Dependence of the photophobic response of the blue-green alga,Phormidium uncinatum,on cations

It is suggested that photophobic responses caused by a sudden step-down in light intensity require the presence of cations in the blue-green alga, Phormidium uncinatum.Drastic removal of cations abolishes the phobic response, which recovers after addition of Ca²⁺ ions. Calcium can be substituted for partially by other cations with an effectivity following the sequence Ca>Mg>Na>Ba>Co=0. During the photophobic response there is a 25% increase in ⁴⁵Ca binding by the cells related to a step-down in light intensity. Three seconds after a light-dark transition there is a sharp increase in the binding of labelled calcium, followed by a subsequent release.Flushing the filaments with high cation concentrations, esp. calcium causes a reversal of movement in the absence of a light stimulus similar to a photophobic reversal. This stimulus could trigger the same sequence of events in the transduction chain bypassing the primary photoresponse.Abbreviations EDTA Ethylene diaminetetraacetic acid - EGTA ethylene glycol-bis (2-aminoethylether) N,N tetraacetic acid     

Gated ion fluxes involved in photophobic responses of the blue-green alga,Phormidium uncinatum

The sensory transduction chain of photophobic responses in the blue-green alga, Phormidium uncinatum seems to involve a gating cation transport through membrane bound ion channels which provides an effective amplification.The calcium conducting ionophore A23187 inhibits the photophobic response totally and induces frequent reversals which resemble phobic responses but occur without any light stimulation. This indicates that the electrogenic ion conductance may depend on a gradient of divalent cations, esp. calcium. The calcium conductance during a photophobic response is further confirmed by the inhibitory effect of ruthenium red and lanthanum, blockers of the electrogenic calcium transport. In the case of lanthanum this inhibition is found at a concentration at which neither the number of motile filaments nor the average speed of movement is impaired.Incorporation of ionophores for monovalent cations (gramicidin and valinomycin) only partially impairs the response. Similarly, inhibition of the Na⁺/K⁺ pump by ouabain is less effective. Thus, the existence of a countercurrent of monovalent cations during the response, which has been described for e.g. ciliates, is yet obscure in blue-green algae.     

Extracellular and intracellular determination of light-induced potential changes during photophobic reactions in blue-green algae

Light-induced potential changes have been measured in the filamentous blue-green alga Phormidium uncinatum both intracellularly and between the two ends of a trichome. There is evidence that these potential changes are correlated with photophobic reactions in this organism.

1. The potential changes follow the light-dark regime with a lag phase of about 10 s. The photophobic reaction time has been found to be about the same length of time.
2. The action spectra of both externally and internally measured light-induced potential changes correspond with the photophobic action spectrum, indicating the participation of the main photosynthetic pigments of Phormidium, chlorophyll a and phycobilins.

A hypothesis is being discussed according to which sensory transduction between photoreceptor and motor apparatus of the cell is mediated by light-induced electrical potential changes.     

Evidence of electrical potential changes in photophobically reacting blue-green algae

The correlation between photophobic responses and light-induced electric potential changes has been studied in the blue-green alga Phormidium uncinatum.

1. The photophobic reaction time depends on both length of preillumination and presentation time of stimulus. Under optimal conditions a reaction time of about 10 s has been determined.
2. Light-induced potential changes can be measured by means of external electrodes with a small gap between them bridged by a population of perpendicularly oriented trichomes. These potential changes follow a light-dark cycle with a lag phase of about 10 s.
3. The amplitude of these light-induced potential changes increases with light intensity until it reaches a saturation value of about 12 mV at 10000 lx. The action spectrum resembles the photophobic action spectrum with peaks in the absorption region of C-phycoerythrin and chlorophyll a.

The significance of light-induced potential changes as a means of sensory transduction for photophobic responses in blue-green algae is being discussed.     

