Aluminum in lake water and organs of a fish Tribolodon hakonensis in strongly acidic lakes with a high aluminum concentration

Aluminum in lake water and in the organs of the fish Tribolodon hakonensis was investigated in Lake Usoriko (pH 3.6), Lake Inawashiroko (pH 5.0), and the Tenryu River (pH 7.7). The concentration of total soluble aluminum in the water was 0.51 mg l⁻¹ in Usoriko, 0.05 mg l⁻¹ in Inawashiroko, and less than 0.01 mg l⁻¹ in the Tenryu. The chemical forms of soluble aluminum in the acid water were characterized as Al³⁺, AlL²⁺, and AlL^≦1+. More than 90% of soluble aluminum in the water of Usoriko was Al³⁺, whereas AlL²⁺ was dominant in the water of Inawashiroko. The aluminum concentration in the organs of T. hakonensis in Usoriko was 42 μg g⁻¹ wet weight in gills, 4.2 μg g⁻¹ in muscle, 6.9 μg g⁻¹ in bone, 12.7 μg g⁻¹ in liver, 6.0 μg g⁻¹ in kidney, and 6.0 μg g⁻¹ in intestine, indicating accumulation of aluminum in the gills. The aluminum concentration in the organs of T. hakonensis living in Inawashiroko was approximately the same, in spite of the difference in water chemistry of the two acid lakes, especially for pH and aluminum. This suggests that aluminum accumulation might be controlled in the fish living in the acid lakes. In contrast, the aluminum concentration in the gills of T. hakonensis from the Tenryu was 2 μg g⁻¹. Received: May 20, 1999 / Accepted: December 10, 1999     

Alterations of sperm DNA integrity during cryopreservation procedure and in vitro incubation of bull semen

An association between sperm DNA integrity and fertility was recently shown for frozen–thawed Norwegian Red (NRF) bull semen diluted in skimmed milk egg yolk (SMEY). In general the fertility of NRF cattle is high, however, in comparison with NRF semen in SMEY, NRF semen diluted in Tris EY based extenders has shown reduced fertility. The aim of the present study was to do a split-sample comparison of sperm DNA integrity of NRF bull semen (n = 20) in SMEY and Triladyl^® (Tris EY based) during routine cryopreservation procedure and during in vitro incubation of frozen–thawed semen in modified synthetic oviduct fluid (mSOF). In contrast to the high fertility of NRF cattle, Holstein cattle are experiencing a marked decline in fertility. Therefore, the present study also aimed to compare sperm DNA integrity of NRF (n = 20) and Holstein (n = 20) semen diluted in Triladyl^® during in vitro incubation. The sperm DNA integrity was measured by susceptibility to in situ acid induced denaturation by the Sperm chromatin structure assay (SCSA). Compared to initial values of frozen neat semen, an increase in DNA damage was observed after dilution and cooling (5 °C) and after freezing–thawing of NRF semen in SMEY, but only after freezing–thawing for NRF semen diluted in Triladyl^®. Sperm DNA damage of NRF semen increased during in vitro incubation in mSOF; the increase in percentage of spermatozoa with DNA damage was more prominent in SMEY than in Triladyl^®, while the degree of damage was higher in Triladyl^®, throughout the incubation period. However, while the correlation between DNA damage and sperm survival was negative in SMEY throughout the incubation period, a positive correlation was observed in Triladyl^® after 9 h of incubation, indicating a higher presence of DNA damage in the live sperm population. In comparison with Holstein spermatozoa, the sperm DNA integrity of NRF semen reflected a better ability to withstand alterations induced during in vitro incubation in mSOF. In conclusion, sperm DNA integrity of NRF bull semen was altered during the cryopreservation procedure and in vitro incubation in mSOF. Dilution in Triladyl^® maintained bull sperm DNA integrity better than dilution in SMEY. Furthermore, alterations in Holstein sperm DNA integrity was more pronounced during in vitro incubation in mSOF compared to NRF bull spermatozoa.     

Effects of varying tissue sizes on the efficiency of baboon ovarian tissue vitrification

Objective

The aim of this study was to detect the effects of varying tissue sizes on the efficiency of baboon ovarian tissue vitrification.

Study design

The percentages of morphologically normal primordial follicles and the follicles expressing bax protein in ovarian tissues after vitrification–warming were measured. Besides, the 17-β estradiol levels in the culture supernatants were measured.

