Pueraria isoflavones inhibit XOD and GLUT9 to decrease uric acid production and promote uric acid excretion, respectively

Objective: To analyze the possible mechanism of Pueraria isoflavones inhibiting XOD and GLUT9 to reduce uric acid production and promote uric acid excretion. Methods: August 2021-April 2022, a total of forty SPF male Kunming mice were divided into the healthy group (carboxymethylcellulose sodium at a dose of 250 mg/kg), the model group (HUA mice were given carboxymethylcellulose sodium at a dose of 250 mg/kg), the low group (HUA mice were given pueraria isoflavone at a dose of 125 mg/kg), HUA mice were given pueraria isoflavones at a dose of 250 mg/kg once d frequency)and the high group (HUA mice were given pueraria isoflavones at a dose of 500 mg/kg once d frequency) dosage groups, with 8 mice in each group. The contents of uric acid (SUA), urea nitrogen (BUN) and creatinine (SCr) in serum and urine of each group were compared before and after intervention (30 d). Statistical differences of xanthine oxidase (XOD) and human glucose transporter 9(GLUT9), cy- clooxygenase- 2(COX-2), tumor necrosis factor (TNF-α) and interleukin-1 (IL-1β) contents in renal tissues of each group after intervention (30 d) were compared. Results: After intervention, kidney inflammatory factors (COX-2, TNF-α and IL-1β) in the model group were compared. Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were higher than those of healthy group(P<0.05). Renal inflammatory cytokines (COX-2, TNF-α and IL-1β) in low, medium and high dose groups;Blood and urine indexes (SUA, BUN, SCr);The contents of XOD and GLUT9 were lower than those of model group, and there were low > medium > high dose groups, the comparison between the two groups had statistical significance(P< 0.05). After intervention, the contents of 3 indicators in blood or urine(COX-2, TNF-α and IL-1β) all decreased compared with before intervention, and the differences in intra-group comparison were statistically significant (P<0.05). Conclusion: Pueraria isoflavones can treat HUA mice by inhibiting the expression of XOD and GLUT9, and then play a role in reducing uric acid pro- duction and promoting uric acid excretion, as well as alleviating the degree of disease inflammation.

ISOFLAVONES FROM ASTRAGALUS MEMBRANACEUS

Two new isoflavones (8, 3'-dihydroxy-7,4'-dimethoxyisoflavone, odoratin-7-0-[3-D-glu-copyranoside) and four known isoflavones (formononetin, 7,3'-dihydroxy-8,4'-dimethoxyisoflavone, calycosin, calycosin-7-0-(3-D-glucopyranoside) were isolated from the roots of Astragalus mem-branaceus (Fisch.) Bunge. Their structures were established by spectral analysis.

Inhibitory Effect of Isoflavones on Prostate Cancer Cells and PTEN Gene

Objective To explore the mechanisms by which genistein and daidzein inhibit the growth of prostate cancer cells. Methods LNCaP and PC-3 cells were exposed to genistein and daldzein and cell viability was determined by MTF assay and cytotoxicity of the drugs by LDH test. Flow cytometry （FCM） was used to assess the cell cycle in LNCaP and PC-3 cells. Reverse transcription-polymerase chain reaction （RT-PCR） was applied to examine the expression of PTEN gene （a tumor suppressor gene）, estrogen receptor alpha gene （ERα）, estrogen receptor beta gene （ERβ）, androgen receptor gene （AR） and vascular endothelial growth factor gene （VEGF）. Results The viability of PC-3 and LNCaP cells decreased with increasing concentrations and exposure time of genistein and daidzein. Genistein increased G2/M phase cells in PC 3 cells while decreased S phase cells in LNCaP cells in a dose-dependent manner. Daidzein exerted no influence on the cell cycle of LNCaP and PC-3 cells, but the apoptosis percentage of LNCaP cells was elevated significantly by daidzein, Genistein induced the expression of PTEN gene in PC-3 and LNCaP cells. Daidzein induced the expression of PTEN gene in LNCaP but not in PC-3 cells. The expression of VEGF, ERα and ERβ genes decreased and AR gene was not expressed after incubation with genistein and daldzein in PC-3 cells. In LNCaP cells, the expression of VEGF and AR gene decreased but there was no change in the expression of ERα and ERβ gene after incubation with genistein and daidzein. Conclusion Genistein and daidzein exert a timeand dose-dependent inhibitory effect on PC-3 and LNCaP cells. The down-regulation of ER gene by daidzein influences the growth of PC-3 ceils directly, The inhibition of PC-3 cells by genistein and that of LNCaP cells by genistein and daidzein may be via Alct pathway that is repressed by PTEN gene, which subsequently down-regulates the expression of AR and VEGF genes. Our results suggest that the expression of PTEN gene plays a key role and several pathways may be involved in the suppression of prostate cancer cells by genistein and daidzein.

