Towards a reporter system to identify regulators of cross-talk between salicylate and jasmonate signaling pathways in Arabidopsis

The plant signaling hormones salicylic acid (SA) and jasmonic acid (JA) are regulators of inducible defenses that are activated upon pathogen or insect attack. Cross-talk between SA- and JA-dependent signaling pathways allows a plant to finely tune its response to the attacker encountered. In Arabidopsis, pharmacological experiments revealed that SA exerts a strong antagonistic effect on JA-responsive genes, such as PDF1.2, indicating that the SA pathway can be prioritized over the JA pathway. SA-mediated suppression of the JA-responsive PDF1.2 promoter was exploited for setting up a genetic screen aiming at the isolation of signal transduction mutants that are impaired in this cross-talk mechanism. The PDF1.2 promoter was fused to the herbicide resistance gene BAR to allow for life/death screening of a population of mutagenized transgenic plants. Non-mutant plants should survive herbicide treatment when methyl jasmonate (MeJA) is applied, but suppression of the JA response by SA should be lethal in combination with the herbicide. Conversely, crucial SA/JA cross-talk mutants should survive the combination treatment. SA effectively suppressed the expression of the PDF1.2::BAR transgene. However, suppression of the BAR gene did not result in suppression of herbicide resistance. Hence, a screening method based on quantitative differences in the expression of a reporter gene may be better suited to identify SA/JA cross-talk mutants. Here, we demonstrate that the PDF1.2::GUS reporter will be excellently suited in this respect.Key words: plant defense, salicylic acid, jasmonic acid, cross-talk, mutant screen, Arabidopsis     

Functional characterization and evolution of PTH/PTHrP receptors: insights from the chicken

ABSTRACT: BACKGROUND: The parathyroid hormone (PTH)-family consists of a group of structurally related factors that regulate calcium and bone homeostasis and are also involved in development of organs such as the heart, mammary gland and immune system. They interact with specific members of family 2 B1 G-protein coupled receptors (GPCRs), which have been characterised in teleosts and mammals. Two PTH/PTHrP receptors, PTH1R and PTH2R exist in mammals and in teleost fish a further receptor PTH3R has also been identified. Recently in chicken, PTHfamily members involved in calcium transport were characterized and specific PTHRs are suggested to exist although they have not yet been isolated or functionally characterized. The aim of this study is to further explore the evolution and function of the vertebrate PTH/PTHrP system through the isolation, phylogenetic analysis and functional characterization of the chicken receptors. RESULTS: Two PTHRs were isolated in chicken and sequence comparison and phylogenetic analysis indicate that the chicken receptors correspond to PTH1R and PTH3R, which emerged prior to the teleost/tetrapod divergence since they are present in cartilaginous fish. The vertebrate PTH2R receptor and its ligand TIP39 have been lost from bird genomes. Chicken PTH1R and PTH3R have a divergent and widespread tissue expression and are also evident in very early embryonic stages of development. Receptor stimulation studies using HEK293 cells stably expressing the chicken PTH1R and PTH3R and monitoring cAMP production revealed they are activated by chicken 1-34 N-terminal PTH-family peptides in a dose dependent manner. PTH-L and PTHrP were the most effective peptides in activating PTH1R (EC50 = 7.7 nM and EC50 = 22.7 nM, respectively). In contrast, PTH-L (100 nM) produced a small cAMP accumulation on activation of PTH3R but PTHrP and PTH (EC50 = 2.5 nM and EC50 = 22.1 nM, respectively) readily activated the receptor. PTHrP also stimulated intracellular Ca2+ accumulation on activation of PTH1R but not PTH3R. CONCLUSION: Two PTHR homologues of the vertebrate PTH1R and PTH3R were isolated and functionally characterized in chicken. Their distinct pattern of expression during embryo development and in adult tissues, together with their ligand preference, suggests that they have acquired specific functions, which have contributed to their maintenance in the genome. PTH2R and its activating ligand, TIP39, are absent from bird genomes. Nonetheless identification of putative PTH2R and TIP39 in the genome of an ancient agnathan, lamprey, suggests the PTH/PTHrP ligand and receptor family was already present in an early basal paraphyletic group of vertebrates and during the vertebrate radiation diverged via gene/genome duplication and deletion events. Knowledge of the role PTH/PTHrP system in early vertebrates will help to establish evolution of function.     

Expression of an apoplast‐localized BURP‐domain protein from soybean (GmRD22) enhances tolerance towards abiotic stress

The BURP‐domain protein family comprises a diverse group of plant‐specific proteins that share a conserved BURP domain at the C terminus. However, there have been only limited studies on the functions and subcellular localization of these proteins. Members of the RD22‐like subfamily are postulated to associate with stress responses due to the stress‐inducible nature of some RD22‐like genes. In this report, we used different transgenic systems (cells and in planta) to show that the expression of a stress‐inducible RD22‐like protein from soybean (GmRD22) can alleviate salinity and osmotic stress. We also performed detailed microscopic studies using both fusion proteins and immuno‐electron microscopic techniques to demonstrate the apoplast localization of GmRD22, for which the BURP domain is a critical determinant of the subcellular localization. The apoplastic GmRD22 interacts with a cell wall peroxidase and the ectopic expression of GmRD22 in both transgenic Arabidopsis thaliana and transgenic rice resulted in increased lignin production when subjected to salinity stress. It is possible that GmRD22 regulates cell wall peroxidases and hence strengthens cell wall integrity under such stress conditions.     

