Distylic Hottonia palustris shows high reproductive success in small populations despite low genetic variability

Hottonia palustris L. (Primulaceae) is characterized by a heteromorphic incompatibility system. The strategy of distylic ramets is believed to promote outcrossing, to maintain overall genetic diversity and to prevent inbreeding depression. In spite of this distyly, an extremely low amount of allozymic diversity was observed in 545 individual ramets from 14 populations in Flanders (Belgium). A possible explanation for such low genetic variation is discussed in relation to the vegetative propagating abilities and the ecological niche width of the species. In contrast to the uniformity in allozymes as well as to the feature of single morph populations, there was a high variability in reproductive success between populations such as the number of seeds per ramet (425–2633), the number of flowers per ramet (9–36) and the mean weight of seeds (0.03–0.17 mg). Small populations and even those consisting of only one style morph may show a high reproductive success. As a whole, H. palustris showed a negative relationship of reproductive success with the surface area of its populations.     

A bacteriophytochrome regulates the synthesis of LH4 complexesin <Emphasis Type="Italic">Rhodopseudomonas palustris</Emphasis>

The non-sulphur purple bacterium Rhodopseudomonas palustris contains five pucAB genes for peripheral light-harvesting complexes. Bacteria grown under high-light conditions absorb at 800 and 850 nm but in low-light the 850 nm peak is almost absent and LH2 complexes are replaced by LH4. The genome contains six bacteriophytochromes (Bph). Bphs sense light in the red/far-red through a reversible Pr to Pfr transformation that controls gene expression. Bph3 (RPA1537) controls the expression of a cluster of photosynthetic genes, however most of the peripheral light harvesting complex genes are outside of this region. The pucAB-d genes encode LH4 peptides and are near two Bphs (RPA3015, RPA3016). We have characterised three Bphs and show that Bph4 RPA3015 and Bph3 RPA1537 have different dark stable states. It is known that Bph3 is active in its red absorbing Pr form and suggests a working hypothesis that Bph4 is active in the Pfr state. We show that LH4 expression can be induced with red light at the Pr absorption maximum (708 nm) of Bph4. The property of light transmission of water maybe an important factor in understanding this adaptation. Bph4 can sense the reduction in light intensity indirectly through an increase in ratio of transmitted red/far-red light. The red right activated Bph4 regulates the synthesis of LH4 which concentrates bacteriochlorophyll a pigment absorption at 800 nm to exploit a recovery in water light transmission in this region.     

Using reverse micelles as microreactor for hydrogen production by coupled systems of Nostoc / R. palustris and Anabaena / R. palustris

Uptake hydrogenase negative mutants of bloom forming cyanobacteria (Nostoc and Anabaena) and the fermentative bacteria Rhodopseudomonas palustris P₄ were used together for producing hydrogen within the reverse micelles fabricated by N-ethyl hexyl sodium sulfosuccinate (AOT) in isooctane and cetyl trimethyl ammonium bromide (CTAB) in benzene. The rate of H₂ production in AOT/isooctane reverse micellar system was found to be more promising in comparison to the CTAB/Benzene reverse micellar entrapment. After mutagenesis in 2.0% (v/v) ethyl methane sulphonate (EMS) mutants of Nostoc and Anabaena were selected on BG-11 plates (containing 2% agar) and then used for analysis of produced hydrogen. In comparison to the unmutated Nostoc with R. palustris (within AOT/isooctane) the coupled system of mutated Nostoc and R. palustris produced H₂ by 3.9-fold higher rate, which is 8.6 mmol H₂/h/mg protein. Whereas, mutated Anabaena coupled with R. palustris produced 4.8 times higher hydrogen production within (AOT)/isooctane reverse micelles in comparison to the unmutated Anabaena with R. palustris. Effect of nitrogen to carbon ratio (N/C) on hydrogen production was studied and Anabaena/R. palustris and Nostoc/R. palustris systems were, respectively, found to generate 11.2 and 9.8 mmol H₂/h/mg protein continuously for 3 days. Effects of temperature and light intensity were also investigated and we found that 32°C temperature and 1,000 Lux light intensity are the optimum values in these systems. Addition of sodium dithionite also resulted in further enhancement of the rate and duration of hydrogen production in both (mutated Nostoc/R. palustris and mutated Anabaena/R.␣palustris) systems.     

