free half-strength MS medium and incubationunder 24-h light. Regenerated plants were established successfullyin the field Cytological examination of root tips of in vitroand control plants showed identical chromosome number (2n =16) Delphinium malabaricum (Huth) Munz, micropropagation, tissue culture, rare plant     

Effects of rotifers and ciliates on the growth and survival of Daphnia

Daphnia can suppress ciliates and rotifers through predationand interference competition, but it is not known whether thisproduces any direct benefit to Daphnia. We conducted survivorshipand cohort lifetable experiments to determine whether Daphniacan utilize ciliates and rotifers as food. Three species ofoligotrich ciliates (Halteria grandinella, Strobilidium gyransand Strobilidiumvelox) and one rotifer (Keratella cochlearis)were used. Lifetable experiments were conducted with a basallevel of algae (Cryptomonas sp.), plus either ciliates or rotifers,while survivorship experiments had only the rotifers or ciliates.Densities of 30 H.grandinella ml^–1, 50 S.gyrans ml^–1and 15 S.velox ml^–1 enhanced Daphnia pulex's populationgrowth rate 35–50% over controls with only algae. TenS.gyrans ml^–1 did not produce a significant change inDaphnia's growth rate. Densities of 100 and 300 K.cochlearis^–1 increased Daphnia population growth rates by II and10%, respectively. Both 10 and 50 S.gyrans ml^–1 enhancedDaphnia's survivorship compared to starved controls, but neither100 nor 300 K.cochlearis l^–1 enhanced its survivorship.The amount of enhancement of Daphnia growth rates by rotifersand ciliates is roughly proportional to the death rates imposedby Daphnia. The death rate imposed by Daphnia on rotifers isa function of both algal density and Daphnia size. Per unitbiomass, neither ciliates nor Keratella appear to be as nutritiousfor Daphnia as is Cryptomonas.     

Callus Culture of Sainfoin (Onobrychis viciifolia) and Plant Regeneration through Somatic Embryogenesis

Callus of sainfoin (Onobrychis viciifolia Scop.) was initiatedfrom stem and root explants which were obtained from seedlingsgrowing in vitro, on Linsmaier Skoog (LS) medium supplementedwith 1 mg l^–1 2, 4-D and 1 mg l^–1 BA or only 1 mgl^–1 BA, and the Vacin and Went medium without hormones.Somatic embryos were formed on LS medium containing 1 m l^–1BA. Embryos developed into complete plants on filter paper saturatedwith hormone-free LS medium. Onobrychis viciifolia, somatic embryogenesis, callus culture, plant regeneration     

Somatic Embryogenesis from Clonal Leaf Tissues of Cassava

Leaf lobes were isolated from palmate leaves of clonal cassava(Manihot esculenta Crantz) material growing in vitro or in glasshouseconditions and subjected to a two-stage culture procedure involvingincubation on Murashige and Skoog (MS2) basal medium supplementedwith 2–12 mg l^–1 2,4-D for 20 d (Stage I) beforetransfer to MS2 basal medium supplemented with 0.01 mg l^–12,4-D and 0.1 mg l^–1 6-benzylamino purine (BAP) (StageII medium). Embryogenetic tissues, foliose structures and somatic embryosdeveloped from leaf lobes at all Stage I 2,4-D concentrations,except on those explants isolated from shoot-tip cultures incubatedon MS2 basal medium supplemented with 0.1 mg l^–1 NAA and1.0 mg l^–1 BAP. Leaf lobes isolated directly from glasshouse plants showed optimalembryogenetic competence when subjected to a Stage I cultureperiod of 17 d, although foliose structure initiation was optimalwith shorter Stage I durations. Leaf lobes of 2–4 mm lengthand those isolated from phyllotaxic leaf numbers 4 and 5 showedthe greatest embryogenetic competence. Manihot esculenta, cassava, somatic embryogenesis, tissue culture, morphogenetic competence     

Micropropagation of Betula celtiberica

Plantlets were successfully regenerated from shoot segmentsof Betula celtiberica excised from young seedlings. Initiationand elongation of multiple shoot-buds were obtained after 20d culture in MS-modified medium plus BAP 0.6 mg l^–1 followedby 20 d culture in the same medium in the presence of a reducedBAP concentration (0.1 mg l^–1). Rooting was achieved 7d after having transplanted the isolated shoots to fresh medium,supplemented with IBA (0.2 mg l^–1). Betula celtiberica, birch, micropropagation, organogenesis     

