Adipokinetic hormone enhances nodule formation and phenoloxidase activation in adult locusts injected with bacterial lipopolysaccharide

Interactions between the locust endocrine and immune systems have been studied in vivo in relation to nodule formation and activation of the prophenoloxidase cascade in the haemolymph. Injection of bacterial lipopolysaccharide (LPS) extracted from Escherichia coli induces nodule formation in larval and adult locusts but does not increase phenoloxidase activity in the haemolymph. Nodule formation starts rapidly after injection of LPS and is virtually complete within 8 h, nodules occurring mainly associated with the dorsal diaphragm on either side of the heart, but sometimes with smaller numbers associated with the ventral diaphragm on either side of the nerve cord. Co-injection of adipokinetic hormone-I (Lom-AKH-I) with LPS stimulates greater numbers of nodules to be formed in larval and adult locusts, and activates phenoloxidase in the haemolymph of mature adults but not of nymphs. The effect of co-injection of Lom-AKH-I with LPS on nodule formation is seen at low doses of hormone; only 0.4 pmol of Lom-AKH-I per adult locust is needed to produce a 50% increase in the number of nodules formed. When different components of LPS from the E. coli Rd mutant are tested, the mono- and the diphosphoryl Lipid A components have similar effects to the intact LPS. Remarkably, detoxified LPS activates phenoloxidase in the absence of Lom-AKH-I, although co-injection with hormone does enhance this response. Both diphosphoryl Lipid A and detoxified LPS induce a level of nodule formation that is enhanced by co-injection of Lom-AKH-I, but monophosphoryl Lipid A does not initiate nodule formation even when injected with hormone. Co-injection of a water-soluble inhibitor of eicosanoid synthesis, diclofenac (2-[(2, 6-dichlorophenyl)amino] benzeneacetic acid), reduces nodule formation in response to injections of LPS (both in the absence and presence of hormone) in a dose-dependent manner, but does not prevent activation of phenoloxidase in adult locusts. It is shown that nodule formation and activation of the prophenoloxidase in locust haemolymph can both be enhanced by Lom-AKH–I, but it is argued that these processes involve distinct mechanisms in which eicosanoid synthesis is important for nodule formation, but not for the increased phenoloxidase activity

Interactions between the endocrine and immune systems in locusts

The prophenoloxidase cascade in the haemolymph of mature adult Locusta migratoria migratorioides (R & F) is activated in response to injection of laminarin, a -1,3 glucan. Co-injection of adipokinetic hormone-I (Lom-AKH-I) and laminarin prolongs the activation of the enzyme in a dose-dependent manner. However, injections of bacterial lipopolysaccharide (LPS) do not activate prophenoloxidase unless AKH is co-injected, when there is a dose-dependent increase in the level of phenoloxidase that persists in the haemolymph for several hours. Even when AKH is co-injected, the highest levels of phenoloxidase activity are always greater after injection of laminarin than after LPS, and these two immunogens must activate the prophenoloxidase cascade by quite distinct pathways. In the present study, interactions between the endocrine and immune systems were examined with respect to activation of prophenoloxidase and the formation of nodules: injection of LPS induces nodule formation in adult locusts. With LPS from Pseudomonas aeruginosa, nodules form exclusively in dense accumulations in the anterior portion of the abdomen on either side of the dorsal blood vessel associated with the dorsal diaphragm. However, with LPS from Escherichia coli, fewer nodules are formed but with a similar distribution, except that occasionally some nodules are aligned additionally on either side of the ventral nerve cord. Co-injection of Lom-AKH-I with LPS from either bacteria stimulates greater numbers of nodules to be formed. This effect of coinjection of AKH on nodule formation is seen at low doses of hormone with only 0.3 or 0.4 pmol of Lom-AKH-1, respectively, increasing the number of nodules by 50%. Injections of octopamine or 5-hydroxytryptamine do not mimic either of the actions of Lom-AKH-I described here. Co-injection of an angiotensin-converting enzyme inhibitor, captopril, reduces nodule formation in response to injections of LPS but has no effect on the activation of phenoloxidase. Co-injection of an inhibitor of eicosanoid synthesis, dexamethasone, with LPS influences nodule formation (with or without AKH) in different ways according to the dose of dexamethasone used, but does not affect activation of prophenoloxidase. Eicosanoid synthesis is important for nodule formation, but not for the activation of the prophenoloxidase cascade in locust haemolymph

A Model for the Development of the Rhizobial and Arbuscular Mycorrhizal Symbioses in Legumes and Its Use to Understand the Roles of Ethylene in the Establishment of these two Symbioses

