Upregulation of two ripening‐related genes from a non‐climacteric plant (pepper) in a transgenic climacteric plant (tomato)

Activities of promoters from the capsanthin/capsorubin synthase and fibrillin genes, which are molecular markers for ripening in the non-climacteric pepper fruits, have been studied in transgenic tomato plants that produce fruits of the climacteric type (characterized by an increase in respiration and ethylene production). The promoters of both genes were strongly upregulated during tomato fruit ripening in a manner similar to the induction of these genes in pepper fruits. Induction occurred at the mature green stage preceding ripening (a stage when ethylene production and respiration are known to rise in tomato fruits). Ethylene positively influenced the expression of both genes in tomato. Other plant growth regulators, namely abscisic acid, auxin and polyamines, did not alter gene expression. In contrast, water loss strongly induced both promoters. This dehydration-mediated gene induction was inhibited by mitochondrial respiration inhibitors (mainly of the alternative oxidase). A slight positive effect with light, apparently not linked to normal photosynthesis but rather to photooxidative stress, was also observed. Taken together, the data indicate that activation of oxidase systems, leading to changes in the cellular redox balance, mediates the induction of both genes in tomato. Various cellular compartments are likely to be contributors to this process, which leads to the developmental regulation of nuclear genes encoding plastid-located proteins.     

Gustatory responses to feeding‐ and non‐feeding‐stimulant chemicals, with an emphasis on amino acids, in rainbow trout

Specific receptor and fibre types of rainbow trout Oncorhynchus mykiss involved in the detection and discrimination of amino acids and a heterogeneous collection of compounds were investigated by recording the electrical activity of the maxillary branch of the facial nerve innervating taste buds inside the upper jaw. Proline (Pro), alanine (Ala), leucine (Leu), betaine (Bet) and 2‐amino‐3‐guanidinopropionic acid (Agp) were the major amino acids detected by the gustatory system. The two experimental approaches, concentration‐response curves and cross‐adaptations, showed that all amino acids were detected by three independent receptor types: Pro ‐, Agp/Bet ‐ and Leu ‐receptors. Bile acids, the most potent stimulants recorded, were detected by a single receptor type independent of those for amino acids, with threshold concentrations of 10⁻¹² M. Strychnine, quinine and tetrodotoxin may have partially shared a single receptor mechanism. The gustatory sensibility narrowly tuned towards the amino acid spectrum compared to those for a diverse array of non‐feeding stimulant chemicals, combined with feeding behaviour triggered primarily by vision and olfaction, suggest that in rainbow trout, and possibly other salmonid species, gustatory chemical cues, in addition to food finding and intake, play an important role in detecting poisonous prey and substances.     

The use of pulse‐echo acoustic microscopy to non‐invasively determine the sex of living larval sea lampreys

A rapid, accurate and non‐invasive method to determine the sex of larval sea lampreys Petromyzon marinus , using wide‐field pulse‐scanning acoustic microscopy, is described. Cross‐sectional pulse‐echo scans were made with a high‐resolution acoustic microscope in 48 larvae (110–130 mm total length, L _T), and the acoustic images generated showed such internal body structures as the gonad, intestine, kidneys, cardinal veins, notochord and musculature. Females were identified by the presence of a relatively large (1–1·5 mm diameter) ovary, which was considerably less reflective to the 15 to 25 MHz acoustic signals than the surrounding kidney tissue. Males were recognized by the lack of the large non‐reflective ovary and, in some cases, the appearance of a small (0·2–0·3 mm) testis with slightly stronger reflective properties than the kidney. Identification of sex was confirmed by optical microscopy following dissection, and in a blind test on an additional 10 specimens (121–168 mm L _T), the acoustic method reliably identified sex in 100% of the larvae. These results indicate that acoustic microscopy can determine the sex of live sea lamprey larvae in c . 30 s per animal, a process which until now required dissection or invasive surgery.     

Jasmonic acid promotes germination and lipase activity in non‐stratified apple embryos

The effects of (−)jasmonic acid (JA) on germination of embryos isolated from dormant seeds of apple ( Malus domestica Borb. cv. Antonówka) cultured in darkness or at 12‐h photoperiod were studied, as well as its effects on the activity of alkaline lipase (AlkL, EC 3.1.1.3) in these embryos. The maximum sensitivity of germination to JA occurred on days 3 and 4 of embryo culture. Both germination and enzyme activity were stimulated by JA, its effect being additive to that of light. Inhibitors of lipoxygenase inhibited embryo germination and AlkL activity, both effects being partially reversed by JA treatment. We suggest that (1) JA is implicated in an endogenous complex controlling apple seed germination, and that (2) it acts independently of the mechanism triggered by light.     