Developmental and Reproductive Toxicity of Soybean Isoflavones to Immature SD Rats

Objective To investigate the dose-dependent toxic effect of soybean isoflavone extracts （SIE） on reproductive development in immature rats. Methods Growing male and female rats （n=50 each, 4 weeks） were divided into five groups fed with a standard cereal-based diet and gastrogavaged daily with 0, 30, 150, 300, and 600 mg SIE / kg body weight, respectively, for 12 weeks. Body weight, organ weights, and serum level of estrogen and testosterone were measured. Results Oral administration of SIE had no effect on food intake but decreased food efficiency ratio （P〈0.01）. Suppression on body weight gain by SIE was dose-dependent and the effect was greater on male than on female rats （P〈0.01）. SIE at high doses exhibited hepatotoxicity by increasing a relative liver weight, and also caused a smaller uterus but a greater relative ovary in female rats, while leading to larger relative testis and epididymis in male rats. SIE could decrease progesterone concentrations in female rats, whereas in male rats it reduced not only total testosterone level but also sperm count compared with the control group （P〈0.05）. Conclusion SIE at a range of 50-1000 times of human intake level affects not only growth but also development of reproductive system in growing rats.

Effects of Dietary Soy Isoflavones on Feed Intake,Growth Performance and Digestibility in Juvenile Japanese Flounder(Paralichthys olivaceus)

An 8-week feeding trial was conducted to investigate the effects of dietary soy isoflavones on feeding intake,growth performance,and digestion of juvenile Japanese flounder(Paralichthys olivaceus).Four isonitrogenous(49% crude protein) and isoenergetic(20.1 MJ kg-1) diets were formulated to contain four graded levels of soy isoflavones,namely,0,1,4 and 8 g soy isoflavones in 1 kg of diet.Each diet was randomly fed to triplicate tanks of fish(Initial average weight:2.58 g ± 0.01 g),and each tank was stocked with 35 fish.No significant difference was observed among diets with levels of 0,1 and 4 g kg-1 soy isoflavones in feed intake,weight gain,feed efficiency ratio(FER),proximate composition of fish whole body and apparent digestibility coefficients(ADC) of nutrients and energy(P>0.05).However,high dietary soy isoflavones level(8 g kg-1) significantly depressed weight gain,FER,whole-body crude lipid content of fish and ADC of nutrients(P<0.05).These results indicate that high level of dietary soy isoflavones(above 4 g kg-1) significantly depresses growth responses and FER of Japanese flounder.However,as the content of soy isoflavones in soybean meal is around 1 to 3 g kg-1,the adverse effects might be neglected when soybean products are used as a fish feed ingredient.

Microwave-assisted Extraction of Isoflavones from Belamcanda chinensis

Microwave-assisted extraction （MAE） of isoflavones from Belamcanda chinensis was studied using ethanol as the solvent. The single factor experiment and the orthogonal method were used to optimize the MAE condition. It was concluded that two doses of intermittent microwave power radiation, 300 W each for 4 rain, were needed for extraction. The mass ratio of solvent to material was 8：1 and the alcohol mass fraction was 80%. The extracted liquor was then concentrated under vacuum and degreased with petroleum ether. The yield of total isofiavones was about 8.8% and the contents of tectoridin and iridin were 67.6% and 16.3% respectively. Compared with direct-heating extraction （DHE）, MAE may shorten extracting duration, reduce solvent consumption, and improve yield and purity of isoflavones from Belamcanda chinensis.