Photophobic responses of the cyanobacterium Anabaena variabilis

The photophobic responses in the Cyanobacterium Anabaena variabilis which belongs to the Nostocaceae have been studied with aid of a population method as well as by single trichome observations. In white light experiments both step-up and step-down photophobic responses were observed. The wavelength dependence was examined at a constant fluence rate. The photophobically active light is absorbed by the photosynthetic pigments, mainly by the phycobiliproteins and chlorohyll a. Above 690 nm only negative reactions were observed, i.e. the trichomes left the light trap. In white light experiments DCMU strongly inhibited the photophobic responses, whereas photokinesis was not affected to the same extent indicating that the reaction is coupled with the non cyclic photosynthetic electron transport. DBMIB impaired the photophobic behaviour only slightly. It seems that the photophobic responses of A. variabilis are controlled by a similar mechanism as in Phormidium uncinatum (Oscillatoriaceae) although the two families and, hence, the two species differ in their movement mechanism as well as in their photoactic behaviour.     

Photobehaviour of Paramecium bursaria infected with different symbiotic and aposymbiotic species of Chlorella

The endosymbiotic unit of Paramecium bursaria and Chlorella spec. shows two types of photobehaviour: 1) A step-up photophobic response which possibly depends on photosensitive agents in the ciliate cell itself — as is also shown by alga-free Paramecium bursaria - and can be drastically enhanced by photosynthetic activity of symbiotic algae; and 2) a step-down photophobic response. The step-down response leads to photoaccumulation of green paramecia. Both types of photobehaviour in Paramecium bursaria do not depend on any special kind of algal partners: The infection of alga-free Paramecium bursaria with different Chlorella species results in new ciliatealgae-associations. They are formed not only by combination of the original symbiotic algae with their host, but also by infection with other symbiotic or free-living (aposymbiotic) chlorellae, respecitively. Systems with other than the original algae are not permanently stable — algae are lost under stress conditions — but show the same types of photobehaviour. Photoaccumulation in general requires algal photosynthesis and occurs only with ciliates containing more than fifty algae/cell. It is not mediated by a chemotactic response to oxygen in the medium, since it occurs at light fluence rates not sufficient for a release of oxygen by the symbiotic system, e.g., below its photosynthetic compensation point. Photoresponses can be inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU). Sensory transduction does not depend on any special symbiotic features of the algae, e.g., sugar excretion. The participation of oxygen in the Paramecium cell, of its cytoplasmic pH and of ions released or taken up by endosymbiotic algae in sensory transduction is discussed.     

Effects of UV-B on motility and photoorientation in the cyanobacterium,Phormidium uncinatum

UV-B irradiation has a detrimental effect on the survival of populations of the filamentous cyanobacterium, Phormidium uncinatum, at levels slightly higher than those currently measured at the surface of the earth. The organisms are not damaged or killed by UV-B radiation at 300 nm of 200 Wm^-2 for up to 20 h; but slightly increased levels of UV-B irradiation (2 h of 200 Wm^-2 at 300 nm) drastically impair motility, phototactic orientation and photophobic responses. These photosynthetic organisms require a narrow light intensity range for growth so that any decrease in their ability to actively search for and move into areas of favorable light conditions is bound to affect the survival of a population. The fluorescence yield of both phycobilins and chlorophyll is not altered even after 20 h of UV-B irradiation (200 Wm^-2 at 270 nm) indicating that UV-B at that dose does not affect the photosynthetic apparatus. The organisms are killed either by too bright intensities which bleach the photosynthetic pigments or by the lack of energy when they are unable to avoid moving into dark areas.     

Fine structure of the developing frontal bones and scales of the cranial vault in the cichlid fish Hemichromis bimaculatus (Teleostei,Perciformes)

The development of the frontal bone and the formation of the first head scales are described during post-embryonic ontogeny of Hemichromis bimaculatus, using light and transmission electron microscopy. The frontal bone originates close to the cartilaginous taenia marginalis in a loose mesenchymal cell condensation (=primordium) lying 1 m from the epidermis with which it establishes no cell contacts. The anlage appears at 4.2 mm standard length (SL) in the form of the membranodermal component of the bone, and extends first over the brain and then over the eye; the neurodermal component forms later to surround the supraorbital canal. The first head scales appear at 10.0 mm SL in a dense cell condensation (papilla) adjoining the epidermal-dermal junction and, once formed, remain in this position. In both organs, the initial matrix is similarly composed of woven-fibred bone that soon mineralizes in a similar manner to other dermal elements. In some areas of the frontal bone, parallel-fibred bone is deposited unequally on both surfaces, whereas isopedine is deposited in scales on the deep surface only. Osteoblastic features confirm this eccentric growth. Differences in the shape, organization and localization of the mesenchymal condensations giving rise to the frontal bone and to the scale reflect the existence of two types of dermal cell condensations. Our data are compared with those available for the post-cranial dermal skeleton of fishes both from a developmental and structural viewpoint. Structural differences in the matrices of the frontal bone and scales are discussed in a phylogenetic perspective.     

