Response of Soybean [Glycine max (L.) Merr.] Cultivars to Genistein-Preincubated Bradyrhizobium japonicum: Nodulation and Dry Matter Accumulation under Canadian Short-Season Conditions

The addition of genistein, a plant-to-bacteria signal molecule, to Bradyrhizobium japonicum cells prior to use as inocula has been shown to increase nodule number and promote soybean N₂ fixation at low root zone temperatures. Previous greenhouse and field experiments involving only two cultivars have indicated that soybean [ Glycine max (L.) Merr.] cultivars can vary in their response to genistein application. The objective of this study was to evaluate a range of soybean cultivars for response to genistein application under short-season cool-spring Canadian conditions. A 2-year field study was conducted in 1997 and 1998 with a range of soybean cultivars recommended for Quebec. The 11 cultivars tested represented a range of yield potentials and maturity groups. They were inoculated with genistein-preincubated B. japonicum inocula or regular inocula, applied into the furrow at the time of planting. The results of these experiments indicated that neither maturity nor yield was correlated with increases in nodulation, biomass, and plant total nitrogen content resulting from genistein treatment and that all maturity groups responded to genistein application in essentially the same way. Thus, response of soybean cultivars to genistein addition is regulated by genotype characteristics other than maturity or yield level.     

合成大豆异黄酮的植物基因工程研究进展

异黄酮主要存在于豆科植物中,是一种具有广谱抗菌活性的黄酮类物质。结合本课题组的工作,综述以基因工程的方法在植物内合成异黄酮的研究历史与进展,重点深入分析和讨论异黄酮生物合成所受到的分子调控。     

染料木素对后期蛋鸡生产性能及蛋品质的影响

【目的】探讨染料木素对产蛋后期蛋鸡生产性能、鸡蛋品质和蛋黄抗氧化指标的影响。【方法】采用单因子设计,将450只490日龄罗曼蛋鸡随机分为3组,每组3个重复,每个重复50只。Ⅰ(对照)、Ⅱ和Ⅲ组日粮中分别添加0,500和1 000mg/kg染料木素,每日记录各组蛋鸡的产蛋量和蛋质量,每周统计1次采食量和料蛋比,计算产蛋率。587日龄时,从每组随机取6枚鸡蛋,测定蛋形指数、蛋比重、蛋壳相对质量、蛋壳厚度、蛋壳强度、哈夫单位及蛋黄中染料木素、维生素A、维生素E和丙二醛(MDA)的含量。【结果】与Ⅰ组相比,Ⅲ组蛋鸡采食量和产蛋率显著提高(P<0.05),料蛋比显著降低(P<0.05);蛋质量、蛋形指数和蛋比重差异均不显著(P>0.05),蛋壳相对质量、蛋壳厚度、蛋壳强度和哈夫单位显著提高(P<0.05);蛋黄中染料木素含量显著提高(P<0.05),MDA含量显著降低(P<0.05),维生素A和维生素E含量差异不显著(P>0.05)。Ⅱ组蛋鸡除蛋壳强度外,其余指标均与Ⅰ组差异不显著。【结论】在产蛋后期蛋鸡日粮中添加染料木素,可提高蛋鸡的产蛋性能及鸡蛋的品质和抗氧化能力。     

Effect of genistein on IGF‐1 and IGFBP‐1 in young and aged female rat ovary

The objective of this study was to assess the effects of genistein (GEN) on expression of insulin‐like growth factor 1 (IGF‐1) and insulin‐like growth factor binding protein 1 (IGFBP‐1) in young and aged rat ovary. Forty young female Sprague Dawley (SD) rats (200 ± 20 g) and forty aged female SD rats (490 ± 20 g) were selected and according to weight, they were divided into the following five groups with eight animals in each: negative control group (NC), low‐dose group (L), middle‐dose group (M), high‐dose group (H) and positive control group (PC). GEN group received GEN of 15, 30, 60 mg/kg respectively. It lasted 30 days. Concentrations of serum hormones, IGF‐1 and IGFBP‐1 were determined by enzyme‐linked immunosorbent assay (ELISA). Gene and protein expressions of IGF‐1 and IGFBP‐1 were determined by real‐time PCR and Western blot respectively. Compared with NC, GEN significantly increased oestradiol‐17β(E₂) level in aged rat, reduced luteinizing hormone (LH) level in young and aged rat. Serum levels of IGFBP‐1 in young rats were significantly higher in GEN groups (p < 0.05). mRNA and protein expression levels of IGF‐1 and IGFBP‐1 were positively correlated with GEN dose. GEN could significantly reduce the ratio of IGF‐1/IGFBP‐1 of aged rats. Multivariate Cox regression analysis result showed IGF‐1 and IGFBP‐1 levels significantly correlated with GEN dose. We speculate that there is an association between the addition of GEN and expression of IGF‐1 and IGFBP‐1, and the relationship between them is different in young and aged rat.     