Estrogen Receptor α and β Expressions in Hypothalamus-pituitary-ovary Axis in Rats Exposed Lactationally to Soy Isoflavones and Bisphenol A

Objectives This paper aims to investigate the uterotrophic activities of lactational exposure to combination of soy isoflavones （SIF） and bisphenol A （BPA） and to examine estrogen receptor α （ERα） and estrogen receptor β （ERβ） expressions in hypothalamus-pituitary-ovary axis and uterus.Methods Maternal rats that were breeding about 8 litters were randomly divided into four groups with seven dams in each group.Dams in different treatment groups received corn oil （control）,150 mg/kg BW of SIF,150 mg/kg BW of BPA or combination of 150 mg/kg BW of SIF and 150 mg/kg BW of BPA,respectively,from postnatal day 5 to 11 （PND5-11） by gavage.On PND12 and PND70,10 female litters were killed and hypothalamus,pituitary,ovary and uterus were collected.ERα and ERβ expressions in these organs were detected with Western blotting assay.And vaginal opening time and estrus cycle were examined in animals fed for PND70.Results On PND12,the relative uterine weight of rats treated with ISF or BPA or their combination was significantly higher than that of untreated rats （P〈0.05）.But the relative uterine weight of rats in the co-exposure group was slightly lower than that in the group only exposed to SIF or BPA.On PND 70,however,the relative uterine weight in each treatment group was not statistically different from that in the control group （P〈0.05）.Vaginal opening time and estrus cycle in groups treated with SIF or BPA or their combination were similar to those in the control group （P〈0.05）.Exposure to SIF or BPA or their combination could up-regulate or down-regulate ERα and ERβ expressions in hypothalamus,pituitary,ovary and uterus on PND12 and PND70.These regulation patterns for ERα and ERβ were different in different organs at different time points.Conclusion Lactational exposure to ISF or BPA or their combination could induce uterotrophic responses in neonate rats,which disappeared in later life.But these data fail to suggest a possibility for synergic actions between SIF and BPA.It was also demonstrated that the uterotrophic effects of SIF and BPA exposure might,at least,involve modification of ERα or ERβ expressions in the hypothalamus-pituitary-ovary axis.

Effects of Soybean Isoflavones on In vitro Antioxidative Capacity of Satellite Cells of Porcine Skeletal Muscles

A synthetic isoflavone （ISO-S） or genistein was added in culture medium at different concentrations （0, 10, 20, 30, 40, and 80 p.mol L^-1） to investigate the effects of soybean isoflavones on antioxidative capacity of porcine skeletal muscle satellite cells. After 48 h incubation, the suspension was cryopreserved for the determination of superoxide dismutase （SOD）, catalase （CAT）, glutathione peroxidase （GSH-Px） activities, and malondialdehyde （MDA） content. The mRNA levels of SOD, CAT, and GSH-Px gene in cells were detected with Taqman fluorescent probe method. The results showed that the content of MDA and the activities and the mRNA levels of SOD of porcine skeletal muscle satellite cells were influenced by supplemented soybean isoflavone （P〈0.05） when adding 10-80 μmol L^-1 ISO-S or genistein in the medium. The MDA contents, SOD and CAT activities and their mRNA expression levels of porcine skeletal muscle cells responded quadratically （P〈 0.05） as the level of ISO-S or genistein increased. Pre-incubation of porcine skeletal muscle satellite cells with ISO-S or genistein at 10-40 pmol L-1 elevated the activities and the mRNA expression levels of SOD and CAT in cells concurrently and decreased the cellular content of MDA （P〈 0.05）. The results indicated that pre-incubation of ISO-S or genistein at 10- 40μmol L^-1 could improve the antioxidative capacity of porcine skeletal muscle satellite cells.

Consumption of soybean,soy foods,soy isoflavones and breast cancer incidence:Differences between Chinese women and women in Western countries and possible mechanisms

Breast cancer is one of the most lethal diseases world-wide.However,there is a large difference in breast cancer incidence among Caucasian,Hispanic,African and Asian(e.g.Chinese)women with Caucasian women being the highest and Asian women being the lowest.It has been suggested that the dietary factors may account for approximately 50%of the breast cancer.One of such dietary components which are typical to Asian but not Caucasian diet is soy foods.A number of epidemiological studies have suggested that increasing soy consumption could be related to the decreased risk of occurrence and/or mortality of breast cancer.In this review,we first described briefly different types of soy products and their nutritional functions and consumption.Then,we described briefly soybean isoflavones,i.e.genistein(GEN),daidzein,glycitein,and presented several lines of evidence to demonstrate the possible association of soy flavone food consumption with incidence and prognosis of breast cancer;finally,we summarized several possible molecular mechanisms,including the effects of GEN as an agonist of ER,epigenetic and genome-wide effects,activation of peroxisome proliferator-activated receptors,induction of apoptosis and stimulation of autophagy,involved in the chemo-preventive effects of GEN on breast cancer.©2013 Beijing Academy of Food Sciences.Production and hosting by Elsevier B.V.All rights reserved.

