Breeding system of the annual Cruciferae, <Emphasis Type="Italic">Arabidopsis kamchatica</Emphasis> subsp. <Emphasis Type="Italic">kawasakiana</Emphasis>

The breeding system of an annual Cruciferae, Arabidopsis kamchatica subsp. kawasakiana, was studied in three natural populations. We applied four experimental treatments, open pollination, bagging, emasculation + bagging, and emasculation + hand-pollination + bagging. None of the emasculated flowers with bags produced fruits but we observed high fruit sets in the other three treatments. The results confirmed that A. kamchatica subsp. kawasakiana is a self-compatible, non-apomictic species that can produce seeds through auto-pollination. Considering the life cycle as an annual, increased reproductive assurance through auto-pollination should be critical for the maintenance of populations of A. kamchatica subsp. kawasakiana.     

Effects of Actinomycin D on Generation Time and Morphogenesis in Paramecium*

SYNOPSIS. The sensitivity of Paramecium tetraurelia (=P. aurelia syngen 4) cells to pulse treatments with various doses of Actinomycin D (AMD) was estimated by comparing the generation times of treated and untreated sister cells. It was found that the delay of division in treated cells depended on the concentration of AMD, on their “age” at the time of the pulse treatment, and on their individual sensitivity. Sensitivity of Paramecium to AMD changes during the cell cycle in a predictable way. About 3 1/2 hr before the normally expected cell fission (total generation time ～ 5 1/2 hr) there is a decrease of sensitivity. Thereafter, the cell enters a new stage with a progressive increase of sensitivity. This 2nd phase ends at the “transition point” (～ 2 hr before cell division), when sensitivity drops abruptly. The division process itself may be altered and slowed down by high concentrations of AMD, even if the drug is applied after the transition point, but this process can never be completely annulled. The impairment of the division mechanism may lead to morphologic anomalies in the offspring. Resorption of oral anlagen in P. tetraurelia probably never occurs during the cell cycle after AMD treatment. The reason for individual variability of the cells, mechanisms controlling development, and the question of an obligate sequence of gene action in each cell cycle are discussed.     

Plasticity of insect reproduction: testing models of flexible and fixed development in response to different growth rates

We tested alternative developmental hypotheses describing when during an insect oviposition cycle reproductive tactics are determined. Newly eclosed adult females of the grasshopper Romalea guttata were raised on eight different feeding treatments consisting of a low food diet, a high food diet, and changes from high to low food, or low to high food, at different times during the first oviposition cycle. When initial food availability was high, a decline in food availability >7 days after adult eclosion produced no significant increase in time to oviposition compared to constant high food. In contrast, when initial food availability was low, an increase in food availability as late as day 14 produced a significant decrease in time to oviposition compared to constant low food. Thus, time to oviposition is determined by feeding rate early in the oviposition cycle, but the time of this determination is later when food availability is lower. Masses of individual eggs were unaffected by these treatments. When initial food availability was high, a decrease in food availability on day 21 produced no significant change in numbers of eggs in a clutch compared to constant high food. In contrast, when initial food availability was low, an increase in food availability after day 7 produced no significant change in number of eggs in a clutch compared to constant low food. Changes in egg production resulted from oocyte resorption, which appeared to become unresponsive to food availability between day 14 and day 21. Our results refute the hypothesis that reproductive tactics are continuously flexible. Development toward oviposition seems to be structured so that reproductive tactics become independent of feeding late during the first oviposition cycle. Reproductive tactics become unresponsive to food at different times for groups initially receiving low or high food, suggesting that a particular developmental state, rather than some absolute time, marks the shift to development that is unresponsive to␣food. Plasticity in reproductive tactics appears to be␣controlled by hormones in a manner similar to the hormonal control of plasticity of metamorphosis in other insects. Received: 21 September 1997 / Accepted: 10 March 1998     

EFFECTS OF HIGH PRESSURE IN THE ARMORED DINOFLAGELLATE SCRIPPSIELLA HEXAPRAECINGULA (PERIDINIALES,DINOPHYCEAE): CHANGES IN THECAL PLATE PATTERN AND MICROTUBULE ASSEMBLY1