金雀异黄素对雌性大鼠体内促性腺激素及胰岛素样生长因子表达的影响

本试验旨在研究金雀异黄素(genistein,GEN)对雌性大鼠体内促性腺激素及胰岛素样生长因子表达的影响。选取40只SD雌性大鼠[体重(200±20)g],随机分为5组,分别为阴性对照(NC)组、GEN低(L)、中(M)、高剂量(H)组及阳性对照(PC)组,每组8只,NC组灌胃花生油(其他组灌胃试剂以此为溶剂);L、M、H组分别灌胃15、30、60 mg/(kg BW·d)GEN,PC组灌胃己烯雌酚0.5 mg/(kg BW·d)。试验期30 d。采用酶联免疫吸附试验(ELISA)法检测血清中卵泡刺激素(FSH)、黄体生成素(LH)、胰岛素样生长因子-1(IGF-1)、胰岛素样生长因子结合蛋白-1(IGFBP-1)含量;实时定量PCR法检测卵巢IGF-1、IGFBP-1 mRNA表达水平。结果表明:与NC组比较,试验组血清中FSH、LH含量有升高趋势,但差异不显著(P0.05),作用效果与PC组一致;试验组血清IGF-1含量略有降低,但差异不显著(P0.05),PC组显著降低(P0.05);试验组血清IGFBP-1含量显著或极显著升高(P0.05或P0.01),PC组显著升高(P0.05);试验组卵巢组织中IGF-1、IGFBP-1 mRNA表达水平均升高,其中M、H组显著升高(P0.05),与PC组变化一致。由此可见,GEN能够提高雌性大鼠血清FSH、LH含量、降低血清IGF-1含量、提高血清IGFBP-1含量,同时提高卵巢中IG FBP-1、IG F-1 mRNA的表达水平,这些指标协同作用于卵巢,能够促进卵泡的成熟,调节卵巢功能。     

Low root zone temperature effects on bean (Phaseolus vulgaris L.) plants inoculated with Rhizobium leguminosarum bv. phaseoli pre-incubated with methyl jasmonate and/or genistein

Abstract

Methyl jasmonate (MeJA) has recently been shown to act as a plant-to-bacteria signal. We tested the hypothesis that pre-induction of Rhizobium leguminosarum bv. phaseoli cells with genistein and/or MeJA would at least partially overcome the negative effects of low root zone temperature (RZT) on bean nodulation, nitrogen fixation and plant growth. Otebo bean plants were grown at constant air temperature (25^oC) and two RZT regimes (25 and 17^oC) and inoculated with R. leguminosarum bv. phaseoli pre-induced with MeJA and/or genistein. Our results indicate that low RZT inhibited nodulation, nitrogen fixation and plant growth. The plants growing at low RZT began fixing nitrogen seven days later compared to those at higher RZT. The low RZT plants had fewer nodules, lower nodule weight, less N fixation, slower plant growth, fewer leaves, smaller leaf area, and less dry matter accumulation comared to plants at a higher RZT. Rhzobium leguminosarum bv. phaseoli cells induced with genistein and/or MeJA enhanced bean nodulation, nitrogen fixation and growth at both optimum and suboptimum RZTs. The results of this study indicate that MeJA improves bean nitrogen fixation and growth at both optimum and suboptimum RZTs, and can be used alone or in combination with genistein to partially overcome the low RZT induced inhibitory effects on nodulation and nitrogen fixation.     

Potassium,Phosphorus, Sulfur,and Boron Fertilization Effects on Soybean Isoflavone Content and Other Seed Characteristics

ABSTRACT

Soybean [Glycine max (L.) Merr.] seeds contain isoflavones that have positive impacts on human health. The objective of this study was to determine the impact of pre-plant mineral fertilization on isoflavone, oil and crude protein concentrations, and seed yield of field-grown soybean. The effects of potassium (0, 50, 100, and 150 kg K ha^?1), phosphorus (0, 25, 50, and 75 kg P ha^?1), sulfur (0, 15, 30, 45 kg S ha^?1), and boron (0, 1.5, 3.0, and 4.5 kg B ha^?1) were tested separately, each with two 00 soybean cultivars (‘Golden’ and ‘Grand Prix’) grown in replicated trials at Sainte-Anne-de-Bellevue, Québec, Canada in 2002/3. Seed total and individual isoflavone concentrations were determined by high-performance liquid chromatography. Seed yield, 100-seed weight, and oil and crude protein (CP) contents were determined concurrently. Across years and cultivars, no fertilizer treatments effects were observed for most variables. This overall lack of response to fertilizers was attributed to the relatively high initial fertility of the sandy loam and sandy clay loam soils used. However, total and individual isoflavone concentrations were significantly affected by year and cultivar. Across experiments, total isoflavone concentration was 33% greater on average in 2003 than in 2002, which was characterized by above-average temperatures and severe drought. Cultivar with the greatest isoflavone concentration varied depending on the year. Fertilization does not appear to be a viable strategy to increase isoflavone concentration of soybean seeds on medium-to high-fertility soils.     