Isoflavones’effects on pharmacokinetic profiles of main iridoids from Gardeniae Fructus in rats

Gardeniae Fructus(GF)and Semen Sojae Praeparatum(SSP)are both medicine food homologies and widely used in Chinese clinical prescriptions together.The research investigated the pharmacokinetics of four iridoids in normal rats and isolfavones-fed rats,which were administered with isolfavones from SSP for 7,14,21 and 28 consecutive days.A validated LC-MS/MS method was developed for determining shanzhiside,genipin-1-gentiobioside,geniposide and their metabolite genipin in rat plasma.Plasma samples were pretreated by solid-phase extraction using paeoniflorin as the internal standard.The chromatographic separation was performed on a Waters Atlantis T3(4.6 mm×150 mm,3 mm)column using a gradient mobile phase consisting of acetonitril and water(containing 0.06%acetic acid).The mass detection was under the multiple reaction monitoring(MRM)mode via polarity switching between negative and positive ionization modes.The calibration curves exhibited good linearity(r>0.997)for all components.The lower limit of quantitation was in the range of 1 e10 ng/m L.The intra-day and inter-day precisions(RSD)at three different levels were both less than 12.2%and the accuracies(RE)ranged fromà10.1%to 16.4%.The extraction recovery of them ranged from 53.8%to 99.7%.Pharmacokinetic results indicated the bioavailability of three iridoid glycosides and the metabolite,genipin in normal rats was higher than that in rats exposed to isoflavones.With the longer time of administration of isoflavones,plasma concentrations of iridoids decreased,while genipin sulfate,the phase II metabolite of genposide and genipin-1-gentiobioside,appeared the rising exposure.The pharmacokinetic profiles of main iridoids from GF were altered by isoflavones.

Two new C-glucofuranosyl isoflavones in puerarin injection

Two new C-glucofuranosyl isoflavones of impurities in puerarin injection were isolated. Their structures were detemained to be 8-C-α-glucofuranosyl-7, 4P-dihydroxyisoflavone and 8-C-β-glucofuranosyl-7, 4＇-dihydroxy- isoflavone by chemical and spectral analysis.

Isoflavones and inflammatory bowel disease

Isoflavones constitute a class of plant hormones including genistein,daidzein,glycitein,formononetin,biochanin A,and irilone,and the major source of human intake is soybeans.Inflammatory bowel disease(IBD)is a chronic recurrent inflammatory disease including ulcerative colitis,Crohn’s disease,and indeterminate colitis,which seriously affects the quality of life of patients and has become a global health problem.Although the pathogenesis of IBD is not very clear,many factors are thought to be related to the occurrence and development of IBD such as genes,immunity,and intestinal flora.How to control IBD effectively for a long time is still a problem for gastroenterologists.Diet has an important effect on IBD.Patients with IBD should pay more attention to diet.To date,many studies have reported that isoflavones have both good and bad effects on IBD.Isoflavones have many activities such as regulating the inflammatory signal pathways and affecting intestinal barrier functions and gut flora.They can also act through estrogen receptors,as they have a similar structure to estrogen.Isoflavones are easy to get from diet for human.Whether they are valuable to be applied to the treatment of IBD is worth studying.This review summarizes the relationship between isoflavones and IBD.

