High frequency somatic embryogenesis with a pampeana-derived genotype of alfalfa (Medicago sativa L.)

Summary Somatic embryos of genotype R11 of the alfalfa variety Pampeana were produced from embryogenic calli derived from leaf sections. They were induced by an auxin shock and its development was attempted on six different media. The best condition for somatic embryo production was inducing callus on MS medium plus 10 M 2,4-D and 4,6 M KIN and transferring them, after the auxin shock, to MS with 10–20 mM NH₄ ⁺ and 30 mM proline. More than 500 somatic embryos per plate were produced. Embryos were grown to plants on MS or half strength MS media and all regenerated plants resembled the original R11 genotype. This technique could be useful in alfalfa Pampeana improvement using genetic modification.     

Effects of glandular trichomes on the development of Phytophthora infestans infection in potato (S. tuberosum)

S. berthaulltii Hawkes, a wild potato species, possesses two types of glandular trichomes on its leaf surface: types A and B wich confer resistance to insects. Preliminary results indicated that the presence of glandular trichomes also confers resistance to Phytophthora infestans, the causal agent of late blight. In this paper we report the evaluation of trichome characteristics (density and exudate activities) performed on 8 lines of potato, including the control varieties Desirée and Tropicana, 5 hybrid lines derived from intercrosses and backcrosses between S. tuberosum and S. berthaultii and one accession of S. berthaultii. Whole plants of the 8 genotypes were inoculated with a spore suspension of Phytophthora infestans in order to evaluate their resistance to late blight. The resultant disease incidence correlated negatively with type A trichome density and with polyphenol-oxidase (PPO) activity of type A trichome glands. The removal of type B trichomes exudate by leaf dipping in ethanol did not influence the disease development after artificial inoculation of detached leaves. Type B trichome exudate was extracted and the fraction corresponding to Fatty Acids Sucrose Esters (FASE) was recovered after Thin Layer Chromatography and tested on the fungal mycelium in vitro. The FASE exhibited a slight inhibitory effect on mycelium growth. The results of the experiments are discussed, and several hypotheses regarding the possible role of types A and B trichomes are formulated. This revised version was published online in July 2006 with corrections to the Cover Date.     

Synthesis and resistance to in vitro proteolysis of transglutaminase cross-linked phaseolin, the major storage protein from Phaseolus vulgaris

The ability of phaseolin to act as an acyl donor and acceptor substrate of transglutaminase was studied by using an enzyme isolated from Streptoverticillium mobarense. Phaseolin, a trimeric storage protein from Phaseolus vulgaris L., was shown to possess both glutamine and lysine residues reactive for the enzyme. The extent of transglutaminase-catalyzed cross-linking has been studied in function of both incubation time and enzyme concentration. Native- and SDS-PAGE demonstrated that phaseolin is intra- and intermolecularly cross-linked by transglutaminase and gives rise to different polymers as well as to modified forms of the protein having a similar molecular weight but lower Stokes radius if compared to unmodified phaseolin. Cross-linked phaseolin was found to be more resistant to proteolytic cleavage than the unmodified counterpart, as demonstrated by in vitro trypsin and pepsin digestion experiments. This behavior could suggest novel possible uses of the transglutaminase-modified phaseolin.     

Diversity characterisation of broccoli (Brassica oleracea L. var. italica Plenck) landraces for their on-farm (in situ) safeguard and use in breeding programs

Brassica oleracea L. is an important species that originated in the eastern Mediterranean area. From there, B. oleracea L. subsp. capitata (L.) DC. convar. botrytis (L.) Alef. var. italica Plenck (broccoli) was introduced into Italy where considerable diversification took place. Several landraces of broccoli, which are highly appreciated by local people for their organoleptic traits, are presently cultivated in both large fields and home gardens. In this study, 10 landraces, 4 synthetics derived from landraces and 4 of the most used F1 hybrids were characterised for morpho-physiological and molecular traits with the principal aim of feeding information into the landrace protection schemes being developed in Italy and into breeding programs. The landraces and derived synthetics are well differentiated from each other and from the F1 hybrids for important agronomic and genetic traits. The principal component analysis based on analysis of morpho-physiological traits showed that overall the 2 main components accounted for 67.74 % of the total variation. The neighbour joining tree, based on 23 microsatellite markers, grouped accessions into 2 main clusters. One includes one synthetic variety and 2 F1 hybrids, while the other cluster includes all the other accessions. There were also several sub clusters. Most of the materials examined showed within-accession diversity. The information gathered in this study can be used to protect landraces through the schemes presently being developed in Italy and to register them in the European common catalogue of “conservation varieties”. In situ (on-farm) conservation of landrace diversity is discussed with reference to landrace use.     

The European seed legislation on conservation varieties: focus, implementation, present and future impact on landrace on farm conservation

After a brief presentation of the European Directives on seed marketing of conservation varieties, data relative to their implementation status are given and discussed in relation to on farm conservation of landraces. Although generally aimed to ensure in situ conservation and the sustainable use of plant genetic resources, the Directives focus on seed production and marketing instead of genetic resource conservation per se. At present, their application has only partially favoured the registration of landraces maintained on farm or preserved in ex situ collections. They can be estimated in thousands in Europe, while only a few landrace with a verified status (51 out of a total of 184 conservation varieties) are presently included in the Common Catalogue. Reasons for this scarce registration are discussed. The European Directives on seed marketing of conservation varieties are a tool to promote on farm conservation of landraces that should be used to a greater extent than at present. Registration of landraces should be strongly and principally promoted by public bodies as a measure to give access to the genetic resources, preserve them on the long term and favour the economic profit of the farmers maintaining them.     

