Performances and Germplasm Evaluation of Quantitative Resistance to Soybean Mosaic Virus in Soybeans

A sample composed of 96 soybean accessions was evaluated for their diseased rate (I),diseased rank (S), latent period (LP) and rate of disease development (R) in order tostudy the quantitative resistance to soybean mosaic virus (SMV) in soybeans. The resultsshowed that the performances of the above four resistance components were significantlydifferent among accessions and that some of the accessions, such as Zhongzihuangdou,Peixian Tianedan, Youbian30 could be infected by four SMV strains, Sa, SC8, N1 and N3,but their I, S, and R were lower and LP longer than most other accessions. These resultsdemonstrated the existence of quantitative resistance to SMV in soybeans. It was foundthat some soybean accessions, such as AGS19 and Lishui Zhongzihuangdou, previouslyidentified as resistant to SMV infection, performed some infection but resistant toexpansion in the present study. In addition, the resistance in Pixian Chadou and HuaiyinQiuheidou might be either qualitative or quantitative. Furthermore, the present studyalso indicated that the resistance spectrum and durability of accessions with quantitativeresistance might be wider and longer than those with qualitative resistance.

Laboratory testing and molecular analysis of the resistance of wild and cultivated soybeans to cotton bollworm, Helicoverpa armigera(Hübner)

Identifying a superior soybean variety with high defoliator resistance is important to avoid yield loss. Cotton bollworm(Helicoverpa armigera Hübner) is one of the major defoliators of soybean(Glycine max [L.] Merr.) worldwide. In this study, we evaluated the effect of H. armigera larvae on ED059, a wild soybean(Glycine soja Sieb. et Zucc.), and three cultivated soybean varieties: Tianlong 2, PI 535807, and PI 533604, in choice and no-choice assays. The percentage of ED059 leaflets consumed by H. armigera was lower than that of the three cultivated soybeans. Larvae that fed on ED059 exhibited low weight gain and high mortality rate.Waldbauer nutritional indices suggested that ED059 reduced the growth, consumption, and frass production of H. armigera larvae. Larvae that fed on ED059 showed lower efficiency of conversion of ingested and of digested food than those that fed on Tianlong 2 and PI 533604.However, they showed statistically similar consumption index and approximate digestibility compared with those fed on the three cultivated soybeans. Quantitative real-time PCR analysis revealed that 24 h after insect attack, ED059 had higher transcript levels of Kunitz trypsin inhibitor 3, Cysteine proteinase inhibitor 2, and Nerolidol synthase 1 but a lower transcript level of Pathogenesis-related protein 1 than Tianlong 2. The gene expression results were consistent with the presence of higher levels of jasmonic acid(JA) and transcript levels of the JA biosynthesis enzyme allene oxide cyclase 3 in ED059 than in Tianlong 2. Our findings indicate that ED059 is a superior soybean line with strong insect resistance that may be mediated via the JA pathway.

Somaclonal variations of Soybeans (<i>Glycine Max.</i>L. Merr) stimulated by drought stress based on random amplified polymorphic DNAs (RAPDs)

In soybeans, drought stress causes 50% yield losses. Breeding for drought tolerance in soybeans has been widely developed using various methods, among which is polyethylene glycol (PEG-6000) induction to simulate drought in vitro. In a previous experiment, three somaclones with different levels of tolerance were generated. The objectives of this research were to determine the RAPD patterns of those somaclones and to investigate the correlation of the RAPD patterns to the drought tolerance characteristics. The results showed eleven RAPD primers capable of amplifying the DNA genome of soybeans, among which four primers were monomorphic and seven were polymorphic. Two of the polymorphic primers, OPK7 and OPK12, are capable of differentiating medium tolerance traits from other traits. Bands that are specific for medium tolerance against drought were 450 bp and 650 bp in size, generated by the OPK7 primer, and the band of 2000 bp, generated by the OPK12 primer. However, there was no band capable of differentiating between sensitive and tolerance varieties/lines, although some changing of the DNA sequence was detected in this research. This indicates that there are other factors responsible for the expression of drought tolerance.