Results

The percentages of morphologically normal primordial follicles in vitrified–warmed ovarian tissues slicing in 0.5–1.5 mm in length and wide, and 1.0 mm in thickness were significantly higher than those slicing in 2.0 mm in length and wide, and 1.0 mm in thickness. Moreover, the follicles expressing bax protein in vitrified–warmed ovarian tissues slicing in 0.5–1.5 mm in length and wide, and 1.0 mm in thickness were significantly lower than those slicing in 2.0 mm in length and wide, and 1.0 mm in thickness. The 17-β estradiol levels in the culture supernatants slicing in 1.0–1.5 mm in length and wide, and 1.0 mm in thickness were significantly higher than those slicing in 0.5 mm or 2.0 mm in length and wide, and 1.0 mm in thickness.

Conclusions

Cortex piece slicing in 1.0–1.5 mm in length and wide, and 1.0 mm in thickness is suitable for baboon ovarian vitrification.     

Effects of fermented oyster mushroom ( Pleurotus ostreats) by-product supplementation on growth performance, blood parameters and meat quality in finishing Berkshire pigs

The objective of the present study was to investigate the effects of fermented oyster mushroom (Pleurotus ostreats) by-production (FOMP) supplementation on the growth performance, blood parameters, carcass traits and meat quality in finishing Berkshire pigs. FOMP was made by mixing oyster mushroom by-production with rice bran and barley bran and this mixture was fermented for 60 days. The experimental diets were 0, 3, 5 and 7% of FOMP added to C, T1, T2 and T3 in the basis diet for 7 weeks. Average daily gain (kg/day) was higher in C and T1 than in T2 and T3 ( P < 0.05). Average daily feed intake (kg/day) and feed conversion increased by the addition of FOMP ( P < 0.05). Total cholesterol and high-density lipoprotein cholesterol were higher in T3 than other treatments ( P < 0.05). Carcass weight (kg) was higher in C and T1 than in T2 and T3 ( P < 0.05). Dressing (%) was higher in C than in T3 ( P < 0.05). Crude protein was lower in T3 than in other treatments ( P < 0.05). Crude fat was higher in T2 and T3 than in C ( P < 0.05). pH₂₄ was higher in C than in other treatments ( P < 0.05). Cooking loss (%) was higher in T1 than T2 ( P < 0.05). Water-holding capacity (%) was higher in C than in T1 ( P < 0.05). In meat colour, CIE a* was lower by the addition of FOMP ( P < 0.05). CIE b* was higher in C than in other treatments ( P < 0.05). In backfat colour, CIE L* was lower in T3 than other treatments ( P < 0.05). CIE b* was lower by addition of FOMP ( P < 0.05). Palmitoleic and oleic acid were higher in T3 than in other treatments ( P < 0.05). Linoleic and arachidonic acids were higher in T2 than in other treatments ( P < 0.05). The results indicate that 3% of FOMP affected the growth performance, carcass traits, meat quality and fatty acid in contrast to addition of 5% of FOMP for Berkshire pigs during the finishing period.     

Photosynthetic acclimation and photoinhibition on exposure to high light in shade-developed leaves of Fagus crenata seedlings

The physiological response of leaves developed in low light (L) on Fagus crenata seedlings exposed to different levels of high light (H: high light, M: medium light) was studied. Measurements were conducted on potted seedlings in the F. crenata forest understory. The seedlings with leaves developed in L were transferred to H (L–H) and M (L–M) in summer. On exposure to high light, the photochemical efficiency of dark-adapted PSII (F_v/F_m) immediately decreased and was followed by a subsequent recovery in both L–H and L–M leaves. The mean value of F_v/F_m in L–H leaves was lower than that in L–M leaves through experiments, indicating that the degree of photoinhibition in L–H leaves was greater than that in L–M leaves. About 1 month after transfer, 37% and 5% of leaves had fallen in L–H and L–M seedlings, respectively. This result also indicated the greater photoinhibition in L–H leaves. Moreover, the photosynthetic capacity (P_Nmax) of L–H leaves decreased. In contrast, the P_Nmax of L–M leaves increased, although the P_Nmax was lower than that of M control leaves. An increase in the xanthophyll cycle pool (VAZ), indicating an increase of the photoprotective function, was found in both L–H and L–M leaves. Especially, the VAZ pool in L–M leaves was higher than that in M leaves by the end of experiments. L–M leaves may avoid photoinhibition effectively by the decrease in excess light with the increase of the P_Nmax or VAZ pool, compared to L–H leaves. Thus, the physiological acclimation on exposure to high light depended on the degree of high light. To achieve successful photosynthetic acclimation with slight photoinhibition, the variation of light intensity before and after exposure to high light would be an important factor because of the difference in excess light.     