Optimum of Mobile Phase Condition for Resolving Isoflavones in RP-HPLC

In reversed-phase high performance liquid chromatography （RP-HPLC）, the mobile phase condition for separating eight isoflavones （daidzin, glycitin, genistin, 6＇-o-acetyl daidzin, 6＇-o-malonyl genistin, daidzein, glycitein and genistein） was optimized using the HCI （High-Purity Separation Laboratory, Department of Chemical Engineering, Inha University） program software. The optimum composition of mobile phase for the separation of the eight isoflavones was obtained. The elution profiles were calculated by the plate theory based on the equations of retention factor, In k=A＋BF＋CF2, where F was the volume percentage of acetonitrile with 0.1% acetic acid （AA）. The first mobile phase composition was water with 0.1% AA/acetonitrile with 0.1% AA （88%/12%, by volume）, followed at 9min later by the second composition of mobile phase which was step-changed to 85%/15%, at 19rain by the third composition which was step-changed to 73%/27%, at 30min when it was changed to 65%/35% and finally it was maintained in isocratic mode to the end of the run time at 50rain. Although, using step gradient mode to separate the isoflavones, the calculated and experimented data were not achieved very good agreement, we could estimate the closed retention time before experiment. And the agreement between the experimental data and the calculated values was relatively good using isocratic separation for eight isoflavones, but the retention time is very long.

Soybean Seed Phenol, Lignin, and Isoflavones and Sugars Composition Altered by Foliar Boron Application in Soybean under Water Stress

Previous research showed that foliar boron (B) application at flowering or seed-fill growth stages altered seed protein, oil, and fatty acids. The objective of this research was to investigate the effects of foliar B fertilizer on seed phenolics (phenol, lignin, and isoflavones) and sugars concentrations. A repeated greenhouse experiment was conducted on soybean [(Glycine max(L.) Merr.)] under watered and water-stressed conditions. Soybean plants were divided into different sets, and each set was subjected to one of the following treatments: W = plants were watered with no foliar B;WB = plants were watered and received foliar B;WS = plants were water-stressed with no foliar B;WSB = plants were waterstressed and received foliar B. Foliar B was applied at rate of 0.45 kg/ha twice at flowering and twice at seed-fill stages. The results showed that total phenol and lignin concentrations were higher in seed collected from water-stressed plants compared with those collected from watered plants whether B was applied or not. The higher total phenol and lignin concentration in seed collected of water-stressed plants may be due to B-deficiency in plant tissues. Application of B resulted in higher concentrations of total seed B and isoflavones under watered and water-stressed plants. Higher cell wall B was higher in water-stressed plants than in watered plants, having an opposite trend to total B. Application of B resulted in higher seed sucrose in watered and water-stressed plants, but raffinose and stachyose were significantly higher under water-stressed plants. The research demonstrated that foliar B fertilizer altered seed phenol, lignin, isoflavones, and sugars, suggesting that B involved in phenolics and sugar metabolism. The higher cell wall B in waterstressed plants than in watered plants supports previous research that B has mainly a structural role. The higher sucrose resulting from foliar B in watered plants is desirable as sucrose contributes to seed quality. The increase of raffinose and stachyose concentrations in seed of water-stressed plants is undesirable as raffinose, and especially stachyose may be involved in water stress/drought tolerance. The current knowledge would help soybean breeders select for higher phenolic compounds and desirable sugars for higher seed qualities under drought conditions.

Effects of soybean isoflavones on reproductive parameters in Chinese mini-pig boars

Background： Soybean isoflavones are structurally similar to mammalian estrogens and therefore may act as estrogen agonists or antagonists. However, it has not been determined if they have any negative effects on reproductive parameters in male livestock. Therefore, the objective of this study was to evaluate the effects of soybean isofiavones on male reproduction using Chinese mini-pig boars as a model. Fifty Xiang boars were randomly divided into five groups and fed diets containing 0, 125, 250, or 500 ppm soybean isoflavones or 0.5 ppm diethylstilbestrol for 60 days. Results： Dietary supplementation with 250 ppm of soy isoflavones markedly increased the testis index （P 〈 0.05）, fructose content （P 〈 0.05）, and o-glycosidase content in testicular tissue （P 〈 0.01）, as well as increased the number of viable germ cells （P 〈 0.01） and the level of Bcl-2 protein （P 〈 0.01）. However, 500 ppm of soybean isoflavones significantly reduced both testis and epididymis indexes （P 〈 0.0S） and lactate dehydrogenase levels （P 〈 0.01）, as well as reduced serum LH and testosterone levels （P 〈 0.05）. High levels of soybean isoflavones also increased malondialdehyde levels （P 〈 0.05）, as well as increased the numbers of early and late apoptotic germ cells （P 〈 0.01） and the level of Bax proteins （P 〈 0.05） in the testis. Conclusions： The results of this study indicate that consumption of soy isoflavones at dietary levels up to 250 ppm did not adversely affect reproductive parameters in Chinese mini-pig boars whereas higher levels of soy isoflavones may adversely affect male reproduction.