We propose a model depicting the development of nodulation and arbuscular mycorrhizae. Both processes are dissected into many steps, using Pisum sativum L. nodulation mutants as a guideline. For nodulation, we distinguish two main developmental programs, one epidermal and one cortical. Whereas Nod factors alone affect the cortical program, bacteria are required to trigger the epidermal events. We propose that the two programs of the rhizobial symbiosis evolved separately and that, over time, they came to function together. The distinction between these two programs does not exist for arbuscular mycorrhizae development despite events occurring in both root tissues. Mutations that affect both symbioses are restricted to the epidermal program. We propose here sites of action and potential roles for ethylene during the formation of the two symbioses with a specific hypothesis for nodule organogenesis. Assuming the epidermis does not make ethylene, the microsymbionts probably first encounter a regulatory level of ethylene at the epidermis–outermost cortical cell layer interface. Depending on the hormone concentrations there, infection will either progress or be blocked. In the former case, ethylene affects the cortex cytoskeleton, allowing reorganization that facilitates infection; in the latter case, ethylene acts on several enzymes that interfere with infection thread growth, causing it to abort. Throughout this review, the difficulty of generalizing the roles of ethylene is emphasized and numerous examples are given to demonstrate the diversity that exists in plants

Test of Host Sanction Hypothesis in Soybean Plants Co-inoculated with Nitrogen Fixing and Non-fixing Bradyrhizobium japonicum

Aims: We tested the proposed mechanism for potential sanctions, that the plant would reduce viability of non-fixing rhizobia inside nodules, performing viable Bradyrhizobium japonicum counts from co-occupied and single-occupied nodules in co-inoculated soybean plants. Study Design: Plants were co-inoculated with two strains of B. japonicum, a highly efficient nitrogen fixing wild-type strain BJD321, and the non-fixing, nifH mutant derivative A3, to produce co-occupied nodules as well as single-occupied nodules. Strain A3 lacks nitrogenase activity but shows similar infection and nodule formation levels respect to the wild-type. As the strains used are equivalent in competitive and nodulation abilities and only differ in the nitrogen fixation ability (by nitrogenase inactivation), and share the same plant, root and even nodule, we can assert that themechanism being tested is plant host sanction, and no other proposed mechanisms like partner choice. Place and Duration of Study: Nitrogen Metabolism Lab, Department of Soil Microbiology and Symbiotic Systems at Zaidín Experimental Station (CSIC State Agency, Granada, Spain). 2010- 2011. Methodology: Axenic seedlings of soybean (Glycine max) cultivar Williams were inoculated with 2 ml of bacterial suspension of BJD321 or A3 strains, alone or in 1:1 mixture and supplied with sterilized N free nutrient solution. Four weeks after inoculation plants of each inoculation treatment (BJD321 + A3, BJD321 only, A3 only) were harvested, nodules were counted and weighed and plated to determine rhizobial strain occupation and population. In the aerial part of plants, determinations of weight, N and C content were done. Results: Co-inoculated plants and plants only inoculated with the BJD321 strain showed a similar nitrogen fixation since they did not differ in dry weight, total N content and total C content. Plants with different inoculation treatments (BJD321 + A3, BJD321 only and A3 only) did not differ in nodule number. In co-inoculated plants, nodule occupation did not differ from the expected among strains (about 33% BJD321 + A3, BJD321 only and A3 only), and the weight of nodules occupied by both strains, BJD321 or A3 alone did not differ. In co-inoculated plants rhizobial viability did not differ between BJD321 and A3 strains, either comparing co-occupied nodules or single-occupied nodules. Nodule size and CFU of rhizobia inside nodules were not correlated, either in coinoculated plants or plants inoculated with BJD321 strain alone. Conclusion: We can conclude that in the soybean-B. japonicum system, plants facing the presence of fixing and non-fixing rhizobial strains do not sanction cheating and can perform as well as plants inoculated with the fixing strain alone.Fil: Marco, Diana Elizabeth. Universidad Nacional de Córdoba. Facultad de Cs.agropecuarias. Area de Producción Organica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Talbi, Chouhra. Universidad Nacional Autónoma de México; MéxicoFil: Bedmar, Eulogio. Consejo Superior de Investigaciones Científicas. Estación Experimental del Zaidín; Españ

Host reaction in paratenic fish hosts against 3rd stage larvae of Anguillicola crassus

Many fish species of Lake Balaton (Hungary) serve as paratenic hosts for the 3rd stage larvae of the eel parasite Anguillicola crassus. The incidence oi different phases of the host reaction varies with paratenic fish host but its nature is basically the same. The appearance of mononuclear cells around the larvae migrating in the tissues or in the abdominal cavity is regarded as a sign of cellular host reaction. The cells adhering to the surface of the larvae gradually assume an epithelioid shape and form a granuloma. With the advancement of the process the larvae and the epithelioid cells surrounding them undergo necrosis, and the granuloma becomes surrounded by a connective tissue capsule. In more chronic cases, the inside of the parasitic nodule surrounded by several layers of connective tissue is filled out by amorphous tissue and parasite debris. In the 'suitable paratenic hosts' Anguillicola sp. larvae not affected by the host's cellular reaction outnumber those affected by it, whereas in the 'less suitable paratenic hosts' the cellular reaction very rapidly forces the larvae into foci and destroys them

Ferromanganese nodules and micro-hardgrounds associated with the Cadiz Contourite Channel (NE Atlantic): Palaeoenvironmental records of fluid venting and bottom currents

