Years: 2010-2012  


This three-year project is aimed to purpose new potential nematicidal formulations, either liquid or solid, through the investigation of the nematicidal properties of extracts and biomasses of these six Medicago spp. (M. sativa, M. arabica, M. arborea, M. hybrida, M. truncatula and M. murex) and of active principles isolated from roots and aerial parts of these materials, against the phytoparatic nematode species economically most relevant for Italian and Polish agriculture as above cited.
Work programme provides the initial cultivation of M. sativa, M. arabica, M. arborea, M. hybrida, M. truncatula and M. murex either in Italy and Poland, in order to verify plant adaptability to the climatic conditions of both countries. In a following step plant extracts will be tested against the root-knot nematode Meloidogyne incognita, the cyst nematode Globodera rostochiensis and the virus-vector Xiphinema index, previously reared on host plants in greenhouse conditions. In particular, aqueous and solvent extracts will be prepared at different concentrations for the treatment of infective larval stages of M.incognita and G. rostochiensis and adult females of X. index. Moreover, hatching tests will be arranged on the egg masses of M. incognita and the cysts of G. rostochiensis, in order to verify the potential ovicidal effects of the extracts. Isolation of saponin components will be conducted by combination of chromatographic techniques in normal and reversed phase mode. Active components of the extracts will be analytically determined by modern spectroscopic methods employed high performance liquid chromatography (HPLC) and ultra performance liquid chromatography (UPLC) both coupled to mass spectrometers (LC-MS) The methods described can be applied both for the quantification of individual glycosides and also for qualitative screening to find species with the highest content of the saponins. Coupled HPLC to mass selective detector is one of the most important techniques of the last decade. The combination offers the possibility of taking advantage of chromatography as separation method and MS as an identification tool. The amazing number of application of mass spectrometric techniques used for the structural analysis and characterization of saponins have been presented and summarized. Each active principle will be separately tested in vitro on the above nematode species, in order to identify the main agents of the nematicidal effect. In a further phase of the project, greenhouse trials in potting mixes will be undertaken with the aim to verify the nematicidal effectiveness of extracts in soil infested by the same nematode species. Extract treatment will be compared with direct soil incorporation of corresponding amounts of plant dry biomass, aiming to verify its potential use as soil organic amendment. All data from the experimental activity will be statistically analysed and then reported in scientific articles.
Timescale of the project:
First year

Production of biomasses of Medicago plants to investigate. Preparation of aqueous or solvent extracts from these plants. Laboratory trials testing the effect of extracts on infective stages of three different nematode species, the root-knot nematode Meloidogyne incognita, the cyst species Globodera rostochiensis and the virus-vector Xiphinema index. Hatching tests with the same extracts on egg masses of M. incognita and cysts of G. rostochiensis.
Second year
Analytical determination of the active components of the extracts by HPLC and UPLC techniques. Structural analysis, quantitative characterization and isolation of saponins from each Medicago species. Laboratory assay of single isolated saponins according to the same experimental programme of the first year.
Third year
Preparation of dehydrated and/or liophilized biomasses from the six Medicago species. Testing of these biomasses at different rates for treatments to soil infested by M. incognita or G. rostochiensis in greenhouse conditions. Determination of nematological parameters on soil and roots from these greenhouse experiments. Statistical analysis and discussion of data from the three-year activity.

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