Variability amongst strains of Fusarium poae assessed by vegetative compatibility and RAPD polymorphism

Vegetative compatibility tests and random amplification of polymorphic DNA (RAPD) were used to assess genetic relationships amongst 54 strains of Fusarium poae obtained from various geographical regions. Twenty-seven strains were assigned to eight multiple member vegetative compatibility groups (VCGs), while the other 27 isolates were found to form single-member VCGs. There was a partial correlation between VCG and geographical origin, but the relationship was not always clear. However, no correlation was observed between the VCG and the host plant of origin. RAPD patterns were closely associated with VCGs in all cases. Members of VCGs that were interconnected by bridging isolates formed common branches in the phenogram constructed on the basis of the RAPD patterns, while strains that belonged to single-member VCGs were scattered throughout the phenogram. These data demonstrate that the combination of traditional and molecular methodologies allows reliable intraspecific subdivisions in an asexual fungus, which is a secondary invader of a wide range of host plants, and so has never been subject to the intense selection pressure of a single host species and lacks pathogenic subgroups.     

空间诱变甜椒M3代果实营养品质的变异分析

试验结果表明:空间诱变甜椒的3个营养品质性状(果实维生素C含量、可溶性固形物含量和糖酸比)在M3代的变异趋于稳定,株系内的株间变异均小于株系间变异,果实维生素C含量和糖酸比在群间的差异不显著,可溶性固形物含量在群间的差异显著;维生素C含量与可溶性固形物含量呈极显著正相关,与糖酸比呈极显著负相关,可溶性固形物含量与糖酸比呈显著正相关。宜筛选单株果实维生素C含量高、可溶性固形物含量和糖酸比适中的株系。优选的株系为A6、A7、C7、B2、B6、B1、A5、C5、B7、C4、A3、B5、A4和C6。     

三个牦牛群体激素敏感脂肪酶（HSL）基因外显子Ⅰ的多态性

 【目的】探究激素敏感脂肪酶（HSL）基因在牦牛内的遗传多态性,为进一步揭示牦牛群体间遗传分化、开展牦牛肉质性状的关联分析以及HSL基因在牦牛物种中的定位、表达调控等研究提供依据。【方法】采用PCR-RFLP方法对九龙牦牛、麦洼牦牛和巴州牦牛共92头牦牛的HSL基因部分外显子I进行SmaI酶切多态性分析;统计基因频率、基因型频率大小,进行卡方（χ2）适合性、独立性检验,并计算纯合度、杂合度、有效等位基因数和多态信息含量等遗传多态参数。【结果】3个牦牛群体在HSL基因外显子I具有SmaI酶切多态性,在该酶切位点存在AA、AB和BB 3种基因型。3个牦牛群体均检测到AA和AB基因型,而BB基因型只在巴州牦牛中检测到。九龙牦牛和麦洼牦牛的AA基因型频率最高,而巴州牦牛AB基因型频率最高;A等位基因频率在3个牦牛群体中均高于B等位基因频率,为优势等位基因。适合性检验表明3个牦牛群体在该酶切位点均处于Hardy-Weinberg平衡状态;独立性检验表明九龙牦牛与麦洼牦牛、巴州牦牛之间表现为差异显著（0.01＜P＜0.05）和差异极显著（P＜0.01）,而麦洼牦牛与巴州牦牛之间表现为差异不显著（P＞0.05）。测序分析表明,HSL基因外显子I第70位（从PCR产物测序片段的5′端计数）发生了单碱基突变（G→A）,并导致了编码氨基酸由甘氨酸（G）变为精氨酸（R）。【结论】牦牛在HSL基因外显子I内具有SmaI酶切多态性,其多态性是因所分析序列内一单碱基突变（G→A）所致,该碱基转换导致了氨基酸由甘氨酸（G）变为精氨酸（R）;在该酶切位点,九龙牦牛、麦洼牦牛和巴州牦牛均处于Hardy-Weinberg平衡状态。九龙牦牛与麦洼牦牛、巴州牦牛之间表现为差异显著（0.01＜P＜0.05）和差异极显著（P＜0.01）,而麦洼牦牛与巴州牦牛之间表现为差异不显著（P＞0.05）。     

Effects of vaccination against brucellosis and clostridia on the intake,performance, feeding behavior,blood parameters,and immune responses of dairy heifers calves

