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[Obstacles of soybean continuous cropping, III. Mechanism of soybean yield increment by marine actinomyces MB-97].

An isolate of marine actinomyces MB-97 identified as Streptomyces microflavus could successfully colonize in the rhizoshpere of soybean, and inhibit Penicillium purpurogenum, a soybean deleterious rhizospheric microorganisms. After applied MB-97, the ratio of bacteria/fungi in the rhizosphere of soybean was increased, and the soil became to be "Bacterial type" from "Fungal type". The populations of P. purpurogenum were apparently suppressed about 80%, and the harm of toxins in soil was weak. The soybean root rot caused by soilborne fungi such as Fusarium was decreased 50%, and MB-97 could stimulate the growth of soybean seedlings. In field study, the mean yield of soybean raised by 15.2%, implying that Streptomyces microflavus was an effective plant growth-promoting rhizobacteria on soybean.

Actinomyces↗

Development of a capillary electrophoresis method for the determination of soybean proteins in soybean-rice gluten-free dietary products.

CE has been applied for the first time to the simultaneous separation of soybean and rice proteins. Treated and untreated capillaries with different effective lengths as well as separation media at different pHs were tested. For that purpose, samples and standard solutions were prepared in 25:75 ACN-water media containing 0.3% v/v acetic acid. The use of an untreated capillary of 50 cm effective length together with an 80 mM borate buffer (pH 8.5) modified with 20% v/v ACN and UV detection at 254 nm were the conditions working the best. These conditions enabled the determination of soybean proteins in gluten-free dietary commercial products elaborated with soybean protein and/or soybean flour and rice flour using the standard additions calibration method. The method was linear up to 26 mg/mL of soybean proteins, the precision (expressed as RSD) was always better than 6%, and recoveries obtained for soybean proteins when spiking commercial products were very close to 100%.

Acetonitriles↗

Endogenous lectins from cultured soybean cells: isolation of a protein immunologically cross-reactive with seed soybean agglutinin and analysis of its role in binding of Rhizobium japonicum.

Incubation of Rhizobium japonicum with the cultured soybean cell line SB-1, originally derived from the roots of Glycine max, resulted in specific adhesion of the bacteria to the plant cells. This binding interaction appears to be mediated via carbohydrate recognition, since galactose can inhibit the heterotypic adhesion but glucose cannot. Affinity chromatography, on a Sepharose column derivatized with N-caproyl-galactosamine, of the supernatant fraction of a SB-1 cell suspension after enzymatic removal of cell wall yielded a single polypeptide (Mr approximately 30,000) on immunoblotting analysis with rabbit antibodies directed against seed soybean agglutinin. Fluorescently labeled rabbit anti-seed soybean agglutinin also yielded specific immunofluorescent staining on the cell wall and plasma membrane of the SB-1 cells. These results suggest that one likely candidate that may mediate the recognition between the Rhizobium and the soybean cells is the endogenously produced SB-1 lectin. This notion is supported by the observation that rabbit anti-seed soybean agglutinin blocked the Rhizobium-soybean cell adhesion, whereas control antibodies did not.

Cell Adhesion↗

Determination of fluazifop-butyl and fluazifop acid in soybeans and soybean oil using liquid chromatography with oxidative amperometric detection.

A new method is described for the determination of the herbicide fluazifop-butyl, and its metabolite fluazifop acid, in soybeans and soybean oil as fluazifop acid. Liquid chromatography with amperometric detection (LC/AD) is used to determine fluazifop acid produced from the metabolism or base hydrolysis of fluazifop-butyl in soybeans and soybean oil. These foods were spiked with fluazifopbutyl at 0.05, 0.10, and 0.50 ppm and hydrolyzed with 0.2N NaOH in methanol. The hydrolysate (adjusted to pH less than or equal to 1) is extracted with dichloromethane and the extract is washed with 1.0% NaHCO3. The NaHCO3 is acidified to pH less than or equal to 1 and extracted with dichloromethane; the partitioning is repeated 2 more times. The dichloromethane is removed, mobile phase solvent is added, and aliquots are injected onto a PRP-1 liquid chromatographic column; fluazifop acid is separated from coextracted compounds and detected at an applied potential of + 1.25 V, using an amperometric electrochemical detector in the oxidation mode. Recoveries ranged from 69 +/- 6.5 to 101 +/- 18% and from 72 +/- 7.5 to 88 +/- 11% for soybeans and soybean oil, respectively. Accuracy of these recoveries was confirmed by use of 14C-radiolabeled fluazifop-butyl and by liquid scintillation spectrometry of the 14C-fluazifop acid released.