Effects of calcium at toxic concentrations of cadmium in plants

Planta - This review provides new insight that calcium plays important roles in plant growth, heavy metal accumulation and translocation, photosynthesis, oxidative damage and signal transduction...     

Effects of intraperitoneal injection of cortisol on non‐specific immune functions of Ctenopharyngodon idella

After grass carps Ctenopharyngodon idellus were injected with cortisol, with (CBC) and without (C) a cocoa butter carrier, the effects of both slowly and rapidly acting exogenous cortisol on their non‐specific immune functions were investigated. On the one hand, after injection with CBC, the cortisol concentration and lysozyme activity in fish serum were enhanced and were sustained at high levels for a long period (30 days). The killing activity in the serum declined with time, and phagocytosis of head kidney macrophages diminished significantly (P < 0·05 or P < 0·01). The leukocrit values in the high dose group (31·8 mg cortisol fish^?1) increased over time, however, with the maximum average being 5·6% at day 30. The spleen mass index in the high dose group was 0·93 × 10^?3 after 30 days, notably lower (P < 0·05) than that in the control group. In addition, a decrease in resistance to Aeromonas hydrophila infection in cortisol‐treated fish was shown, with the final cumulative mortalities being 54·5 and 66·7% in the low and high dose groups, respectively. On the other hand, there was a decrease in both serum cortisol concentration and lysozyme activity of the experimental fish within 2 weeks after injection with C, where plasma bactericidal activities in the high dose group (31·8 mg cortisol fish^?1) were remarkably lower (P < 0·01) than those in the control group at each sampling, but were increased slightly over time. The results of which were different from those in the CBC trial. Phagocytic activity of head kidney macrophages and spleen mass index decreased significantly (P < 0·05), while there were increases in leukocrit value and cumulative mortality due to A. hydrophila. The results of which were similar to those in the CBC trial. This study indicated that the injection of cortisol depressed the non‐specific immune functions of the grass carp and increased its susceptibility to disease.     

Magnesium ions promote assembly of channel‐like structures from beticolin 0, a non‐peptide fungal toxin purified from Cercospora beticola

Beticolins are toxins produced by the fungus Cercospora beticola. Using beticolin 0 (B0), we have produced a strong and Mg²⁺-dependent increase in the membrane conductance of Arabidopsis protoplasts and Xenopus oocytes. In protein-free artificial bilayers, discrete deflexions of current were observed (12 pS unitary conductance in symmetrical 100 mM KCl) in the presence of B0 (approximately 10 μM) and in the presence of nominal Mg²⁺. Addition of 50 μM Mg²⁺ induced a macroscopic current which could be reversed to single channel current by chelating Mg²⁺ with EDTA. Both unitary and macroscopic currents were ohmic. The increase in conductance of biological membranes triggered by B0 is therefore likely to originate from the ability of this toxin to organize itself into transmembrane pores in the presence of Mg²⁺. The pore is poorly selective, displaying permeability ratios P_Cl/P_K, P_Na/P_K and P_Ca/P_K close to 0.3, 0.65 and 0.4, respectively. Such channel-like activity could be involved in the deleterious biological activity of the toxin, by causing the collapse of ionic and electrical gradients through biological membranes together with Ca²⁺ influx and scrambling of cellular signals.     

A non‐invasive stress assay based upon measurement of free cortisol released into the water by rainbow trout

A procedure previously used for sex steroids was adapted to extract free cortisol and cortisone from water samples taken from rainbow trout Oncorhynchus mykiss tanks. Both corticosteroids could be readily detected by radioimmunoassay (RIA), with cortisol being predominant. All stages of the sampling, extraction and RIA procedure were validated for cortisol. An intermittent problem with poor replication was traced to the use of diethyl ether during the extraction procedure, and was overcome by the use of ethyl acetate. Other modifications were also introduced to speed up the procedure. The concentration and time course of release of both corticosteroids were shown to be related to the degree of stress that the fish had been subjected to. It was confirmed that cortisol concentrations in water and estimated cortisol release rates increased in response to handling stress, and that both were correlated with plasma cortisol concentrations. The potential for using water cortisol concentration and release rates to assess the primary stress response of fishes as a non‐invasive alternative to blood sampling is discussed.     

