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PubMed1991

Characterization of a human trypsin resistant to Kunitz soybean trypsin inhibitor. Studies of duodenal juices after tube instillation of raw soybean extract.

Human duodenal juices collected during tube instillation of raw soybean extract into the duodenum contained free trypsin and free Kuntiz soybean trypsin inhibitor (KTI) in the simultaneous presence of trypsin-KTI complexes. It has previously been suggested that this KTI-non-inhibitable trypsin has a general resistance to serine protease inhibitors. Four different trypsin forms have been found and partly characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and isoelectric focusing followed by Western immunoblotting or enzyme staining. In addition, crossed immunoelectrophoresis and affinity chromatography with antibody-coupled gels have been used for identification of free and inhibitor-complexed trypsin.

Blotting, Western↗
PubMed2000

Absorption of calcium, magnesium, phosphorus, iron and zinc in growing male rats fed diets containing either phytate-free soybean protein or soybean protein isolate or casein.

The effect of dietary phytate-free soybean protein (PFS) on intestinal mineral absorption and retention was examined in growing male rats using a three-day mineral balance technique. The rats were fed diets containing PFS, soybean protein isolate (SPI) or casein at a 20% level for 5 wk. Total calcium (Ca), magnesium (Mg), phosphorus (P), iron (Fe) and zinc (Zn) contents in diets were adjusted to 0.35, 0.05, 0.7, 0.0035 and 0.003%, respectively, by supplementation of the diet with their salts. Mineral absorption and retention ratios in rats fed the PFS diet were significantly higher than those in rats fed either the SPI or casein diet. These results suggest that PFS may be a promising dietary protein source for improving the mineral bioavailability in humans.

Animals↗
PubMed1994

Interaction of the soybean (Glycine max) seed lectin with components of the soybean protein body membrane.

In Leguminosae, lectins occur in the protein bodies which are membrane-lined storage organelles. Studies of the interaction between lectins and other protein body components could provide information about the internal organization of protein bodies and give indication of the biological function of lectins. From soybean (Glycine max) seeds, protein bodies and protein body membranes therefrom were isolated. By a technique similar to the ELISA procedure, it was shown that the membranes interact with the soybean lectin, SBA. This interaction can be inhibited by the haptenic monosaccharides specific for SBA, galactose and N-acetylgalactosamine, and is dependent on pH and ionic strength. By thin layer chromatography and lecting specific staining, it was shown that lipids from the protein body membrane are SBA-reactive.

Acetylgalactosamine↗
PubMed1980

Ultrastructural localization of soybean agglutinin on thin sections of Glycine max (soybean) var. Altona by the gold method.

Soybean agglutinin (SBA) has been localized in Glycine max (soybean) var. Altona at the ultrastructural level by the gold method. SBA was detected by marking thin sections of different part of the seed with gold granules (12 nm in size) labelled with anti-SBA antiserum. Upon examination by transmission electron microscopy, the lectin was found uniformly distributed in most of the protein bodies of the cotyledon. SBA was also present in the embryo axis.

Gold↗
PubMed2006

The characterization of the soybean polygalacturonase-inhibiting proteins (Pgip) gene family reveals that a single member is responsible for the activity detected in soybean tissues.

Polygalacturonase-inhibiting proteins (PGIPs) are leucine-rich repeat (LRR) proteins that inhibit fungal endopolygalacturonases (PGs). They are encoded by multigene families whose members show functional redundancy and subfunctionalization for recognition of fungal PGs. In order to expand the information on the structure and functional features of legume PGIP, we have isolated and characterized four members of the soybean Pgip gene family and determined the properties of the encoded protein products. Sequence analysis showed that these genes form two clusters: one cluster of about 5 kbp containing Gmpgip1 and Gmpgip2, and the other containing Gmpgip3 and Gmpgip4 within a 60 kb fragment of a separate BAC clone. Sequence diversification of the four members resides mainly in the xxLxLxx region that includes residues forming the beta-sheet B1. When compared with other legume Pgip genes, Gmpgip3 groups with the bean genes Pvpgip1 and Pvpgip2, suggesting that these genes are closer to the ancestral gene. At the protein level, only GmPGIP3 shows the capability to inhibit fungal PGs. The spectrum of inhibition of GmPGIP3 against eight different fungal PGs mirrors that of the PGIP purified from soybean tissues and is similar to that of the bean PvPGIP2, one of the most efficient inhibitors so far characterized. We also report that the four Gmpgip genes are differentially regulated after wounding or during infection with the fungal pathogen Sclerotinia sclerotiorum. Following fungal infection Gmpgip3 is up regulated promptly, while Gmpgip2 is delayed.

