紫花苜蓿细胞质雄性不育恢复基因的初步定位

以紫花苜蓿(Medicago sativa)不育系MS-GN-1A为母本,恢复系MS178为父本组合构建BSA分离群体,获得的F1代均表现为雄性可育,在大田种植了221株F2代群体单株,盛花期将花粉颗粒染色后,显微镜下观察统计出,不育的F2代株数为57,可育F2代株数164,并没有观察到半不育植株。将所有植株划分为可育和不育两组,并构建可育和不育DNA混池,混池DNA从可育和不育植株组DNA中各随机抽取20个样品,以此对恢复基因定位。随机挑选160对已知的四倍体苜蓿SSR引物扩增基因池DNA,获得2个具有多态性的分子标记,分别是Mt2c12、AW166,初步定位Rf基因在类群Composite5上。将类群Composite5上的所有引物进行合成,进一步进行引物筛选,最终获得4个具有多态性的标记BI68、Mt2c12、BG267和AW776153,遗传距离分别为19.0、20.9、44.6和72.1cM。     

Impact of cold-season grazing and supplementary feeding on rumen metabolic parameters and micro-organisms in the ewes of Gansu Alpine Fine Wool Sheep北大核心CSCD

An experiment was conducted to scientifically identify the appropriate feed for Gansu Alpine Fine Wool ewes in order to change the animals' breeding mode, which currently can only be mated at the age of 2.5 years due to insufficient nutritional supply. The experiment selected 48 12-month-old ewes and randomly divided them into group a (Diet 1), group b (Diet 2), group c (Diet 3), and a control group with no supplementary feeding. The feed test was run for 90 days. The rumen pH of groups a and b were very significantly higher (P<0.01) or group c significantly lower (P<0.05) than control group. The rumen ammonia nitrogen, protein nitrogen and total nitrogen of groups a, b and c were all higher than control group (P<0.01). The total volatile fatty acids of group a and b were all higher than the control group (P<0.01), group b significantly higher than group c (P<0.05). The molar ratio of acetic acid and propionic acid in group a was very significantly higher than control group (P<0.01), group b and c were significantly higher than control group (P<0.05). The molar ratio of butyric acid in group a was very significantly higher than group c (P<0.01). The ratio of acetic acid and propionic acid in group b was significantly lower than control group (P<0.05). The number of several rumen fiber-degrading bacteria and protozoa in groups a, b and c were very significantly (P<0.01) or significantly (P<0.05) higher than the control. In conclusion, this study shows that supplementary feeding can positively affect the rumen metabolic parameters and the number of several rumen microorganisms in Gansu Alpine Fine Wool ewes. In general, the effects of Diet 1 were the strongest, while Diet 2 also produced notable improvements. © 2018, Editorial Office of Acta Prataculturae Sinica. All rights reserved.     

湖南省某生猪批发市场疫病传播风险评估

为评估湖南省某生猪批发市场猪群疫病的发生风险,找出可能的风险因素,便于提出针对性预防措施,采用自行设计的调查问卷,结合访谈和现场调查,对27户经营户进行了调查,并将调查结果和调运数据相结合进行了疫病传播风险定性评估。结果显示:该市场疫病传入的可能性"高",内部传播的可能性"中等",疫病传出的可能性"高"。生猪调运频次多、数量大,经纪人行为复杂,运输车辆清洗消毒不彻底是可能的风险因素。评估结果提示,应尽量减少经纪人行为影响,重点加强对运输车辆的清洗消毒。     

农五师葡萄钾素动态变化及调节措施研究

通过叶分析研究了不同葡萄品种叶柄全钾5～9月含量动态变化规律.结果表明:葡萄叶柄钾含量在生育期内整体呈波浪式下降趋势,前期下降迅速,中期平缓,进入浆果成熟期后又趋于下降；强势品种在整个生长季节内对钾的吸收大于弱势品种；早熟品种叶柄内钾含量变化趋势比晚熟品种提前.根据新疆农五师葡萄园区土壤有效钾含量分布现状,建议生产中应根据葡萄钾元素动态变化特点,不同团场应结合当地土壤有效钾含量相应补充.     