The role of respiration in the activation of photosynthesis upon illumination of dark adapted Chlamydomonas reinhardtii

It is reported that O₂ is required for the activation of photosynthesis in dark adapted Chlamydomonas reinhardtii in State 1, under low light intensity. The concentration of dissolved O₂ of ca. 9 µM is sufficient to saturate the requirement. When the concentration of O₂ is 3 μM or below, the activation of photosynthesis is strongly inhibited by myxothiazol, a specific inhibitor of the mitochondrial cytochrome bc₁. The effect of this inhibitor decreases as the O₂ concentration is raised, to disappear completely above 50 μM. Low concentrations of uncouplers delay the activation of photosynthesis, but do not inhibit it when steady state is reached. It is concluded that in State 1 C. reinhardtii mitochondrial respiration is required for the activation of photosynthesis upon illumination of dark adapted cells only when the concentration of O₂ is too low (less than 5 μM) to allow an appreciable activity of the Mehler reaction. The role of respiration does not seem to be due to the synthesis of ATP by oxidative phosphorylation, because photosynthesis activation is not sensitive to oligomycin.     

Potential respiration is a better respiratory predictor than biomass in young Artemia salina

These experiments test whether respiration can be predicted better from biomass or from potential respiration, a measurement of the mitochondrial and microsomal respiratory electron transport systems. For nearly a century Kleiber's law or a similar precursor have argued the importance of biomass in predicting respiration. In the last decade, a version of the Metabolic Theory of Ecology has elaborated on Kleiber's Law adding emphasis to the importance of biomass in predicting respiration. We argue that Kleiber's law works because biomass packages mitochondria and microsomal electron transport complexes. On a scale of five orders of magnitude we have shown previously that potential respiration predicts respiration as well as biomass in marine zooplankton. Here, using cultures of the branchiopod, Artemia salina and on a scale of less than 2 orders of magnitude, we investigated the power of biomass and potential respiration in predicting respiration. We measured biomass, respiration and potential respiration in Artemia grown in different ways and found that potential respiration (Ф) could predict respiration (R), both in µlO₂ h⁻¹ (R = 0.924Φ + 0.062, r² = 0.976), but biomass (as mg dry mass) could not (R = 27.02DM + 8.857, r² = 0.128). Furthermore the R/Ф ratio appeared independent of age and differences in the food source.     

A new role for the SHATTERPROOF genes during Arabidopsis gynoecium development

Gynoecium development is a complex process which is regulated by key factors that control the spatial formation of the apical, medial and basal parts. SHATTERPROOF1 (SHP1) and SHP2, two closely related MADS-box genes, redundantly control the differentiation of the dehiscence zone and promote the lignification of adjacent cells. Furthermore, SHP1 and SHP2 have shown to play an important role in ovule identity determination. The present work identifies a new function for these two genes in promoting stigma, style and medial tissue development. This new role was discovered by combining the shp1 shp2 double mutant with the aintegumenta (ant) and crabs claw (crc) mutants. In quadruple mutant flowers, the inner whorl is composed of unfused carpels which lack almost completely apical and medial tissues, a phenotype similar to the previously reported fil ant and lug ant double mutants.     

