Engineering modified Bt toxins to counter insect resistance

The evolution of insect resistance threatens the effectiveness of Bacillus thuringiensis (Bt) toxins that are widely used in sprays and transgenic crops. Resistance to Bt toxins in some insects is linked with mutations that disrupt a toxin-binding cadherin protein. We show that susceptibility to the Bt toxin Cry1Ab was reduced by cadherin gene silencing with RNA interference in Manduca sexta, confirming cadherin's role in Bt toxicity. Native Cry1A toxins required cadherin to form oligomers, but modified Cry1A toxins lacking one alpha-helix did not. The modified toxins killed cadherin-silenced M. sexta and Bt-resistant Pectinophora gossypiella that had cadherin deletion mutations. Our findings suggest that cadherin promotes Bt toxicity by facilitating toxin oligomerization and demonstrate that the modified Bt toxins may be useful against pests resistant to standard Bt toxins.     

Soil genesis and heterogeneity of phosphorus forms and carbon below mounds inhabited by primary and secondary termites

Termite activities are known to significantly influence small-scale soil properties in tropical savannas. The lateral and vertical extent of the alterations to the nest's surrounding, and particularly resulting impacts on diagnostic soil horizons remain largely unresolved until today. We examined the effects of mound-building termites on soil genesis and constitutive chemical soil properties in and below their nests. Two transects to a soil depth of 100 cm were dug below three younger mounds of Cornitermes silvestrii (the primary nest builder), three older mounds in which C. silvestrii had died out and which were secondarily colonized mainly by Nasutitermes kemneri, and three reference sites in the Brazilian Cerrado. The samples were characterized by standard procedures for soil classification; in addition, phosphorus extractions were conducted on selected samples using NaHCO₃ for labile P forms, and concentrated HCl for stable P forms. This data set was then used to build calibration models for the prediction of labile and stable inorganic (P_i) and organic (P_o) P forms, as well as for contents of organic carbon (OC), for the remaining samples applying mid-infrared spectroscopy in combination with partial least squares regression (MIRS-PLSR). We can show that the termite influence on the soil was sufficiently large to change diagnostic characteristics of the soils under termite mounds. The MIRS-PLSR predictions were suitable for quantifying organic carbon and most of the labile and stable phosphorus fractions. They showed an enrichment of OC, NaHCO₃-P_o and NaHCO₃-P_i contents in nests inhabited by primary and secondary termites by factors of 1.6–2.0 and 1.4–1.5, respectively. The soils surrounding the nests had higher contents of OC and NaHCO₃-P under both nest types vertically down to 30 cm below the lower nest border, and OC and NaHCO₃-P_i contents were elevated at minimum to a lateral distance of 60 cm away from the nest border. As the pattern of HCl_conc-P_i, which comprised 95% of total P, showed no variations, we conclude that the higher NaHCO₃-P_i amount was formed in termite nests by changing the availability of the more stable HCl_conc-P_i. In contrast to the contents, the OC and NaHCO₃-P stocks below the mounds inhabited by primary termites were comparable to those inhabited by secondary ones, because the bulk density of the secondarily inhabited nests was elevated. This was due to a transport of clay-rich material from the subsurface argic horizons into the nests. Here, the secondary termites even reverted the lessivation observed in the reference soils and under mounds inhabited by primary termites, thus causing the soil types to change from Alisols and Acrisols to the properties of Umbrisols.     

Genetic parameters and comparison between native and local landraces of Eucalyptus globulus Labill. ssp. globulus growing in the central highlands of Colombia

Aims

The objective of this study was to compare the merit of the Colombian landrace relative to the various Australian native races of Eucalyptus globulus Labill. ssp. globulus and study the genetic control of key traits such as growth, wood density, and leaf phase change in the unique conditions of the Colombian highlands.

Methods

The genetic study was based on open-pollinated families from native Australian and Colombian landrace origin, tested across four trials spanning two generations of breeding. A multisite mixed linear model with genetic groups was fitted to the data to estimate race merit and the variance and covariances between traits, ages, and sites.

Results

Race effects for growth were small and only significant at the older site. In contrast, races differ significantly for height to phase change and density. The Colombian landrace and South and NE Tasmania races changed leaf type at a higher tree height. King Island and Recherche Bay had low density values, whereas the Colombian landrace had the highest. Heritability was moderate for growth traits (between 0.09 and 0.40), high for height to phase change (between 0.42 and 0.69), and moderate for wood density (0.28). The genetic correlation between growth and height to phase change was in general positive, but variable across sites. There was no correlation between growth and density. Despite large differences in growth between trials, pairwise genetic correlations suggest that genotype-by-site interaction is negligible. However, there was a poor correspondence between first-generation (ex-native stands) and second-generation families (ex-multi-provenance progeny trial). This suggests that breeding value estimates based on native open-pollinated material are unreliable.

Conclusion

Results confirm negligible race differences for growth, but not for wood density. Future breeding efforts should include the various genetic backgrounds including the Colombian landrace which constitutes a very important source, especially because of its high density. Despite a low Genotype by Environment interaction, the poor genetic correspondence between the first- and second-generation trials indicates that selections for growth should be based on information coming mainly from the advanced-generation trials.     

