大麦EMS突变体库构建和突变体筛选

为研究大麦重要功能基因并给大麦遗传育种提供新的种质资源,通过甲基磺酰乙酯(EMS)诱变处理优质、高抗黄花叶病大麦品种苏啤6号,从2400个M_2代株系中筛选出152个突变株系,突变频率为6.33%。其中,幼苗习性突变株系36个,突变率为1.50%;叶部突变株系34个,突变率为1.42%;茎部突变株系33个,突变体率为1.38%;穗部突变株系22个,突变率为0.92%;成熟期和育性突变株系27个,突变率为1.13%。经过M_3代验证,获得可稳定遗传的各类突变株系43个。     

脂肪含量对再制片状干酪品质的影响

研究不同脂肪含量和脂肪替代物对再制片状干酪品质的影响。对5 种不同脂肪含量再制片状干酪的基本理化指标、色泽、质构和感官品质进行测定和评估。结果表明：5 种不同脂肪含量的再制片状干酪pH值、蛋白质和脂肪含量存在显著性差异，酶解浓缩黄油的添加对再制片状干酪亮度值、红度值和黄度值均有明显影响；用酶解浓缩黄油替代全部黄油的减脂50%干酪全质构特性与其他组样品有差异，酶解浓缩黄油部分替代黄油制备的4号减脂干酪全质构特性除凝聚性、回复性外，与1～3号干酪无显著差异；通过感官评定发现，酶解浓缩黄油替代全部黄油的减脂50%干酪（5号）感官评分最低，酶解浓缩黄油部分替代黄油的减脂50%干酪（4号）风味和滋味有所增强，总体得分与全脂干酪产品无显著差异。因此通过降低黄油含量，并添加酶解浓缩黄油可提升减脂再制片状干酪的品质。     

H3N2犬流感病毒M1蛋白的原核表达及鉴定

为制备犬流感病毒(H3N2) M1蛋白纯品,针对M1基因序列设计引物,用聚合酶链式反应(PCR)扩增目的基因片段,扩增产物克隆至表达载体pET-SUMO中并转化至宿主菌BL21(DE3),诱导表达目的蛋白,探索纯化工艺,制备目的蛋白,并用Western blot检测纯化的M1目的蛋白。通过PCR成功扩增出大小为771 bp的M1基因,成功构建p ET-SUMO-M1表达载体,表达的融合蛋白相对分子量为41 kD,主要以可溶形式表达,纯化后获得蛋白纯品,Western blot检测显示用M1蛋白(28 k D)免疫小鼠制备的多抗能与制备的蛋白纯品发生特异性反应,从而证明蛋白纯品为M1目的蛋白。试验制备出的M1蛋白纯品可为进一步制备通用型抗犬流感病毒抗体提供纯品抗原。     

伐地那非增加EGCG-β-乳球蛋白纳米粒对肝癌细胞的抑制作用

较高浓度的EGCG才能抑制癌细胞的增殖,通过纳米化和EGCG与其他药物的联合使用是提高EGCG生物活性的重要策略。本研究将EGCG和伐地那非（VD）同时包埋于β-乳球蛋白（β-Lg）纳米载体中,制备出EGCG-VD-β-Lg纳米粒（EVβ-NPs）,体外试验证实,EVβ-NPs能提高人肝癌细胞（HepG2细胞）中Caspase-3活性,使HepG2细胞在S期产生明显的阻滞,诱发细胞核分裂,从而导致HepG2细胞凋亡。研究结果表明,将EGCG与微量的VD联合使用,并通过纳米化包埋可以显著提高EGCG的抗癌活性。这一方法在EGCG抗癌制品的开发方面具有潜在的价值。     

H9N2禽流感病毒全基因组密码子使用偏好性及影响因素分析

【目的】探究H9N2禽流感病毒(AIV)全基因组的密码子使用偏好性及影响因素。【方法】选取2010—2018年国内H9N2 AIV流行毒株的全基因组为研究对象,分析其碱基组成特性、最优密码子、密码子使用偏好性的影响因素以及病毒对宿主密码子使用模式的适应性。【结果】H9N2 AIV的全基因组中AU含量高于GC。大部分最优密码子以A或U结尾,有效密码子数(ENC)平均值为52.86,提示存在密码子使用偏好性但偏好性较低。密码子使用偏好性主要受到突变压力和自然选择的共同作用,其中自然选择(所占比例为61.79%～76.15%)作用大于突变压力(所占比例为23.85%～38.21%)。H9N2 AIV对人Homo sapiens的密码子适应指数平均值为0.739～0.741,提示H9N2AIV禽流感病毒可能已适应人类的密码子使用模式。【结论】本研究为H9N2 AIV的基因进化分析、已有疫苗的密码子优化和新型疫苗(密码子去优化疫苗)研制提供了理论依据。     