白藜芦醇和褪黑素对绵羊胎儿成纤维细胞转染及卵母细胞体外成熟的影响

试验旨在利用白藜芦醇(Re)和褪黑素(MT)提高绵羊胎儿成纤维细胞电穿孔转染(电转)效率、优化卵母细胞体外成熟体系,以提高转基因动物生产效率。将绵羊胎儿成纤维细胞分为6组:对照组、添加白藜芦醇高、中、低(R^-5、R^-6、R^-7)3种浓度组及白藜芦醇和褪黑素联合添加组(R^-5M^-5、R^-6M^-7),通过电转效率和细胞凋亡情况确定电转体系优化方案;通过对细胞周期、线粒体膜电位及活性氧(ROS)的检测研究其作用机制。将绵羊卵母细胞分为对照组及白藜芦醇和褪黑素(R^-5M^-5、R^-6M^-7)联合添加组,通过绵羊卵母细胞体外成熟后卵丘细胞扩展、第一极体排出及体外受精胚胎的卵裂率检测其对卵母细胞体外成熟的作用。研究结果显示,分离的绵羊胎儿成纤维细胞P3代增殖旺盛,细胞生长曲线呈典型的S型;与对照组相比,所有试验组胎儿成纤维细胞的电转效率均显著提高(P<0.05),R^-5M^-5组电转效率最高;R^-6M^-7组显著降低细胞凋亡率(P<0.05),R^-5、R^-6、R^-5M^-5和R^-6M^-7组均显著促进细胞增殖(P<0.05);R^-6、R^-7、R^-5M^-5和R^-6M^-7组G1期细胞增加,S期细胞相对减少,但差异均不显著(P>0.05);R^-6、R^-7、R^-5M^-5和R^-6M^-7组成纤维细胞内ROS水平均显著降低(P<0.05),R^-6、R^-5M^-5和R^-6M^-7组线粒体膜电位均显著增加(P<0.05);R^-6M^-7组显著促进绵羊卵丘细胞扩展、提高卵母细胞第一极体排出率和体外受精胚胎发育能力(P<0.05)。综上,白藜芦醇和褪黑素可以优化绵羊胎儿成纤维细胞电转体系,提高卵母细胞体外成熟效率,为白藜芦醇和褪黑素在转基因动物生产中的应用提供参考依据。     

2014年全国桑蚕新品种实验室鉴定（陕西点）报告

2014年春季对来自镇江蚕种场、苏州大学、无锡西漳蚕种场、山东广通蚕种集团有限公司及山东农业大学的5对桑蚕新品种,秋季对来自苏州大学、西南大学、中国农业科学院蚕业研究所的3对桑蚕新品种进行了实验室鉴定。通过龄期观察和生命率、茧质、产茧量调查与分析,客观反映参鉴品种在陕西试验点的饲育表现和性状特点。     

Copper‐Induced Oxidative Stress and Responses of the Antioxidant System in Roots of Medicago sativa

The effects of various copper (Cu) concentrations on the antioxidative system in the roots of Medicago sativa were explored. The results indicated that the Cu content of the roots reached a value of 854 μg g^?1 DW at 10 μm Cu and a value of 4415 μg g^?1 DW at 100 μm Cu, suggesting that M. sativa has better ability to tolerate and accumulate Cu than other Cu‐bioaccumulators, and is a potential plant for phytoremediation. Treatment with Cu resulted in a significant increment in the levels of H₂O₂, O₂˙^? and OH˙. The reduced form of ascorbate and glutathione reached a peak at 30 μm Cu, and was followed by a sharp depletion to a lower level than that of the control. In contrast, the levels of the oxidised forms of ascorbate and glutathione showed a progressive increment with increasing Cu concentrations, suggesting that the antioxidant system was unable to cope with Cu stress at higher Cu levels. Under the Cu concentrations tested, the activity of catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APX, EC 1.11.1.11) and glutathione reductase (GR, EC 1.6.4.2) increased at lower Cu concentrations, and then decreased, reaching a maximum at 30 μm of Cu for APX and GR, at 10 μm for CAT, whereas the activities of guaiacol peroxidase (POD, EC 1.11.1.7) were gradually increased with increasing Cu concentrations. PAGE analysis of superoxide dismutase (SOD, EC 1.1.5.1.1) revealed that one band is a Mn‐SOD and five bands are identified as Cu, Zn‐SOD, whereas Fe‐SOD isoforms were not found in the roots of alfalfa. Cu at 10–100 μm increased the intensity of constitutive isozymes of CAT, APX and POD, whereas it decreased the intensity of isozymes of glucose‐6‐phosphate dehydrogenase (G6PDH, EC 1.1.1.49) significantly. The activities of lipoxygenases (LOX, EC 1.13.11.12) were gradually augmented with increasing Cu concentrations, demonstrating that LOXs are probably involved in production of lipid hydroperoxides and superoxide anion. There was a continuous and pronounced enhancement in the activity of esterase (EST, EC 3.1.1.1) in roots treated with 10–30 Cu μm , whereas EST activity in roots exposed to above 30 μm Cu declined, suggesting that EST plays a protective role under lower Cu concentrations stress.     