Reversing agriculture from intensive to sustainable improves soil quality in a semiarid South Italian soil

Intensive agriculture (IA) is widespread in South Italy, although it requires frequent tillage, large amounts of fertilizers and irrigation water. We have assessed the efficacy of reversing IA to sustainable agriculture (SA) in recovering quality of a typical South Italy soil (Lithic Haploxeralf). This reversion, lasting from 2000 to 2007, replaced 75% of nutrients formerly supplied inorganically by farmyard manuring and reduced the tillage frequency. Several chemical and biochemical properties, functionally related to C and N mineralisation–immobilisation processes and to P and S nutrient cycles, were monitored annually from 2005 to 2007 in the spring. Reversing IA to SA decreased soil bulk density, almost doubled the soil organic matter (SOM) as favoured the immobilisation of C and N, increased most soil microbial indicators but decreased contents of nitrate, mineral N and K₂SO₄-extractable C. The K₂SO₄-extractable C/K₂SO₄-extractable organic N ratio suggested that substrate quality rather than the mass of readily available C and N affected biomass and activity of soil microflora. Also, the largely higher 10-day-evolved CO₂–C-to-inorganic N ratio under SA than IA indicated that higher C mineralisation, associated with higher microbial biomass N immobilisation, occurred under SA than IA. Decreases in most soil enzyme activities under IA, compared to SA, were much higher than concomitant decreases in SOM content. Soil salinity and sodicity were always higher in IA than SA soil, although not critically high, likely due to the intensive inorganic fertilisation as irrigation waters were qualitatively and quantitatively the same between the two soils. Thus, we suggest that the cumulative small but long-term saline (osmotic) and sodic (dispersing) effects in IA soil decreased the microbial variables more than total organic C and increased soil bulk density.     

Mitigation of olive mill wastewater toxicity

The toxicity of olive mill wastewaters (OMW) is commonly attributed to monomeric phenols. OMW were treated in an aerated, stirred reactor containing agricultural soil, where the oxidative polymerization of phenols took place. In 24 h, OMW monomeric phenols decreased by >90%. This resulted in a corresponding reduction in phytotoxicity, as measured by germination tests with tomato and English cress seeds, and in microbial toxicity, as measured by lag phase duration in Bacillus cereus batch growth. Soil germination capability after irrigation with OMW was assessed in long-term pot experiments. The relative germination percentage of tomato was higher when the soil was irrigated with treated OMW rather than with untreated ones, although it was lower than the control (e.g., soil irrigated with distilled water). At longer incubation times, a complete recovery of the soil germination capability was achieved with treated, but not with untreated, OMW.     

Reconstruction of Microraptor and the evolution of iridescent plumage

Iridescent feather colors involved in displays of many extant birds are produced by nanoscale arrays of melanin-containing organelles (melanosomes). Data relevant to the evolution of these colors and the properties of melanosomes involved in their generation have been limited. A data set sampling variables of extant avian melanosomes reveals that those forming most iridescent arrays are distinctly narrow. Quantitative comparison of these data with melanosome imprints densely sampled from a previously unknown specimen of the Early Cretaceous feathered Microraptor predicts that its plumage was predominantly iridescent. The capacity for simple iridescent arrays is thus minimally inferred in paravian dinosaurs. This finding and estimation of Microraptor feathering consistent with an ornamental function for the tail suggest a centrality for signaling in early evolution of plumage and feather color.     

Effect of air-drying treatment on enzymatic activities of soils affected by anthropogenic activities

The influence of air-drying on dehydrogenase, invertase, -glucosidase, urease, phosphatase, arylsulphatase and phenoloxidase activities was measured in three soils affected by anthropogenic activities and in control soils sampled from neighbouring areas. The air-drying changed enzyme activity, but the behaviour was neither site-specific nor enzyme-sensitive. Canonical discriminant analysis of enzymatic activities of both moist fresh and air-dried samples was able to discriminate between altered and unaltered soils irrespective of the site. Numerical alteration indices were developed as a linear function of the seven enzyme activities and the one obtained from fresh sample data was more successful. Air-drying apparently alters in unpredictable ways enzyme activities of soils, which could lead to misinterpretation of results.     

Controlled-release systems for the delivery of cyromazine into water surface

Controlled-release systems (CRSs) for the insect growth regulator cyromazine (Neporex), as a larvicide against Culex pipiens (Culicidae) mosquitoes, are under development. Despite promising results obtained previously for both in vitro (dissolution tests) and in vivo systems (mosquito larvae floating on water surfaces), it was evident that the CRSs did not remain afloat for sufficiently long times. The present study was thus conducted to determine the relationship between the process parameters and the potential contact time of the active ingredient with the mosquito larvae. A series of CRSs were prepared by an extrusion process, alone or in combination with a supplementary coating. The active ingredient was incorporated into the matrix or the coating. The matrix comprised low-density polyethylene 600 and perlite, and the coating, a polyurea with or without the addition of paraffin wax. The study showed that the cumulative release of the active ingredient into water could be controlled by manipulating the preparation techniques, the types and concentrations of inert materials, and the concentration of the active ingredient.