Antifungal effects of iron sulfate on grapevine fungal pathogens

The present study aimed to determine the most efficient experimental conditions of iron sulfate use leading to optimal inhibition in the development of fungal pathogens. Assays have been focused on fungal species inducing severe grapevine diseases. FeSO₄ directly inhibited the in vitro mycelial growth of Botrytis cinerea, Eutypa lata, Phaeomoniella chlamydospora, Phaeoacremonium aleophilum, Diplodia seriata, and Neofusicoccum parvum with variable efficiency in the range of 0.5–10 mM. The development was always completely inhibited at 20 mM. This inhibitory effect was greatly increased at acidic pH values. The anionic moiety of the molecule was of importance since bromide, chloride and sulfate were highly active, whereas acetate and oxalate showed a small effect. Electron microscope observations on E. lata and B. cinerea showed that a treatment with FeSO₄ induced dramatic changes in the hyphal organization leading to cell death. No toxicity was observed on grapevine leaves following repeated FeSO₄ sprays in the antifungal concentration range. Therefore, FeSO₄ may be proposed to effectively replace the long-term pollutant use of CuSO₄ as an antifungal agent, with the additional advantage of iron being an important plant micronutrient.     

Winter variation in xylem sap pH of walnut trees: involvement of plasma membrane H+-ATPase of vessel-associated cells

We studied seasonal variation in xylem sap pH of Juglans regia L. Our main objectives were to (1) test the effect of temperature on seasonal changes in xylem sap pH and (2) study the involvement of plasma membrane H+-ATPase of vessel-associated cells in the control of sap pH. For this purpose, orchard-grown trees were compared with trees grown in a heated (> or = 15 degrees C) greenhouse. During autumn, sap pH was not directly influenced by temperature. A seasonal change in H+-ATPase activity resulting from seasonal variation in the amount of protein was measured in orchard-grown trees, whereas no significant seasonal changes were recorded in greenhouse-grown trees. Our data suggest that H+-ATPase does not regulate xylem sap pH directly by donating protons to the xylem, but by facilitating secondary active H+/sugar transport, among other mechanisms.     

Cross-protection of Salmonella abortusovis, S. choleraesuis, S. dublin and S. gallinarum in mice induced by S. abortusovis and S. gallinarum: bacteriology and humoral immune response

Cross-protection induced by primary infection with Abortusovis and Gallinarum was examined against challenge injection with these Salmonella serotypes as well as with Dublin and Choleraesuis, the other virulent serotypes. Abortusovis induced efficient protection against the other Salmonella. Gallinarum was ineffective against Choleraesuis. Even with low multiplication in mice, the Gallinarum J91 strain induced a weak but significant protection against Dublin (same O group serotype). The antibodies in the blood of mice were tested with ELISA specific for the Salmonella antigens used to prime or to challenge animals. The Gallinarum J91 strain was detected to be more antigenic in ELISA than the other Salmonella antigens. It is difficult to conclude on a correlation between IgM or IgG antibodies and induction of protection, because of the variability in immune response according to the different serotype used. Nevertheless, the negative linkage between a number of bacteria in the spleen of mice challenged with Gallinarum and Dublin, and the level of IgM and IgG antibodies specific for the challenging serotype, showed that humoral immune response could be one element of cross-protection, mainly by the immune response against the same O serotype.     

Spatial activity and expression of plasma membrane H+-ATPase in stem xylem of walnut during dormancy and growth resumption

Plasma membrane H+-ATPase (PM H+-ATPase) plays a key role in nutrient transport, stress responses and growth. To evaluate proton motive force differences between apical and basal parts of acrotonic 1-year-old shoots of walnut (Juglans regia L. cv 'Franquette') trees, spatial and seasonal changes in PM H+-ATPase were studied in mature xylem tissues. During both the dormancy and growth resumption periods, and in both the apical and basal parts of the stem, PM H+-ATPase activity showed positive correlations with the amount of immunodetectable protein. In spring, at the time of growth resumption, higher activities and immunoreactivities of PM H+-ATPase were found in the apical part of the stem than in the basal part of the stem. In spring, the decrease in xylem sugar concentration reflected the high sugar uptake rate. Our data suggest that PM H+-ATPase plays a major role in the uptake of carbohydrates from xylem vessels during growth resumption. These results are discussed in the context of the acrotonic tendency of walnut shoots.     