The possible role of cortical microtubules in dinoflagellates was studied using high‐pressure treatments applied to nonmotile cells (just after ecdysis) of Scrippsiella hexapraecingula T. Horig. et Chihara. Whereas considerable disorganization of cortical microtubules was observed when cells were exposed to high‐pressure treatments of 98 MPa or more for 5–15 min, they were mostly intact in cells exposed to a pressure of <98 MPa for 5 min. After nonmotile cells were exposed to high‐pressure treatments sufficient to disorganize the cortical microtubules, they produced new motile cells with thecal plate patterns that differed considerably from the pattern known for this species. Increasing the intensity of high pressure applied to nonmotile cells resulted in an increase in the number of cells that exhibited disorganized cortical microtubules as well as a change in their thecal plate pattern, suggesting that high pressure disorganizes cortical microtubules leading to a change in the thecal plate pattern.     

Nodulation ofMedicago sativa in solution culture

Summary When nitrate was maintained in continuous supply in solution cultures at concentration 0.02 mM to 2 mM it reduced the numbers of curled root hairs and of nodules. But the addition of indole-3-acetate with the inoculum overcame the nitrate-inhibition of curling without affecting nodulation; and there was no quantitative connection between numbers of curled hairs and of nodules when nitrate treatments of limited duration were applied at different times in relation to inoculationNitrate reduced nodule number to some extent even if present only before inoculation, or only on the first, second or third day after inoculation, so that reduction in nodule number was difficult to attribute solely or chiefly to interference with any one phase of the nodulation process. However, nitrate inhibited formation of infection threads and augmented the proportion of arrested infection threads, to such an extent that this could be important in limiting nodule number.Nitrate also delayed nodulation. The delay occurred early after inoculation and corresponded with delay in the appearance of infection threads.     

The use of chemicals, antagonists, repellents and physical barriers for the control of Lycoriella auripila (Diptera: Sciaridae), a pest of the cultivated mushroom Agaricus bisporus

The sciarid, Lycoriella auripila, is a serious pest of commercial mushroom production. A series of trials demonstrated that the use of early, specifically-targeted, treatments of insecticides and/or antagonists and repellents, which distance treatment time from crop harvest, have the potential to play a useful part in the control of initial and subsequent generations of this pest. Of the treatments examined, those involving a drench treatment of the compost at filling (before pasteurisation) proved to be the most effective. Cyromazine and diflubenzuron were the most active insecticides tested, with cyromazine achieving a superior level of control of the initial infestation. Repellents and antifeedants were also effective, with calcium oxalate and sinapic acid both achieving about 50% control when applied at filling. Treatments applied later during the production cycle, unless in combination with a treatment at filling, were progressively less effective at controlling both the initial sciarid infestation and later generations of larvae. Multiple treatments caused greater reductions in fly populations than did the single treatments and continued to do so throughout the cropping cycle, the greatest reduction in the initial generation (79%) occurring with a triple treatment of cyromazine. With the exception of some diflubenzuron treatments, those that were effective resulted in increases in yield. The use of a physical paper barrier caused significant increases in both fly numbers and total yield.     

A density-dependent model of Cirsium vulgare population dynamics using field-estimated parameter values

Two versions of a stage-structured model of Cirsium vulgare population dynamics were developed. Both incorporated density dependence at one stage in the life cycle of the plant. In version 1 density dependence was assumed to operate during germination whilst in version 2 it was included at the seedling stage. Density-dependent parameter values for the model were estimated from annual census data in a factorial grazing experiment. Version 1 of the model produced significant estimates of density dependence under field conditions. The estimated values, when included in a simulation of the dynamics, produced two-point limit cycles under conditions of hard grazing. The limit cycles were most pronounced at the early rosette stage. Comparison of the effects of density dependence at the two different stages in the life cycle revealed a strong difference in predicted dynamics. This emphasizes the importance of determining where density dependence operates under field conditions and the potential problems of arbitrarily assigning it to particular life-history stages. Version 1 of the model produced a good prediction of observed mean plant density across the different grazing treatments (r ²=0.81, P<0.001).     

