杂交玉米新组合金单999的育苗移栽制种技术

金单999是贵州省旱粮研究所选育,适宜南方丘陵和山区种植的高产、稳产、多抗玉米新品种。本文对金单999制种过程中的隔离、肥料、播种、管理、去杂、去雄等环节作了详细的阐述和要求。     

Targeted Knock-in of a Fluorescent Protein Gene into the Chicken Vasa Homolog Locus of Chicken Primordial Germ Cells using CRIS-PITCh Method

In chickens, primordial germ cells (PGCs) are effective targets for advanced genome editing, including gene knock-in. Although a long-term culture system has been established for chicken PGCs, it is necessary to select a gene-editing tool that is efficient and precise for editing the PGC genome while maintaining its ability to contribute to the reproductive system. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) and CRISPR-mediated precise integration into the target chromosome (CRIS-PITCh) methods are superior as the donor vector is easier to construct, has high genome editing efficiency, and does not select target cells, compared to the homologous recombination method, which has been conventionally used to generate knock-in chickens. In this study, we engineered knock-in chicken PGCs by integrating a fluorescent protein gene cassette as a fusion protein into the chicken vasa homolog (CVH) locus of chicken PGCs using the CRIS-PITCh method. The knock-in PGCs expressed the fluorescent protein in vitro and in vivo, facilitating the tracking of PGCs. Furthermore, we characterized the efficiency of engineering double knock-in cell lines. Knock-in cell clones were obtained by limiting dilution, and the efficiency of engineering double knock-in cell lines was confirmed by genotyping. We found that 82% of the analyzed clones were successfully knocked-in into both alleles. We suggest that the production of model chicken from the knock-in PGCs can contribute to various studies, such as the elucidation of the fate of germ cells and sex determination in chicken.     

凝胶渗透色谱净化-气相色谱分离同时测定饲料中30种有机磷农药残留

文中建立了凝胶渗透色谱净化-气相色谱法同时测定饲料中敌敌畏、乐果等30种有机磷农药残留的方法。饲料样品用乙腈体系提取,采用凝胶色谱柱净化,毛细管色谱柱分离(DB-50+,30 m×0.25 mm×0.25μm),火焰光度检测器(FPD)检测,外标法定量。其方法检测限为5.6~54μg/kg,三水平添加回收率为71.3%~114.1%,RSD为0.3%~14.8%。方法简便、准确,并能满足饲料中多种有机磷农残测定的需要。     

基于实时荧光定量PCR技术的猪伪狂犬病活疫苗含毒量评价

为定量分析贵州省临床用猪伪狂犬病(pseudorabies,PR)活疫苗中病毒含量,根据GenBank中公布的伪狂犬病病毒(pseudorabies virus,PRV)gB基因序列(登录号:M17321.1)设计1对特异性引物,建立实时荧光定量PCR方法,并对来自6个不同厂家的猪伪狂犬病活疫苗进行含毒量分析。结果显示,试验成功建立了可用于PRV检测的实时荧光定量PCR方法,标准曲线中标准品各稀释度质粒拷贝数与Ct值呈良好的线性关系,标准曲线方程为:Y=-0.97X+33.66(R^2=0.999),该方法最低检测病毒含量为3.19×10~1拷贝/μL,敏感性高且具有良好的重复性和特异性。应用所建立的实时荧光定量PCR方法对贵州省临床用6个厂家生产的猪伪狂犬病活疫苗的病毒含量进行检测,结果显示,6种市售猪伪狂犬病活疫苗(A～F)中病毒含量分别为1.67×10~7、4.83×10~5、2.64×10~6、4.27×10~7、3.39×10~6和3.68×10~5拷贝/μL,来自不同厂家的疫苗病毒含毒量存在明显差异,最大差异可达116倍,其中来自A、D厂家的两种猪伪狂犬病活疫苗具有较高的含毒量。本试验所建立的实时荧光定量PCR方法可用于猪伪狂犬病活疫苗病毒含量的快速检测和评价,对猪伪狂犬病活疫苗的质控和临床用疫苗选择具有一定的指导意义。     

苍耳对不同植物幼苗的化感作用研究

在室内采用离体生物测定法,测定了苍耳水浸提物对小麦、高粱、黄瓜、油菜和萝卜的化感效应。结果表明,苍耳水浸提液对5种植物种子的萌发和生长均有较强的抑制作用,且相同浓度处理时,对胚根的生长抑制作用高于对胚芽的抑制作用。5种受体植物中,以油菜最为敏感,其次为萝卜。本研究还通过测定化感物质对植物光合作用的影响及MDA含量的变化,初步探讨了苍耳化感物质的作用特征。在0.025 g/mL浓度下,苍耳对去胚乳小麦幼芽、幼根的抑制率分别为58.33%和67.31%;同样浓度下,苍耳对黑暗条件下培养的高粱幼芽、幼根的抑制率分别为5.58%和32.34%;可以看出,苍耳对植物光合作用的影响明显高于对非光合作用的影响。化感物质处理后黄瓜和萝卜幼苗MDA含量均高于未加化感物质的对照处理,且化感物质浓度越高MDA含量越高。由此可以看出,苍耳对植物的化感作用与抑制植物的光合作用并使植物的MDA含量升高有关,对植物的非光合作用亦有不同程度的影响。     