Nodule Formation and Development in Soybeans(Glycine max L.) in Response to Phosphorus Supply in Solution Culture

Phosphorus (P) is necessary for growth and nitrogen fixation, and thus its deficiency is a major factor limiting legume production in most agricultural soils. The effect of phosphorus supply on nodule development and its role in soybeans (Glycine max L.) was studied in a nutrient solution. Plants were inoculated with Bradyrhizobium japonicum and grown for 35 days in a glasshouse at a day and night temperature of 25℃and 15℃, respectively. Although increasing P supply increased the concentrations of P and N in the shoots and roots, the external P supply did not significantly affect the P concentration in the nodules, and the N fixed per unit nodule biomass decreased with increasing P supply. The nitrogen content in the shoots correlated well with the P content (r = 0.92**). At an inoculation level of 102 cells mL-1, the P supply did not affect the number of nodules; however, at inoculation levels of 103.5 and 105 cells mL-1, increasing P supply increased both the number and size of nodules. Irrespective of the inoculation level, increasing P supply increased the nodule biomass relative to the biomass of the host plant. It is suggested that the P deficiency specifically inhibited the nodule development and thereby the total N2 fixation.

Breeding Soybeans for Resistance to Physiological Race 4 of Cyst Nematode

Soybean cyst nematode causes serious damage to soybean production. In 1991, we started breeding studies on the resistance of soybeans to the cyst nematode. We found that near the Beijing area the dominant race of the cyst nematode was race 4. We made more than 50 combinations of cross. The best combination was Dan 8 X PI 437654 which resulted in marked segregation in plant height, pod habit, resistance to cyst nematode and maturity. We obtained many new soybean lines highly resistant to the cyst nematode through the pedigree method of selection, enlarging the number of plants of good combinations, alternative breeding in the North and in the South, and identification at an early generation. We now have released three soybean cultivars, Zhonghuang 12, Zhonghuang 13 and Zhonghuang 17 with moderate resistance to the cyst nematode in Beijing, Anhui, Tianjin and Northern China. In addition, we obtained many lines which were highly resistant to the cyst nematode.

Relationships Between C_4 Enzyme Activities and Yield in Soybeans (Glycine max (L.) Merr.)

To study the relationships between C4 enzyme activities and yield, C4 enzyme activities （phosphoenolpyruvate carboxylase （PEPCase）, NADP-malate dehydrogenase （NADP-MDH）, NADP-malic enzyme （NADP-ME）, and pyruvate phosphate dikinase （PPDK）） in different organs of ten soybean cultivars with different yields were measured at different growth stages in China. The result showed that four enzyme activities in C4 pathway were obviously different among cultivars, especially PPDK activity was not detected in the leaves of Dongnong 1567 and Dongnong 1068 and the young leaves of Gongjiao 9107-1 and Dongnong 97-172, but there were weak activities in pod coats. The order of C4 enzyme activities is young leaves 〈 old leaves 〈 pod coats. The correlation coefficients between PEPCase activity and yield and between NADP-MDH activity at blooming stage and yield were 0.6979 and 0.6565, respectively, and both reached the significant level （5%）, and PEPCase activity kept significant positive correlation with plant photosynthetic rate. There was a negative correlation between NADP-ME activity and yield, and no correlation was found between PPDK activity and yield.

Detection of RAPD Markers Linked to Gene lx_1 in Soybeans

Near-isogenic lines of soybean lipoxygenase, which contain genes Lox, lx1, lx2, lx3, lx1.3, lx2.3,respectively, were used for polymorphic analysis by RAPD technique.520 10-mer-oligonucleotideprimers were screened, and thirteen primers showed polymorphism among near-isogenic lines.There were six primers showed special polymorphic bands among lines lx1 and lx1.3. Especially,primer S352 presented the stable results in which a 900 bp band was found in the lines lx1 andlx1.3, and primer S352900 was detected with F2 generation of cross 96P11×Century-1, indicatedprimer S352900 could be identified as a RAPD marker linked to gene lx1 in soybeans, the distanceof linkage was 7.6 cM.