Chemical Phenomena↗

Influence of the application of three different elicitors on soybean plants on the concentrations of several isoflavones in soybean seeds.

Soybean [Glycine max (L.) Merr.] is a rich source of isoflavones that are often affected by biotic and abiotic factors. The objectives of this study were to evaluate the effect of various concentrations of three natural elicitors applied at different soybean growth stages on isoflavone content and to compare the efficiency of several solvent systems in isoflavone extraction and quantification. The isoflavones extracted from R96-3444 soybean using eight solvent systems were separated, identified, and quantified by a high-performance liquid chromatography (HPLC) procedure. The soybean plants were sprayed with salicylic acid, methyl salicylate, or ethyl acetate at 0, 10(-6), 10(-3), and 10(-1) M at R1 (blooming) or R4 (full pods) growth stage. Results showed that 10(-3) M ethyl acetate sprayed at the R1 stage significantly increased total isoflavone content and the levels of some individual isoflavones in soybean seeds. With all the elicitors that were tested, concentration was a more important factor than application time with respect to isoflavone content with lower concentrations being more effective on most isoflavones. A 53% acetonitrile solvent system was the best solvent system for extracting total isoflavone, malonyl glucosides, genistein, glycitin, genistin, acetyl-daidzin, and acetyl-genistin. The results of this study will be useful for increasing the isoflavone content in desirable soybean varieties and improving isoflavone concentration during extraction.

Acetates↗

Soybean protein isolate and soybean lectin inhibit iron absorption in rats.

Inhibitory effects of soybean protein isolate (SPI) and soybean lectin on the intestinal absorption of nonheme iron were investigated by in vivo studies in rats. Rats fed the SPI-based diet absorbed significantly less iron than did control rats fed the casein-based diet. Supplementing the SPI diets with 8% D-galactose significantly increased the incorporation of iron into liver ferritin, although D-galactose did not significantly increase iron absorption. Heat treatment of SPI significantly increased iron absorption. Ascorbate did not enhance iron absorption in rats fed the SPI-based diet. The presence of lectin in an aqueous extract of SPI was suggested by hemagglutination activity as well as by immunoreactivity with soybean lectin antibody. Soybean lectin introduced into ligated segments of the upper small intestine of rats inhibited ferrous iron absorption. This inhibitory effect, especially in the mucosal uptake, was significantly improved by addition of N-acetyl-D-galactosamine to soybean lectin. Soybean lectin had no effect on ferric iron absorption. Our results suggest that a portion of the reduction in iron absorption in rats fed SPI may be due to lectins.

Animals↗

Analysis of expressed sequence tags from roots of resistant soybean infected by the soybean cyst nematode.

The soybean cyst nematode (SCN) Heterodera glycines is the most devastating pest of soybean in the U.S.A. The resistance response elicited by SCN in soybean is complex, and genes involved in the response to a large extent are unknown and not well characterized. We constructed cDNA libraries made from mRNA extracted from roots of the resistant soybean Glycine max L. Merr. 'Peking' at 12 h, 2 to 4 days, and 6 to 8 days post inoculation with the soybean cyst nematode, population NL1-RHp, similar to race 3. Expressed sequence tag analysis of the libraries provides rapid discovery of genes involved in the response of soybean to the nematode. A total of 3454 cDNA clones were examined from the three libraries, of which 25 cDNAs were derived from nematode RNA. The levels of certain stress-induced genes such as SAM22 and glutathione S-transferase (GST8) were elevated in the SCN-infected roots relative to uninoculated roots. Early defense response genes, particularly ascorbate peroxidase and lipoxygenase, were abundant in the 12-h library. By 6-8 days, the expression of most of those genes was not as abundant, whereas genes coding for unknown proteins and stress-induced proteins continued to be highly expressed. These ESTs and associated information will be useful to scientists examining gene and protein interactions between nematodes and plants.

Animals↗

Specific immunoglobulins to soybean hull allergens in soybean asthma.