Clinical and immunological heterogeneity of canine subepidermal blistering dermatoses with anti‐laminin‐332 (laminin‐5) auto‐antibodies

Laminin‐332 (laminin‐5) is a basement membrane heterotrimeric protein composed of alpha‐3, beta‐3 and gamma‐2 laminin chains. Laminin‐332 polypeptides are targeted by auto‐antibodies in human patients with mucous membrane (cicatricial) pemphigoid or, more rarely, subepidermal vesicular diseases that resemble epidermolysis bullosa acquisita (EBA) or bullous pemphigoid (BP). The objectives of this report were to characterize the clinical, histopathological and immunological characteristics of nine dogs with auto‐antibodies targeting laminin‐332. Immunological investigations consisted of direct immunofluorescence (IF), indirect IF with intact and salt‐split canine gingival, and salt‐split normal or laminin‐332‐deficient human skin, immunoblotting with purified human laminin‐332 and immunoblotting with recombinant NC1 domain of human collagen VII. All dogs exhibited varying degrees of skin blistering and ulceration associated with microscopic subepidermal vesiculation with or without inflammatory cells. Indirect IF established that circulating IgG auto‐antibodies bound the dermal side of salt‐split canine lip and human skin. In five dogs, IgG variably recognized the basement membrane of laminin‐332‐deficient human skin (three dogs negative, two dogs positive). In all nine dogs, IgG auto‐antibodies detected purified human laminin‐332 by immunoblotting. In two dogs, additional targeting of collagen VII‐NC1 was present. These observations establish laminin‐332 as a novel basement membrane antigen in dogs with autoimmune blistering diseases with variable clinical phenotypes. The names ‘acquired junctional epidermolysis bullosa’, ‘anti‐laminin‐332 mucous membrane pemphigoid (MMP)’ and ‘mixed auto‐immune subepidermal blistering dermatosis’ are proposed for dogs with clinical signs reminiscent of EBA, MMP or BP respectively.     

苜蓿中草酸含量的测定及煮制对其含量影响研究

紫花苜蓿营养丰富,越来越受到大家的青睐,但苜蓿中的草酸会对人体健康产生间接影响.本文通过对比实验研究了煮制对苜蓿中草酸含量的影响情况.结果显示:新鲜苜蓿中含有一定量的草酸,但煮制后能去除约75％的草酸,故水煮苜蓿是有效去除草酸并保留营养成分的首选方法.     

万州区农业产业化、农村工业化、农村城镇化发展不协调的表现研究

通过大量调查，分析了万州区农业产业化、农村工业化、农村城镇化发展不协调的表现。具体表现在：农业产业化与农村工业化发展不协调，工业化程度偏低，农业产业化程度不高，没有形成规模经济和产业链；农业产业化与农村城镇化发展不协调，影响市场容量的扩大，制约农业自身的发展，制约第三产业发展和农民进入城镇就业步伐，城镇化辐射带动能力弱；农村工业化与农村城镇化发展不协调，城镇对生产要素的集聚功能差，农村工业布局较分散且技术落后，“空城现象”明显；农业产业化、农村工业化、农村城镇化与区域经济发展不协调，区域经济发展水平不高，区域经济结构运行质量不高，区域经济的市场竞争力不强，区域经济的发展环境不优。     

大骨鸡MSTN基因的表达检测

以大骨鸡为试材,采用RT-PCR法检测了MSTN基因在大骨鸡不同器官中的表达情况。结果表明,MSTN基因在大骨鸡骨骼肌中表达水平较高,在心肌、肾脏、脑、肠、舌中也有微量表达,但在肺、肝脏中未检测出MSTN mRNA,而且在14日龄时表达水平高于35和56日龄。     

玉米种子培养皿法发芽试验误差分析

培养皿法是玉米种子发芽试验常采用快捷方便的方法,但此方法的发芽率比较低、变异系数较大,不能反映种子本来固有的发芽水平。对此,笔者进行了培养皿法发芽试验的研究,结果表明,在培养皿内种胚向下是造成发芽率低和变异系数大的原因,若将种胚全部向上置放,就能减少发芽率的误差,体现出玉米种子的正常发芽水平。