Daphnia populations in three interconnected lakes with roach as the principal planktivore

Data on two co-existing Daphnia species, D.cucullata (G.O.Sars)and D.hyalina (Leydig), from three neighboring and interconnectedlakes in northern Poland, revealed an extremely confined rangeof population density fluctuations throughout summer in eachof the two species in each of the three lakes, with D.cucullatabeing an order of magnitude more abundant than D.hyalina (meanof 11.5 ind. l^–1 for D.cucullata and 1.1 ind. l^–1for D.hyalina with 95% confidence limits of ±2.5 and±0.5 ind. l^–1, respectively), in spite of markedlychanging fecundity. There was no apparent phase of decline orincrease that could be related to the distinct phases of lowor high fecundity resulting from different food levels. Analysisof the gut contents of roach (Rutilus rutilus), the dominantplanktivore in the three lakes, revealed identical selectivityfor each of the two Daphnia species, suggesting that the lowerdensity of the D.hyalina population was compensated for by thegreater conspicuousness of individuals of this species. It isconcluded that the population density of these Daphnia speciesremains far below the carrying capacity of the habitat and doesnot depend on food levels. Food availability merely sets therate of population increase, while the actual population densityin the lakes studied reflects the species' vulnerability topredation by planktivorous fish.     

Differentiation of Shoot Buds in vitro in Tissue 1Sisymbrium irio L.

Shoot bud formation was induced in the stem callus of Sisymbriumirio L., a Cruciferous plant. The callus was established onMurashige and Skoog medium with IAA (1?0 mg l^–1) and kinetin(0?5 mg l^–1). The effect of three purines (kinetin, 6-benzylaminopurine,and 6-methylaminopurine) incorporated singly along with IAAin MS medium was investigated. It was found that kinetin orMAP (3–5 mg l^–1) along with IAA (0?5 mg l^–1)were the most effective in inducing shoot bud formation. Adeninesulphate (10 mg l^–1) with kinetin (1?0 mg l^–1) alsoinduced bud differentiation. The morphogenetic potential of the callus to differentiate shootbuds was seemingly lost in 2 year old callus cultures. However,on successively subculturing on a regeneration medium shootbuds differentiated and the number of buds formed improved onfurther subculture. Two types of meristematic outgrowths were recognized: (i) arisingfrom superficial cells and (ii) arising from deep-seated cellsin the vicinity of tracheidal elements. However, both typesformed meristematic nodules on the surface of which shoot budsdifferentiated. Some embryoids were also recognized arisingsuperficially.     

Effects of Gibberellic Acid and Naphthaleneacetic Acid on Petiole Senescence and Subtended Peduncle Growth of Cyclamen persicum Mill

Removal of the blade from the leaf subtending the first flowerbud on Cyclamen persicum ‘Swan Lake’ plants causedthe petiole of that leaf to senesce, but had no effect on thegrowth of the flower peduncle in the debladed petiole's axil.A 10 mg NAA l^–1 application generally had no effect onpetiole senescence, peduncle elongation or flowering date whenapplied to the cut end of the petiole after blade removal. A25 mg GA₃ l^–1 application or a combination of 25 mg GA₃l^–1 application or a combination of 25 mg GA₃ l^–1plus 10 mg NAA l^–1 delayed petiole senescence and enhancedpeduncle elongation and subsequent flowering. No treatment significantlyaltered peduncle length at the time of flowering. Cyclamen persicum Mill, ‘Swan Lake’, tissue receptivity, flowering, GA₃, NAA     

The Effect of Inorganic Nutrients on the Hormonal Requirements of Normal, Habituated, and Crown-gall Tissue 4

The addition of Braun and Wood's inorganic supplements (845mg l^–1 KCl, 1800 mgl^–1 NaNO₃, 300 mg l^–1 NaH₂PO₄.2H₂O,790 mg l^–1 (NH₄)₂SO₄) to White's medium caused markedincreases in the growth of normal tissues of Helianthus annuus,Nicotiana rustica, Daucus carota, and Vinca rosea and crown-galltumour tissues of H. annuus. However, no evidence was obtainedwhich suggested that the presence of these extra salts markedlyinfluenced the essential requirements of normal callus for auxinsand kinetin. In contrast their presence significantly influencedthe hormonal requirements of certain habituated cultures ofH. annuus and V. rosea. These habituated cultures had specificauxin requirements on White's medium while either an auxin orkinetin was sufficient on high-salts medium. These results arediscussed in relation to previous reports which suggested thatthe biosyntheses of auxins and other growth factors in normaland crown-gall cultures are specifically activated by certaininorganic ions.     