Soybean asthma, which occurred as an epidemic among patients in Barcelona, Spain, is associated with specific IgE to soybean hull allergens. The purpose of this study was to investigate the possible role of specific IgG, IgG subclasses, IgA, and IgM in the pathogenesis of soybean asthma. We studied 3 groups of subjects from Barcelona: group 1, 12 asthmatic epidemic patients; group 2, 23 asthmatic nonepidemic patients; and group 3, 32 nonallergic subjects. Specific IgE was determined by radioimmunoassay and specific IgG, IgG subclasses (1, 2, 3, and 4), IgA, and IgM by amplified enzyme-linked immunosorbent assay. Cross-inhibition studies were performed for specific IgE and IgG4. We partially characterized the soybean hull allergens that bind specific IgE, IgG, and IgG4 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/Western blot. Percentage of positive results for the assays of the 8 Igs are as follows: for group 1, 100% (IgE), 75% (IgG), 16.6% (IgG1), 8.3% (IgG2), 0% (IgG3), 66.6% (IgG4), 25% (IgA), and 25% (IgM); for group 2, 4.3% were positive for specific IgE only; and for group 3, 0% (IgE), 0% (IgG), 6.2% (IgG1), 9.4% (IgG2), 9.4% (IgG3), 9.4% (IgG4), 6.2% (IgA), and 6.2% (IgM). The correlation between the specific IgE and the other specific Igs was significant between IgE and IgG4 in group 1 only (r = 0.752, p < 0.01). Cross-inhibition studies demonstrated a higher inhibitory capacity for IgG4 than for IgE. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/ Western blot demonstrates three low molecular weight protein bands that bind specific IgE, IgG, and IgG4. This study suggests that specific IgG4 to soybean hull allergens plays a role in the pathogenesis of soybean asthma and corroborates the role of specific IgE in the same disease.

Allergens↗

Cross-resistance and resistance longevity as induced by bean leaf beetle, Cerotoma trifurcata and soybean looper, Pseudoplusia includens herbivory on soybean.

Cross-resistance, and longevity of resistance, induced by the bean leaf beetle, Cerotoma trifurcata, was studied IN the soybean PI 227687 that exhibited induced response in earlier studies. Bean leaf beetle adults and soybean looper, Pseudoplusia includens, larvae were used to induce resistance and to determine beetle feeding preference. Beetles were collected from soybean fields 2 to 5 days prior to the feeding preference test. The level of cross-resistance induced by soybean looper herbivory to subsequent bean leaf beetle feeding was higher when compared to cross-resistance induced by bean leaf beetle herbivory against subsequent feeding by soybean looper. Further, herbivory by the bean leaf beetle also induced resistance against soybean looper feeding. In the longevity study, leaflets from treated plants were collected 5, 10, 12, 14, 16, 20 and 25 days after initiation of feeding. Pairwise comparisons of leaflets from plants treated by bean leaf beetle herbivory with untreated plants revealed that induced responses were highest 14 and lowest 25 days after initiation of feeding. On other sampling days, levels of induced response varied with the sampling day.

Animals↗

Evaluation of low-oligosaccharide, low-phytate whole soybeans and soybean meal in canine foods.

Eight mature dogs (19.3 +/- 0.1 kg) were used in an experiment to compare the effects of feeding soybeans containing low concentrations of oligosaccharides and phytate on nutrient availability in complete foods fed to dogs. All foods were formulated to be isonitrogenous and contained low-oligosaccharide, low-phytate soybean meal (LLM); conventional soybean meal (SBM); low-oligosaccharide, low-phytate whole soybeans (LLB); or conventional whole soybeans (WSB) as the protein source. Daily DMI averaged 287 +/- 4 g/d. Fecal outputs were greatest for LLB and WSB, averaging 48.2 g DM/d. Small intestinal DM digestibility ranged from 80.9% (LLM) to 74.0% (LLB) but was unaffected by treatment. Large intestinal DM digestibility did not differ among treatments (P = 0.652). Total-tract DM digestibility was higher (P = 0.020) for LLM (87.0%) than for SBM (84.8%). No difference in total-tract DM digestibility was observed for the two WSB foods (average = 83.3%; P = 0.286). Nitrogen retention did not differ among foods containing LLM and SBM (1.2 g of N/d; P = 0.486) or LLB and WSB (0.9 g of N/d; P = 0.225). Small intestinal N digestibility did not differ among LLM- and SBM-containing foods (80.6%; P = 0.190) or LLB and WSB (69.3%; P = 0.640). Total-tract N digestibility did not differ among foods containing LLM and SBM (83.5%; P = 0.627) or LLB and WSB (76.8%; P = 0.968). Tryptophan digestibility was higher for SBM than for LLM (P = 0.039). Histidine and tryptophan digestibilities were higher in WSB compared with LLB (P = 0.049 and P < 0.001, respectively). No differences in nonessential AA digestibility were observed when comparing soy-based foods. The results of this study demonstrate that LLM, SBM, LLB, and WSB can be effective sources of protein for canine foods and have a high digestibility. Differences in small intestinal digestibility of tryptophan and histidine may require consideration when formulating diets using low-oligosaccharide, low-phytate soybeans or meal.