Amino Acid Sequence↗
PubMed2005

Laser capture microdissection (LCM) and expression analyses of Glycine max (soybean) syncytium containing root regions formed by the plant pathogen Heterodera glycines (soybean cyst nematode).

Roots of soybean, Glycine max cv. Kent L. Merr., plants susceptible to the soybean cyst nematode (SCN), Heterodera glycines Ichinohe, were inoculated and allowed to develop feeding sites (syncytia) for 8 days. Root samples enriched in syncytial cells were collected using laser capture microdissection (LCM). RNA was extracted and used to make a cDNA library and expressed sequence tags (ESTs) were produced and used for a Gene Ontology (GO) analysis. RT-PCR results indicated enhanced expression of an aquaporin (GmPIP2,2), alpha-tubulin (GmTubA1), beta-tubulin (GmTubB4) and several other genes in syncytium-enriched samples as compared to samples extracted from whole roots. While RT-PCR data showed increased transcript levels of GmPIP2,2 from LCM tissue enriched in syncytial cells, in situ hybridization showed prominent GmPIP2,2 hybridization to RNA in the parenchymal cells tightly juxtaposed to the syncytium. Immunolocalization indicated stronger alpha-tubulin signal within the syncytium as compared to surrounding tissue. However, alpha-tubulin labeling appeared diffuse or clumped. Thus, LCM allowed for the isolation of tissue enriched for syncytial cells, providing material suitable for a variety of molecular analyses.

Amino Acid Sequence↗
PubMed1990

Effects of the photobleaching herbicide, acifluorfen-methyl, on protoporphyrinogen oxidation in barley organelles, soybean root mitochondria, soybean root nodules, and bacteria.

The photobleaching herbicide, acifluorfen-methyl (AFM), has been reported to be an inhibitor of the heme and chlorophyll biosynthetic enzyme protoporphyrinogen oxidase (Protox) in several plant species. However, AFM had no effect on the levels of Protox activity measured in a mitochondrial fraction from soybean roots. In contrast, AFM inhibited Protox activity in etioplasts from barley leaves and in mitochondria from barley roots, but the extent of inhibition varied depending upon the assay conditions and was maximal only in the presence of 5 mM dithiothreitol (DTT). AFM inhibition was enhanced by preincubation of barley organelle extract in the presence of DTT. Preincubation of barley extract with DTT and AFM together (but not with AFM alone) caused extensive enzyme inhibition which was not reversible by dialysis. These findings have implications for the mechanism of AFM action and for the differential effect of these herbicides on crop and weed species. AFM had no effect on the Protox activity of membranes from free-living bacterial cell of Bradyrhizobium japonicum or Escherichia coli, or on the high levels of Protox activity associated with the plant-derived membrane surrounding the symbiotic bacteria within the soybean root nodule.

Bacteria↗
PubMed1984

Detection of genomic-length soybean mosaic virus RNA on polyribosomes of infected soybean leaves.