Electrochemical changes in media due to microbial

Microbial metabolism affected the electrical impedance parameters of a two terminal-measuring cell-containing growth media. The relationship between microbial growth and relative changes in both The capacitive and resistive parts of impedance was examined. Both components of impedance were shown to be indicative of bacterial growth. In low conductivity media the change in the conductance of the media (G_sol) clearly correlated to bacterial growth. In more conductive media the relative changes in G_sol were smaller, and in these media measurements of the changes of polarization capacitance (C_pol) were useful for monitoring bacterial growth.Yeast growth in two media resulted in large changes in C_pol (20–100%) while the changes in G_sol were very small (1–4%). This result indicated that, for some combinations of microorganisms and media, measuring C_pol might be preferable over G_sol for the detection of microbial growth.Microbial metabolism resulted in a change of 2–2.5 units in pH. This pH change resulted in a 40% change in C_pol but less than a 14% change in G_sol.     

Studies on the activity of brown adipose tissue in suckling,pre-obese, obob mice

The properties and activity of brown adipose tissue have been investigated in suckling, pre-obese, $\text{ob}\text{ob}$ mice in order to determine whether decreased thermogenesis in the tissue precedes the development of obesity in this mutant. At 14 days of age there was no difference between the $\text{ob}\text{ob}$ and normal animals in the total amount of interscapular brown adipose tissue, and the DNA content, protein content, and cytochrome oxidase activity of the tissue were similar in the two groups of mice. Respiration rates of brown adipose tissue mitochondria in the presence of albumin were, however, greater in the normal than the $\text{ob}\text{ob}$ animals, although after the addition of GDP to recouple the mitochondria there was no difference between the two groups. The mitochondrial membrane potential, measured with [³H]methyltriphenylphosphonium, was less affected by exogenous GDP in $\text{ob}\text{ob}$ mice than in normal animals. GDP binding to brown adipose tissue mitochondria, an index of the proton conductance pathway, was much greater in normal than in $\text{ob}\text{ob}$ mice at both 10 and 14 days of age; the decreased GDP binding in the mutant animals was found to result from a reduction in the number of binding sites. It is concluded that brown adipose tissue mitochondria of pre-obese $\text{ob}\text{ob}$ mice are more tightly coupled than those of normal siblings, and that the activity of the ‘thermogenic’ proton conductance pathway is lower in the mutant animals. A decrease in thermogenesis in brown adipose tissue is therefore an early event in the development of the $\text{ob}\text{ob}$ mouse and precedes the appearance of obesity.     

Effects of Botrytis cinerea and Pseudomonas syringae infection on the antioxidant profile of Mesembryanthemum crystallinum C3/CAM intermediate plant

Mesembryathemum crystallinum plants performing C₃ or CAM (crassulacean acid metabolism) appear to be highly resistant to Botrytis cinerea as well as to Pseudomonas syringae. Fungal hyphae growth was restricted to 48 h post-inoculation (hpi) in both metabolic types and morphology of hyphae differed between those growing in C₃ and CAM plants. Growth of bacteria was inhibited significantly 24 hpi in both C₃ and CAM plants. B. cinerea and P. syringae infection led to an increase in the concentration of H₂O₂ in C₃ plants 3 hpi, while a decrease in H₂O₂ content was observed in CAM performing plants. The concentration of H₂O₂ returned to the control level 24 and 48 hpi. Changes in H₂O₂ content corresponded with the activity of guaiacol peroxidase (POD), mostly 3 hpi. We noted that its activity decreased significantly in C₃ plants and increased in CAM plants in response to inoculation with both pathogens. On the contrary, changes in the activity of CAT did not correlate with H₂O₂ level. It increased significantly after interaction of C₃ plants with B. cinerea or P. syringae, but in CAM performing plants, the activity of this enzyme was unchanged. Inoculation with B. cinerea or P. syringae led to an increase in the total SOD activity in C₃ plants while CAM plants did not exhibit changes in the total SOD activity after interaction with both pathogens. In conclusion, the pathogen-induced changes in H₂O₂ content and in SOD, POD and CAT activities in M. crystallinum leaves, were related to the photosynthetic metabolism type of the stressed plants rather than to the lifestyle of the invading pathogen.     