The aim of this study was to identify possible effects of different vaccination strategies (concomitantly or not) against brucellosis and clostridia on intake, performance, feeding behavior, blood parameters, and immune responses of dairy heifers calves. Fifty heifers calves were enrolled [38 Gyr (Zebu, Bos taurus indicus) and 12 5/8 Holstein Gyr]. At 120 d of age, animals were randomly distributed among 3 groups: B (n = 18), vaccinated against brucellosis; C (n = 14), vaccinated against clostridia and CB (n = 18), vaccinated concomitantly for both. Rectal and thermographic temperatures were evaluated on days 1, 0, 1, 2, 3, 5, 7,10, 14, and 28 relatives to the vaccination day. Feed and water intake, body weight (BW), and feeding behavior were monitored daily by an electronic feeding system. Blood was sampled on days 0, 3, 7, 14, and 28, relative to the vaccination day for determination of glucose and -hydroxybutyrate (BHBA) concentrations. Blood sampled on day 0 (prevaccination) and on days 28 and 42 were used to evaluate the immune response against Brucella abortus and clostridia. There was an increase in rectal temperature between the first and the third day postvaccination in the 3 groups. The thermography revealed an increase of local temperature for 7 d on groups B and CB. Group C had increased local temperature for a longer period, lasting for up to 14 d. Dry mater intake was reduced for groups B and CB, but no alteration was observed for group C. No alterations regarding initial BW, final BW, average daily weight gain, and feed efficiency were observed. No differences were observed for the 3 vaccination groups for blood parameters throughout the evaluation period. The concomitant vaccination against brucellosis and clostridia led to lower neutralizing antibody titers against epsilon toxin of Clostridium perfringens and botulinum toxin type C of C. botulinum (C > CB > B). When cellular proliferation assay and serological tests to B. abortus were evaluated, no differences were observed between groups B and CB. The present results indicate that the concomitant vaccination against brucellosis and clostridia has no relevant impact on the intake, performance, and feeding behavior of dairy calves. However, the concomitant vaccination of vaccines against these 2 pathogens impacts animal immunity against clostridial infections.     

牦牛StAR基因克隆及其生物信息学与组织表达特性的研究

旨在克隆获得牦牛StAR基因编码序列（CDS）并进行生物信息学分析,探究其mRNA组织表达特性。本研究以屠宰场采集的成年母牦牛心、肝、脾、肺、肾、卵巢、输卵管、子宫组织（n=5）,不同年龄（胎牛、1岁、2岁）牦牛的卵巢（n=3）,不同发情周期（卵泡期、黄体期）的牦牛卵巢（n=3）,黄体期黄牛的卵巢（n=3）及实验室冻存的牦牛颗粒细胞为研究材料。以牦牛黄体期卵巢cDNA为模板,用逆转录PCR克隆StAR基因,并使用MEGA7.0和ExPASy-ProtParam等软件分析其生物信息学特性;采用实时荧光定量PCR技术分析牦牛StAR基因组织表达特性。结果发现,StAR基因CDS区长858 bp,编码285个氨基酸,StAR蛋白总体带正电荷,属于碱性亲水稳定蛋白,无跨膜结构及信号肽,主要存在于细胞质和线粒体; StAR基因具有较高的保守性,符合物种进化规律。牦牛StAR基因在卵巢表达水平最高（P<0.01）,且2岁时卵巢表达水平极显著高于胎牛和1岁龄（P<0.01）,黄体期卵巢表达水平极显著高于卵泡期（P<0.01）;黄体期黄牛卵巢中StAR基因的表达量极显著高于牦牛（P<0.01）;在颗粒细胞的体外培养过程中StAR基因表达量逐渐上升,在培养24 h时达到高峰（P<0.01）,随后显著降低。综上所述,StAR基因序列较为保守,在牦牛卵巢组织中表达最高,且表达水平随年龄与卵巢周期而变化,提示StAR基因可能参与牦牛卵巢及黄体功能相关的繁殖调控。     

妊娠后期营养限制对蒙古绵羊胎儿生物原胺类神经递质的影响

本试验旨在研究妊娠后期营养限制对蒙古绵羊胎儿生物原胺类神经递质的影响。选择健康、体况相近的蒙古绵羊18只,对其进行同期发情、配种后,从妊娠第90天开始,随机分为3个组:营养限制1组[NG1组,n=6,代谢能(ME)=0.175 MJ/(kg BW 0.75·d)]、营养限制2组[NG2组,n=6,ME=0.330 MJ/(kg BW 0.75·d)]和对照组[CG组,n=6,ME=0.670 MJ/(kg BW 0.75·d)],在妊娠第140天时屠宰,取各组胎儿及其脑组织和血液。结果显示:NG1组胎儿的脑重较CG组显著增高(P<0.05),并且NG1组胎儿脑中5-羟色胺(5-HT)(P<0.01)、肾上腺素(EPI)(P<0.05)含量显著或极显著高于CG组,去甲肾上腺素(NA)的含量极显著低于CG组(P<0.01)。NG2组胎儿脑中5-HT含量极显著高于CG组(P<0.01),而NA(P<0.01)、多巴胺(DA)(P<0.05)含量显著或极显著低于CG组。另外,NG1组胎儿血浆中5-HT(P<0.01)、EPI(P<0.01)、NA(P<0.05)、DA(P<0.01)含量显著或极显著低于CG组;NG2组胎儿血浆中5-HT(P<0.01)、NA(P<0.01)、DA(P<0.05)含量显著或极显著低于CG组。由此得出,妊娠后期营养限制使得蒙古绵羊胎儿脑中5-HT、EPI含量升高和NA含量降低,血浆中5-HT、EPI、NA、DA含量降低。     