Antibiotic resistance and detection of β-lactamase in bacterial strains of Staphylococci and Escherichia coli isolated from foodstuffs

Seventy strains of Staphylococcus spp. and Escherichia coli (35 each) were isolated from various foodstuffs and identified on the basis of cultural, morphological and biochemical characteristics and were further tested for their antibiotic susceptibility with commonly used antibiotics/drugs. 69.2% of the strains of Staphylococci were resistant to co-trimazine and 34.6% were resistant to penicillin-G. 19.2% of the staphylococcal isolates exhibited resistance to cloxacillin, nalidixic acid, methicillin and tetracycline whereas 15.3% of the staphylococcal isolates were resistant to amoxycillin and nitrofurantoin. The isolated E. coli strains exhibited sharp peaks of resistance to antimicrobial agents such as tetracycline (72%), doxycycline (60%) and nalidixic acid (48%). Forty-four percent of the E. coli strains were resistant to nitrofurantoin and penicillin-G respectively. Among the 13 antibiotics/drugs tested for resistance, six different resistance patterns were observed in staphylococcal isolates and seven different resistance patterns were observed in the E. coli isolates from various foodstuffs. Bacterial strains exhibiting MIC values 100 g/ml for ampicillin and cloxacillin were screened for -lactamase activity and out of 10 staphylococcal isolates, seven were found to be positive for -lactamase, whereas out of 13 E. coli isolates tested for -lactamase production, only three were found to be positive.     

Qualitative and quantitative composition of pigments in Phaeodactylum tricornutum (Bacillariophyceae) stressed by iron

The effect of Fe(III) deficiency on qualitative and quantitative changes in pigment composition in Phaeodactylum tricornutum Bohlin was demonstrated by HPLC and AAS. Maximum content of pigments showed the diatom cells incubated at the optimum iron concentration, i.e., 10 M. The contents of chlorophyll a, chlorophyll c ₁+c ₂, fucoxanthin, diadinoxanthin and ,-carotene were 109.99, 20.16, 40.39, 1.29 and 1.48 fg per cell, respectively. The results obtained showed that Fe(III) affected qualitative and quantitative pigment composition in P. tricornutum. The content of individual pigments, proportions between accompanying pigments and their ratios to chlorophyll a were important indicators of phytoplankton response to iron stress. The strong reduction in ,-carotene content, several times (2–5) increase in diadinoxanthin level as compared to ,-carotene, and high amount of diadinoxanthin in relation to chlorophyll a were observed in algae growing at very low Fe(III) concentrations, 0.001 and 0.01 M. The data suggested that phytoplankton pigments could be a potential physiological marker.     

Phototaxis in the flagellates,Euglena gracilis and Ochromonas danica

Oriented movement with respect to laterally impinging white light of the flagellates Euglena gracilis and Ochromonas danica has been analyzed in an individual cell study with a microvideographic technique. Using the deviation of track segments (in given time intervals of 1 s) from the light direction as raw data allowed a computer based analysis of the direction distribution. A number of statistical methods employed to test the significance of the obtained results demonstrated an obvious phototactic orientation in Ochromonas which was positive (toward the light source) in low illuminance (1.25 lx=5.3×10^-3 Wm^-2) and negative in higher illuminance (>12.5 lx=5.3×10^-2 Wm^-2). Since in this flagellate the threshold for negative phototaxis is much lower than that for the step-up photophobic response, the hypothesis that negative phototaxis may be brought about by repetitive step-up phobic responses can be rejected for at least this organism. In Euglena positive phototaxis was observed in 50 lx (=0.21 Wm^-2), while an illuminance of 500 lx (=2.1 Wm^-2) caused a negative phototaxis.The experiments were carried out in this laboratory     