Detection and inactivation of allergens in soybeans:A brief review of recent research advances

In the last decade,soybean allergies have been on the increases to such an extent that they have now become a public health issue thus prompting more studies and researches on the topic.The allergenicity of soybean is attributed to its protein fraction.The best way to prevent hypersensitive patients from ingesting allergenic compounds is to exclude such soybean allergens from their diet.As a result,it is essential to provide detailed and reliable knowledge of food ingredients.Therefore,precise and reliable approaches for detecting soybean allergens found in various food products must be used.The main way to reduce allergy risk is the identification of allergenic sites in food and their inactivation by various food-processing methods.It has been reported that food processing may lead to the modification of conformational structure of the protein or protein distortion that inhibit the binding of immunoglobulin E(Ig E)to epitopes on food allergens and also the mechanism of allergic reactions.Food processing technologies employed for inactivating allergenic epitopes used thermal and nonthermal techniques.Currently,several detection methods including protein-based and DNA-based approaches using analytical techniques such as enzyme-linked immunosorbent assay(ELISA),enzyme allergosorbent test(EAST),radioallergosorbent test(RAST),lateral flow immunoassay(LFIA),immunoblotting,realtime polymerase chain reaction(PCR),mass spectrometry and biosensors have been improved for identifying and quantifying these epitopes.This research focused on allergenic proteins of soybean,the most modern approaches for detecting and quantifying these allergens,and finally,the various methods used to inactivate these proteins and their effects on soy allergenicity.

The identification of presence/absence variants associated with the apparent differences of growth period structures between cultivated and wild soybeans

The cultivated soybean（Glycine max（L.） Merr.） was distinguished from its wild progenitor Glycine soja Sieb.＆ Zucc.in growth period structure,by a shorter vegetative phase（V）,a prolonged reproductive phase（R） and hence a larger R/V ratio.However,the genetic basis of the domestication of soybean from wild materials is unclear.Here,a panel of 123 cultivated and 97 wild accessions were genotyped using a set of 24 presence/absence variants（PAVs） while at the same time the materials were phenotyped with respect to flowering and maturity times at two trial sites located at very different latitudes.The major result of this study showed that variation at PAVs is informative for assessing patterns of genetic diversity in Glycine spp.The genotyping was largely consistent with the taxonomic status,although a few accessions were intermediate between the two major clades identified.Allelic diversity was much higher in the wild germplasm than in the cultivated materials.A significant domestication signal was detected at 11 of the PAVs at 0.01 level.In particular,this study has provided information for revealing the genetic basis of photoperiodism which was a prominent feature for the domestication of soybean.A significant marker-trait association with R/V ratio was detected at 14 of the PAVs,but stripping out population structure reduced this to three.These results will provide markers information for further finding of R/V related genes that can help to understand the domestication process and introgress novel genes in wild soybean to broaden the genetic base of modern soybean cultivars.

Variable Responses to CO2 of the Duration of Vegetative Growth and Yield within a Maturity Group in Soybeans

Prior experiments in indoor chambers and in the field using free-air carbon dioxide enrichment (FACE) systems indicated variation among soybean cultivars in whether and how much elevated CO2 prolonged vegetative development. However, the cultivars tested differed in maturity group, and it is not known whether variation exists in CO2 effects on the duration of vegetative growth within a maturity group. In these experiments, a total of five soybean cultivars of maturity group IV were grown at ambient and elevated CO2 in the field in Maryland, USA using FACE systems, over three years. The time of first flowering, the time of the first open flowers at the apex of the main stem, the total number of main stem nodes at maturity, and seed yield were recorded. In each year of the study, there were cultivars in which elevated CO2 did not affect the duration of vegetative growth or the main stem node number, and other cultivars in which elevated CO2 prolonged vegetative growth and increased the number of main stem nodes and seed yield at maturity. The stimulation in yield by elevated CO2 was highly correlated with the increase in the number of main stem nodes, indicating that CO2 effects on the duration of vegetative growth may be important in adapting soybean to higher atmospheric CO2.

