银狐垂体细胞系构建研究

分别采用组织块法和消化法2种接种方法进行银狐垂体细胞系构建研究。在消化法中使用不同浓度的胰蛋白酶来检验效果,同时在原代培养期使用反复差速贴壁法纯化,传代培养期结合使用反复差速贴壁以及两步消化法进行处理。结果表明：组织块法接种后成纤维细胞生长明显,原代培养后期成纤维细胞已为主要细胞;降低胰蛋白酶浓度能降低对细胞损伤,但消化时间会增长;采用消化法接种在抑制成纤维细胞生长方面效果好于组织块法。纯化效果上,结合反复差速贴壁法与两步消化法处理后获得了较好的效果,在原代培养时期能较好去除成纤维细胞并通过持续使用该方法可在传二代后获得较高纯度的细胞,纯度能达到80%以上,能够建立细胞系。     

北京油鸡胚胎成纤维细胞系建立与生物学特性研究

采取组织块直接培养法，对北京油鸡胚胎组织进行原代和继代培养，成功地建立了成纤维细胞系。并对培养细胞进行了形态学、细胞生长动力学观察，以及核型和乳酸脱氢酶、苹果酸脱氢酶的同工酶分析。结果表明，该细胞系的群体倍增时间(PDT)为24h；细胞染色体中二倍体占主体，为76％～88％；乳酸脱氢酶、苹果酸脱氢酶同工酶电泳图谱与本室其它细胞系有明显差别；细菌、真菌、病毒、支原体检测呈阴性。该细胞系的建立，使北京油鸡这一国家重要种质资源在细胞水平上保存下来，也为基因组文库和体细胞克隆等研究提供了理想的生物材料。     

南农选系草地早熟禾POD同工酶的酶谱特征及与其它品种的差异

利用聚丙烯酰胺垂直板凝胶电泳技术对南农选系及16 个当前生产上应用较多的草地早熟禾品种的过氧化物酶同工酶进行了研究。结果表明,供试材料共在18 个位点表现酶带。南农选系表现出13条酶带,除与其它16个品种所共有的6 条主酶带外,还出现了其特有的7 条酶带。对不同品种POD同工酶酶谱信息进行相异性类平均聚类分析,17 个供试品种(系)聚为3 类,其中南农选系、美国品种“Merit”及其它15个品种分别各成一类。据此推测,南农选系与其它供试品种的亲缘关系相对较远,为一个相对独立的品系。     

苜蓿雄性不育系组织培养技术的研究

以苜蓿雄性不育系Ms-4幼嫩茎段、叶片、叶柄为外植体,诱导苜蓿雄性不育系再生植株.结果表明:叶柄是诱导愈伤组织的良好外植体,体细胞胚诱导率为90%,再生率49%.苜蓿雄性不育系插条生根的最佳处理为接种前用水进行预处理0.5h,外植体为完整插条,培养基为A1、A2,其生根率为97%,生根系数为5.3.     

The effect of hindclaw height differential and subsequent trimming on lameness in large dairy cattle herds in Canterbury,New Zealand

Abstract

AIMS: To determine if inequality in height between claws within each hindlimb of dairy cattle is a risk factor for the development of lameness and to determine the effect of preventive trimming of these claws on the subsequent risk of lameness.

METHODS: Cows (n=2,695) on three dairy farms in Canterbury, New Zealand, were examined and the height differential between their hind claws on each limb was measured. Cows were blocked by age, then breed and grouped by height differential, before being randomly assigned to either treatment or control groups. Cows in the control group were not trimmed, whilst cows in the treatment group were all trimmed according to the ‘Dutch Trimming’ method by an experienced foot trimmer. All subsequent clinical incidences of lameness were identified, recorded and classified according to five categories. The overall incidence of lameness, the effect of claw height differential on the incidence of lameness and the effect of trimming on the incidence of lameness were assessed using both GEE modelling and survival analysis.