Influence of Isoflavones on Cadmium-induced Adverse Effects in Vascular Endothelial Cells (ECV 304)

Objective To study the possible intervention of isoflavones in cytotoxicity induced by cadmium in vascular endothelial cells. Methods An ECV 304 cell line derived from human umbilical vein endothelial cells was adopted. Genistein / daidzein was added prior to or simultaneously with CdCl2, cell viability was determined by MTT assay, and metallothionein mRNA expression was monitored by RT-PCR method. Results Cell viability was higher in isoflavone and CdCl2 co-treated groups than that in CdCl2 treated group, with CdCl2 concentration at 10, 20, 40, and 80 μmol/L, respectively. However this increase was not observed in the group treated with CdCl2 at a concentration of 60 μmol/L. Isoflavones (10-10 mol/L to 10-5 mol/L) were added 24 h before cells were challenged with 80 μmol/L CdCl2 for 24 h or simultaneously with 80 μmol/L CdCl2. Genistein increased cell viability only at 10-5 mol/L, while daidzein caused a dose-dependent increase from 10-10 mol/L to 10-5 mol/L in co-treatment with CdCl2. In pre-treatment, genistein (10-7 to 10-5 mol/L) increased cell viability whereas only 10-5 mol/L of daidzein exerted protection. Apparent protection could be found when the cells were pre-treated with 10-5 mol/L isoflavones for over 12 h, whereas 24 h incubation was required in such a co-treatment, with the exception of daidzein that had a significant protection in only 3 h. Isoflavones (10-6 mol/L) incubated for 3 h to 24 h, increased MT IIA and MT IF mRNA expression, but the induction could not last for more than 24 h. Co-treatment with isoflavones could induce an additional induction of MT IIA mRNA expression in cells exposed to cadmium. However, the additional induction of MT IIA and MT IF mRNA was not seen when pre-treatment was carried out with isoflavones, with the exception of an increase in MT IIA mRNA expression in the daidzein pre-treated group. Conclusion Genistein/daidzein could reverse the cytotoxicity of cadmium either in pre-treatment or in co-treatment. The protection is the strongest in 10-5 mol/L of isoflavones with a dose-dependent pattern. There are differences between genistein and daidzein in their protective effects. Whether the protection of isoflavones is related to their capacity of inducing MT mRNA expression remains to be elucidated.

Subchronic Toxicity Study on Soy Isoflavones in Rats

Objective To investigate the subchronic toxicity of soy isoflavones （SIF） in male rats. Method Fifty Sprague-Dawley rats were randomly divided into 5 groups, 10 rats per group. SIF were given to rats in different groups by gavage at dose of 0, 0.2, 0.5, 1.5, and 4.5 g/kg bw, respectively for 13 weeks. Clinical manifestations, body weight, and food consumption were observed weekly. At the end of the study, urinalysis, hematology, clinical chemistry, total testosterone, and follicle-stimulating hormone were tested, and histopathological examinations were performed. Results No mortality, ophthalmic abnormalities or treatment-related clinical signs were identified during the study. As compared with the control group, significantly lower body weights and food consumption were observed in 1.5 and 4.5 g/kg bw groups. In clinical chemistry tests, triglyceride was significantly decreased and high-density lipoprotein cholesterol was significantly increased in all SIF-treated groups. Total testosterone levels were significantly lower in 0.50, 1.50, and 4.5 g/kg bw dose groups than in the control group. Microscopic examination showed that the mammary glands exhibited hyperplasia and excreted latex in rats of the 4.5 g/kg bw group. No changes attributable to treatment of SIF in other parameters were found. Conclusion SIF at high dosages caused significant endocrine disruption in male rats. The no observed adverse effect level （NOAEL） of SIF to male rats in this study is considered to be 0.20 g/kg bw.