Establishment and application of an accurate identification method for fragrant soybeans

In order to screen the aroma characteristics of soybean,a new method was established which can quickly quantify the content of 2-acetyl-1pyrroline(2-AP),an important compound related to soybean aroma,using gas chromatography-mass spectrometry(GC-MS).Based on peak profile,total peak area and retention time as test indexes,an accurate identification method for fragrant soybeans was established.The optimum parameters of the protocol consisted of column temperature70℃,sample injector temperature 180℃,optimum extraction alcohol content 1 m L,Na Cl content 0.1 g,ultrasonication time10 min,and extraction time 1 h,which were established by using the orthogonal test of single factors and three factors with four levels(L_9(3)^4).2-AP content of leaves had significant correlations with seeds,which were easier to measure.The protocol was simple and easy to carry out,consumed only small amounts of reagents,and provided accurate and reliable results with good reproducibility.A total of 101 soybean genotypes from different geographical sources were analyzed using this protocol.The results showed that the average content of 2-AP was 0.29 mg L^(–1),ranging from 0.094 to 1.816 mg L^(–1),and the genetic diversity index was 0.54.Among all genotypes-tested,they were classified into three grades,including seven elite genotypes identified as"grade one fragrant soybeans",which were Zhonglong 608,Heinong 88,Ha13-2958,Hongmiandou,Heinong 82,Huangmaodou,and Jiyu 21.These results provide both an identification technique and several elite aroma genotypes for gene discovery and good quality breeding in soybean.

Hypothetical Commercialization of Biotech Soybeans in China： Impacts on Domestic Markets and International Trade

China is the leading importer of soybeans in the world. If China adopts high yield biotech soybean varieties, China＇s soybean production would increase significantly and this would impact global soybean markets. This paper assesses the possible effects of adoption of herbicide-tolerant, biotech soybeans in China and its impact on the world soybean trade. Results indicate that under a low technology fee and high adoption rate, China would be able to satisfy its soybean food demand. Urban consumers＇ strong preference for non-biotech soybean oil may lead to increase in imports of non-biotech soybeans.

Bioinformatics Analysis of Disease Resistance Gene PR1 and Its Genetic Transformation in Soybeans and Cultivation of Multi-resistant Materials

In agricultural production,a single insect-resistant and disease-resistant variety can no longer meet the demand.In this study,the expression vector pCAMBIA-3301-PR1 containing the disease-resistant gene PR1 was constructed by means of genetic engineering,and the PR1 gene was genetically transformed to contain the PR1 gene through the pollen tube method.In CryAb-8Like transgenic high-generation T7 receptor soybean,a new material that is resistant to insects and diseases is obtained.For T2 transformed plants,routine PCR detection,Southern Blot hybridization,fluorescence quantitative PCR detection,indoor and outdoor pest resistance identification and indoor disease resistance identification were performed.The results showed that there were 9 positive plants in the routine PCR test of T2 generation.In Southern Blot hybridization,both PR1 and CryAb-8Like genes are integrated in soybeans in the form of single copies.Fluorescence quantitative PCR showed that the expression levels of PR1 and CryAb-8Like genes are different in different tissues.The average expression levels of PR1 gene in plant roots,stems,and leaves are 2.88,1.54,and 5.26,respectively.CryAb-8Like genes are found in roots,stems,and leaves.The average expression levels were 1.36,1.39,and 4.25,respectively.The insectivorous rate of the CryAb-8Like gene in outdoor plants with positive insect resistance identification was 3.78%.The disc partition method was used indoors for pest resistance identification,and the bud length of transformed plants increased significantly.The average mortality rate of untransformed plants in indoor disease resistance identification was as high as 56.66%,and the average mortality rate of plants transformed with PR1 gene was 10.00%,and disease resistance was significantly improved.Therefore,a new material with resistance to diseases and insects is obtained.