不同时段伊犁马行为特征的差异性分析

【目的】 研究马的行为规律及变化过程,为马匹的选择、训练及健康评估等提供理论参考。【方法】 选取健康状况良好的伊犁马母马12匹,采集视频数据,记录马匹在各时间段摄食、静息、探究、修饰、排泄和其他等各种行为指标,比较分析各时段行为的差异性。【结果】 舍饲伊犁马在行为规律上全天时间摄食>静息>探究>其他>修饰,不同时段伊犁马行为时间分配上,咀嚼、伸展时间下午时间段极显著大于早、晚时间段(P<0.01),卧息、站息时间晚上极显著大于早上、下午时间段(P<0.01),站立、警觉时间早上、下午时间段均极显著大于晚上时间段(P<0.01),舔食时间早上时间段极显著大于晚上时间段(P<0.01),啃痒时间早上时间段极显著大于下午时间段(P<0.01),频次指标中,咀嚼次数下午时间段极显著大于晚上时间段(P<0.01),显著大于早上时间段(P<0.05),舔食、摇头次数早上时间段极显著大于晚上时间段(P<0.01),卧息次数晚上时间段均极显著大于早上、下午时间段(P<0.01),警觉、站立次数早上、下午时间段均极显著大于晚上时间段(P<0.01),啃痒、伸展早上时间段显著大于下午时间段(P<0.05),排便次数早上、下午、晚上各时间段均存在极显著差异(P<0.01)。【结论】 舍饲伊犁马全天中用于摄食的时间最长,其后依次是静息、探究、其他、修饰。咀嚼、舔食多集中在早上、下午各时间段;蹭痒多集中在早上时间段;卧息、站息、伸展多集中于晚上时间段;站立、警觉在白天各时间段均较集中;在嗅探、蹭痒、哈欠、刨地、抬蹄、排尿上没有发现差异,通过观察不同时段所表现的行为指标对马匹行为进行监测,可为实际生产过程中马匹的管理提供客观参考指标,减少对马的伤害,防止恶癖的发生。     

年龄、温度、膘情对伊犁马卵泡发育的影响

【目的】 研究新疆昭苏地区伊犁马发情周期内卵泡发育变化规律,分析年龄、温度、膘情对伊犁马母马卵泡发育的影响,为掌握伊犁马母马卵泡发育规律提供理论依据。【方法】 选取伊犁哈萨克自治州昭苏县康苏马业配种站内,同一草场放牧情况下84匹母马作为研究对象监测发情期内卵泡发育,分析对比不同年龄、温度、膘情对伊犁马卵泡发育时长的差异性。【结果】 年龄Ⅱ组TF3-F5显著高于年龄Ⅲ组(P＜0.05)。年龄Ⅱ组TF4-5显著高于年龄Ⅲ组(P＜0.05)。平均高温对TF1-F2具有显著影响(P＜0.05),平均高温Ⅰ组TF1-F2显著高于平均高温Ⅱ组(P＜0.05)。平均低温对TF1-F2具有极显著影响(P＜0.01),平均低温Ⅰ组TF1-F2极显著高于平均低温Ⅱ组(P＜0.01)。积温对TF1-F2具有显著影响(P＜0.05),积温Ⅱ组TF1-F2极显著高于积温Ⅰ组(P＜0.01)。膘情对TF1-F2、TF3-F4、TF1-F5、TF2-F5、TF3-F5均具有极显著影响(P＜0.01),对TF2-F3具有显著影响(P＜0.05)。膘情Ⅰ组TF1-F2极显著高于膘情Ⅲ组(P＜0.01)。膘情Ⅰ组TF2-F3显著高于膘情Ⅲ组(P＜0.05)。膘情Ⅲ组TF3-F4极显著低于膘情Ⅰ组(P＜0.01),膘情Ⅲ组TF3-F4显著低于膘情Ⅰ组、膘情Ⅱ组(P＜0.05)。膘情Ⅰ组TF1-F5极显著高于膘情Ⅱ组、膘情Ⅲ组(P＜0.01),膘情Ⅱ组TF1-F5显著高于膘情Ⅲ组(P＜0.05)。膘情Ⅰ组TF2-F5极显著高于第Ⅲ组(P＜0.01),膘情Ⅰ组TF2-F5显著高于膘情Ⅱ组、膘情Ⅲ组(P＜0.05)。【结论】 年龄、温度、膘情均一定程度的影响伊犁马母马的卵泡发育,其中膘情对卵泡发育有极显著影响,体况较差可抑制母马卵泡发育。     