Lavandula angustifolia essential oil as a novel and promising natural candidate for tick (Rhipicephalus (Boophilus) annulatus) control

Lavandula angustifolia is a well known herbal medicine with a variety of useful properties, including its acaricidal effect. This experiment was carried out to study the bioacaricidal activity of L. angustifolia essential oil (EO) against engorged Rhipicephalus (Boophilus) annulatus (Acari; Ixodidae) females. For this purpose six serial concentrations (0.5, 1.0, 2.0, 4.0, 6.0 and 8.0% w/v) of L. angustifolia EO were used. There was considerable mortality in concentrations more than 4.0% although there were no differences between 6.0 and 8.0% in all measured criteria. The mortality rate 24 h after inoculation was 73.26 and 100% in groups treated with 4.0 and 8.0% EO, respectively. Lavender EO also reduced tick egg weight in a concentration-dependent manner. The amount of eggs produced varied from 0.12 g (at 0.5% EO) to 0.00 g (at 8.0% EO) but did not differ statistically from the control. L. angustifolia EO caused 100% failure in egg laying at 6.0 and 8.0% whereas this value in the control group was zero. A positive correlation between L. angustifolia EO concentration and tick control, assessed by relative mortality rate and egg-laying weight, was observed by the EO LC/EC₅₀, which, when calculated using the Probit test, was 2.76-fold higher than the control. Lavender is a promising acaricidal against R. (B.) annulatusin vitro.     

Seasonal growth dynamics of Laurencia papillosa and Gracilaria coronopifolia from a highly eutrophic reef in southern Taiwan: temperature limitation and nutrient availability

Both field and laboratory studies were used to investigate the effects of temperature limitation and nutrient availability on seasonal growth dynamics of Laurencia papillosa and Gracilaria coronopifolia from a nearshore coral reef in the southern tip of Taiwan during 1999-2000. L. papillosa was a summer blooming alga abundant in August-November and G. coronopifolia was abundant year round except April-May. L. papillosa blooms in the summer were attributed to its preference for high temperatures and highly sensitivity to low temperatures. A wider temperature range and a significant stimulation of growth by high N inputs can explain the appearance of G. coronopifolia year round and also its maximum growth in November-March. Levels of dissolved inorganic nitrogen (DIN) and soluble reactive phosphorus (SRP) in water column were extremely high, but the growth of these two rhodophytes still suffered nutrient limitation that the type and severity of nutrient limitation were variable over time and also between two species. The growth of L. papillosa was limited by P in the early growth stage (August-September) as indicated by decreased tissue P contents, increased C/P and N/P molar ratios and increased alkaline phosphatase activity (APA) and in the later growth stage, it was subjected to N-limitation, evidenced by decreased tissue N contents and C/P and N/P molar ratios and increased tissue P contents. The growth of G. coronopifolia was also P-limited as indicated by increased tissue N contents and concomitantly decreased tissue P contents, while marked drops in tissue P contents below the subsistence level in mid September and December 1999 reveal severe P limitation, which was supported by increased alkaline phosphatase activity. Higher critical nutrient contents and nutrient thresholds for maximum growth of G. coronopifolia suggest that G. coronopifolia faced more frequent nutrient limitation compared to L. papillosa. In conclusion, the results from these laboratory and field studies provide evidence that the seasonal abundance of L. papillosa and G. coronopifolia from southern Taiwan was determined by seasonal variations in seawater temperatures and nutrient concentrations as well as different physiological growth strategies. Seawater temperature and nutrient availability were important determinants of seasonal abundance of L. papillosa while the seasonal abundance of G. coronopifolia was influenced by nutrient availability.     

Primary production,respiration and calcification of the temperate free-living coralline alga Lithothamnion corallioides

Calcification and primary production responses to irradiance in the temperate coralline alga Lithothamnion corallioides were measured in summer 2004 and winter 2005 in the Bay of Brest. Coralline algae were incubated in dark and clear bottles exposed to different irradiances. Net primary production reached 1.5 μmol C g⁻¹ dry wt h⁻¹ in August and was twice as high as in January–February. Dark respiration showed significant seasonal variations, being three-fold higher in summer. Maximum calcification varied from 0.6 μmol g⁻¹ dry wt h⁻¹ in summer 2004 to 0.4 μmol g⁻¹ dry wt h⁻¹ in winter 2005. According to P–E curves and the daily course of irradiance, estimated daily net production and calcification reached 131 μg C g⁻¹ dry wt and 970 μg CaCO₃ g⁻¹ dry wt in summer 2004, and 36 μg C g⁻¹ dry wt and 336 μg CaCO₃ g⁻¹ dry wt in winter 2005. The net primary production of natural L. corallioides populations in shallow waters was estimated at 10–600 g C m⁻² y⁻¹, depending on depth and algal biomass. The mean annual calcification of L. corallioides populations varied from 300 to 3000 g CaCO₃ m⁻². These results are similar to those reported for tropical coralline algae in terms of carbon and carbonate productivity. Therefore, L. corallioides can be considered as a key element of carbon and carbonate cycles in the shallow coastal waters where they live.     