Exogenous inorganic carbon sources for photosynthesis in seawater by members of the Fucales and the Laminariales (Phaeophyta): ecological and taxonomic implications

Summary Characteristics of inorganic carbon assimilation by photosynthesis in seawater were investigated in six species of the Fucales (five Fucaceae, one Cystoseiraceae) and four species of the Laminariales (three Laminariaceae, one Alariaceae) from Arbroath, Scotland. All of the algae tested could photosynthesise faster at high external pH values than the uncatalysed conversion of HCO ₃ ^- to CO₂ can occur, i.e. can use external HCO ₃ ^- . They all had detectable extracellular carbonic anhydrase activity, suggesting that HCO ₃ ^- use could involve catalysis of external CO₂ production, a view supported to some extent by experiments with an inhibitor of carbonic anhydrase. All of the algae tested had CO₂ compensation concentrations at pH 8 which were lower than would be expected from diffusive entry of CO₂ supplying RUBISCO as the initial carboxylase, consistent with the operation of energized entry of HCO ₃ ^- and / or CO₂ acting as a CO₂ concentrating mechanism. Quantitative differences among the algae examined were noted with respect to characteristics of inorganic C assimilation. The most obvious distinction was between the eulittoral Fucaceae, which are emersed for part of, or most of, the tidal cycle, and the other three families (Cystoseiraceae, Laminariaceae, Alariaceae) whose representatives are essentially continually submersed. The Fucaceae examined are able to photosynthesise at high pH values, and have lower CO₂ compensation concentrations, and lower K_1/2 values for inorganic C use in photosynthesis, at pH 8, than the other algae tested. Furthermore, the Fucaceae are essentially saturated with inorganic C for photosynthesis at the normal seawater concentration at pH 8 and 10°C. These characteristics are consistent with the dominant role of a CO₂ concentrating mechanism in CO₂ acquisition by these plants. Other species tested have characteristcs which suggest a less effective HCO ₃ ^- use and CO₂ concentrating mechanism, with the Laminariaceae being the least effective; unlike the Fucaceae, photosynthesis by these algae is not saturated with inorganic C in normal seawater. Taxonomic and ecological implications of these results are considered in relation to related data in the literature.     

Root traits, nutrient uptake, multi-location grain yield and benefit–cost ratio of two lentil (Lens culinaris, Medikus.) varieties

Lentil is a protein-rich pulse, grown mainly in developing countries as a rain-fed crop in nutrient-poor soils. Hence, the importance of root traits for efficient capture of soil nutrients and water can be crucial to its economical yield. Little is known about the lentil root system and even less about its relationship to grain yield. We compared the root system of two Bangladeshi lentil varieties, Barimasur-3 (BM-3) and Barimasur-4 (BM-4), in a pot experiment and related it to their multi-location grain yield in the fields. BM-4 maintained faster root development both at an early growth stage (20days after sowing) and at flowering (60days) compared to BM-3. The roots of BM-4 penetrated the 25cm depth of the soil profile after 19±1days and while those of BM-3 took 24±2days to reach the same depth. The roots of BM-4 were covered with denser (26±3mm^–1) and longer (0.48±0.11) root hairs than BM-3 (density 17±2mm^–1, length 0.32±0.09mm). The differential presence of root hairs increased the effective length of root system of BM-4 by 12 times and that of BM-3 by five times. The lentil varieties did not differ in their ability to induce pH change and acid phosphatase activity in rhizosphere. In the pot experiment, the uptake of macro-nutrients (K, P, Ca, and Mg) as well as micro-nutrients (Fe, Mn, Zn, Cu, B and Mo) by BM-4 was significantly higher, compared to BM-3. The varieties produced the same amount of shoot biomass. At five of six agro-ecological distinct field locations in Bangladesh, BM-4 gave significantly higher (10–20%) grain yield than BM-3. Linked with the higher grain yield, the benefit-cost ratio (BCR) of BM-4 was 3.14 and that of BM-3 were 2.62, indicating that BM-4 provided better return per unit investment, compared to BM-3, supported by the better root morphology and higher nutrient uptake. This may be one of the reasons supporting the popularity and preferred adoption of BM-4 among the Bangladeshi farmers, who grow lentil mainly on nutrient-poor soils. The results indicate the benefits of selection and breeding for superior root traits for better agro-economics.     