Effect of Energy Under-Reporting on Secular Trends of Dietary Patterns in a Mediterranean Population

BackgroundDiet is an important factor in the prevention of chronic diseases. Analysis of secular trends of dietary patterns can be biased by energy under-reporting. Therefore, the objective of the present study was to analyse the impact of energy under-reporting on dietary patterns and secular trends in dietary patterns defined by cluster analysis.ResultsThree clusters, “healthy”, “mixed” and “western”, were identified for both surveys. The “mixed” cluster was the predominant cluster in both surveys. Excluding EUR reduced the proportion of the “mixed” cluster up to 6.40% in the 2000 survey; this caused secular trend increase in the prevalence of the “mixed” pattern. Cross-classification analysis of all participants and PER’ data showed substantial agreement in cluster assignments: 68.7% in 2000 and 84.4% in 2005. Excluding EUR did not cause meaningful (≥15%) changes in the “healthy” pattern. It provoked changes in consumption of some food groups in the “mixed” and “western” patterns: mainly decreases of unhealthy foods within the 2000 and increases of unhealthy foods within the 2005 surveys. Secular trend effects of EUR were similar to those within the 2005 survey. Excluding EUR reversed the direction of secular trends in consumption of several food groups in PER in the “mixed” and “western” patterns.ConclusionsEUR affected distribution of participants between dietary patterns within and between surveys, secular trends in food group consumption and amount of food consumed in all, but not in the “healthy” pattern. Our findings emphasize threats from energy under-reporting in dietary data analysis.     

Effect of estrogen on the expression of GnRH and kisspeptin in the hypothalamus of rats during puberty

The aim of this study was to assess whether changes in kisspeptin and GnRH levels could be attributed to sex steroids at puberty onset. We used the ovariectomy (OVX) model in rats treated with 17β-estradiol (E₂; OVX + E₂), or oil (OVX + oil), and in intact rats treated with E₂ (intact + E₂) or oil only (intact + oil) to determine gene expression changes of Kiss1 and Gnrh1 in the hypothalamus and protein expression of kisspeptin and GnRH in the different areas of the hypothalamus. In the intact + E₂ and OVX + E₂ rats on the day of the onset of puberty, GnRH-immunoreactive (ir) cell numbers decreased (P < 0.05) in the arcuate nucleus but were increased in the preoptic area; Kisspeptin-ir cells increased (P < 0.05) in the arcuate nucleus, periventricular nucleus, and preoptic area; no difference (P > 0.05) was found in the paraventricularis nucleus for GnRH-ir or kisspeptin-ir cells. Additionally, levels of Kiss1 and Gnrh1 messenger RNA in the hypothalamus were significantly higher (P < 0.05) in the OVX + E₂ or intact + E₂ rats than in the OVX + oil or intact + oil animals, respectively. In the OVX + oil rats, OVX significantly increased (P < 0.05) levels of Gnrh1 and Kiss1 messenger RNA and the expression of GnRH and kisspeptin in the hypothalamus compared to intact + oil animals. These results suggest that kisspeptin and GnRH play major roles in modulating the activity of estrogen circuits at the onset of puberty.     

Cultivation and carrageenan yield and quality ofKappaphycus alvarezii in the waters of Vietnam

The carrageenan-producing red algaKappaphycus alvarezii (Doty) Doty was brought to Vietnam from Japan in 1993. Branch fragments of this species were cultivated in a pond, lagoon, inlet and offshore in Vietnam for the first time. The best daily growth rate (DGR) of plants grown in the lagoon area attained 9–11 % day^–1 in May to June (cold season). The water temperature and salinity in this area ranged from 27.2–32.4 °C and 31.4–33.7 °C, respectively. DGR of plants grown in the inlet ranged from 7 to 9% day^–1 in June. Grazing by fish has been observed to occur in this area. The DGR of plants grown in the pond ranged from 5–6% in January–July, but decreased to less than 4% day^–1 in August (hot season). K. alvarezii in Vietnam showed a carrageenan yield of 18.8–24.6% and gel strength of 1566–1712 g cm^–2. These values are similar ones obtained fromK. alvarezii cultivated in the Philippines and Indonesia.     

