Objective To isolate and characterize symbiotic microorganisms from adesertified Mediterranean ecosystem and to develop macromolecularmethods for their identification. To also select effectivecultures from existing collections and to employ these methodsfor their characterisation, screening and to monitor the fate ofthe introduced microbial inoculants within a mixed population ofmicrosymbionts.The site for field work is a desertified ecosystem in Almeria, SESpain (230mm annual rainfall). Arbuscular-mycorrhizal fungi(AMF), Rhizobium bacteria and plant-growth promoting bacteria(PGPR) will be isolated from the roots of shrub legumes. Thediversity of AMF at the site will be assessed through the year.Pot cultures will be established and characterized on sporemorphology, anatomy of colonisation, and isoenzyme profiles.Bacteria will be characterized by conventional moleculartechniques. Ecological characteristics of the AMF will bestudied in laboratory microcosms. Their reproductive andspreading strategy will be evaluated by measuring the levels ofcolonization relative to production of extraradical mycelium, andmetabolic activity of mycelium using image analysis.Greenhouse studies will compare indigenous AMF populations indesertified soils with exotic isolates. Inoculant AMF in tandemwith effective rhizobial strains will also be assessed forstimulation of nitrogen fixation using 15N methodologies, and Ntransfer will also be ascertained by isotopic dilutiontechniques. Bacterial isolates will be marked genetically forresistance to antibiotics via selection of spontaneous mutants todetect them in field soils.Immunological, electrophoretic and other methods will bedeveloped to compare the spread of introduced and indigenous AMFusing rhizoboxes where hyphae can be separated from roots.Isozyme profiles of AMF will be screened for potential to monitorwhich AMF is present. Competition between isolates will also bestudied using monoclonal antibodies (Ab) using hyphae asimmunogens to gather data on the spread of AMF by measuring theproportion of the total extramatrical mycelium belonging toindividual AMF. Antibodies will be screened against the otherAMF isolates in order to find Ab able to differentiate AMF. Insurveys of mixed populations in rhizoboxes, the mycelium will beextracted and assayed using fluorescent stain-conjugated Ab or inan ELISA format utilizing Ab coupled to enzymes which catalyze aspecific quantitative colour reaction. The immunofluorescentstaining of the AMF will allow the observation and identificationof hyphae in situ, this will be combined with activity stainingusing an image analysis system to quantify the differences influorescence.The selected organisms will be used as inoculants to enhance theestablishment of three pioneer shrubs in a desertifiedMediterranean ecosystem, thus accelerating its sustainablerehabilitation. The inoculant microsymbionts will be madeavailable for the target desertified ecosystem. Small-scalefield plots of pioneer legumes inoculated with the recommendedmicrosymbionts will be established and monitored for successfulestablishment. Fields of science natural sciencesbiological sciencesmicrobiologybacteriologynatural sciencesbiological sciencesmicrobiologymycologynatural sciencesbiological sciencesecologyecosystemsnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymesagricultural sciencesagriculture, forestry, and fisheriesagriculturegrains and oilseedslegumes Programme(s) FP3-ENV 1C - Specific research and technological development programme (EEC) in the field of the environment, 1990-1994 Topic(s) 0403 - Desertification in the Mediterranean area Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator University of Kent at Canterbury Address CT2 7NJ Canterbury United Kingdom See on map EU contribution € 0,00 Participants (3) Sort alphabetically Sort by EU Contribution Expand all Collapse all CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS Spain EU contribution € 0,00 Address 1,profesor albareda 1 18008 Granada See on map FORESTRY COMMISSION RESEARCH AGENCY United Kingdom EU contribution € 0,00 Address Northern research station EH25 9SY Roslin-midlothian See on map TECHNICAL UNIVERSITY OF MUNICH Germany EU contribution € 0,00 Address Alte akademie 12 85350 Freising See on map