Irrigation Level Affects Isoflavone Concentrations of Early Maturing Soya Bean Cultivars

Field experiments were conducted in 2003/2004 in Québec to determine the effects of irrigation levels (none, low and high) and cultivars (AC Orford, AC Proteina and Golden) on soya bean [Glycine max (L.) Merr.] isoflavone concentrations and yields. Seed yield, yield components, and oil and crude protein (CP) concentrations were concurrently determined. Response to irrigation was greater in 2003, which was substantially warmer and drier than in 2004. In both years, most responses were observed with the lower of the two irrigation levels evaluated, which increased total isoflavones concentration by an average of 45 % compared with a non‐irrigated control. Cultivars, however, responded differently to irrigation. In 2003, response of AC Proteina was greater than that of AC Orford, while Golden did not respond. In 2004, some responses were observed with AC Proteina and Golden but none with AC Orford. Overall, in both years, AC Proteina had the greatest isoflavone concentrations and AC Orford the lowest. Responses of seed yield and yield components depended on the year and were also greater in 2003. Both irrigation treatments generally increased seed yield and yield components compared with a non‐irrigated control; the response was greater with the higher irrigation level. Irrigation had no effect on oil and CP concentrations. Finally, isoflavone yield response to irrigation was again greater in 2003, and depended on the cultivar. Results thus demonstrate that specific soil moisture levels will maximize soya bean isoflavone concentrations, excess irrigation sometimes negating any potential benefits.     

大豆异黄酮在雏鸡机体中吸收、转运及沉积的观察

9日龄雏鸡128只按初重随机分为4组:饲喂基础日粮对照组(C组)、基础日粮 3 mg/kg大豆黄酮组(3Da组)、基础日粮 6mg/kg大豆黄酮组(6Da组)和基础日粮 染料木素组(Ge组),试验期持续2周。与对照组相比,3Da组、6Da组和Ge组的日增重有增加趋势,料重比有所下降;空肠食糜、肝脏和肌肉中6Da组大豆黄酮含量分别增加288.15%(P<0.01),81.48%(P<0.05),155.97%(P<0.01);Ge组分别增加415.64%,172.48%,164.18%。食糜、肝脏中染料木素含量,6Da组分别增加154.16%和150.51%;Ge组分别增加238.13%和148.48%。肌肉内未检测出染料木素。上述结果提示,大豆黄酮和染料木素在雏鸡体内吸收、转运,并在肝脏和肌肉内代谢。两者可相互转化,仅大豆黄酮在肌肉内沉积。     

染料木黄酮对3T3-L1细胞增殖与分化的影响

试验旨在研究染料木黄酮对3T3-L1细胞增殖与分化的影响以及对分化过程中相关基因表达的影响。以浓度为0、10、50、100、200μmol/L的染料木黄酮处理细胞,测定不同天数3T3-L1细胞相对增殖量、细胞合成脂肪量以及第8天与脂肪合成有关的基因PPARγ、FASN、LPL、ACC、HSL的相对表达量。结果表明：在增殖方面,染料木黄酮能显著抑制3T3-L1前脂肪细胞的增殖（P〈0.05）。随着染料木黄酮浓度的增加,细胞增殖的抑制作用越强。当浓度为200μmol/L时,细胞停止增殖。在分化方面,染料木黄酮能显著减少3T3-L1前脂肪细胞分化合成脂肪的量（P〈0.05）。随着染料木黄酮浓度的增加,细胞分化合成脂肪量减少。染料木黄酮能显著减少3T3-L1细胞与脂肪合成有关的基因PPARγ、FASN、LPL、ACC、HSL的相对表达量（P〈0.05）。在浓度为50μmol/L时,PPARγ基因相对表达量最低;在浓度为200μmol/L时,FASN基因相对表达量最低;在浓度为10μmol/L时,LPL基因相对表达量最低;在浓度为200μmol/L时,ACC基因相对表达量最低;在浓度为10μmol/L时,HSL基因相对表达量最低。分析推测,染料木黄酮通过抑制与脂肪合成相关基因的表达来减少脂肪合成量。