A soft technology to improve survival and reproductive performance of Litopenaeus stylirostris by counterbalancing physiological disturbances associated with handling stress

The consequences of handling stress (fishing, transfer, eyestalk ablation) on shrimp broodstock are poorly documented. The weakness of farmed shrimp, Litopenaeus stylirostris, during winter is a major problem in New Caledonia, because of seasonal climate (tropical–sub-temperate). The transfer of broodstock in winter from earthen outdoor ponds to indoor maturation tanks in the hatchery (T = 20 °C, Salinity = 35‰, fed shrimp) usually leads, after 48 h, to high mortality (up to 70%). Eyestalk ablation to induce ovarian maturation in females leads to further mortality.

Starting from a background analysis of physiological disturbances (initial osmoregulatory imbalance) associated with handling stress (Wabete, N., Chim, L., Lemaire, P., Massabuau, J.-C., 2004. Caractérisation de problèmes de physiologie respiratoire et d'échanges ioniques associés à la manipulation chez la crevette pénéide Litopenaeus stylirostris à 20 °C. Styli 2003. Trente ans de crevetticulture en Nouvelle-Calédonie. Ed. Ifremer. Actes Colloq. 38, 75-84.), we developed a protocol using a soft technology, based on modifications of water salinity, temperature and feeding regime. The aim was to minimize problems of osmoregulatory imbalance and associated mortalities. The protocol we developed, called the LSD OT protocol (Low Salinity and Diet, Optimal Temperature), was first evaluated on sub-adult shrimp (20–25 g) and then applied to broodstock. Survival after transfer and following eyestalk ablation, as well as reproductive achievement (spawning rate, nauplii number) was considerably improved when shrimps were transferred under “physiological comfort” i.e. warmed isosmotic water (26 °C and 26‰) and unfed for 3 d. This new handling protocol, based on a better control of salinity, temperature and feeding conditions, has been transferred successfully to private hatcheries and already contributes to an increased profitability of New-Caledonian shrimp industry.     

Immunological detection of Phaeoacremonium aleophilum,a fungal pathogen found in esca disease

Phaeoacremonium aleophilum is a fungal pathogen implicated in esca, a devastating disease affecting grapevines around the world. We have previously reported that the fungus secreted in its culture medium a variety of polypeptides, which allowed us to develop a serological method of detection. In this method, rabbit antibodies raised against the polypeptide fraction recognized secreted fungal proteins with high sensitivity (commonly 1 ng). These antibodies were specific since they cross-reacted with polypeptides secreted by various strains of Pm. aleophilum, but not with the proteins secreted by many other fungal pathogens implicated in other grapevine infections. Importantly, as shown by ELISA tests and immunolocalization, they did not cross-react with the secreted polypeptides of Eutypa lata (agent of Eutypa dieback), Neofusicoccum parvum (found in Black Dead Arm), and Phaeomoniella chlamydospora (also found in the fungal complex inducing esca). This serological approach permitted the development of a reliable dot-blot method to detect the presence in woody members of Pm. aleophilum in selectively infected trunk and canes. This method of diagnosis is rapid, easy to perform and non destructive for grapevines. Advantages and drawbacks of the method are discussed.     