Soil bacterial communities under slash and burn in Mozambique as revealed by a metataxonomic approach

The slash-and-burn system is a subsistence agronomical practice widespread in tropical areas worldwide. This system has been extensively studied,especially for its impacts on agronomical aspects and soil physicochemical properties; however, knowledge of soil microbial diversity under slash and bum is scarce. In this study, for the first time, soil bacterial diversity of three locations from Central Mozambique, where slash and burn has been practiced for different durations of the forest fallow p...     

Hydrocarbon and fatty acid composition of cheese as affected by the pasture vegetation type

The determination of the geographical origin of dairy products is an ongoing issue. In this paper the effects of botanical diversity of two pastures on the hydrocarbon and fatty acid composition of cheese fat were studied, over 2 years of experimentation. Two areas in the Italian southwestern Alpine region, dominated by Trifolium alpinum (T) and Festuca nigrescens (F) vegetation, respectively, were chosen, and milk obtained from cows grazing on these pastures was used to produce a semihard traditional cheese. Cheese samples showed a significantly different composition of most linear hydrocarbons, odd-chain (C15, C17, and C17:1) and unsaturated (trans-11,cis-15-C18:2, C18:3, C20:4n-6, C20:4n-3, and 20:5n-3) fatty acids, according to pasture type. The ratio between C(29) and C(27) linear hydrocarbons, unlike the absolute content of the single molecules, showed a good discriminating ability between the two pastures and was little affected by the natural variability due to the climatic and environmental factors.     

Effect of commercial lignosulfonate-humate on Zea mays L. metabolism

Lignosulfonate-humate a and lignosulfonate-humate b, derived by an industrial process from lignin, were studied chemically and biologically, and their effects on maize metabolism compared with the responses induced by humic substances obtained from leonardite. Lignosulfonate-humate a and lignosulfonate-humate b elicited hormonelike activity and leonardite displayed giberellin properties. To improve our understanding of their biological action, lignosulfonate-humate a, lignosulfonate-humate b and leonardite were supplied to maize plants and their effect was studied on growth, nitrogen metabolism and photosynthesis. All products increased root and leaf growth. Glutamine-synthetase, glutamate-synthase enzyme activities and protein content were all increased. The treatments also increased chlorophyll content, glucose, fructose and rubisco enzyme activity, suggesting a positive role of lignosulfonate-humate a, lignosulfonate-humate b and leonardite in the photosynthetic process. In addition, an increase in phenol content was observed. In light of these results, being environmentally friendly products, lignosulfonate-humate a and lignosulfonate-humate b could be used to increase crop yield.     

Effects of pig slurry application on soils and soil humic acids

The effect of three annually consecutive additions of pig slurry at two rates (90 and 150 m3 x ha(-1) x year(-1) on soils and soil humic acids (HAs) was investigated in a field experiment under semiarid conditions. Soils and pig slurries were analyzed by standard methods. The HAs were isolated from soils and pig slurry by a conventional procedure based on alkaline extraction, acidic precipitation to pH 1, purification by repeated alkaline dissolutions and acidic precipitations, water washing, dialysis, and final freeze-drying. The HAs obtained were analyzed for elemental (C, H, N, S, and O) and acidic functional group (carboxylic and phenolic) composition, and by UV-vis, FT-IR, fluorescence, and ESR spectroscopies. With respect to the control soil, the pig slurry amended soils had greater pH and electrical conductivity, slightly larger total N content, and smaller values of C/N ratio. A decrease of total organic C was observed only in soils amended for 2 and 3 years at the higher slurry rate. With respect to control soil HA, pig slurry HA was characterized by larger contents of S- and N-containing groups, smaller acidic functional group and organic free radical contents, a prevalent aliphatic character, extended molecular heterogeneity, and smaller aromatic polycondensation and humification degrees. Amendment with pig slurry HA determines a number of modifications in soil HAs, including increase of C, S, and COOH contents, C/N ratios, and aliphaticity and decrease of extraction yields and N, O, phenolic OH, and organic free radical contents. These effects are generally more evident after the first year of slurry application and tend to disappear with increasing number of treatments. Most probably, over the years the slightly humified slurry HA is mineralized through extended microbial oxidation, whereas only the most recalcitrant components, such as S-containing, phenolic, and aliphatic structures, are partially accumulated by incorporation into soil HA.     

Biodegradability of Disinfectants in Surface Waters from Buenos Aires: Isolation of an Indigenous Strain Able to Degrade and Detoxify Benzalkonium Chloride

Biodegradability of chlorhexidine (CH), triclosan (TC), and benzalkonium chloride (CBA) has been tested in 18 surface water sampling points in the urban area of Buenos Aires. Sampling points were located in both the Reconquista and the Matanza-Riachuelo basins as well as in the La Plata River. High tolerance to the three disinfectants was found and indigenous strains capable of degrading CBA and TC were isolated. Neither tolerance nor biodegradation were correlated with sewage pollution. A strain that degrades CBA was identified as belonging to the genus Pseudomonas using the API20NE system and 16SRNA sequencing. In batch assays, the strain was capable of degrading 100, 200, and up to 500 mg L^?1 of CBA in 10, 25, and 46 h respectively with specific growth rates (μ) of 0.56, 0.30, and 0.14 h^?1. The efficiency of the process was between 99.5–98.0% in terms of compound removal and between 93.8–89.1% in terms of chemical oxygen demand (COD). The detoxification of the compound as a result of the biodegradation was assessed using Pseudokirchneriella subcapitata, Vibrio fischeri, and Lactuca sativa as test organisms.