Changes of amino acid sequence in PEST-like area and QEEET motif affect degradation rate of D1 polypeptide in photosystem II

The degradation rate of the D1 polypeptide was measured in threeSynechocystis PCC 6803 mutantsin vivo. Mutations were introduced into a putative cleavage area of the D1 polypeptide (QEEET motif) and into the PEST-like area. PEST sequences are often found in proteins with a high turnover rate. The QEEET-motif mutants are CA1 [(E242-E244);Q241H] and E243K, and the third mutation, E229D, was directed to the PEST-like area. During high-light illumination (1500 mol photons m^-2s^-1) that induced photoinhibition of photosystem II (PSII), the half-life time of the D1 polypeptide in mutant E229D (t _1/2=35 min) was about twice as long as in AR (control strain) cells (t _1/2=19 min). In growth light (40 mol photons m^-2s^-1), the degradation rate of the D1 polypeptide in E229D and AR strains was the same (t _1/25 h). In growth light the D1 polypeptide was degraded faster in both QEEET-motif mutants than in the AR strain, but in photoinhibitory light the degradation rates were similar. According to these results, the highly conservative QEEET motif as such is not required for the proteolytic cut of the D1 polypeptide, but it does affect the rate of degradation. No simple correlation existed between the degradation rate of the D1 polypeptide and the susceptibility of PSII to photoinhibition in mutant and AR cells under our experimental conditions.     

Photosensory transduction in Euglena gracilis: Effect of some metabolic drugs on the photophobic response

We have investigated the effect of some metabolic drugs, 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), 2,4-dinitrophenol (DNP), sodium azide (NaN₃), on the photobehavior of single cells of Euglena gracilis, in order to clarify the relevance of different metabolic pathways in the process of photoperception and sensory transduction in this alga. The results obtained show that the photophobic response of Euglena is not affected by the action of these drugs. This suggests that neither the photosynthetic process nor oxidative phosphorylation play a significant role in the phenomenon of photosensory transduction in Euglena.List of Abbreviations DNP 2,4-dinitrophenol - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - PSI Photosystem I - PSII Photosystem II     

Involvement of actin filaments in chloroplast division of the algaClosterium ehrenbergii

Summary Studies have been made of whether actin filaments and microtubules are involved in the chloroplast division ofClosterium ehrenbergii (Conjugatae). Fluorostaining with rhodamine-phalloidin showed 5 types of localization of F-actin: (1) cables of actin filaments running in the cortical cytoplasm along the cell's long axis, (2) condensed actin filaments at the septum, (3) perinuclear distribution of actin filaments, (4) F-actins in a marking pin-like configuration adjacent to the nucleus of semicells just before completion of chloroplast kinesis, and (5) actin filaments girdling the isthmus of the constricted and dividing chloroplasts. Cytochalasin D (CD) at a concentration of 6 to 25 M caused significant disruption of actin filaments and the arrest of chloroplast kinesis, nuclear division, septum formation and cytoplasmic streaming within 3 to 6h. Chloroplast kinesis and cytoplasmic streaming recovered when cells were transferred to the medium without CD after CD treatment, or were subjected to prolonged contact with CD for more than 9h. In these cells there was a coincidental reappearance of actin filaments. A tubulin inhibitor, amiprophos-methyl at 330 M, did not inhibit chloroplast kinesis but did inhibit division and positioning of the nucleus. These results suggest that actin filaments do play a role in the mechanism of chloroplast kinesis but that microtubules do not appear to be involved in the process.Abbreviations APM amiprophos-methyl - CD cytochalasin D - DAPI 4,6-diamidino-2-phenylindole - DIC Nomarski differential interference contrast - DMSO dimethyl sulfoxide - Rh-Ph rhodamine-phalloidin     

Phytochrome action in light-grown plants: the influence of light quality and fluence rate on extension growth in Sinapis alba L.