Non-symbiotic nitrogen fixation in the rhizospheres of rice,maize and different tropical grasses

Nitrogenase activity estimated in the rhizospheres of rice, maize and different tropical grasses grown under controlled laboratory conditions was shown to depend upon plant species. High nitrogenase activity (2000–6000 nmoles C₂H₄ h^?1 g^?1 dry root) occurred in rice rhizosphere, this activity being only 10 times lower than that of symbiotic systems; in the rhizosphere of many other grasses grown in a similar way nitrogenase activity was as low as 10 nmoles C₂H₄ h^?1 g^?1 dry root. The influence of soil type on nitrogenase activity was impressive; but the exact nature of the factors implicated could not be established. A rather weak flush of nitrogenase activity in the rhizosphere occurred in the early stage of the plant growth; it was probably due to the exudation of compounds from the seed and lasted 2 or 4 days according to the size of the seed. When the plant entered into its intense photosynthetic phase, the nitrogenase activity gradually increased. When the shoots were severed, nitrogenase activity in the rhizosphere ceased. Nitrogenase activity in the rhizosphere responded greatly to light intensity. Extrapolation of these laboratory findings to the field is discussed.     

Adaptation of Medicago sativa cv. Gabès to long‐term NaCl stress

The perennial Medicago sativa cv. Gabès is widely grown on saline soils in Tunisian oases. The mechanisms by which this NaCl‐tolerant cultivar maintains a positive growth balance were analyzed. In this plant of considerable agronomic interest, biochemical analyses were conducted in order to study the effects of salinity on mature leaves. Free‐radical detoxification mechanisms and changes induced by the accumulation of reactive oxygen species (ROS) in response to the NaCl stress were compared between the upper (young) and lower (old) carbohydrate source leaves. Long‐term NaCl (150 mM) treatment significantly reduced the size of source leaves supporting growth. Salinity damage was greater in the lower than in the upper leaves. This damage was associated with a high Na⁺ : K⁺ ratio and a decrease in the activity of H₂O₂‐scavenging enzymes, leading to lipid peroxidation. In lower source leaves that were mainly affected by ionic stress, superoxide dismutase (SOD) was overexpressed and guaiacol peroxidase (GPX) activity increased. In contrast, in upper source leaves that were mainly exposed to water deficit, catalase and ascorbate peroxidase (APX) activities increased whereas GPX activity was unchanged. The upper source leaves maintained adequate ionic and water status and an efficient ROS detoxification, allowing sinks to be supplied with photoassimilates and maintaining a positive growth balance in this cultivar of alfalfa.     

The reproductive brain in fish

In fish as in other vertebrates, the brain is actively involved in the control of reproduction, first by participating, under the influence of external factors, in the establishment of an appropriate endocrine status, but also by allowing synchronization of the partners by the time of spawning. It is now well established that the pituitary gonadotropic function is controlled by multiple stimulatory and inhibitory factors, originating mainly from the preoptic region and the mediobasal hypothalamus, both target regions for sexual steroids. Little is known about the mechanisms involved in the mediation of external and internal factors, however there is indication that internal factors, such as androgens and melatonin, known to trigger particular behavioural and endocrine responses, act both at the level of neuroendocrine territories, but also on sensorial systems, which are the actual sites of action for external factors. This paper represents an attempt to summarize and integrate the recent literature devoted to the different aspects of the brain as a major participant in the complex endocrine and behavioural mechanisms of reproduction in fish.     

Somatic embryogenesis, micropropagation and plant regeneration of "Early Mature" walnut trees (Juglans regia) that flower in vitro

Some walnut trees (Juglans regia L.) originating from central Asia display an early flowering phenotype. These "Early Mature" (EM) trees may produce flowers within months of germination. Secondary flowering waves are also observed within a growing season. Inflorescences may carry male, female and hermaphrodite flowers. Progeny obtained from selected EM trees were cultured in vitro to initiate clonal propagation of these genotypes. Embryogenic lines were established through the culture of immature zygotic embryos. Microshoot lines were obtained from germinated somatic or zygotic embryos. Plants showing EM phenotypes were recovered through direct conversion of somatic embryos or adventitious rooting of microcuttings. During the in vitro propagation phase, flower buds were observed on microshoots after three to six subcultures. Histological analysis showed that most of these flowers were hermaphrodite. In vitro apical buds were used to clone the walnut orthologous cDNAs of the AGAMOUS and APETALA 3 MADS-box genes. Northern blots revealed a preferential expression of both of these homeotic genes in flowers. The results highlight the usefulness of EM lines to study the genetic cues controlling flowering and sexual maturity in woody perennials.