Soybean mosaic virus (SMV)-related RNAs were examined in both polyribosomal and nonpolyribosomal fractions of systemically infected soybean leaves. Viral RNAs were detected by Northern blot hybridization analysis using two cloned SMV-cDNAs representing different regions of the viral genome as hybridization probes. Genomic length SMV-RNA (Mr of 3.3 X 10(6] was found in specific association with EDTA-sensitive polyribosomes of infected leaves, indicating that it functions as a messenger RNA in these cells. A smaller SMV-related RNA (Mr of 1.6 X 10(6] was sometimes detected in the polyribosomal fraction; however, reconstruction experiments indicate that this RNA is a breakdown product of the genomic-length RNA, generated during cell fractionation or RNA extraction. Two other SMV-related RNAs with Mr of 2.0 and 0.78 X 10(6) were sometimes detected in infected cells and were not generated from genomic SMV-RNA or intact virus particles in reconstruction experiments. However, these RNAs were exclusively associated with the EDTA-resistant, nonpolyribosomal fraction of infected cells. These data suggest that genomic-length SMV-RNA is the only viral RNA which is translated in these infected plants.

Cloning, Molecular↗
PubMed2005

Amino acid, carbohydrate, and fat composition of soybean meals prepared at 55 commercial U.S. soybean processing plants.

To quantify variation in U.S. soybean meals (SBM), samples were collected from 55 U.S. soybean (SB) processing plants located in seven of the geographic SB maturity zones at three time points. These samples were analyzed for crude and acid-hydrolyzed fat, oligosaccharide, and amino acid concentrations. Acid-hydrolyzed fat concentrations were poorly correlated (r = 0.28) to crude fat concentrations and were higher for SBM prepared in the southern zones (V-VII) as compared with the northern zones (I and II). Raffinose and verbascose concentrations were lowest (P < 0.05) for SBM prepared in northern maturity zones, while stachyose concentrations were highest for SBM prepared in central maturity zones (III and IV). Total essential, total nonessential, and total amino acid concentrations were lowest for SBM prepared in northern zones. There was variation in oligosaccharide and amino acid concentrations over time, probably due to variation in composition of SB arriving at the plants within maturity zone.

Amino Acids↗
PubMed2003

A catalogue of molecular, physiological and symbiotic properties of soybean-nodulating rhizobial strains from different soybean cropping areas of China.

We have analysed 198 fast-growing soybean-nodulating rhizobial strains from four different regions of China for the following characteristics: generation time; number of plasmids; lipopolysaccharide (LPS), nodulation factors (LCOs) and PCR profiles; acidification of growth medium; capacity to grow at acid, neutral, and alkaline pH; growth on LC medium; growth at 28 and 37 degrees C; melanin production capacity; Congo red absorption and symbiotic characteristics. These unbiased analyses of a total subset of strains isolated from specific soybean-cropping areas (an approach which could be called "strainomics") can be used to answer various biological questions. We illustrate this by a comparison of the molecular characteristics of five strains with interesting symbiotic properties. From this comparison we conclude, for instance, that differences in the efficiency of nitrogen fixation or competitiveness for nodulation of these strains are not apparently related to differences in Nod factor structure.

Bacterial Proteins↗
PubMed1999

Comparison of Aspergillus niger phytase and Trichoderma reesei phytase and acid phosphatase on phytate phosphorus availability in pigs fed on maize-soybean meal or barley-soybean meal diets.

The efficacy of Aspergillus niger (APhy) phytase, Trichoderma reesei (TPhy) phytase and acid phosphatase (TAcPh) preparations in improving the utilization of phytin-phosphorus in the maize-soybean meal (SBM) or barley-SBM (800:200 g kg-1) diets was studied in two separate digestibility and balance trials with ten growing pigs using 5 x 5 Latin square designs. The positive control diet contained a total phosphorus (P) of 6.5 g kg-1, while the negative control as well as the APhy, TPhy and TAcPh supplemented diets which did not contain additional inorganic-P, had a total P of 4.1 g kg-1. The APhy and TPhy supplements provided phytase activity of 1000 PU g-1 together with AcPh of 8000 HFU g-1. TAcPh at a level of 8000 HFU g-1 was the only addition to one diet. The intrinsic phytase activity of barley was 355 PU g-1 while maize and soybean meal showed no phytase activity. Phytase supplements of the APhy and TPhy sources increased ash digestibility in both diets but had only a minor effect on nitrogen utilization. The addition of phytase improved absorption of P by 21%-units in barley-SBM diet and 29%-units in maize-SBM diet, without any difference between the two phytase sources. The retained P in diets with phytase was higher than in diets without phytase, 4.4 (APhy), 4.5 (TPhy) vs. 2.9 g d-1 in barley-SBM-diets and 3.7 (APhy), 4.0 (TPhy) vs. 1.8 g d-1 in maize-SBM-diets. No difference was found between the two sources of phytase. TAcPh without additional phytase did not show any effect on P absorption or retention. Ca absorption and retention were improved due to the phytase treatments. Supplementing pig diets with either APhy or TPhy sources seems to be equally effective in enhancing the availability of phytate-P. Consequently, these supplements can reduce the P-excretion of pigs by 32-40% as compared with the diet supplemented with inorganic-P.