Inhibition of viral adhesion and infection by sialic-acid-conjugated dendritic polymers

Multiple sialic acid (SA) residues conjugated to a linear polyacrylamide backbone are more effective than monomeric SA at inhibiting influenza-induced agglutination of red blood cells. However, "polymeric inhibitors" based on polyacrylamide backbones are cytotoxic. Dendritic polymers offer a nontoxic alternative to polyacrylamide and may provide a variety of potential synthetic inhibitors of influenza virus adhesion due to the wide range of available polymer structures. We evaluated several dendritic polymeric inhibitors, including spheroidal, linear, linear-dendron copolymers, comb-branched, and dendrigraft polymers, for the ability to inhibit virus hemagglutination (HA) and to block infection of mammalian cells in vitro. Four viruses were tested: influenza A H2N2 (selectively propagated two ways), X-31 influenza A H3N2, and sendai. The most potent of the linear and spheroidal inhibitors were 32-256-fold more effective than monomeric SA at inhibiting HA by the H2N2 influenza virus. Linear-dendron copolymers were 1025-8200-fold more effective against H2N2 influenza, X-31 influenza, and sendai viruses. The most effective were the comb-branched and dendrigraft inhibitors, which showed up to 50000-fold increased activity against these viruses. We were able to demonstrate significant (p < 0.001) dose-dependent reduction of influenza infection in mammalian cells by polymeric inhibitors, the first such demonstration for multivalent SA inhibitors. Effective dendrimer polymers were not cytotoxic to mammalian cells at therapeutic levels. Of additional interest, variation in the inhibitory effect was observed with different viruses, suggesting possible differences due to specific growth conditions of virus. SA-conjugated dendritic polymers may provide a new therapeutic modality for viruses that employ SA as their target receptor.     

A potential role for invasive sunbleak in the further dissemination of a non‐native parasite

Since its first introduction to English waters in the mid 1980s, sunbleak Leucaspius delineatus has spread widely and it is shown in the present study that this non‐native cyprinid is a new host for Neoergasilus japonicus . This copepod parasite from South East Asia has a limited host range and distribution in the U.K. Rapid dispersal of sunbleak in the U.K. raises concerns for further dispersal of this introduced parasite.     

Assessment and control of non‐indigenous brown bullhead Ameiurus nebulosus populations using fyke nets in shallow ponds

In order to evaluate fyke nets as a tool for population assessment and mass removal of brown bullhead Ameiurus nebulosus populations, small, shallow ponds were sampled during late summer in 1999 and 2000. Catch efficiency, species and size selectivity of fyke nets were investigated with the mark‐recapture method. Two different strategies were applied: (1) on six occasions, the mark‐recapture method was applied to entire populations, and (2) in six other cases, sampling was done in two replicated enclosed areas (625 m²). When applied to entire populations, fyke nets were found to be size selective, and capture efficiency significantly increased with standard length. Conversely, no size selectivity of fyke nets was found in enclosed areas. Sampling in replicated enclosures yielded reproducible estimates of population density and biomass. The results suggest that double fyke nets, when combined with the mark‐recapture technique, are a very useful tool for the efficient and reliable assessment of brown bullhead populations. The results of the study also suggest that double fyke nets may potentially be a cost‐effective tool for the mass removal of non‐indigenous brown bullhead populations.     

Differential lifetime success and performance of native and non‐native Atlantic salmon examined under communal natural conditions

The lifetime success and performance characteristics of communally reared offspring of wild native Burrishoole (native), ranched native (ranched) and non‐native (non‐native) Atlantic salmon Salmo salar from the adjacent Owenmore River were compared. Non‐native 0+ year parr showed a substantial downstream migration, which was not shown by native and ranched parr. This appears to have been an active migration rather than competitive displacement and may reflect an adaptation to environmental or physiographic conditions within the Owenmore River catchment where the main nursery habitat is downstream of the spawning area. There were no differences between native and ranched in smolt output or adult return. Both of these measures, however, were significantly lower for the non‐native group. A greater proportion of the non‐native Atlantic salmon was taken in the coastal drift nets compared to the return to the Burrishoole system, probably as a result of the greater size of the non‐native fish. The overall lifetime success of the non‐native group, from fertilized egg to returning adult, was some 35% of native and ranched. The ranched group showed a significantly greater male parr maturity, a greater proportion of 1+ year smolts, and differences in sex ratio and timing of freshwater entry of returning adults compared to native, which may have fitness implications under specific conditions.     