Irradiance and nutrient limitation of Dicytota spp. populations on Conch Reef, Florida Keys, USA

The photosynthetic performance, pigmentation and response to nutrient enrichment of a Dictyota community were studied over a 32 m depth gradient on Conch Reef, Florida Keys, USA. Dictyota spp. was the dominant space occupier on Conch Reef. During summer months from 1994 to 2001, mean percent cover was 43% at 7 and 21 m depths. Percent cover of Dictyota spp. was markedly lower at 32 m relative to shallower sites. The metabolism of Dictyota menstrualis and Dictyota pulchella were negatively impacted by attenuation of irradiance with increased depth such that ETR_max and P/R ratio decreased by 85% and 47%, respectively in samples from 7 to 32 m. Decreased cover of Dicytota spp. at 32 m relative to shallower sites may be the result of the inability of this species complex to acclimate to low irradiance levels as indicated by a lack of change in α and photosynthetic pigment content with increased depth.The response of D. menstrualis populations on Conch Reef to nutrient enrichment was variable. During August 2000, a natural enrichment experiment was conducted as D. menstrualis photosynthetic performance was surveyed both during and for days after a period of cool, nutrient-rich internal tidal bores bathed the 21 m site. No changes in in situ photosynthetic performance were observed either during or for 2 to 4 days after this natural event suggesting no nutrient limitation. In August 2001, a manipulative nutrient enrichment experiment was conducted with D. menstrualis from 7, 21 and 32 m. Increases in Φ_II were found in individuals exposed to nutrient enrichment from all depths, indicating at that time D. menstrualis was nutrient-limited on Conch Reef. Variation in the physiological response to nutrient enrichment may be the result of the frequency of internal tidal bores on Conch Reef in the months prior to our experiments. Variation in the responses by Dictyota spp. to irradiance and nutrient enrichment suggests that long-term monitoring over relevant temporal and spatial scales is necessary to accurately characterize limits on productivity and spread of this weedy species assemblage.     

Screen of Bacillus thuringiensis toxins for transgenic rice to control Sesamia inferens and Chilo suppressalis

Transgenic rice to control stem borer damage is under development in China. To assess the potential of Bacillus thuringiensis (Bt) transgenes in stem borer control, the toxicity of five Bt protoxins (Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ba and Cry1Ca) against two rice stem borers, Sesamia inferens (pink stem borer) and Chilo suppressalis (striped stem borer), was evaluated in the laboratory by feeding neonate larvae on artificial diets containing Bt protoxins. The results indicated that Cry1Ca exhibited the highest level of toxicity to both stem borers, with an LC₅₀ of 0.24 and 0.30 μg/g for C. suppressalis and S. inferens, respectively. However, S. inferens was 4-fold lower in susceptibility to Cry1Aa, and 6- and 47-fold less susceptible to Cry1Ab and Cry1Ba, respectively, compared to C. suppressalis. To evaluate interactions among Bt protoxins in stem borer larvae, toxicity assays were performed with mixtures of Cry1Aa/Cry1Ab, Cry1Aa/Cry1Ca, Cry1Ac/Cry1Ca, Cry1Ac/Cry1Ba, Cry1Ab/Cry1Ac, Cry1Ab/Cry1Ba, and Cry1Ab/Cry1Ca at 1:1 (w/w) ratios. All protoxin mixtures demonstrated significant synergistic toxicity activity against C. suppressalis, with values of 1.6- to 11-fold higher toxicity than the theoretical additive effect. Surprisingly, all but one of the Bt protoxin mixtures were antagonistic in toxicity to S. inferens. In mortality-time response experiments, S. inferens demonstrated increased tolerance to Cry1Ab and Cry1Ac compared to C. suppressalis when treated with low or high protoxin concentrations. The data indicate the utility of Cry1Ca protoxin and a Cry1Ac/Cry1Ca mixture to control both stem borer populations.     