配施有机肥条件下油菜化肥氮减施潜力研究

探明湖南省油菜种植区配施有机肥条件下化肥氮减施潜力,为该地区油菜生产中氮肥减施增效提供参考。田间试验在湖南省郴州市进行,供试油菜品种为湘油420,采用裂区设计,研究配施等量(2250 kg/hm²)有机肥条件下不同氮肥用量(0、90、135、180、225、270 kg/hm²)对油菜生长、产量、理论产油量和经济效益的影响。结果表明,所有氮肥水平下配施有机肥均显著提高油菜产量,且在低氮水平下增产效果更加明显,有机肥的增产效果主要通过增加单株角果数和每角果粒数来实现。在不施与施有机肥条件下,施氮量分别为213和199 kg/hm²时油菜籽产量最高,分别为1754和2514 kg/hm²,氮肥减少了6.6%、产量提高了43.3%;施用有机肥条件下达到不施有机肥的最高产量值仅需127 kg/hm²氮肥,减少了44.1%。所有供氮水平下,施用有机肥可显著增加氮素积累量、理论产油量和经济效益,包括有机肥氮的氮肥偏生产力也呈增加趋势,而包括有机肥氮的氮肥农学效率和氮肥表观利用率变化较小;与不施有机肥相比,施有机肥的最高理论产油量施氮量减少了3.0%,最高效益施氮量减少了20.0%,而理论产油量和效益分别增加了41.1%和50.0%。     

高寒草甸土壤微生物功能多样性对氮肥添加的响应

采用Biolog-ECO生态板法研究了4个施N水平[0 g/m²（CK）,10 g/m²（N₁₀）,20 g/m²（N₂₀）,30 g/m²（N₃₀）]下土壤理化性质和微生物功能多样性的变化规律。结果表明:中高水平氮肥添加显著增加了速效氮、速效磷含量和根土比,但降低了土壤含水量。施N肥0-10 cm土层平均颜色变化率降低,而10-20 cm土层则被提高,且N₂₀最大。施N肥后降低了0-10 cm土层土壤微生物Shannon-Wiener指数、Pielou指数和McIntosh指数,但10-20 cm土层则被提高。PCA分析表明,施N肥改变了土壤微生物代谢功能类型,糖类、氨基酸、酸类是土壤微生物主要利用的碳源类型。RDA分析表明,在0-10 cm土层,土壤含水量和全氮是影响土壤微生物功能多样性的主要因子,而10-20 cm土层主要受土壤含水量和速效磷的影响。综上,施N肥增加了土壤的有效养分含量,进而改变土壤微生物功能多样性、碳源利用能力和数量。     

沥青混凝土心墙抗水力劈裂可靠度分析

沥青混凝土心墙抗水力劈裂是高心墙堆石坝的关键技术问题,其可靠度计算在国内尚属空白,本文提出了非线性有限元与响应面法、JC法相结合的可靠度分析方法,构建了沥青混凝土心墙抗水力劈裂的功能函数,以目前亚洲最高的沥青混凝土心墙堆石坝--冶勒为例,根据非线性有限元对各随机变量波动的计算结果,并应用JC法计算了其结构可靠度指标.结果表明,沥青混凝土心墙β值为5～7,总体水平满足规范要求β低值区主要分布在沥青混凝土心墙中下部,其成果与传统方法计算出的安全系数以及沥青混凝土心墙温度监测分析结果一致,心墙无破坏.该方法计算效率高,实用性强,成果合理、可信,具有借鉴推广应用价值.     

等壳碾压混凝土重力坝层间抗滑稳定可靠度分析

针对以有限元为基础分析碾压混凝土重力坝层间可靠度其功能函数不能显式表达的特点,采用响应面法、JC法、窄界限法原理相结合的可靠度计算方法。由三维非线性有限元对等壳水电站碾压混凝土重力坝各随机变量分位点值的计算结果,采用抗剪断公式构建该电站非溢流坝段层面的功能函数,求得响应面方程,并应用JC法和窄界限法计算层间单元可靠度指标及层间体系可靠度指标,进一步分析材料的变异系数对体系可靠度的影响。结果表明,φ是影响体系可靠度指标的最敏感因子,碾压各层β值一般为4.16～5.70,层间体系可靠度β'为4.12,满足工程要求。该方法计算效率高,实用性强,用于碾压混凝土重力坝层间抗滑稳定可靠度分析是可行的,值得推广。