Asymmetric re‐entrainment and phase response of the circadian activity rhythm of a nocturnal marsupial (Petaurus breviceps: Phalangeridae)

Abstract

Sugar Gliders (Petaurus breviceps) re‐entrain faster after 8‐h delay shifts of an LD 12:12 and an LD 8:16 (31–56:0.3 lux each) than after 8‐h advance shifts of these Zeitgeber cycles. In order to test whether this asymmetric re‐entrainment behavior is related to, or even caused by the phase response characteristics of the circadian system, the phase response of the activity rhythm to short and long light pulses was studied. Short light pulses (15 min of 31–56 lux against a background intensity of 0.3 lux) caused only relatively small delay shifts when applied around the onset, and more pronounced advance shifts when given at the end of the activity time (α). Onset and end of activity shifted by different amounts. Long light pulses produced by 8‐h advances and delays of one single lighttime of an LD 12:12 elicited pronounced phase delays when applied at the beginning of the activity time, but only minor phase advances when given at the posterior part of α. These results indicate that in Petaurus breviceps the phase response characteristics to long light pulses exerting parametric effects of light are responsible for the pronounced asymmetry effect in re‐entrainment. Differing phase responses of onset and end of activity point to a two‐oscillator structure of the circadian pacemaker system in this marsupial.     

Effects of sequential stimuli on Halobacterium salinarium photobehavior.

We analyzed the motor photoresponses of Halobacterium salinarium to different test stimuli applied after a first photophobic response produced by a step-down of red-orange light (prestimulus). We observed that pulses given with a suitable delay after the prestimulus produced unusual responses. Pulses of blue, green, or red-orange light, each eliciting no response when applied alone, produced a secondary photophobic response when applied several seconds after the prestimulus; the same occurred with a negative blue pulse (rapid shut-off and turning on of a blue light). Conversely, no secondary photophobic response was observed when the test stimulus was a step (a step-up for red-orange light, a step-down for blue light) of the same wavelength and intensity. When the delay was varied, different results were obtained with different wavelengths; red-orange pulses were typically effective in producing a secondary photophobic response, even with a delay of 2 s, whereas the response to a blue pulse was suppressed when the test stimulus was applied within 5 s after the prestimulus. The secondary photophobic response to pulses was abolished by reducing the intensity of the prestimulus without affecting the primary photophobic response. These results, some of which were previously reported in the literature as inverse effects, must be produced by a facilitating mechanism depending on the prestimulus itself, the occurrence of reversals being per se ineffective. The fact that red-orange test stimuli are facilitated even at the shortest delay, whereas those of different wavelengths become effective only after several seconds, suggests that the putative mechanism of the facilitating effect is specific for different signaling pathways.     

Controlling invasive Arrhenatherum elatius and promoting native prairie grasses through mowing

Abstract. Control of invasive plants is a key element of conservation and restoration efforts. We report results from a five‐year field experiment in western Oregon, USA that evaluates the effects of different mowing regimes on the non‐native and invasive perennial grass Arrhenatherum elatius, the native perennial prairie grasses Danthonia californica and Festuca roemeri, and groups of other native and non‐native grasses and forbs. Eight treatments were designed to test hypotheses about the role of mowing height and time of application on the plant community. Differences among treatments emerged only after two or three years of treatment. This delay in response reinforces the need for long‐term studies. Annual mowing was most effective at reducing Arrhenatherum cover and flowering when applied in late spring or early summer, the time of Arrhenatherum flowering and expected maximum above‐ground allocation. Double mowing and mowing at 15 cm were more effective in reducing Arrhenatherum cover than were single mowing and mowing at 50 cm. All treatments increased the cover and flowering of Danthonia. Statistical model analysis showed that increases in cover and flowering of the native grass Danthonia were caused by its release from suppression by Arrhenatherum. Fouryears of the most effective treatment, mowing at 15 cm in late spring, converted an Arrhenatherum‐dominated site to a prairie dominated by native grasses. This is one of the few documented cases of pest plant control causing an increase in native plant abundance. These results show that mowing, properly applied, can be an effective tool for restoring degraded, Arrhenatherum‐dormnated prairies.     