6-Phytase↗
PubMed2004

Enumerative and binomial sequential sampling plans for soybean aphid (Homoptera: Aphididae) in soybean.

Since the discovery of the soybean aphid, Aphis glycines Matsumura, in midwestern U.S. soybean, Glycine max L., in 2000, the aphid has become a significant economic pest. Basic information about estimating population density within fields is unknown. Therefore, we developed two sampling plans to efficiently characterize A. glycines densities. Enumerative and binomial sequential plans were developed using 89 data sets collected from 10 commercial fields sampled during 2001-2003. Re-sampling software was used to validate the enumerative plan on whole plant counts, based on Taylor's power law parameters (a = 9.157 and b = 1.543). For research applications, the enumerative plan was modified to provide an actual precision level of 0.10 (SE/mean), which resulted in an average sample number of 310 individual plants. For integrated pest management (IPM) purposes, we developed an enumerative plan with an actual precision of 0.25, which resulted in an average sample number of 38 individual plants. For IPM applications, the binomial plan will likely be more practical. Binomial plans were developed using two tally thresholds at five action thresholds. Final analysis of the operating characteristic curve for each plan indicated that the tally threshold of > or = 40 aphids per plant, and an action threshold of 0.837 (84% of the plants infested) provided the most correct treat (4%) and no-treat (95%) decisions, with very low incorrect treat (0.5%) and no-treat (0.5%) decisions. A tally threshold of > or = 40 aphids per plant and action thresholds of 84% of plants infested is equivalent to a mean density of 250 aphids per plant, a recently recommended economic threshold. Using this threshold, the minimum required sample number for the binomial plan was 11 plants.

Agriculture↗
PubMed2006

Genetic analysis of resistance to soybean mosaic virus in j05 soybean.

Soybean cultivar J05 was identified to be resistant to the most virulent strain of soybean mosaic virus (SMV) in northeastern China. However, the reaction of J05 to SMV strains in the United States of America is unknown, and genetic information is needed to utilize this germplasm in a breeding program. The objectives of this study were to determine the reaction of J05 to all US strains of SMV (G1-G7), the inheritance of SMV resistance in J05, and the allelic relationship of resistance genes in J05 with other reported resistance genes. J05 was crossed with susceptible cultivar Essex (rsv) to study the inheritance of SMV resistance. J05 was also crossed with PI 96983 (Rsv1), L29 (Rsv3), and V94-5152 (Rsv4) to test the allelism of resistance genes. F(2) populations and F(2:3) lines from these crosses were inoculated with G1 or G7 in the greenhouse. Inheritance and allelism studies indicate that J05 possesses 2 independent dominant genes for SMV resistance, one at the Rsv1 locus conferring resistance to G1 and necrosis to G7 and the other at the Rsv3 locus conditioning resistance to G7 but susceptibility to G1. The presence of both genes in J05 provides resistance to G1 and G7. J05 is unique from the previous sources that carry 2 genes of Rsv1Rsv3 and will be useful in breeding for SMV resistance.

Alleles↗
PubMed1998

Pharmacokinetics of soybean isoflavones in plasma, urine and feces of men after ingestion of 60 g baked soybean powder (kinako).