Using light-grown plants of Sinapis alba an analysis has been made of the effect on extension growth of adding far red light to a background photosynthetic source. It has been possible to distinguish between the increase in fluence rate and the reduction of the amount of phytochrome present as Pfr, which are both consequences of the addition of supplementary far red light, and to determine that the response of increased extension growth is due only to the latter. It is shown that the degree of fluence rate dependency varies with photoequilibrium and the significance of this interaction is discussed in terms of the mode of action of phytochrome and of its role in the natural light environment.Abbreviations PAR photosynthetically active radiation - SAN 9789 4-chloro-5-(methylamino)-2-(,,-trifluoro-m-tolyl)-3(2H) pyridazinone - Pfr far-red absorbing form of phytochrome - Pr red-absorbing form of phytochrome - LER logarithmic extension rate     

CALCIUM RELEASE FROM INTRACELLULAR STORES IS NECESSARY FOR THE PHOTOPHOBIC RESPONSE IN THE BENTHIC DIATOM NAVICULA PERMINUTA (BACILLARIOPHYCEAE)1

Complex photoreceptor pathways exist in algae to exploit light as a sensory stimulus. Previous studies have implicated calcium in blue‐light signaling in plants and algae. A photophobic response to high‐intensity blue light was characterized in the marine benthic diatom Navicula perminuta (Grunow) in van Heurck. Calcium modulators were used to determine the involvement of calcium in the signaling of this response, and the fluorescent calcium indicator Calcium Crimson was used to image changes in intracellular [Ca²⁺] during a response. A localized, transient elevation of Calcium Crimson fluorescence was seen at the cell tip at the time of cell reversal. Intracellular calcium release inhibitors produced a significant decrease in the population photophobic response. Treatments known to decrease influx of extracellular calcium had no effect on the population photophobic response but did cause a significant decrease in average cell speed. As the increase in intracellular [Ca²⁺] at the cell tip corresponded to the time of direction change rather than the onset of the light stimulus, it would appear that Ca²⁺ constitutes a component of the switching mechanism that leads to reversal of the locomotion machinery. Our current evidence suggests that the source of this Ca²⁺ is intracellular.     

Dynamics of the cytoskeleton of epidermal cells in situ and in culture

Summary The cytoskeleton of primary tissue-culture cells from the epidermis of Xenopus laevis tadpoles was investigated by phase-contrast, immunofluorescence, and electron microscopy. The connection between the arrangement of different types of filaments and the mechanical properties of the epidermis is discussed. The bilayered epidermis attains stability from thick bundles of tonofilaments interconnecting the basal desmosomes. Twisting of tonofilaments around each other can explain the occurrence of elastic filamentous curls forming a meshwork braced between rows of small desmosomes in the apical region of the epidermis. Actin is arranged as a diffuse meshwork and sometimes forms bundles intermingling with tonofilament bundles. Surface membranes and rows of small desmosomes are delineated by actin and contain -actinin. Actin raises the tension for rounding and spreading of cells. Microtubules stabilize already well-developed lamellae.     

C-banding patterns in the AustralianBulbine (Liliaceae): The perennial group,B. bulbosa s. lato

C-banding studies support earlier evidence thatB. bulbosa, as a previously circumscribed, is heterogeneous, consisting of three distinct entities: (1) theB. bulbosa complex (B. bulbosa s. str.) at 4x (2n = 24), 8x (2n = 48) and 12x (2n = 72) ploidy levels, (2) the rock lily and (3) the Kroombit population (both 2n = 46). Each of these three main groups has a distinctive banding profile, though centromeric and telomeric dot bands, variably expressed, are common to all. In theB. bulbosa complex, substantial heterochromatin development, apart from bands associated with the NORs on chromosomes 1 L, 2 S and 3 L, occurs only at the terminal regions of the short arms of the large and middlesized acrocentric chromosomes, with considerable polymorphic and polytypic variation in the number and size of the heterochromatic blocks, especially at the 4x level. Queensland 8xB. bulbosa populations differ in having terminal heterochromatin, probably associated with NORs, on 11 S and 12 S, and in having some strong interstitial bands. The differences appear to correlate with attributes relating to flower morphology, and may have systematic significance. The karyotypes of rock lily and Kroombit are somewhat similar but the former has a characteristic C-band profile with multiple interstitial bands on chromosomes 1–5 and 7–9, whereas the latter has only one interstitial band on chromosome 9.First contribution of a series on cytoevolution in the AustralianBulbine. Two introductory papers in Austral. J. Bot.34 (2)