Analysis of Low Melting Point Fat Constituents of Black Soybeans by HS-SPME-GC-MS

[Objectives] To analyze the low melting point fat constituents in the black soybeans with green and yellow heart and their relative content,and compare the differences in the low melting point fat constituents between different kinds of black soybeans. [Methods] Using HS-SPME-GC-MS,the qualitative analysis was performed on the low melting point fat constituents of black soybeans; using peak area normalization method,the relative content of constituents was calculated. [Results]A total of 42 peaks were identified from the low melting point fat constituents of the black soybeans with yellow heart,and 18 kinds of chemical constituents were identified,accounting for 81.39% of total relative content of low melting point fat constituents; a total of 37 peaks were identified from the low melting point fat constituents of the black soybeans with green heart,and 15 kinds of chemical constituents were identified,accounting for 83.24% of total relative content of low melting point fat constituents. There were 9 kinds of common chemical constituents for the two kinds of black soybeans,and 5-allylguaiacol had the highest relative content,followed by hexanol. [Conclusions] There was no significant difference in the low melting point fat constituents between two kinds of black soybeans.

Activity Of Bacterial Proteolytic Enzymes on Antinutritional Factorsin Soybeans and the Effecton Growth and Organ Weights of Piglets

A significant reduction of trypsin inhibitory activity by selected bacterial proteolytic enzymes was demonstrated in vitro.Two trials were conducted to examine the capacity of the tested enzymes to inactivate soybean ANFs in vivo.In trial I,twenty four piglets weaned at four weeks of age were assigned in replicate groups of 4 piglets per pen to one of three dietary treatments:(1)control;(2)Enzyme 1 supplemented(E 1);(3)Enzyme 2 supplemented (E 2).In trial II,twenty piglets weaned at five weeks of age were alloted to five treatment diets:(1)contro,l:(2)0.1% P4 supplemented;(3)0.5% P4 supplemented;(4)0.1% P7 supplemented;(5)0.5% P7 supplemented.The optimum pH for hydrolysis was 8 for E ,9 11 for E 2,8.5 for P4 and nuctral for P7.After 17 days of the trial,daily gain of piglets on enzymes E 1 and E 2 was 36% and 18% more than that in the control group,although the difference was not significant.The animals on the treated groups had a tendency to have lighter heart(7.8 and 5.9%),spleen(11.1 and 7.4%) and pancreas(16.7 and 12.5% for E 1 and E 2 respectively)in relation to empty body weight than those in the control.The small intestine of pigs on the treated groups was significantly lighter(18.9 for E 1 and 7.7% for E 2) than that in the control(P<0.05).The stomach (26.4 and 24%,p=0.198) and cecum(21.9 and 9.4%,p=0.114) also showed the same pattern.The growth depression was attributed to reduced feed intake caused by antinutritional factors in soybeans.It is concluded that supplements of proteolytic enzymes E 1 or E 2 had a positive effect on growth and efficiency and caused much less reaction in the gut as manifested by the weight of the tract and of its accessory organs.Dietary saupplements of P4 or P7 had no significant effect on growth,but reduced reaction of soybean antinutritional factors in the gut,especialy P4 in dose of 0.5%.The growth depression was attributed to low feed intake caused by antinutritional factors in soybeans.

Activity of Peroxidase Enzyme in Grains of Some Varieties of Soybeans Grown in Uzbekistan

The article provides information on the activity of peroxidase enzyme in the composition of the seeds and bark of some varieties of soy cultivated in Uzbekistan and the importance of this enzyme in plant life. As is known, soybeans are one of the main nutritional plants of Agriculture. Orzu and Vilana varieties are resistant to other varieties, and the main thing is that they can be planted in crop fields in all regions of the Republic, achieve high fertility and thereby satisfy the demand of the population for feed products.