The effects of organic and inorganic fertilizer applications to Miscanthus×giganteus,Arundo donax and Phalaris arundinacea,when grown as energy crops in Wales,UK

Two pot trials and one field trial were established to investigate the effects of organic and inorganic fertilizer applications to energy crops grown in mid‐Wales. Chicken litter and sewage cake applied at a high level in excess of MAFF recommendations produced an increased yield response in Miscanthus and Arundo plants. Miscanthus plants exhibited an increased growth response to all fertilizers applied in its second year. Fertilizer applications in accordance with MAFF recommendations produced no significant differences in yields for Miscanthus or Arundo potted plants. In the field there was an increased yield response of Miscanthus to inorganic nitrogen applications compared with organic manures, but not with control plots. Analysis of the Miscanthus plant material at harvest showed significant differences in the nitrogen, potassium and copper content between treatments. No mineral content differences were shown for Miscanthus rhizome material or Arundo plant material. The Phalaris plants did not exhibit significant differences in growth or yield parameters, but their plant matter showed differences in nitrogen, phosphorus, potassium, calcium, sulphur and boron content between treatments.     

Biosensor online control of citric acid production from glucose by Yarrowia lipolytica using semicontinuous fermentation

Our study aimed at the development of an effective method for citric acid production from glucose by use of the yeast Yarrowia lipolytica. The new method included an automated bioprocess control using a glucose biosensor. Several fermentation methodologies including batch, fed‐batch, repeated batch and repeated fed‐batch cultivation were tested. The best results were achieved during repeated fed‐batch cultivation: Within 3 days of cycle duration, approximately 100 g/L citric acid were produced. The yields reached values between 0.51 and 0.65 g/g and the selectivity of the bioprocess for citric acid was as high as 94%. Due to the elongation of the production phase of the bioprocess with growth‐decoupled citric acid production, and by operating the fermentation in cycles, an increase in citric acid production of 32% was achieved compared with simple batch fermentation.     

Research note: Irradiance of photosynthetically active radiation determines ultraviolet-susceptibility of photosynthesis in Ulva lactuca L.(Chlorophyta)

The respective ratio of photosynthetically active to ultraviolet radiation is of crucial importance to results obtained in ultraviolet (UV)‐research on photoautotrophic organisms. Specimens of the green macroalga Ulva lactuca L. were exposed to a constant irradiance of UV‐radiation at increasing irradiances of photosynthetically active radiation (PAR). The effects of experimental irradiance and spectral composition on photoinhibition of photosynthesis and its recovery were monitored by chlorophyll fluorescence measurements and the activity of the xanthophyll cycle was assessed by high performance liquid chromatography‐(HPLC) based pigment analysis. Results indicate a UV‐induced delay in recovery from PAR‐induced photoinhibition and a deceleration of violaxanthin conversion within the xanthophyll cycle due to the presence of UV‐radiation. Also the concentration of the protective pigment lutein increased considerably and could be indicative of the existence of an additional light‐protective mechanism, as, for example, the lutein‐epoxid cycle in Ulva. In total, results clearly show that the extent of UV‐induced inhibition of photosynthesis to be found in UV‐exposure experiments is highly dependent on the irradiance of background photosynthetically active radiation: with increasing irradiance of PAR the UV‐effects were diminished. Exemplified by the green algae Ulva lactuca this study demonstrates the crucial importance of the ratios of PAR:UV applied in UV‐research, particularly when conducting laboratory experiments in an ecological context.     