Can phaeopigments be used as markers for Daphnia grazing in Lake Constance?

The formation of chlorophyll a degradation products was measuredwith natural phytoplankton from Lake Constance and Daphnia magnaand native Daphnia as grazers in grazing experiments duringspring bloom conditions using high-pressure liquid chromatography(HPLC). Chlorophyll a start concentrations were between 1.2and 16.3 µg l^–1; phaeopigment weights constituted5% of chlorophyll a weight. Only phaeophorbide a was a markerfor Daphnia grazing; concentrations of other phaeopigments (phaeophytina, chlorophyllide a and two unidentified phaeopigments) didnot increase during Daphnia grazing. Conversion efficiencies(chlorophyll a to phaeophorbide a) were between 0 and 43% ona weight basis, and between 0 and 65% on a molar basis. Conversionefficiencies were highest at high grazer density (40 Daphnial^–1) and after a 24 h exposure time. Grazing by microzooplanktonprobably led to the formation of the two unidentified phaeopigments.In Lake Constance, Daphnia density was significantly positivelycorrelated with the phaeophorbide a/chlorophyll a ratio whenit was <5000 Daphnia m^–3. However, when higher Daphniadensities were included in calculations, then Daphnia densitywas positively, but insignificantly, correlated with the phaeophorbidea/chlorophyll a ratio. This suggests that when the level offood per Daphnia is low, then grazing is more efficient withless production of phaeophorbide a and a higher production ofcolourless products.     

Somatic Embryogenesis and Its Hormonal Regulation in Tissue Cultures of Freesia refracta

Somatic embryogenesis can be induced in tissue cultures of Freesiarefracta either directly from the epidermal cells of explants,or indirectly via intervening callus. These two pathways ofsomatic embryogenesis can be controlled and regulated by varyingthe combinations and levels of exogenous hormones. When younginflorescence segments were cultured in vitro on modified N₄(MN₄) medium supplemented with 2 mg l^–1 indoleacetic acid(IAA) and 3 mg l^–1 6-benzylaminopurine (BAP), some ofthe epidermal cells began to exhibit the features of embryogeniccells. These cells produced embryoids and developed into newplants through direct somatic embryogenesis. If the same explantswere placed on Murashige and Skoog's (MS) medium containing2 mg l^–1 IAA, 05 mg l^–1 BAP and 05 mg l^–1naphthaleneacetic acid (NAA), pale-yellow translucent nodularcalluses appeared on the surface of the explants. When thiskind of callus was transferred to MN6 medium with 2 mg l^–1IAA and 3 mg l^–1 BAP, embryoids formed which further developedinto plantlets. The regenerated plants were morphologicallynormal and possessed the normal diploid chromosome number of2n = 22. A similar result has also been obtained with youngleaf explants of this plant. The early segmentations of embryogeniccells and the development of embryoids were studied using histologicaland scanning electron microscopic techniques, and the resultshave been discussed in association with the ontogeny and originof the embryoids. Freesia refracta Klatt, somatic embryogenesis, plant regeneration, exogenous hormones     

Effects of a Range of O2 Concentrations on Porosity of Barley Roots and on their Sugar and Protein Concentrations

Barley was grown at a range of oxygen concentrations (0.5–9mg l^–1), in nutrient solutions. Growth of both shootsand seminal roots was restricted by O₂ concentrations lowerthan 2–3 mg l^–1) but nodal root growth was not. Root porosities were increased even at those O₂ concentrationswhich did not restrict growth, and were inversely proportionalto the protein levels of the roots. Sugar concentrations increasedappreciably only at those O₂ concentrations which also restrictedgrowth. Hordeum vulgare L., barley, root porosity, sugar, protein, oxygen concentration     

Herbivorous nutrition of Cyclops vicinus: the effect of a pure algal diet on feeding, development, reproduction and life cycle