Changes of the Content and Antioxidant Activity of Natural Peptides in Germinated Soybeans

[ Objective] This study aimed to investigate the content and antioxidant activity of natural peptides in various soybean sprouts during the germination process. [Method] By using black soybean sprouts, yellow soybean sprouts and green soybean sprouts as raw materials, the content, DPPH ·clearance rate, OH · clearance rate and reduction capacity of natural peptides in various soybean sprouts at different germination time were determined, to evaluate the antioxidant capacity of natural peptides and compare it with that of Vc. [ Result] The results showed that the contents of natural peptides in black soybean sprouts, yellow soybean sprouts and green soybean sprouts varied at different germination time. To be specific, the content of natural peptides in black soybean sprouts reached the maximmn of 0. 194 mg/g at 2 d post-germination; the content of natural pepfides in yellow soybean sprouts reached the maximum of 0.231 mg/g at 2 d post-germination; the content of natural peptides in green soybean sprouts exhibited a negative correlation with the germination time, which reached the maximum of 0. 164 mg/g at 1 d post-germination. [ Conclusion] According to the determination of DPPH ·clearance rate, OH ·clearance rate and reduction capacity of natural peptides in three varieties of soybean sprouts, natural peptides in yellow soybean sprouts possess the highest antioxidant activity, followed by those in black soybean

GmSTF accumulation mediated by DELLA protein GmRGAs contributes to coordinating light and gibberellin signaling to reduce plant height in soybean

Plant height influences plant architecture,lodging resistance,and yield performance.It is modulated by gibberellic acid(GA)metabolism and signaling.DELLA proteins,acting as central repressors of GA signaling,integrate various environmental and hormonal signals to regulate plant growth and development in Arabidopsis.We examined the role of two DELLA proteins,GmRGAa and GmRGAb,in soybean plant height control.Knockout of these proteins led to longer internodes and increased plant height,primarily by increasing cell elongation.GmRGAs functioned under different light conditions,including red,blue,and far-red light,to repress plant height.Interaction studies revealed that GmRGAs interacted with the blue light receptor GmCRY1b.Consistent with this,GmCRY1b partially regulated plant height via GmRGAs.Additionally,DELLA proteins were found to stabilize the protein GmSTF1/2,a key positive regulator of photomorphogenesis.This stabilization led to increased transcription of GmGA2ox-7b and subsequent reduction in plant height.This study enhances our understanding of DELLA-mediated plant height control,offering Gmrgaab mutants for soybean structure and yield optimization.

A Novel Approach for Evaluation of Food Functions and Safety Applied in RR GM Soybeans

[Objective] This study aimed to evaluate tbe healthy risk of genetically modified （ GM ） soybeans by using a novel approach for functions and safety of food. [ Me^od] Different from traditional evaluation of substantial equivalence, three great innovations were performed in this study, involving in basic diet, evalu- ation approaches and principle, as well as the clarification of connotation differences between absolute and relative mass of organs. Hence a novel BDI-GS （Bendib Damage Index and General Score） evaluation approach was established and applied in comparative evaluation between RR GM and natural soybeans. Healthy male ICR mice during linear growth were selected; experimental mice were fed with 15% RR GM soybeans and 15% natural soybeans blending maize meal diets, and control mice were fed with single maize meal diet for 13 d; the mice were dissected after collecting blood samples and perfectly obtained nine organs or tissues to re- cord their masses and conduct statistical analyses. [Result] Plenty of matching information was obtained through simple design. The growth performance of treated mice was markedly of individual differences, some mice were thwarted due to regular intake of RR soybeans. Meanwhile, the functions and safety of RR soybeans were markedly lowered in overall nutritional and healthy effects than those of natural soybeans expressed in GS values, and presents some declines in nutrition and health of thymus, pancreas and spermary; especially, it can make thymus immune （P 〈0.05） in markedly lower level than that of natural soybeans. [ Conclusion] Therefore, major troubles and risks of RR soybeans intake are of personal risks in different degrees, in addition, it may increase sub-health and related chronic epi- demics risks, and herein it will presents certain safety issues. The creation of this novel evaluation system provides a simple and available evaluation approach for functions and potential risks revelation of food effects, and will yield far-reaching influences to safety evaluation and healthy development of GM foods, as well as public health.