Auxin distribution is differentially affected by nitrate in roots of two rice cultivars differing in responsiveness to nitrogen

Background and Aims

Although ammonium (NH₄⁺) is the preferred form of nitrogen over nitrate (NO₃⁻) for rice (Oryza sativa), lateral root (LR) growth in roots is enhanced by partial NO₃⁻ nutrition (PNN). The roles of auxin distribution and polar transport in LR formation in response to localized NO₃⁻ availability are not known.

Methods

Time-course studies in a split-root experimental system were used to investigate LR development patterns, auxin distribution, polar auxin transport and expression of auxin transporter genes in LR zones in response to localized PNN in ‘Nanguang’ and ‘Elio’ rice cultivars, which show high and low responsiveness to NO₃⁻, respectively. Patterns of auxin distribution and the effects of polar auxin transport inhibitors were also examined in DR5::GUS transgenic plants.

Key Results

Initiation of LRs was enhanced by PNN after 7 d cultivation in ‘Nanguang’ but not in ‘Elio’. Auxin concentration in the roots of ‘Nanguang’ increased by approx. 24 % after 5 d cultivation with PNN compared with NH₄⁺ as the sole nitrogen source, but no difference was observed in ‘Elio’. More auxin flux into the LR zone in ‘Nanguang’ roots was observed in response to NO₃⁻ compared with NH₄⁺ treatment. A greater number of auxin influx and efflux transporter genes showed increased expression in the LR zone in response to PNN in ‘Nanguang’ than in ‘Elio’.

Conclusions

The results indicate that higher NO₃⁻ responsiveness is associated with greater auxin accumulation in the LR zone and is strongly related to a higher rate of LR initiation in the cultivar ‘Nanguang’.     

Differential susceptibility to oxidative stress of two scleractinian corals: antioxidant functioning of mycosporine-glycine

This study examined the importance of mycosporine-glycine (Myc-Gly) as a functional antioxidant in the thermal-stress susceptibility of two scleractinian corals, Platygyra ryukyuensis and Stylophora pistillata. Photochemical efficiency of PSII (F_v/F_m), activity of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT), and composition and abundance of mycosporine-like amino acids (MAAs) in the coral tissue and in symbiotic zooxanthellae were analyzed during 12-h exposure to high temperature (33 °C). After 6- and 12-h exposures at 33 °C, S. pistillata showed a significantly more pronounced decline in F_v/F_m compared to P. ryukyuensis. A 6-h exposure at 33 °C induced a significant increase in the activities of SOD and CAT in both host and zooxanthellae components of S. pistillata while in P. ryukyuensis a significant increase was observed only in the CAT activity of zooxanthellae. After 12-h exposure, the SOD activity of P. ryukyuensis was unaffected in the coral tissue but slightly increased in zooxanthellae, whereas the CAT activity in the coral tissue showed a 2.5-fold increase. The total activity of antioxidant enzymes was significantly higher in S. pistillata than in P. ryukyuensis, suggesting that P. ryukyuensis is less sensitive to oxidative stress than S. pistillata. This differential susceptibility of the corals is consistent with a 20-fold higher initial concentration of Myc-Gly in P. ryukyuensis compared to S. pistillata. In the coral tissue and zooxanthellae of both species investigated, the first 6 h of exposure to thermal stress induced a pronounced reduction in the abundance of Myc-Gly but not in other MAAs. When exposure was prolonged to 12 h, the Myc-Gly pool continued to decrease in P. ryukyuensis and was completely depleted in S. pistillata. The delay in the onset of oxidative stress in P. ryukyuensis and the dramatic increase in the activities of the antioxidant enzymes in S. pistillata, which contains low concentrations of Myc-Gly suggest that Myc-Gly provides rapid protection against oxidative stress before the antioxidant enzymes are induced. These findings strongly suggest that Myc-Gly is functioning as a biological antioxidant in the coral tissue and zooxanthellae and demonstrate its importance in the survival of reef-building corals under thermal stress.     