Ferromanganese nodule fields and hardgrounds have recently been discovered in the Cadiz Contourite Channel in the Gulf of Cadiz (850–1000 m). This channel is part of a large contourite depositional system generated by the Mediterranean Outflow Water. Ferromanganese deposits linked to contourites are interesting tools for palaeoenviromental studies and show an increasing economic interest as potential mineral resources for base and strategic metals. We present a complete characterisation of these deposits based on submarine photographs and geophysical, petrographic, mineralogical and geochemical data. The genesis and growth of ferromanganese deposits, strongly enriched in Fe vs. Mn (av. 39% vs. 6%) in this contourite depositional system result from the combination of hydrogenetic and diagenetic processes. The interaction of the Mediterranean Outflow Water with the continental margin has led to the formation of Late Pleistocene–Holocene ferromanganese mineral deposits, in parallel to the evolution of the contourite depositional system triggered by climatic and tectonic events. The diagenetic growth was fuelled by the anaerobic oxidation of thermogenic hydrocarbons (δ13CPDB=−20 to −37‰) and organic matter within the channel floor sediments, promoting the formation of Fe–Mn carbonate nodules. High 87Sr/86Sr isotopic values (up to 0.70993±0.00025) observed in the inner parts of nodules are related to the influence of radiogenic fluids fuelled by deep-seated fluid venting across the fault systems in the diapirs below the Cadiz Contourite Channel. Erosive action of the Mediterranean Outflow Water undercurrent could have exhumed the Fe–Mn carbonate nodules, especially in the glacial periods, when the lower core of the undercurrent was more active in the study area. The growth rate determined by 230Thexcess/232Th was 113±11 mm/Ma, supporting the hypothesis that the growth of the nodules records palaeoenvironmental changes during the last 70 ka. Ca-rich layers in the nodules could point to the interaction between the Mediterranean Outflow Water and the North Atlantic Deep Water during the Heinrich events. Siderite–rhodochrosite nodules exposed to the oxidising seabottom waters were replaced by Fe–Mn oxyhydroxides. Slow hydrogenetic growth of goethite from the seawaters is observed in the outermost parts of the exhumed nodules and hardgrounds, which show imprints of the Mediterranean Outflow Water with low 87Sr/86Sr isotopic values (down to 0.70693±0.00081). We propose a new genetic and evolutionary model for ferromanganese oxide nodules derived from ferromanganese carbonate nodules formed on continental margins above the carbonate compensation depth and dominated by hydrocarbon seepage structures and strong erosive action of bottom currents. We also compare and discuss the generation of ferromanganese deposits in the Cadiz Contourite Channel with that in other locations and suggest that our model can be applied to ferromanganiferous deposits in other contouritic systems affected by fluid venting

<i>Medicago truncatula</i> functional genomics: an invaluable resource for studies on agriculture sustainability

Legume functional genomics has moved many steps forward in the last two decades thanks to the improvement of genomics technologies and to the efforts of the research community. Tools for functional genomics studies are now available in Lotus japonicus, Medicago truncatula and soybean. In this chapter we focus on M.truncatula, as a model species for forage legumes, on the main achievements obtained due to the reported resources and on the future perspectives for the study of gene function in this species

GuaB Activity Is Required in Rhizobium tropici During the Early Stages of Nodulation of Determinate Nodules but Is Dispensable for the Sinorhizobium meliloti–Alfalfa Symbiotic Interaction

The guaB mutant strain Rhizobium tropici CIAT8999-10T is defective in symbiosis with common bean, forming nodules that lack rhizobial content. In order to investigate the timing of the guaB requirement during the nodule formation on the host common bean by the strain CIAT899-10.T, we constructed gene fusions in which the guaB gene is expressed under the control of the symbiotic promoters nodA, bacA, and nifH. Our data indicated that the guaB is required from the early stages of nodulation because full recovery of the wild-type phenotype was accomplished by the nodA-guaB fusion. In addition, we have constructed a guaB mutant derived from Sinorhizobium meliloti 1021, and shown that, unlike R. tropici, the guaB S. meliloti mutant is auxotrophic for guanine and induces wild-type nodules on alfalfa and Medicago truncatula. The guaB R. tropici mutant also is defective in its symbiosis with Macroptilium atropurpureum and Vigna unguiculata but normal with Leucaena leucocephala. These results show that the requirement of the rhizobial guaB for symbiosis is found to be associated with host plants that form determinate type of nodules.Fil: Collavino, Mónica Mariana. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Botánica del Nordeste. Universidad Nacional del Nordeste. Facultad de Ciencias Agrarias. Instituto de Botánica del Nordeste; ArgentinaFil: Riccillo, Pablo M.. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; ArgentinaFil: Grasso, Daniel Horacio. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Crespi, Martín. Centre National de la Recherche Scientifique; FranciaFil: Aguilar, Orlando Mario. Universidad Nacional de La Plata. Facultad de Ciencias Exactas; Argentin