Reduction of ultraviolet-induced mitotic delay by caffeine in G2-phase irradiated plasmodia of Physarum polycephalum

Synchronously mitotic surface Plasmodia ofPhysarum polycephalum were ultra-violet-irradiated at different times during G2-phase (—4 h to —20 min with respect to metaphase), and treated immediately thereafter with varying concentrations of caffeine. It was observed that ultraviolet-induced mitotic delay is reduced significantly by this methylxanthine. In plasmodia irradiated between —4 and —1 h with respect to metaphase, the effect was concentration-dependent and the need for a certain threshold dose for obtaining the reduction in delay was apparent. However, higher doses than this were fairly toxic when applied at this part of the cycle and led to more mitotic delay than that obtained with UV alone. The most striking observation made during this study was the phase-specific precipitous effect seen in those plasmodia irradiated at about 20 min before mitosis which almost eliminated the long delay due to ultraviolet-irradiation. These results are discussed in the context of some of the known effects of ultraviolet and caffeine on a mitosis-promoting factor. It is proposed that the significant reduction of ultraviolet-induced mitotic delay reported here is due to the reactivation of the ultraviolet-inactivated mitosis-promoting factor by caffeine. Alternatively, it is possible that caffeine may prevent the inactivation of this factor by ultraviolet.     

Comparison of the effects of Altozar and DDT on females ofIxodes persulcatus Sch. during and after feeding

Ixodes persulcatus females were treated with Altozar and DDT at one of six consecutive time-points during the period beginning with attachments and ending with the commencement of oviposition.The moment of tick attachment (point 1) appeared to be the most sensitive time for the action of Altozar; a delay of detachment of some of the engorged females was observed, as well as a decrease in their mean weight and the non-detachment of a considerable number of the females over a long period. Similar effects were observed when Altozar was applied on the first and third days after attachments (points 2 and 3) although in the first case they were less pronounced. Altozar treatments on the fifth day of feeding, shortly before oviposition, and especially immediately after detachment (points 4,5 and 6, respectively) mainly resulted in non-viability of the eggs.DDT was ineffective when it was applied at point 1. DDT treatments of ticks at points 3 and 5 were highly effective; this was expresseds by a reduction in weight of the engorged females and a suppression of their fecundity at point 3, and death of all the larvae at different times after hatching at point 5. Many larvae also perished when the ticks were treated with DDT at point 6. Ticks were quite tolerant to DDT action when they were treated at points 2 and 4.The action of both compounds ultimately resulted in the inhibition of tick reproduction, but the rate and method depended on the compound applied as well as on the time of treatment. Altozar effects after treatment at points 1–3 may be considered to be hormonal.     

Coordination of DNA damage tolerance mechanisms with cell cycle progression in fission yeast

DNA damage tolerance (DDT) mechanisms allow cells to synthesize a new DNA strand when the template is damaged. Many mutations resulting from DNA damage in eukaryotes are generated during DDT when cells use the mutagenic translesion polymerases, Rev1 and Polζ, rather than mechanisms with higher fidelity. The coordination among DDT mechanisms is not well understood. We used live-cell imaging to study the function of DDT mechanisms throughout the cell cycle of the fission yeast Schizosaccharomyces pombe. We report that checkpoint-dependent mitotic delay provides a cellular mechanism to ensure the completion of high fidelity DDT, largely by homology-directed repair (HDR). DDT by mutagenic polymerases is suppressed during the checkpoint delay by a mechanism dependent on Rad51 recombinase. When cells pass the G2/M checkpoint and can no longer delay mitosis, they completely lose the capacity for HDR and simultaneously exhibit a requirement for Rev1 and Polζ. Thus, DDT is coordinated with the checkpoint response so that the activity of mutagenic polymerases is confined to a vulnerable period of the cell cycle when checkpoint delay and HDR are not possible.     

