Nitrogen lowers the sulfur amino acid content of soybean (Glycine max [L.] Merr.) by regulating the accumulation of Bowman-Birk protease inhibitor

Soybeans in general contain 35-40% protein. Efforts are underway to increase further this protein content, thus enhancing their nutritive value. Even though higher protein is a desirable characteristic, whether such an increase will be accompanied by enhanced protein quality is not known. Soybean protein quality could be significantly improved by increasing the concentration of the sulfur-containing amino acids, cysteine and methionine. To ascertain if a correlation existed between protein quantity and quality, a comparison of the amino acids of soybeans differing in protein content was made. Soybeans with higher protein content had a significantly lower percentage of sulfur amino acids, while those with lower protein exhibited a higher content of cysteine and methionine. Nitrogen application elevated the protein content but lowered that of the sulfur amino acids. Transmission electron microscopy examination of thin sections of low protein soybean seeds revealed several protein storage vacuoles that were partially filled with storage proteins. Fluorescence two-dimensional difference gel electrophoresis of soybean seed proteins revealed that nitrogen application favored the accumulation of the beta-subunit of beta-conglycinin while decreasing the accumulation of Bowman-Birk protease inhibitor (BBI), a protein rich in cysteine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 60% 2-propanol-extracted proteins showed a drastic reduction in the accumulation of BBI with increasing protein content. Northern blot analysis indicated that nitrogen had a negative influence on the expression of the BBI gene. Our results indicate that the negative correlation between total protein and sulfur amino acid content is mostly mediated by the differential accumulation of BBI.     

Effect of six decades of selective breeding on soybean protein composition and quality: a biochemical and molecular analysis

To evaluate the extent of the genetic change and its effects on the seed protein composition of soybean cultivars released during the past 60 years, representative ancestral cultivars and those derived from selective breeding were grown in a side-by-side comparison. Total seed protein content, determined by combustion analysis of nitrogen, revealed a decline in the protein content after decades of selection and breeding. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis comparison of protein profiles of the soybean cultivars indicated that relative expression of most of the seed storage proteins had not varied substantially from the ancestral lines to the present commercial cultivars. There was noticeably less beta-subunit of beta-conglycinin, a protein devoid of sulfur amino acids, in the modern cultivars represented by Mustang, Pioneer 93B09, and Asgrow 3602. Comparison of the amino acid profiles of soybean seed, a benchmark of the protein's nutritional quality, revealed that the ancestral progenitor, G. soja, was significantly higher in cysteine, glutamic acid, histidine, and arginine than either the ancestral or the modern cultivars. Selective breeding over the past 60 years minimally affected the overall amino acid composition. The degree of divergence in the DNA sequence of the genes encoding glycinin and beta-conglycinin in the ancestral and modern cultivars was investigated using Southern hybridization and the polymerase chain reaction. Even though some restriction fragment polymorphisms could be detected, overall, the banding patterns were remarkably similar among the ancestral cultivars and those derived from them, suggesting a high degree of conservation of seed-storage protein genes. The results of our study suggest that selection and breeding for yield during the past 60 years had no major influence on the protein composition, ostensibly because of limited genetic diversity among the parental lines.     

Identification, characterization, epitope mapping, and three-dimensional modeling of the alpha-subunit of beta-conglycinin of soybean, a potential allergen for young pigs

Soybean meal (SBM), the major byproduct of soybean oil extraction, is the main protein source for swine diets globally. In the United States, 8.6 million metric tons of SBM was used in swine rations in 2004. The pathological effect and immunological response of SBM feeding have been demonstrated in swine. In this study, we have utilized plasma collected from piglet feed with SBM in immunoblot analysis to detect proteins that elicited antigenic responses. We have identified soybean beta-conglycinin alpha-subunit as being a potential allergen for young piglets. Characterization of this protein indicated that deglycosylation and pepsin digestion did not eliminate immunoreactivity of this protein. Epitope mapping utilizing planar cellulose supports technology (SPOT) showed that three peptides spanning amino acids S185-R231 were critical for the allergenicity. A computer-generated three-dimensional structure model of the alpha-subunit of beta-conglycinin indicated that the antigenic epitopes were located on the surface of the protein. Information from this study may assist in the construction of recombinant nonallergenic soybean protein useable for both immunotherapy and the potential production of hypoallergenic soybean plants.     