To take advantage of the various pharmacologic activities of soy bean isoflavones, more detailed studies of the absorption and excretion rates of these compounds in humans and subsequent evaluation of their bioavailabilities are required. We conducted a pharmacokinetic study of soybean isoflavones in seven healthy male volunteers. After ingestion of 60 g of kinako (baked soybean powder, containing 103 micromol daidzein and 112 micromol genistein), changes of the isoflavone and metabolite concentrations in plasma, urine and feces were measured by gas chromatography-mass spectrometry. The plasma concentration of genistein increased after 2 h and reached its highest value of 2.44 +/- 0.65 micromol/L 6 h later. The plasma concentration of daidzein peaked at 1.56 +/- 0.34 micromol/L at the same time, but it was always lower than that of genistein. Peak plasma concentration of O-desmethylangolensin (O-DMA) and equol appeared after the daidzein peak in four and two subjects, respectively. In contrast with plasma, daidzein was the main component in urine. Urinary daidzein excretion started to increase shortly after the rise in its plasma concentration and reached 2.4 micromol/h 8 h after ingestion of kinako. Genistein excretion in urine paralleled that of daidzein, but the value at 6 h was about half (1.1 micromol/h). The majority of ingested isoflavones after ingestion of kinako were recovered on d 2 or 3 in the feces. Total recovery of daidzein, O-DMA and equol from urine and feces was 54.7%, calculated from daidzein intake; 20.1% of administered genistein was recovered as genistein. The half-lives of plasma genistein and daidzein were 8.36 and 5.79 h, respectively. The individual plasma and urinary concentrations of equol and O-DMA were quite variable; subjects were classified as high and low metabolizers. The high plasma concentration of isoflavones for at least several hours after a single ingestion of soy protein suggests that these compounds may interact with macromolecules and have biological effects.

Absorption↗
PubMed1999

Nutrient content and protein and energy digestibilities of ethanol-extracted, low alpha-galactoside soybean meal as compared to intact soybean meal.

A study was conducted with adult Leghorn roosters to determine TMEn, dry matter digestibility, amino acid content and digestibility, and cellulose and hemicellulose content and digestibility of ethanol-extracted, low alpha-galactoside soybean meal (SBM). Results were compared to that of commercially available 47% CP SBM. Twenty-one roosters were utilized; seven roosters were precision-fed each of the two soybean meal treatments, and seven served as feed-deprived controls. Excreta were collected for 48 h after feeding. A second experiment compared the TMEn, as determined with roosters, to AMEn, as determined with male broiler chicks, of 44% and 47% CP SBM and three different ethanol extraction procedures to remove alpha-galactosides. The ethanol extraction procedure concentrated CP, gross energy, and TMEn. Dry matter digestibility of SBM increased from 52.1% (control SBM) to 63.3% (ethanol-extracted SBM) because of the ethanol extraction procedure. The average amino acid digestibility of ethanol-extracted SBM was 91.6%, compared to 88.0% for control SBM. The digestibility of methionine, alanine, valine, and lysine was improved by the ethanol extraction procedure. Total cellulose and hemicellulose digestion, measured as digestion of specific monosaccharide components, was increased by the ethanol extraction. The TMEn and AMEn values for the different control and extracted SBM were similar. These data indicate that the ethanol extraction of SBM to remove alpha-galactosides resulted in a product that is more protein and energy dense than commercial SBM. Advantages in digestion of amino acids and fiber may contribute to the increase in TMEn observed in both young and adult birds.

Animal Feed↗
PubMed2004

Real-time PCR detection of lactic acid bacteria in cecal contents of eimeria tenella-lnfected broilers fed soybean oligosaccharides and soluble soybean polysaccharides.