Persistent effects of seedling treatment on growth of sugar beet in pots

Further evidence is provided that the environment of sugar-beet seedlings, or growth substances applied to seedlings, continues to influence growth when the plants are later in other environments. Sugar-beet seeds were germinated at 20 °C in 8, 16 or 24 h photoperiods of constant light intensity, i.e. with different amounts of total radiation. When the seedlings had two leaves (15–18 days old) they were transferred to large pots in the glasshouse. Some seedlings were treated with (2-chloroethyltrimethylammonium chloride) either sprayed on the leaves or applied to the soil, at different times. The treatments affected areas of individual leaves throughout the growing period; plants raised in 24 h photoperiod had the largest leaves, and those in 8 h photoperiod the smallest. Consequently, 24 h plants had most dry matter and 8 h plants least. Plants given most radiation produced leaves fastest and CCC applied early increased the rate, but as the leaves were smaller, except late in 1967, and died sooner, the leaf area duration was less and so yields were less. CCC applied later did not affect leaf production. There was no interaction between amount of radiation and CCC treatment. Twenty-four hour plants had the greatest net assimilation rate (E) early. CCC decreased E early, but increased it later and more when sprayed on the leaves than when applied to the soil. Some factor, possibly pot size, eventually restricted growth and probably diminished the effect of the treatments applied to the seedlings.     

Potentiation by p-fluorophenylalanine of the division-delaying effects of high hydrostatic pressure in synchronized Tetrahymena pyriformis.

Cultures of heat-synchronized Tetrahymena pyriformis, growing in a proteose peptone medium, were subjected to short pulses of the amino acid analogue, p-fluorophenylalanine, and high hydrostatic pressure. The pulses of these agents were chosen so that, when applied individually, they did not appreciably delay cell division. However, combined treatments, analogue pulse followed by pressure pulse, produced a pronounced synergism. The results are interpreted as further evidence to support the inclusion of analogue division proteins in pressure labile assemblies during the progression of Tetrahymena into division.     

Effect of phosphorus source and rate of application on VAM fungal infection and growth of maize (Zea mays L.)

The effects of two phosphorus (P) sources (triple superphosphate and Ghafsa phosphate rock), applied at rates equivalent to 44kg ha^-1 and 22 kg ha^-1, on vesicular-arbuscular mycorrhizal (VAM) fungal infection in roots, dry matter yield and nutrient content of maize grown in an oxisol and an alfisol, were investigated in a growth cabinet. The application of 44 kg P ha ^-1 resulted in root infection by VAM fungi not was significantly different (P<-0.01) from when no P was applied. Root infection was significantly greater when P was applied as triple superphosphate at the rate of 22 kg ha^-1 the higher rate. Phosphate rock treatments at both rates of application resulted in significantly greater root infection than in controls with no P or when triple superphosphate was applied at 44 kg ha^-1. Plant P uptake increased in all soils with the different P treatments compared with the control. No direct effects of the treatments on the aluminium and zinc contents of maize plants were observed. In the gleyic alfisol, reduced Mn uptake as a result of increased infection of plants with the superphosphate treatments was observed. Higher Mn was also found in plants with the higher rate of superphosphate treatment than with the phosphate rock treatments in the haplustox, although infection rates in plants with the latter treatments were higher. With the exception of plants with the phosphate rock treatment applied at 22kg ha^-1, dry matter yields of plants with all P sources were significantly greater than the controls.     

Cell proliferation and cell cycle alterations in oesophageal p53‐mutated cancer cells treated with cisplatin in combination with photodynamic therapy

Objectives: The major goal of anti‐cancer therapies is selective destruction of tumour cells with minimum side effects on normal cells. Towards this aim, combination of different therapeutic modalities has been evaluated for improving control of neoplastic diseases and quality of life for the patient. Photodynamic therapy (PDT) is a procedure for treatment of various types of cancer, but its combination with other established treatments has not been evaluated in detail. We have used KYSE‐510 cells from a human oesophageal carcinoma as an in vitro model to investigate whether cisplatin (CDDP) could be combined with PDT to increase cell death with respect to single treatments. Materials and methods: p53‐mutated KYSE‐510 cells were treated with CDDP alone or in combination with PDT. Analyses of cell viability, cell cycle progression and apoptosis induction were carried out at specific times after treatments. Results: Decrease in cell viability, cell cycle arrest at the G₂/M‐ and S‐phases boundary, and apoptosis induction were observed after single and combined treatments. Conclusions: Our results show that low CDDP doses (0.25–1 μm ) induce cell mortality and cell cycle perturbation, which were more evident when given in combination with PDT, but in contrast to work of other authors no synergistic activity was found. Apoptosis occurred via intrinsic pathways in treated cells, although it did not represent the predominant mode of cell death.