Effect of hydrogen peroxide and ozone treatment on improving the malting quality

The effect of hydrogen peroxide (HP) and ozone (O₃) treatment during barley steeping on the quality of malt produced from two barley varieties (GrangeR and AC Metcalfe) by micro-malting was investigated. The two steeping oxidation treatments that was observed to promote barley acrospire growth. Ozone treatment improved the malt enzyme activity of endo-protease, α-amylase, free beta-amylase and total limit dextrinase to differing extents, with GrangeR improving to a greater degree. HP treatment contributed to the increase of α-amylase, β-glucanase and endo-protease. Surprisingly, HP or ozone oxidation during malting resulted in different and novel outcomes for total beta-amylase in GrangeR and AC Metcalfe. In GrangeR, total beta-amylase activity reduced with respect to the control in both treatments. In comparison with AC Metcalfe there was a substantial increase of 78% with HP and 90% O₃ in total beta-amylase activity. Malt quality including wort free amino nitrogen, β-glucan, turbidity and diastatic power was differentially increased by the oxidation induction treatment during steeping in malting. Gene expression analysis indicated that the effects of the steep oxidation treatments on enzyme and malt quality were putatively linked with the up-regulation of certain genes involved in GA synthesis (GA20ox1) and ABA catabolism (ABA8′OH). Barley grain germination assay results also showed that moderate HP induction could improve barley germination tolerance to the ABA effect. Malting including steep oxidation induction was shown to be beneficial to malt quality by improving the resultant wort quality and the efficiency of the beer brewing process. These observations point the way towards improving malt quality and the efficiency of the malting process.     

Using vibrational molecular spectroscopy to detect moist heating induced carbohydrates structure changes in cool-climate adapted barley grain

There is limited research to study how moist heating affects internal structure of barley grain on a molecular basis. The objectives of this study were to use vibrational molecular spectroscopy: 1) to determine the moist heating induced changes of barley carbohydrate (CHO) structure on a molecular basis, 2) to study the effects of moist heating on CHO chemical profiles, Cornell Net Carbohydrate and Protein System (CNCPS) subfractions, in situ rumen degradation, and predicted intestinal carbohydrate supply of barley grain; and 3) to reveal the association between molecular structure spectral features and CHO related metabolic characteristics. Barley samples (CDC cowboy) were collected from Kernen Crop Research Farm (Saskatoon, Canada) during two consecutive years. Half of each sample was kept as raw barley and the other half underwent moist heating (autoclaving at 120 °C for 60 min). The molecular spectroscopy (attenuated total reflectance-fourier transform infrared, ATR-FTIR) was used to detect the barley CHO related molecular structure spectral features. Moist heating did not affect carbohydrate related chemical profiles and CNCPS subfractions but it decreased rumen degradable carbohydrate. Rumen undegradable and intestinal digestion of CHO subfractions were not affected by moist heating. The advanced vibrational molecular spectroscopy can be used to detect carbohydrate molecular spectral features. Nutrient utilization prediction using molecular spectral characteristics is warranted and further investigation is encouraged.     

Periplaneta americana peptide decreases apoptosis of pig-ovary granulosa cells induced by H2O2 through FoxO1

Oxidative stress can induce apoptosis of granulosa cells and lead to follicular atresia, thereby reducing the number of pigs giving birth. The aim of this study was to investigate the protective effect of Periplaneta americana peptide (PAP) on the apoptosis of the granulosa cells of pig ovaries (PGCs) induced by hydrogen peroxide (H₂O₂) via FoxO1. PGCs were treated with H₂O₂ to establish a cell apoptosis model. Cell viability was measured using the cell counting kit-8 (CCK-8) assay, and cell apoptosis was detected using flow cytometry. The malondialdehyde (MDA) level and nitric oxide (NO) content were detected to reflect the oxidative stress. Western blotting, qRT-PCR and overexpression were undertaken to determine the expression of FoxO1 and caspase-3, and immunofluorescence was used to detect FoxO1 in the nucleus and cytoplasm. PGCs were treated with 100 μM H₂O₂ for 6 hr, which resulted in oxidative damage and apoptosis and an apoptosis rate for PGCs of 32.95%. Next, PGCs were treated with 400 μg/ml PAP for 24 hr to repair the apoptosis induced by H₂O₂. PAP improved cell viability in H₂O₂-stimulated PGCs, the increased MDA level and NO content caused by H₂O₂ stimulation were reversed and the apoptotic rate of PGCs was reduced. The qRT-PCR and Western blotting results indicated that PAP decreased the H₂O₂-induced apoptosis and the expression of FoxO1 and caspase-3 in PGCs. The effect of PAP was the same following FoxO1 overexpression. FoxO1 was expressed in the nucleus when stimulated by H₂O₂ or overexpression; however, it migrated to the cytoplasm following PAP treatment. PAP decreased the apoptosis of PGCs induced by H₂O₂ by regulating FoxO1 expression and nuclear translocation.