Amino Acids↗

[Effects of selenium on soybean chloroplast ultra-structure and microelement content of soybean leaves under continuous cropping stress].

Pot and field experiments were carried out to study the effect of low Se concentration on soybean chloroplast ultra-structure and on Mg, Fe, Mn and contents in soybean leaves under continuous cropping stress. The results showed that Se could certainly increase the contents of Mg, Fe and Mn in soybean leaves when the application rate of Se was below 0.50 mg.kg-1 in pot experiment, and below 300 g.hm-2 in field experiment. In pot experiment, Se concentration positively correlated with Mg and Fe contents in soybean leaves, but the relationship between Mn and Se was hardly to be quantified. Transmission electron microscopic observation indicated that under continuous cropping stress, application of Se could hold the soybean chloroplast membrane structure in good condition. When Se was not applied, the chloroplast swelled, and its grana disappeared, and even, transformed into etioplast. These experiments indicated that application of low Se could alleviate peroxidative stress resulting from continuous cropping, and keep the soybean grow normally.

Chloroplasts↗

Chemical and nutritional characteristics of United States soybeans and soybean meals.

Two experiments were conducted to determine U.S. soybean meal (SBM) variation. In experiment 1, SBM from 55 U.S. processors was evaluated. Significant (P < 0.05) but numerically small differences were detected in dry matter (DM) and organic matter (OM) concentrations. Crude protein (CP) concentrations (51.6-54.6%) were higher (P < 0.05) in SBM produced in the southern U.S. Lipid and total dietary fiber concentrations also differed (P < 0.05). The protein dispersibility index was affected (P < 0.05) by the source of SBM. In experiment 2, soybeans and resultant SBMs were obtained from 10 U.S. processing plants. Soybean DM, OM, and CP concentrations differed (P < 0.05). Soybean meal varied (P < 0.05) in CP (48.2-56.2%), acid-hydrolyzed fat (3.3-9.2%), total dietary fiber (17.0-20.7%), and lysine concentrations. Soybean meal carbohydrate composition was also affected by processing conditions. These results indicate a significant variation in chemical and nutritional characteristics of U.S. SBM from different sources.

Amino Acids↗

The ribosomal protein P0 of soybean (Glycine max L. Merr.) has antigenic cross-reactivity to soybean seed lectin.

Soybean (Glycine max L. Merr.) mutants lacking the ability to produce the lectin normally found in soybean seeds (SBL) are designated Le-. A protein of higher molecular weight that cross-reacts with antibodies raised to SBL was found at nearly equivalent levels in roots, hypocotyls, and leaves, and at lower levels in cotyledons and dry seeds of both Le+ and Le- soybean cultivars. Earlier work suggested that this protein was a novel lectin. Clones isolated from a Le- soybean root cDNA library produced a cross-reacting protein of the same size in Escherichia coli. Sequence analysis of these clones revealed a high degree of similarity to the ribosomal protein P0. The cross-reacting protein co-purified with ribosomes, and a monoclonal antibody raised to purified brine shrimp P0 cross-reacted to the same protein. The protein showed no lectin activity in a hemagglutination assay, nor did it bind to an N-acetyl-D-galactosamine affinity column. On the basis of this evidence, we conclude that the SBL-cross-reacting protein is not a lectin but a homologue of the ribosomal protein P0. Consequently, Le- soybeans must produce a lectin that is dissimilar to SBL at both the DNA and amino acid levels and we suggest that it is this lectin which is involved in nodulation.

Amino Acid Sequence↗

Identification and characterization of a soybean ethylene-responsive element-binding protein gene whose mRNA expression changes during soybean cyst nematode infection.