Conformation of Some Hexuronic Acids and d-Galactopyranosiduronic Acid Moiety in the Peracetylated and Permethylated Derivatives of Orange Pectic Acid in Solutions

1. The 1C conformation was estimated for α-d-galactopyranosiduronic acid moiety of pectic acid in the permethylated derivative dissolved in 1 n NaOD-D₂O and in the peracetylated derivative dissolved in dimethyl sulfoxide-d₆, and the C1 conformation was estimated for some derivatives of d-galactopyranuronic acid in chloroform-d by NMR spectroscopy.

2. Random conformation of the whole macromolecule was estimated for pectic acid in water on the basis of no appearance of any induced Cotton effects in the 200 ~ 700 mμ region in the ORD spectra of pectic acid-anionic dye complexes.

3. The conformation was supported by the fact that the rate of periodate oxidation of pectic acid at 5° was slightly decreased in comparison with that of amylase in 7 m urea solution.

     

In vitro protein synthesis capacities in a cold stenothermal and a temperate eurythermal pectinid

The translational system was isolated from the gills of the Antarctic scallop Adamussium colbecki (Smith) and the European scallop Aequipecten opercularis (Linnaeus) for in vitro protein synthesis capacities (g protein mg FW^–1 day^–1) and the translational capacities of RNA (k_{RNA in vitro} mg protein mg RNA^–1 day^–1). In vitro protein synthesis capacity in the cold-adapted pectinid at 0 °C was similar to the one found in the temperate scallop at 25 °C. These findings might reflect cold compensated rates in Adamussium colbecki, partly explainable by high tissue levels of RNA. Cold-compensated in vitro protein synthesis capacities may further result from increments in the translational capacity of RNA. The thermal sensitivity of the translation machinery was slightly different in the two species, with significantly lower levels of Arrhenius activation energies E_a and Q₁₀ in Adamussium colbecki in the temperature range 0–15 °C. Reduced protein synthesis and translational capacities were found in vitro in gills of long-term aquarium-maintained Adamussium colbecki and were accounted for by a loss of protein synthesis machinery, i.e. a reduction in RNA levels, as well as a decrease in the amount of protein synthesized per milligram of RNA (RNA translational capacity, k_{RNA in vitro}). Such changes may involve food uptake or mirror metabolic depression strategies, like those occurring during winter. Consequences of high in vitro RNA translational capacities found in the permanently cold-adapted species are discussed in the context of seasonal food availability and growth rates at high latitudes.Abbreviations DPM disintegrations per minute - DTT dithiothreitol - E_a Arrhenius activation energy - k_s fractional protein synthesis rate - k_{RNA in vivo} translational efficiency - k_{RNA in vitro} translational capacity - PCA perchloric acid - Phe phenylalanine - PLA phospho-L-arginine - PSU practical salinity units - RNAse ribonuclease - TCA trichloroacetic acidCommunicated by G. Heldmaier     

Allozyme polymorphism of Mdh and α-Gpdh in Ixodes ricinus populations: comparison of borreliae-infected and uninfected ticks

Ixodes ricinus Linnaeus (Acari: Ixodidae) ticks are vectors of numerous infectious diseases in humans and animals. The allozyme variability of MDH and α-Gpdh was detected by native polyacrylamide gel electrophoresis in I. ricinus natural populations in three localities in Serbia. Four alleles of Mdh locus (MDH 1, MDH 2, MDH 3 and MDH X) and four alleles of α-Gpdh locus (VS, S, F and VF) were detected. Interpopulation differences in Mdh and α-Gpdh allele frequencies were statistically insignificant. Significant difference in α-Gpdh allele frequencies between males and females was recorded in the largest sample only. Differences in allele frequencies, detected between borreliae-infected and uninfected I. ricinus ticks, were close to the level of statistical significance, especially for α-Gpdh locus. Clear significant difference appeared in females when sexes were tested separatelly (P = 0.037). It is interesting that genotypes containing rarer alleles (MDH 1 and S) were infected in higher proportion in comparison to other genotypes. Our results point towards a possible role of Mdh and α-Gpdh loci in I. ricinus ticks in the determination of energy requirements for host seeking. Sex differences in α-Gpdh allele frequencies suggest that selective pressure, concerning efficiency of reserve materials utilisation, points to α-Gpdh rather than to Mdh locus.     