INTERPOPULATION VARIATION IN JUVENILE SURVIVAL AND GROWTH RATES OF LYMNAEA PEREGRA (GASTROPODA: PULMONATA); TEMPERATURE AT RECRUITMENT AS A SELECTION PRESSURE?

Three neighbouring populations of Lymnaea peregra had recruitmentin the summer (June and July), but one population (Sheaf) hada second recruitment in September and October. We hypothesizedthat juveniles of the Sheaf population would be subject to selectionunder both ‘summer’ and ‘winter’ conditions,and thus should be more resistant to low-temperature stressthan juveniles of the other populations. The hypothesis wassupported by the findings that Sheaf juveniles survived andgrew better over a wider range of temperatures (2, 10, 15 and20°C )while juveniles of the other two populations wereadapted only to higher temperatures (15 and 20°C). Therewas evidence that some of these traits were genetically fixed. ^*Present address: Department of Applied Science, City Polytechnicof Hong Kong, Mongkok, Kowloon, Hong Kong. (Received 15 September 1988; accepted 16 December 1988)     

The unconventional P‐loop NTPase OsYchF1 and its regulator OsGAP1 play opposite roles in salinity stress tolerance

YchF proteins are a group of mysterious but ubiquitous unconventional G‐proteins found in all kingdoms of life except Archaea. Their functions have been documented in microorganisms, protozoa and human, but those of plant YchF homologues are largely unknown. Our group has previously shown that OsYchF1 and its interacting protein, OsGAP1, play opposite roles in plant defense responses. OsGAP1 was found to stimulate the GTPase/ATPase activities of OsYchF1 and regulate its subcellular localization. In this report, we demonstrate that both OsYchF1 and OsGAP1 are localized mainly in the cytosol under NaCl treatment. The ectopic expression of OsYchF1 in transgenic Arabidopsis thaliana leads to reduced tolerance towards salinity stress, while the ectopic expression of OsGAP1 has the opposite effect. Similar results were also obtained with the Arabidopsis homologues, AtYchF1 and AtGAP1, by using AtGAP1 overexpressors and underexpressors, as well as an AtYchF1‐knockdown mutant. OsYchF1 and OsGAP1 also exhibit highly significant effects on salinity‐induced oxidative stress tolerance. The expression of OsYchF1 suppresses the anti‐oxidation enzymatic activities and increases lipid peroxidation in transgenic Arabidopsis, and leads to the accumulation of reactive oxygen species (ROS) in tobacco BY‐2 cells, while the ectopic expression of OsGAP1 has the opposite effects in these two model systems.     

Notes on the genus Sinocapritermes (Isoptera: Termitidae) from China, with description of a new species

Abstract. The genus Sinocapritermes is briefly reviewed, and a new species described. A key is also provided to the twelve species of Sinocapritermes recorded from China to date.     

DIFFERENCES IN THE SHELL SHAPE OF LYMNAEA PEREGRA (MULLER) (GASTROPODA: PULMONATA) FROM LOTIC AND LENTIC HABITATS; ENVIRONMENTAL OR GENETIC VARIANCE?

Lymnaea peregra from lotic habitats (running waters) had largeraperture-length/shell-length ratios than contemporaries fromlentic sites (standing waters). This pattern was observed infield samples from two separate years (1985 and 1986). Laboratoryculture experiments suggested that shell shape could be geneticallydetermined in at least some populations. We hypothesize thatlotic snails possess bigger feet, and hence larger aperturesthan lentic individuals as an adaptation to withstand watercurrent ^*Present address Departments of Biology, King's College (KQC),University of London, Campden Hill Road, London W3 7AH, England (Received 14 September 1987;     

Chick embryo chorioallantoic membrane (CAM): an alternative predictive model in acute toxicological studies for anti-cancer drugs

The chick embryo chorioallantoic membrane (CAM) is a preclinical model widely used for vascular and anti-vascular effects of therapeutic agents in vivo. In this study, we examine the suitability of CAM as a predictive model for acute toxicology studies of drugs by comparing it to conventional mouse and rat models for 10 FDA-approved anticancer drugs (paclitaxel, carmustine, camptothecin, cyclophosphamide, vincristine, cisplatin, aloin, mitomycin C, actinomycin-D, melphalan). Suitable formulations for intravenous administration were determined before the average of median lethal dose (LD₅₀) and median survival dose (SD₅₀) in the CAM were measured and calculated for these drugs. The resultant ideal LD₅₀ values were correlated to those reported in the literature using Pearson’s correlation test for both intravenous and intraperitoneal routes of injection in rodents. Our results showed moderate correlations (r²=0.42 − 0.68, P<0.005–0.05) between the ideal LD₅₀ values obtained using the CAM model with LD₅₀ values from mice and rats models for both intravenous and intraperitoneal administrations, suggesting that the chick embryo may be a suitable alternative model for acute drug toxicity screening before embarking on full toxicological investigations in rodents in development of anticancer drugs.