Nutritional requirements, functional response, development andreproduction of Cyclops vicinus were studied with exclusivelyalgal food. Phytoflagellates were found to be adequate foodresources for both juvenile development and egg production.Ingestion measurements were performed with Chlamydomonas reinhardii.The functional response data give evidence for low feeding efficiency,especially for the naupliar stages. A difference between naupliiand older instars was also found in their quantitative foodneeds. A higher threshold food concentration was observed fornaupliar development (0.4 mg C l^–1) than for copepoditedevelopment (0.2 mg C l^–1). The calculation of assimilationefficiencies suggests that the high food requirements are dueto low specific ingestion rates rather than poor assimilationefficiency. Development time decreased as algal density increasedand males developed more quickly than females at all food concentrations.Body size and carbon content increased with increasing foodconcentration. Continuous egg production was observed abovea food concentration of 0.5 mg C l^–1. The results haveimplications for the life cycle of C. vicinus. Summer diapauseis interpreted as a strategy to avoid starvation of the juvenilestages. The facultative herbivory of the adults might be anadvantage when competing against other more carnivorous cyclopoidcopepods. ¹Present address: Agricultural University of Wageningen, Departmentof Mathematics, Dreijenlaan 4, NL-6703 HA Wageningen, The Netherlands     

Experimental studies of food and suspended sediment effects on growth and reproduction of six planktonic cladocerans

Life-long somatic growth and egg production responses of Daphniabarbata, Daphnia gibba, Daphnia laevis, Daphnia pulex, Moinamicrura and Diaphanosoma excisum were examined in relation todifferent food and suspended sediment levels, in factorial experimentsat 20°C. Food treatments involved additions of 0, 100, 500and 2500 µg C l^–1 of Selenastrum to 20 µmfiltered water, generally from source lakes. These food levelswere tested at two suspended sediment levels in all species,viz. (i) natural source-lake nephelometric turbidities of 7–65nephelometric turbidity units (NTU) ({small tilde}10–70mg l^–1 TSS) and (ii) {small tilde}3- to 4-fold enrichmentof natural levels (8-fold in D.excisum). A third sediment-freetreatment was tested in all species of Daphnia other than D.gibba.Growth in body length and reproductive output were assesseddaily for individual animals. Main and interactive effects offood and sediment on growth were assessed at selected ages fromday 3 through subsequent life using analysis of variance (ANOVA).Strong and highly significant positive food effects were apparentat all ages in all species examined, with the curious exceptionof D.excisum. The onset of maturity and fecundity were alsopositively related to food level. Sediment effects on growth,age of maturity and fecundity were variable, but tended to bemuch weaker than food effects. Both stimulatory (positive, especiallyat low food levels) and inhibitory (negative) effects of sedimenton growth were apparent, both between and within species. Performancein sediment-free water was very poor, except in D.pulex. Food-sedimentinteraction effects were variable. Generally, turbidwater specieswere influenced less adversely by sediment than clear-waterspecies, indicating the existence of environmentally appropriateadaptive responses. Overall, the findings indicate that lightlimitation of algal resources by suspended sediments, ratherthan the condition of high turbidity per se, is responsiblefor the apparently negative impact of suspended sediments frequentlyobserved on natural populations of cladoceran zooplankters.     

Fish-induced changes in zooplankton grazing on phytoplankton and bacterioplankton: a long-term study in shallow hypertrophic Lake Sobygaard