副猪嗜血杆菌单克隆抗体的制备及其B细胞抗原表位的鉴定

为筛选具有临床诊断价值的副猪嗜血杆菌(H.parasuis)单克隆抗体(MAb),本研究采用H.parasuis 5型强毒株(HPS-5)免疫BALB/c小鼠,利用常规细胞融合技术制备了一株稳定分泌MAb的杂交瘤细胞株。Western blot结果表明该MAb能够特异性识别所有H.parasuis标准菌株;免疫共沉淀技术与蛋白质谱分析显示该MAb所识别的蛋白为ATP结合盒转运蛋白(ABCT)家族的寡肽透过酶(OppA);利用MAb对噬菌体12肽库进行淘选,8个阳性噬菌体克隆展示有"KTPAE-R"保守序列,对应于H.parasuis SH29755株和SH0165株的469位~475位的氨基酸残基。对OppA氨基酸序列比对结果表明H.parasuis与其它10种猪常见细菌病病原序列一致性为9.1%~74.5%,本研究结果为进一步研究H.parasuis感染的病原学诊断建立了良好的基础。     

黄芩地膜露头栽培密度研究

目的:探索地膜露头栽培模式下陇西同类地区黄芩栽培的最佳密度。方法:采用35 cm黑膜覆盖露头栽培模式,在控制统一行距的条件下,设计4种不同的株距进行田间栽培试验,通过观测黄芩地上部分、地下药用部位的生长量、试验区药材产量和种子产量等生物经济指标,结合经济效益分析,优选黄芩地膜露头栽培的最佳密度。结果:株距5 cm栽培的鲜药产量较高,但药材商品规格、产籽量均低,增产不增收;株距15 cm栽培的鲜药商品规格较高,但产量和产籽量均偏低,经济效益低;株距10 cm栽培的出苗率、鲜药产量、产籽量、田间长势、药材商品规格表现好。结论:黄芩35 cm黑膜覆盖露头栽培,在陇西同类地区的最佳密度为行距30 cm、株距10 cm,即亩保苗22 000-22 500株。     

5-烯醇式丙酮酰-莽草酸-3-磷酸合成酶(EPSPS)基因AM79 aroA的活性位点分析

AM79 aro A(WO/2009/059485)基因是从草甘膦高度污染的土壤中克隆的、具有我国自主知识产权的新型草甘膦抗性基因,具有重要应用价值。为进一步解析AM79 EPSPS蛋白的作用机制,本研究对此蛋白的活性位点进行了研究。进化树分析显示AM79 EPSPS属于Ⅰ型EPSPS。序列比对结果表明AM79 EPSPS中第107位的丙氨酸(Ala),第114位的苯丙氨酸(Phe),第355位的Ala和第356位的组氨酸(His)是特异的。采用重叠延伸PCR介导的核苷酸定点突变技术,对这些保守的氨基酸位点进行定点突变。将AM79 aro A及其突变基因转入大肠杆菌(Escherichia coli)aro A缺陷型菌株ER2799中,并进行草甘膦抗性鉴定。结果显示,Phe114、Ala355和His356等氨基酸的突变,使AM79 EPSPS蛋白的草甘膦抗性降低,表明这些氨基酸位点对于维持AM79 EPSPS高抗草甘膦的能力是必需的。同源建模结果表明,这些保守氨基酸位点的突变,使AM79 EPSPS蛋白的结构发生变化,这可能是导致突变蛋白草甘膦抗性能力降低的原因。两个Ⅰ型EPSPS(荧光假单杆菌(Pseudomonas fluorescens)G2EPSPS和拟南芥(Arabidopsis thaliana)EPSPS)对应氨基酸突变后,其草甘膦抗性没有明显变化,表明AM79 EPSPS中几个特异的氨基酸位点的功能在Ⅰ型EPSPS中并不是保守的。本研究结果为解析AM79 EPSPS高抗草甘膦的作用机理及对AM79 EPSPS进行定向进化提供了理论依据。