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Wheat alpha-amylase inhibitors represent an important tool in engineering crop plants against bean bruchids. Because Acanthoscelides obtectus is a devastating storage bean insect-pest, we attempted to purify and characterize its gut alpha-amylases, to study their interaction with active proteinaceous inhibitors. Two digestives alpha-amylases (AoA1 and AoA2) were purified from gut larvae, showing molecular masses of 30 and 45 kDa for each one, respectively. The stoichiometry interaction between these alpha-amylases with two wheat inhibitors (0.19 and 0.53) showed a binding complex of 1:1 enzyme:inhibitor. In vivo activities of these inhibitors against A. obtectus were also evaluated using a rich ammonium sulfate inhibitor fraction (F(20)(-)(40)) and purified inhibitors after reversed phase high-performance liquid chromatography columns. Incorporation of three different inhibitor concentrations (0.25, 0.5, and 1.0% w/w) into artificial seeds showed that addition of the purified 0.19 inhibitor at the highest concentration (1.0%) reduced the larval weight by 80%. Similar data were observed when 0.53 inhibitor was incorporated at 0.5%. When the concentration of purified 0.53 was enhanced to 1.0%, no larvae or adult emergence were observed. Our data suggest that these alpha-amylase inhibitors present great potential for use in Phaseolus genetic improvement programs.  相似文献   
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The benefits of manure as a source of nutrients for plants and to improve soil quality are well-known. Monitoring of manure application is needed if environmental issues are to be prevented. In particular, the availability and accumulation of phosphorus (P) has to be subject to rigorous monitoring. This study aims to both evaluate the efficacy of the resin method in extracting inorganic labile P in soils under the long-term application of dairy liquid manure (DLM), and verify the influence of DLM on the recovery of applied P and soil legacy P. To realize our objectives, two long-term field experiments were established under a no-tillage system with annual DLM application on sandy clay loam (sandy Oxisol) and clayey (clayey Oxisol) soils. Treatment consisted of DLM applications (0, 60, 120, 180 m3 ha−1 year−1), independent of mineral fertilizer. Soil samples were taken from the 0–5, 5–10 and 10–15 cm layers after 10 years from the beginning of the trial. A single extraction with resin underestimated inorganic labile P; however, successive extractions were able to take up 43% and 26% of the total P in sandy and clayey Oxisols, respectively, whereas in a single extraction the take-up was 17% and 8% from the same soils, respectively. The resin method was more effective in extracting P from the sandy Oxisol. Thus, when interpreting soil P contents for fertilizer planning, the soil texture should be taken into account. DLM application decreased P recovered from applied P, ranging from 54% to 83% (sandy), and 43% to 67% (clayey), and substantially increased soil legacy P.  相似文献   
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