This experiment was conducted to test whether dietary soybean meal oligosaccharides (SMO) and water-soluble polysaccharides (SMP) can assist broiler chickens in resisting Eimeria tenella, and to determine the survival of lactic acid bacteria in cecal contents postinfection. All birds received a soybean meal-free diet. The 6 experimental treatments were as follows: positive (COR) and negative (COW) control groups, 2 groups fed diets containing either 1% SMO or 0.5% SMP from 1 to 11 d of age; a vaccinated group (VAC), and an anticoccidial medicated group (ANT). Chickens of all treatments except COW were orally infected with 1000 sporulated oocysts of E. tenella on d 15. Fecal oocyst shedding was monitored per treatment group between d 5 and 13 postinfection. Lactic acid bacteria (LAB) in cecal contents were evaluated by a real-time PCR technique on d 7 postinfection. The results showed that the SMO and SMP groups had a lower number of oocysts per gram of feces during the monitoring period than the COR group. Threshold cycles were 22.21, 27.68, 13.99, 14.92, 12.97, and 14.85, for COW, COR, SMO, SMP, VAC, and ANT groups, respectively; specific PCR products were confirmed by the results of melting curve analysis and agarose gel electrophoresis. The results suggest that these LAB communities were promoted by SMO and SMP and have a competitive exclusion function when broiler chickens are infected with E. tenella.

Animal Feed↗
PubMed2005

Relative bioavailability of phosphorus and true amino acid digestibility by poultry as affected by soybean extraction time and use of low-phytate soybeans.

The objectives of this study were to determine if lengthening the time that soybeans (SB) spend in the extractor during preparation of soybean meal (SBM) results in increased relative bioavailability of phosphorus without negatively impacting true amino acid digestibilities, and to compare those modified SBM with that produced from a low-phytate SB. Three SBM were prepared under uniform conditions with the exception of the length of time SB spent in the extractor [45 min (300 rpm), 60 min (225 rpm), or 90 min (150 rpm)]. A SBM prepared from low-phytate SB was obtained for comparison. Relative phosphorus bioavailability in chicks and true amino acid digestibilities by cecectomized roosters were determined. Increasing the length of time that SB spent in the extractor from 45 to 90 min resulted in lower phytate phosphorus and increased phosphorus bioavailability from 34 to 56%. However, this increase came at the expense of available lysine status, with the SBM extracted for 90 min containing less total lysine and less digestible lysine than the SBM extracted for 45 min (traditional extraction time). Phosphorus bioavailability from SBM prepared from low-phytate SB was 1.5 times higher than for SBM extracted for 45 min. Increasing the length of time that SB spend in the extractor led to an increase in bioavailable phosphorus but a decrease in bioavailable lysine, potentially negating the positive effect on phosphorus.

Amino Acids↗
PubMed2005

Performance and nutrient utilization of laying hens fed low-phosphorus corn-soybean and wheat-soybean diets supplemented with microbial phytase.

Two experiments were conducted with laying hens (Lohmann Brown) in an individual cage system and with single feeding conditions. Experiment 1 (n = 24) was a performance trial (22 to 61 wk) to evaluate phytase effects on performance and nutrient utilization in corn-soybean meal (CSM1) and wheat-soybean meal (WSM1) basal diets (0.12% NPP; 3.1% Ca) supplemented (300 U/kg) with an experimental microbial phytase (CSM2 and WSM2) or 1.5 g/kg inorganic P (CSM3 and WSM3). Experiment 2 (n = 16) was also conducted as a performance trial (22 to 61 wk) only using CSM diets with dietary treatments similar to those in experiment 1. In addition, parallel N and P balance experiments in 2 age periods (26 and 33 wk, respectively) were conducted. In experiment 1, no significant (P < 0.05) differences in mortality, feed intake, egg production, egg weight, or body weight were observed. Tibia bone mineral composition was significantly affected by microbial phytase. Microbial phytase in the low-P CSM diet significantly (P < 0.05) improved the feed conversion ratio. In experiment 2, only feed conversion ratio was significantly improved by microbial phytase. The phytase supplementation had no significant effect on P excretion, P balance, P utilization, N balance, N utilization, or AMEn in the balance experiments.

6-Phytase↗