Involvement of Ca2+-calmodulin in Cd2+ toxicity during the early phases of radish (Raphanus sativus L.) seed germination

The toxicity of Cd²⁺in vivo during the early phases of radish (Raphanus sativus L.) seed germination and the in vitro Cd²⁺ effect on radish calmodulin (CaM) were studied. Cd²⁺ was taken up in the embryo axes of radish seeds; the increase in fresh weight of embryo axes after 24 h of incubation was inhibited significantly in the presence of 10 mmol m^?3 Cd²⁺ in the external medium, when the Cd²⁺ content in the embryo axes was c. 1.1 μmol g^?1 FW. The reabsorption of K⁺, which characterizes germination, was inhibited by Cd²⁺, suggesting that Cd²⁺ affected metabolic reactivation. The slight effect of Cd²⁺ on the transmembrane electric potential of the cortical cells of the embryo axes excluded a generalized toxicity of Cd²⁺ at the plasma membrane level. After 24 h of incubation, Cd²⁺ induced no increase in total acid-soluble thiols and Cd²⁺-binding peptides able to reduce Cd²⁺ toxicity. Ca²⁺ added to the incubation medium partially reversed the Cd²⁺-induced inhibition of the increase in fresh weight of embryo axes and concomitantly reduced Cd²⁺ uptake. Equilibrium dialysis experiments indicated that Cd²⁺ bound to CaM and competed with Ca²⁺ in this binding. Cd²⁺ inhibited the activation of Ca²⁺-CaM-dependent calf-brain phosphodiesterase, inhibiting the Ca²⁺-CaM active complex. Cd²⁺ reduced the binding of CaM to the Ca²⁺-CaM binding enzymes present in the soluble fraction of the embryo axes of radish seeds. The possibility that Cd²⁺ toxicity in radish seed germination is mediated by the action of Cd²⁺ on Ca²⁺-CaM is discussed in relation to the in vivo and in vitro effects of Cd²⁺.     

Within-canopy variation in the rate of development of photosynthetic capacity is proportional to integrated quantum flux density in temperate deciduous trees

The present study was undertaken to test for the hypothesis that the rate of development in the capacity for photosynthetic electron transport per unit area (J_max;A), and maximum carboxylase activity of Rubisco (V_cmax;A) is proportional to average integrated daily quantum flux density (Q_int) in a mixed deciduous forest dominated by the shade‐intolerant species Populus tremula L., and the shade‐tolerant species Tilia cordata Mill. We distinguished between the age‐dependent changes in net assimilation rates due to modifications in leaf dry mass per unit area (M_A), foliar nitrogen content per unit dry mass (N_M), and fractional partitioning of foliar nitrogen in the proteins of photosynthetic electron transport (F_B), Rubisco (F_R) and in light‐harvesting chlorophyll‐protein complexes (V_cmax;A ∝ M_AN_MF_R; J_max;A ∝ M_AN_MF_B). In both species, increases in J_max;A and V_cmax;A during leaf development were primarily determined by nitrogen allocation to growing leaves, increases in leaf nitrogen partitioning in photosynthetic machinery, and increases in M_A. Canopy differences in the rate of development of leaf photosynthetic capacity were mainly controlled by the rate of change in M_A. There was only small within‐canopy variation in the initial rate of biomass accumulation per unit Q_int (slope of M_A versus leaf age relationship per unit Q_int), suggesting that canopy differences in the rate of development of J_max;A and V_cmax;A are directly proportional to Q_int. Nevertheless, M_A, nitrogen, J_max;A and V_cmax;A of mature leaves were not proportional to Q_int because of a finite M_A in leaves immediately after bud‐burst (light‐independent component of M_A). M_A, leaf chlorophyll contents and chlorophyll : N ratio of mature leaves were best correlated with the integrated average quantum flux density during leaf development, suggesting that foliar photosynthetic apparatus, once developed, is not affected by day‐to‐day fluctuations in Q_int. However, for the upper canopy leaves of P. tremula and for the entire canopy of T. cordata, there was a continuous decline in N contents per unit dry mass in mature non‐senescent leaves on the order of 15–20% for a change of leaf age from 40 to 120 d, possibly manifesting nitrogen reallocation to bud formation. The decline in N contents led to similar decreases in leaf photosynthetic capacity and foliar chlorophyll contents. These data demonstrate that light‐dependent variation in the rate of developmental changes in M_A determines canopy differences in photosynthetic capacity, whereas foliar photosynthetic apparatus is essentially constant in fully developed leaves.