Perform Stability of Isoflavones of Soybean Cultivar Evaluated by Genotype-genotype×environment(GGE) Biplot

As one of the secondary metabolites,the isoflavones formed during the development of soybean[Glycine max(L.)Merr.]seeds.The total and individual isoflavone contents,a typical quantitative trait,were affected by significant genotypes of environments(GE)interaction and controlled by many genes with main or minor effects.In the present study,99 soybean cultivars,collected from northeastern China,were used to analyze the isoflavone performances.Genotype-genotype×environment(GGE)biplot software demonstrated an ability to provide information on genetic main effects than solely on phenotypic perform.Highperformance liquid chromatography(HPLC)system was used to extract and determine the isoflavone contents.The results indicated that most genotypes significantly varied among six tested environments.P40(Xiaolimoshidou)was the best-performed genotype with mean performance and stability for glycitein content across six different environments.P88(L-59Peking)was the super genotype with mean performance and stability on each tested environment for daidzein,genistein and the total isoflavone.E5(Gongzhuling in 2016)was the best environment for optimal environmental factor mining.P70(Charleston),P67(Baichengmoshidou)and P50(Jiunong 20)were the optimal genotypes with the highest field among 99 cultivars on each tested environment for genistein.P70(Charleston),P67(Baichengmoshidou)and P14(Hefeng 25)were the optimal genotypes with the highest field among 99 cultivars on each tested environment for daidzein.P40(Xiaolimoshidou),P45(Jinshanchamodou),P33(Dongnong 48)and P56(L-5)were the optimal genotypes with the highest field among 99 cultivars on each tested environment for glycitein.P70(Charleston)and P67(Baichengmoshidou)were the optimal genotypes with the highest field among 99 cultivars on each tested environment for the total isoflavone.GGE biplot was a rational method for stability and adaptation evaluation of soybean isoflavones,and could assist soybean breeder to select a good culture and a suitable tested site.It provided a scientific basis for the establishment of a breeding site and a selection site of soybean isoflavones.This study was valuable to identify genotypes with stable performances of isoflavones of these 99 cultivars for developing new cultivars.

A value-added cooking process to improve the quality of soybean:Protecting its isoflavones and antioxidant activity

This study investigated the effects of domestic cooking process on the variations of soybean isoflavones,aiming at understanding the conversion of-glucosides and aglycones isoflavones during the process and the relation with antioxidant activity.It was found that-glucosides isoflavones was significantly increased from 223.01(raw)to 727.29 mg/g(frying at 160◦C for 2 min),but boiling showed only a slight increase to 258.14 mg/g.The process for the mixed cooking of soybeans with vegetables was also evaluated,which is quite common in home cuisine.The results showed all bioactive ingredients were aggressively destroyed by over processing,but interestingly,green pepper and kelp exhibited isoflavones generation potentials for soybean.In addition,cooking from 60 to 160◦C for 2 or 5 min,showed a significantly decrease on FRAP.However,in the case of fried soybeans which treated at 120◦C or 160◦C,when extending the heating time to 5 min,their FRAP activity got a significant increase.The present study may provide a practical guidance for healthy soybean cooking,by using frying around 120◦C for 5 min and mixed with some vegetables such as green peppers or kelp.

Evaluation of Isoflavones as Allelochemicals with Strong Allelopathic Activities of Kudzu Using Protoplast Co-Culture Method with Digital Image Analysis

The inhibitory allelopathic activity of Pueraria montana (Kudzu), and activities of two putative allelochemical isoflavones, puerarin and daidzein, were evaluated using the protoplast co-culture method with digital image analysis using lettuce as a recipient (DIA-PP method). Cotyledon protoplasts of Kudzu were isolated using Cellulase R10 and Driselase 20 in 0.6 M mannitol solution. Optimal hormonal condition and density for growth of Kudzu protoplasts were surveyed. Medium for co-culture of Kudzu or isoflavones with lettuce protoplasts was 50 μl liquid MS basal medium containing 1 μM 2,4-dichlorophenoxyacetic acid, 0.1 μM benzyladenine, 3% sucrose, and 0.4 M or 0.6 M mannitol. Protoplast division of lettuce was strongly inhibited by Kudzu at a low density (104/ml). Slightly less inhibition by Kudzu on cell wall formation and yellow pigment accumulation stages of lettuce growth was also observed. Puerarin did not inhibit the growth of lettuce protoplasts at three growth stages but slightly stimulated growth at high concentrations. By contrast, daidzein, aglycon of puerarin, inhibited growth at three stages of lettuce protoplast growth and strongly inhibited cell division at 100 μM. Daidzein might be one cause of the strong inhibitory allelopathic activity of Kudzu. Grade of inhibitory activities was compared with that of other allelopathic plants including an invader plant and their allelochemicals studied using the DIA-PP method.