RESULTS: The incidence of lameness identified in any untrimmed hindlimb irrespective of diagnosis, measured at a limb level and assessing the first incidence of lameness only, was 10.0% for a 6-month period from November to drying off. The predominant lesion identified was white line disease (WLD; 7.9% incidence and 78.6% of all lameness). The overall incidence of hindlimb lameness measured at cow level for the same period was 14.5%. The risk of lameness was influenced by breed (p=0.016), age (p=0.002) and claw height differential (p=0.026). There were no significant interactions. The risk of lameness was lowest where the claw height differential was 0–2 mm. The incidence of lameness in the first 70 days following trimming was higher in older cows than in younger cows (p<0.001). Trimming did not affect lameness incidence (p=0.185). The Cox proportional hazard model demonstrated that farm (p<0.001), trimming group (p=0.021); and age (p=0.021) significantly affected the median days to lameness in the first 70 days following trimming.

CONCLUSIONS AND CLINICAL RELEVANCE: Hind feet with lower height differentials between claws had a lower incidence of lameness. Trimming of these feet to remove the height differential did not reduce their overall incidence of lameness, but did increase days to first lameness.     

水稻两用核不育系育性转换光温反应型的分类研究

 在人工控制的9种光温处理下，对76份水稻两用核不育系的育性转换光温反应进行了研究。根据自交结实率的光周期效应、温度效应和光温互作效应的分析，供试的两用核不育系中，占96%的不育系的育性转换光温反应型可归为以下3种：（1）光周期敏感不育型，其统计学特征是育性转换的光温互作效应和光周期效应显著而温度效应不显著；（2）温度敏感不育型，其统计学特征是育性转换的温度效应显著而光周期效应不显著、光温互作效应显著或不显著；（3）光温互作不育型，其统计学特征是育性转换单独的光周期效应和温度效应不显著，仅光温互作效应显著。在供试的粳型不育系中，有34.8%、13.0%和47.8%的材料分属上述3种光温反应型;在供试的籼型不育系中,有73.6%和22.6%的材料分属温敏型和光温互作型,没有一份材料为光敏型。结果表明现有的两用核不育系育性转换光温反应是多型性的。文中还对实用性籼型两用核不育系育性转换光温反应型的选择问题作了讨论。     

猪原始生殖嵴细胞(PGCs)建系因素的研究

从五指山猪(WSZP)近交系第8～13代培育群中，先后选用21头5～10月龄青年母猪，分别于授精后25～30d采集胎儿106个，进行原始生殖嵴(PGCs)细胞分离、培养等建系技术研究。以DMEM F10(1:1)为基础培养液，按添加或不添加生长因子，将培养液分为A、B、C3种，并以STO细胞作饲养层，在38℃、5.0％CO2和湿润的气相中进行培养建系。结果获得胚胎生殖嵴细胞(EG)细胞系6个细胞株，其中1个EG细胞株传至11代、2个传至5代、1个传至4代、2个传至3代冻存。并进行了AKP染色、体外分化、冷冻-解冻复苏和嵌合体制作等鉴定研究。研究发现：不同胚龄对EG细胞建系具有一定影响，不同培养液对EG细胞建系效果不同，STO细胞饲养层的质量是建株、传代、冷冻-解冻复苏的关键因素之一。EG细胞系的初步建立，为今后筛选进入种系的EG细胞系、实施体外基因操作提供了可能。     

花粉管通道法在玉米自交系改良中的应用

就几年来花粉管通道法在玉米自交系改良中的应用情况加以概括,提出了导入方法的适宜细则,对后代变异情况加以论述,并对今后该项工作的开展提出几点建议.     

鲁西黄牛体细胞克隆保种技术的研究

以体外培养的鲁西黄牛耳皮肤成纤维细胞作核供体，研究利用体细胞克隆技术保存我国地方黄牛优良品种的技术方法。通过组织块培养法建立的鲁西黄牛（1♂，4♀）成纤维细胞系，作为核移植供体细胞，利用屠宰场母牛卵巢卵母细胞经体外培养成熟后作为核受体进行核移植，试验结果表明：重构胚的融合率为62．5％（242／387），分裂率为63．6％（154／242），体外培养第7天囊胚发育率为42．9％（66／154）。体外培养第7天的囊胚的内细胞团细胞（ICM）与滋养层细胞数平均为37和47，ICM占44．2％。体外发育到7d的囊胚新鲜胚胎的移植妊娠率（新鲜胚胎）移植受体10头，60d妊娠率为20％（2／10）。