Microsensor study of photosynthesis and calcification in the scleractinian coral, Galaxea fascicularis: active internal carbon cycle

Sources of inorganic carbon (Ci) for photosynthesis and calcification and the mechanisms involved in their uptake in scleractinian corals were investigated in microcolonies of Galaxea fascicularis. Direct measurements of Ca²⁺, pH and O₂ on the surface and inside the polyp's coelenteron were made with microsensors. Gross photosynthesis (Pg) and net photosynthesis (Pn) were measured on the surface. Light respiration (LR) was calculated from Pg and Pn. The effect of light/dark and dark/light switches on Ca²⁺ and pH dynamics on the surface and inside the coelenteron were followed. To evaluate the different sources of Ci for photosynthesis and calcification, Ci-free seawater and 6-Ethoxyzolamide and Acetazolamide, inhibitors for carbonic anhydrase (CA) were used.In normal seawater, Pg was about seven times higher than Pn, the LR was ca. 80-90% of the Pg. Thus, most of the O₂ produced in Pg are immediately consumed in respiration, indicating the presence of a highly active internal C-cycle. As the internal C-cycle is highly active, a large part of the Ci for calcification will have passed through the metabolism of the symbiont. The high LR provides ATP for energy requiring processes in light.Ci for photosynthesis and calcification can come from seawater in the form of free Ci, respiration of photosynthates (internal C-cycle) or respiration of the ingested plankton. These sources form a common carbon pool (C-pool) that is used for the different processes.In Ci-free seawater, Pg decreased by about 12.5%, indicating that most of the photosynthetically fixed Ci can temporarily be supplied from internal sources. The initial decalcification, observed directly upon the switch to Ci-free seawater, showed that the Ca-pools in the coral are exchangeable. Part of the Pg in Ci-free seawater may depend on this decalcification for its Ci supply.Three localities of CA were defined. One on the surface facing seawater and one on endodermal cells facing the coelenteron, while the third is intracellular. The inhibition of CA decreased Pg by about 30%, while it increased the concentration of Ca²⁺ as a result of a decrease in its precipitation. The reduction of photosynthesis and calcification by CA inhibition demonstrated that both processes need the enzyme for the supply of Ci. The pH on the surface and inside the coelenteron decreased upon 6-Ethoxyzolamide addition indicating a role of CA in pH control.     

Methylviologen and dibromothymoquinone treatments of pea leaves reveal the role of photosystem I in the Chl a fluorescence rise OJIP

The effects of dibromothymoquinone (DBMIB) and methylviologen (MV) on the Chl a fluorescence induction transient (OJIP) were studied in vivo. Simultaneously measured 820-nm transmission kinetics were used to monitor electron flow through photosystem I (PSI). DBMIB inhibits the reoxidation of plastoquinol by binding to the cytochrome b₆/f complex. MV accepts electrons from the FeS clusters of PSI and it allows electrons to bypass the block that is transiently imposed by ferredoxin-NADP⁺-reductase (FNR) (inactive in dark-adapted leaves). We show that the IP phase of the OJIP transient disappears in the presence of DBMIB without affecting F_m. MV suppresses the IP phase by lowering the P level compared to untreated leaves. These observations indicate that PSI activity plays an important role in the kinetics of the OJIP transient. Two requirements for the IP phase are electron transfer beyond the cytochrome b₆/f complex (blocked by DBMIB) and a transient block at the acceptor side of PSI (bypassed by MV). It is also observed that in leaves, just like in thylakoid membranes, DBMIB can bypass its own block at the cytochrome b₆/f complex and donate electrons directly to PC⁺ and P700⁺ with a donation time τ of 4.3 s. Further, alternative explanations of the IP phase that have been proposed in the literature are discussed.     