Accumulation of Cadmium and Antioxidant and Hormonal Responses in the Indian Major Carp <Emphasis Type="Italic">Cirrhinus mrigala</Emphasis> During Acute and Sublethal Exposure

Acute (24 h) and sublethal (35 days) effects of cadmium chloride (CdCl₂) were examined in Cirrhinus mrigala using various endpoints (accumulation pattern, thyroid hormones (THs), and antioxidants). The mean concentrations of CdCl₂ for 24 and 96 h were found to be 35.974 and 22.387 mg L^?l, respectively. LC50 concentration of CdCl₂ for 24 h (35.97 mg L^?l) was used for the acute study. For the sublethal studies, fish were exposed to 3.59 mg L^?1 (Treatment I) and 7.19 mg L^?1 (Treatment II) corresponding to 1/10th and 1/5th of 24 h LC50 of the CdCl₂. During acute exposure, higher accumulation of CdCl₂ was noticed in the gill, liver, and kidney of C. mrigala, which is found in the order gill > liver > kidney tissues. Similarly, in sublethal treatments (Treatment I and II), a concentration and time-dependent increase of CdCl₂ accumulation was noticed in the order of gill > liver > kidney. GSH, GST, and GPx activities were found to be relatively lower from the treated groups in both acute and sublethal treatments. However, LPO activity was significantly increased in CdCl₂-treated fish C. mrigala. Further, plasma T₃ reduction was more pronounced than T₄ in acute study. During sublethal treatments, both T₄ and T₃ levels showed a continuous decrease as the exposure period extended. All the values in this study were statically significant (P < 0.01 and P < 0.05).     

Influence of Damping on the Bending and Twisting Modes of Flax Fibre-reinforced Polypropylene Composite

The effects of damping on the bending and twisting modes of flax fibre-reinforced polypropylene composites are investigated. The laminate was manufactured by a vacuum bagging process; its dynamic behaviour was then found from the vibration measurements of a beam test specimen using an impulse hammer technique to frequencies of 1 kHz. The frequency response of a sample was measured, and the bending and twisting responses at resonance were used to estimate the natural frequency and loss factor. The single-degree-of-freedom circle-fit method and Newton’s divided differences formula were used to estimate the natural frequencies as well as the loss factors. The damping estimates were also investigated using a “carpet” plot. The results show significant variations in loss factors depending on the type of mode. The loss factor generally lies in the range of 1.7-2.2 % for the bending modes, while 4.8 % on average for the twisting modes. Numerical estimates of the response, and in particular the natural frequencies, were made using a Mechanical APDL (ANSYS parametric design language) finite element model, with the beam being discretised into a number of shell elements. The natural frequencies from the finite element analysis show reasonably good agreement (errors < 5 %) with the measured natural frequencies.     

Effects of mutual shading of tree crowns on prediction of photosynthetic light-use efficiency in a coastal Douglas-fir forest

Gross primary production (GPP) is often expressed as the product of absorbed photosynthetically active radiation and the efficiency (epsilon) with which a plant community uses absorbed radiation in biomass production. Light-use efficiency is affected by environmental stresses, and varies diurnally and seasonally. Uncertainty about epsilon can be a serious limitation when modeling GPP. An important determinant of epsilon is the amount and type of solar radiation incident on a canopy, because an abundance of light can trigger a photo-protective reaction, diminishing GPP. The radiation regime in a forest canopy is determined by the predominant sky conditions and by mutual shading of tree crowns. Shading effects, producing shifts in the amount of incident direct and diffuse solar radiation, have been largely ignored, however, because they depend on forest structure and are difficult to measure. We describe a new approach for estimating changes in mutual canopy shading throughout the day and year based on a canopy structure model derived from light detection and ranging (LiDAR). Proportions of canopy shading were then combined with eddy covariance data to assess the explanatory power for variance in epsilon by regression tree analysis over half-hourly, daily and weekly time scales. The approach explained between 75 and 97% of variance in epsilon, representing an increase of between 5 and 16% compared with models driven solely by meteorological variables.