Early gene expression response of barley root tip to toxic concentrations of cadmium

Plant Molecular Biology - Already a short-term Cd treatment induces changes in gene expression in barley root tips via IAA and ROS signaling during mild and severe Cd stress, respectively. Even a...     

A non‐invasive method to assess the impact of electronic tag insertion on stress levels in fishes

The present study investigated, for the first time, a non‐invasive technique for monitoring cortisol levels in fishes that had been implanted with electronic transmitters. This technique involved the measurement of cortisol released into the water by individual fish. Carp Cyprinus carpio and roach Rutilus rutilus that had been tagged with miniature dummy acoustic transmitters responded to the surgical implantation of the tags with an immediate (1 to 4 h) increase in cortisol concentrations in the water. These amounts of cortisol were higher than in 'handled only' and 'handled and anaesthetized' groups. Estimated cortisol release rates for both species correlated well with the direct measurements in the water. In both species, water cortisol concentration and cortisol release rates returned to pre‐stress levels within 12 h of tag‐insertion, and remained low for the remainder of the experiment. The calculation of cortisol release rates highlighted a difference of a factor of about four between peak release rates in carp and roach. There was no evident long‐term cortisol stress response to the presence of a tag in the body cavity in either species.     

Coupling non‐invasive physiological assessments with telemetry to understand inter‐individual variation in behaviour and survivorship of sockeye salmon: development and validation of a technique

Approximately 200 km from the mouth of the Fraser River, British Columbia, Canada, adult sockeye salmon Oncorhynchus nerka , were gastrically implanted with radio transmitters without anaesthetic. Subsets of the transmitter implanted fish were also biopsied which included drawing blood from the caudal peduncle (3 ml), removal of gill tissue (0·03 g) and quantification of energetic status using a microwave fat meter. Several experiments were used to test the hypothesis that the biopsy had a negligible effect on the subsequent survival and migratory behaviour of transmitter implanted fish. In the first experiment, no difference was found in the survival (both 100%) or tag retention (both 100%) between the two treatment groups (transmitter implanted with and without biopsy) when fish were held in pens for 24 h in the marine environment. Similarly, in other experiments where fish were released to the ocean to resume their migratory journey, no statistical differences were found in the travel times of fish in the two treatment groups, or in the proportion of fish that passed in‐river telemetry checkpoints. These results indicated that the handling and biopsy methods produced similar levels of mortality and tag retention as the telemetry treatment alone and that any changes in behaviour between the two treatment groups did not adversely affect migration time. Based upon the evidence provided from the biotelemetry of >300 adult sockeye salmon, it was felt that this general type of approach could be applicable to other fish species.     

Inhibition of DNA demethylation enhances plant tolerance to cadmium toxicity by improving iron nutrition

Although the alteration of DNA methylation due to abiotic stresses, such as exposure to the toxic metal cadmium (Cd), has been often observed in plants, little is known about whether such epigenetic changes are linked to the ability of plants to adapt to stress. Herein, we report a close linkage between DNA methylation and the adaptational responses in Arabidopsis plants under Cd stress. Exposure to Cd significantly inhibited the expression of three DNA demethylase genes ROS1/DML2/DML3 (RDD) and elevated DNA methylation at the genome-wide level in Col-0 roots. Furthermore, the profile of DNA methylation in Cd-exposed Col-0 roots was similar to that in the roots of rdd triple mutants, which lack RDD, indicating that Cd-induced DNA methylation is associated with the inhibition of RDD. Interestingly, the elevation in DNA methylation in rdd conferred a higher tolerance against Cd stress and improved cellular Fe nutrition in the root tissues. In addition, lowering the Fe supply abolished improved Cd tolerance due to the lack of RDD in rdd. Together, these data suggest that the inhibition of RDD-mediated DNA demethylation in the roots by Cd would in turn enhance plant tolerance to Cd stress by improving Fe nutrition through a feedback mechanism.