Filtration and respiration of the deep living bivalve Acesta excavata (J.C. Fabricius, 1779) (Bivalvia; Limidae)

Here we report the first study of clearance rate and respiration rate of a deep living bivalve, Acesta excavata (J.C. Fabricius, 1779) (Mollusca: Limidae). We found that A. excavata had extreme values both for clearance and respiration rates compared to other bivalves. It has the second largest clearance rate ever reported, 13.36 l h^− 1 g^− 1, and the second lowest value of respiration rate, 0.12 ml O₂ h^− 1 g^− 1. The gill area of 7063 mm² g^− 1 is one of the largest found in bivalves so far. We suggest that these values indicate a physiological adaptation to the low and irregular food supply in the deep sea rather that a specific adaptation to depth.     

The role of ammonium in photoprotection against high irradiance in the red alga Grateloupia lanceola

Combined and/or interactive effects of inorganic nitrogen (as ammonium) and irradiance on the accumulation of nitrogenous compounds, like UV-absorbing mycosporine-like amino acids (MAAs), chlorophyll a and phycobiliproteins, were examined in the red alga Grateloupia lanceola (J. Agardh) J. Agardh in a high irradiance laboratory exposure and a subsequent recovery period under low light. Also, photosynthetic activity as in vivo chlorophyll fluorescence of photosystem II, i.e. optimum quantum yield (F_v/F_m), electron transport rate (ETR) and quantum efficiency, were examined. Photosynthetic activity, phycobiliproteins and internal nitrogen content declined during the 3-day PAR (photosynthetically active radiation; 600 μmol s⁻¹ m⁻²) and PAR + UVR (ultraviolet radiation; UVB 280–315 nm 0.8 W m⁻², UVA 315–400 nm 16 W m⁻²) exposure. Ammonium supplied in the culture medium (0, 100 and 300 μM NH₄Cl) modified the responses of the alga to high irradiance exposures in a concentration dependent manner, mainly with respect to recovery, as the highest recovery during a 10-day low light period was produced under elevated concentration of ammonium (300 μM). The recovery of photosynthetic activity and phycobiliproteins was enhanced in the algae previously incubated under PAR + UVR as compared to exposure to only PAR, suggesting a beneficial effect of UVR on recovery or photoprotective processes under enriched nitrogen conditions. However, the content of MAAs did not follow the same pattern and thus it could not be concluded as the cause of observed enhanced recovery.     

Expression and characterization of the chitinases from Serratia marcescens GEI strain for the control of Varroa destructor, a honey bee parasite

Serratia marcescens GEI strain was isolated from the gut of the workers of Chinese honey bee Apis cerana and evaluated in the laboratory for the control of Varroa destructor, a parasite of western honey bee A. mellifera. The supernatant and the collected proteins by ammonium sulfate from the bacterial cultures showed a strong miticidal effect on the female mites, with 100% mite mortality in 5 days. Heat (100 °C for 10 min) and proteinase K treatment of the collected proteins destroyed the miticidal activity. The improved miticial activity of this bacterial strain on chitin medium indicated the involvement of chitinases. The expressed chitinases ChiA, ChiB and ChiC1 from S. marcescens GEI by recombinant Escherichia coli showed pathogenicity against the mites in the laboratory. These chitinases were active in a broad pH range (5-9) and the optimum temperatures were between 60 and 75 °C. Synergistic effects of ChiA and ChiB on the miticidal activity against V. destructor were observed. The workers of both honey bee species were not sensitive to the spraying and feeding chitinases. These results provided alternative control strategies for Varroa mites, by formulating chitinase agents and by constructing transgenetic honey bees.