The impact of fish-mediated changes on the structure and grazingof zooplankton on phytoplankton and bacterioplankton was studiedin Lake Søbygaard during the period 1984–92 bymeans of in vitro grazing experiments (¹⁴C-labelled phytoplankton,³H-labelled bacterioplankton) and model predictions. Measuredzooplankton clearance rates ranged from 0–25 ml l^–1h^–1 on phytoplankton to 0–33 ml l^–1 h^–1on bacterioplankton.The highest rates were found during thesummer when Daphnia spp. were dominant. As the phytoplanktonbiomass was substantially greater than that of bacterioplanktonthroughout the study period, ingestion of phytoplankton was26-fold greater than that of bacterioplankton. Multiple regressionanalysis of the experimental data revealed that Daphnia spp.,Bosmina longirostris and Cyclops vicinus, which were the dominantzooplankton, all contributed significantly to the variationin ingestion of phytoplankton, while only Daphnia spp. contributedsignificantly to that of bacterioplankton. Using estimated meanvalues for clearance and ingestion rates for different zooplankters,we calculated zooplankton grazing on phytoplankton and bacterioplanktonon the basis of monitoring data of lake plankton obtained duringa 9 year study period. Summer mean grazing ranged from 2 to4% of phytoplankton production and 2% of bacterioplankton productionto maxima of 53 and 88%, respectively. The grazing percentagedecreased with increasing density of planktivorous fish caughtin August each year using gill nets and shore-line electrofishing.The changes along a gradient of planktivorous fish abundanceseemed highest for bacterioplankton. Accordingly, the percentagecontribution of bacterioplankton to the total ingestion of thetwo carbon sources decreased from a summer mean value of 8%in Daphnia-dominated communities at lower fish density to 0.7–1.1%at high fish density, when cyclopoid copepods or Bosmina androtifers dominated. Likewise, the percentage of phytoplanktonproduction channelled through the bacteria varied, it beinghighest (5–8%) at high fish densities. It is argued thatthe negative impact of zooplankton grazing on bacterioplanktonin shallow lakes is highest at intermediate phosphorus levels,under which conditions Daphnia dominate the zooplankton community.     

Somatic Embryogenesis and Plant Regeneration from Inflorescence Culture of Java Citronella: (Cymbopogon winterianus)

Embryogenic callus was induced from immature inflorescence segmentsof Java citronella (Cymbopogon winterianus) and maintained for2 years on Murashige and Skoog's medium supplemented with 2,4-D(l mg l^–1). The callus cells retained the original chromosomenumber of 2n = 20. The somatic embryos germinated into plantletson MS basal medium or medium with IAA, NAA, BAP or KN individually(l mg l^–1). The regenerated plantlets developed a goodroot system on full strength solid MS inorganics medium withIAA (1 mg l^–1). The regenerated plants were similar tothe donor plant in morphology and had the same chromosome number,but showed some variation in the essential oil content. Java citronella, Cymbopogon winterianus, somatic embryogenesis, regeneration, inflorescence culture     

In Vitro Regeneration from Seedling Organs of Brassica oleracea var. italica Plenck cv. Green Comet I. Effect of Plant Growth Regulators

The calabrese cultivar Brassica oleracea var. italica cv. GreenComet was used in a study of the effects of exogenous hormoneson the growth and differentiation of seedling organs in vitro.Four types of explants were tested: hypocotyl segments, rootsegments, primary leaf discs and cotyledon discs. These explantswere incubated on media containing factorial combinations ofBAP x IBA, BAP x NAA, KN x IBA and KN x NAA (all at 0, 0.1,10 and 10.0mg l^–1). Hypocotyls were the most regenerativeexplants; shoot production was favoured by cytokinin: auxinratios greater than one and was decreased by IBA at 10 mg l^–1when callus was produced. Shoot formation from root explantsoccurred either in the absence of hormones or with low concentrations;no shoot was produced when any hormone was present at 10 mgl^–1. In contrast, shoot production from primary leaf diseswas favoured by high concentrations of both auxin and cytokininwith the combination of BAP and IBA the most effective. Shootproduction from cotyledon discs was sporadic with no consistentresponse on any auxin/cytokinin combination. After further experimentson the optimization of hormone concentration, the followingcombinations were chosen as allowing reliable regeneration:0.1 mg l^–1 BAP+0.1mg l^–1 IBA for hypocotyl segments,0.075 mg l^–1 KN +0.025 mg l^–1 IBA for root segments,and 5.0 mg l^–1 BAP+5.0 mg l^–1 IBA for leaf discs. Brassica oleracea var. italica, calabrese, tissue culture, seedling, auxin, cytokinin     

Multiplication In Vitro of Limonium estevei Fdez. Casas

Plantlets of Limonium estevei Fdez. Casas, an endangered Spanishspecies, were successfully regenerated from nodal segments excisedfrom young seedlings. Initiation of multiple adventitious budswere obtained in MS modified medium plus 1 mg l^–1 IBAand 0·1 mg l^–1 BAP. Rooting was achieved by transferof the isolated shoots to fresh MS medium without plant growthregulators. Fully grown plants were established in a pottingmix and are growing well in a greenhouse. Limonium estevei, in vitro multiplication, adventitious regeneration