Ethylene-responsive element-binding proteins (EREBPs) are members of a family of plant transcription factors. Conserved EREBP domains of these proteins bind to the GCC box, an ethylene-responsive promoter element found in many pathogenesis-related (PR) genes. Using degenerate primers to the EREBP domain from diverse plant species, an EREBP homolog was isolated from a soybean cDNA library. Gel mobility-shift assays revealed that the translation product of this cDNA bound specifically to GCC box sequences. We, therefore, named this gene Glycine max ethylene-responsive element-binding protein 1 (GmEREBP1), i.e., a gene coding for the first confirmed GCC box-binding protein of soybean. GmEREBP1 mRNA abundance was analyzed by RNA blot hybridizations in soybean roots and shoots of cultivars Corsoy 79 and Hartwig, which are susceptible and resistant, respectively, to the soybean cyst nematode (Heterodera glycines). These analyses revealed that GmEREBP1 is expressed in a root-preferential manner and that GmEREBP1 mRNA abundance is changed after H. glycines infection. GmEREBP1 mRNA abundance decreased in infected (susceptible) 'Corsoy 79' roots, whereas it increased in abundance in infected (resistant) 'Hartwig' roots. Furthermore, ethephon treatment repressed GmEREBP1 mRNA accumulation in both cultivars, whereas wounding increased expression in both cultivars. These changes in mRNA steady-state levels suggest that GmEREBP1 plays a role in soybean-H. glycines interactions.

Amino Acid Sequence↗

High-efficiency induction of soybean hairy roots and propagation of the soybean cyst nematode.

Cotyledon explants of 10 soybean [Glycine max (L.) Merr.] cultivars were inoculated with Agrobacterium rhizogenes strain K599 with and without binary vectors pBI121 or pBINm-gfp5-ER possessing both neomycin phosphotransferase II (nptII) and beta-glucuronidase (gus) or nptII and green fluorescent protein (gfp) genes, respectively. Hairy roots were produced from the wounded surface of 54-95% of the cotyledon explants on MXB selective medium containing 200 microg ml(-1) kanamycin and 500 microg ml(-1) carbenicillin. Putative individual transformed hairy roots were identified by cucumopine analysis and were screened for transgene incorporation using polymerase chain reaction. All of the roots tested were found to be co-transformed with T-DNA from the Ri-plasmid and the transgene from the binary vectors. Southern blot analysis confirmed the presence of the 35S-gfp5 gene in the plant genomes. Transgene expression was also confirmed by histochemical GUS assay and Western blot analysis for the GFP. Attempts to induce shoot formation from the hairy roots failed. Infection of hairy roots of the soybean cyst nematode (Heterodera glycines Ichinohe)-susceptible cultivar, Williams 82, with eggs of H. glycines race 1, resulted in the development of mature cysts about 4-5 weeks after inoculation. Thus the soybean cyst nematode could complete its entire life cycle in transformed soybean hairy-root cultures expressing GFP. This system should be ideal for testing genes that might impart resistance to soybean cyst nematode.

Animals↗

Identification of QTLs associated with resistance to soybean cyst nematode races 2, 3 and 5 in soybean PI 90763.

Soybean cyst nematode (SCN) is a major soybean pest throughout the soybean growing regions in the world, including the USA. Soybean PI 90763 is an important SCN resistance source. It is resistant to several SCN populations including races 2, 3 and 5. But its genetics of resistance is not well known. The objectives of this study were to: (1) confirm quantitative trait loci (QTLs) for resistance to SCN race 3 in PI 90763 and (2) identify QTLs for resistance to SCN races 2 and 5. QTLs were searched in Hamilton x PI 90763 F(2:3)populations using 193 polymorphic simple sequence repeats (SSRs) covering 20 linkage groups (LGs). QTLs for resistance to SCN were identified on LGs A2, B1, E, G, J and L. The same QTL was suggested for resistance to different SCN races where their 1-LOD support intervals of QTL positions highly overlapped. The QTL on LG G was associated with resistance to races 2, 3 and 5. The QTL on LG B1 was associated with resistance to races 2 and 5. The QTL on LG J was associated with resistance to races 2 and 3. The QTLs on LGs A2 and L were associated with resistance to race 3. The QTL on LG E was associated with resistance to race 5. We conclude that LGs A2 and B1 may represent an important distinction between resistance to SCN race 3 and resistance to SCN races 2 and 5 in soybean.

Animals↗