哺乳动物性别决定机理及性别鉴定方法研究进展

哺乳动物的性别决定是由SRY基因为调控中心、多基因参与的级联调控过程。本文综述了性别决定基因及其功能、性别决定的分子模型及性别鉴定的方法等方面的研究进展。     

家畜繁殖生物技术研究现状及趋势

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。     

Quick detection of sex determining region protein gene in mouse fetuses

AIM: To detect quickly the Y-chromosome specific sex determining region protein (Sry) gene in mouse fetuses on embryonic day 14.5 with a PCR method. METHODS: We designed specific primers with the OLIGO 5. 0 software. Templates were prepared in 30 minutes by the following way. About 1 mg embryonic tissue but not fetal liver was suspended, and treated with 200μL of lysis buffer, consisting of PCR buffer containing 20 mg/L proteinase K, 0. 5% NP-40, and 0.05% Tween 40, at 60°C for 15 minutes, heated for 5 minutes at 100 °C, 10μL was used as template. The PCR react ion was performed in 50μL, using two sets of primers specific for Sry gene (chromosome Y) and IL-3 gene (chromosome 11) . PCR conditions and cycle numbers were optimized. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The specificity of the method was conf irmed by fluorescent in situ hybridization (FISH) using a specific male probe on embryonic tissue cells. RESULTS: Electrophoresis showed that PCR product of male control DNA consisted of a 649 bp product representing the IL-3 gene and a 444 bp product representing the Y-specific Sry gene, female control DNA only one 649 bp product. Fetuses with two bands matching those as seen inmale control DNA are the presumpt ive male fetuses. Fetuses, only the IL-3-associated 649 bp band, are the presumptive female fetuses. These were confirmed by FISH. The ent ire procedure took <3. 5 h. CONCLUSION: The established PCR assay offers a quick, simple, accurate, and sensitive detection of sex determining region protein gene in mouse fetuses. This method allowed the preparation and culture of pure male and female hematopoietic stem cells from fetal tissue.     

草菇厚垣孢子的研究

研究了草菇厚垣孢子在不同的C/N比的培养基中产生的数量。结果表明 ,不同的C/N比对草菇厚垣孢子的形成有一定的影响 ,其最佳的C/N比为 30～ 4 0。草菇品种间产生厚垣孢子的数量的差异并不显著。另外 ,本试验首次定量分析了厚垣孢子的产量数 ,对常规性分析方法进行了改进。     

罹患口腔炎乌苏里蝮的相关生理学指标分析

口腔炎是人工养殖蛇类的常见病之一,为了深入了解和尽早做好预防工作,对罹患口腔炎的成年雄蛇乌苏里蝮(Gloydius ussuriensis)进行解剖观察,并检测了相关器官的系数、血液学指标和激素等。结果表明:整体肥满度、躯体肥满度、肾系数等数值罹患口腔炎蛇略高于健康蛇;而肝系数、精巢系数和脂肪体系数等罹患口腔炎的蛇低于健康雄蛇,且前两者呈显著性差异,后者呈极显著性差异。性激素检测结果显示罹患口腔炎的蛇血液睾酮和雌二醇含量均低于健康蛇,且均具极显著性差异。血细胞指标中,红细胞数量、红细胞分布均差、白细胞总数、淋巴细胞数量和淋巴细胞比率等均为罹患口腔炎的蛇高于健康雄蛇,且具显著性差异,但嗜中性粒细胞和淋巴细胞的比值(NLR)、中性粒细胞数量和比率、中间细胞数量及其比率均为患口腔炎的雄蛇低于健康蛇,且具显著性差异。结论:口腔炎不仅影响了主要代谢器官的器官系数的变化,而且血液多项指标和性激素水平也有明显差异。性激素含量的变化将对乌苏里蝮的正常生殖繁育产生不可忽视的影响。另外,NLR和红细胞分布宽度2个血常规指标可作为早期诊断炎症发生的首选参数。     

文冠果实时荧光定量PCR内参基因的筛选

为筛选适用于文冠果(Xanthoceras sorbifolium Bunge)性别分化过程研究和不同器官的内参基因,本研究以文冠果性别分化关键期的顶芽和侧芽、根、茎、叶、种仁、雌能花和雄能花为材料,通过实时荧光定量PCR(qRT-PCR)鉴定7个候选内参基因(Actin,UBQ,EF-1α,TUB,eIF-4α,18S rRNA和GAPDH)在各样品中的表达量,并评价7个基因的表达稳定性。结果表明,7个候选内参基因在不同样品中的表达量存在明显差异,各基因在雌能花中的表达均显著高于其他材料,在雄能花中的表达处于较低水平;适用于文冠果性别分化过程的最优内参基因为UBQ和eIF-4α,适用于不同器官的最优内参基因为EF-1α和GAPDH。研究结果将对今后进行文冠果性别分化过程研究和不同器官中的基因表达分析提供参考。     

半胱胺对育成期水貂生长和饲料转化率的影响

选择健康育成期水貂208只,其中200只为试验组,8只为对照组,采用二因子试验设计:一个因子为半胱胺的不同添加量,分别按每千克体质量添加10、40、70、100、130 mg半胱胺;另一因子为半胱胺的不同添加间隔时间,分别为1、3、5、7、9 d 5个时间间隔.通过饲养试验研究了半胱胺对育成期水貂体质量增加、饲料转化率...     

分子标记技术在工业大麻性别分化研究中的应用进展

工业大麻性别表达影响着工业大麻种子、纤维和药用成分产量,其性别表达除了受到遗传物质控制外,还受到外界环境条件的影响,这种复杂性使得性别研究一直是工业大麻研究的难点和热点。分子标记技术能在DNA水平上鉴定工业大麻性别,开发与工业大麻性别紧密连锁的分子标记可为大麻性别早期鉴定和特定性别分子辅助选择育种提供理论依据。本文就分子标记在工业大麻性别研究中的应用进展进行了总结,并对应用前景进行了展望。     

Relationship between corn yield and chemical composition of the ear leaf measured with different regression models

Abstract

Corn yields and leaf samples were obtained from experimental plots receiving varying rates of N, P and K. Yields were regressed on leaf levels of N, P, K, Ca, Mg, B, Cu, Fe, Mn and Zn as independent variables. Various polynomial regressions were fitted to the yields and “goodness of fit”; of particular mathematical models was used as a basis for evaluating particular biological and statistical concepts.

It was found that no regression, where chemical elements were used as ratios, fit the observations as well as a quadratic polynomial or its square root transformation. This suggests that within the set of data used, emphasis on particular cation or anion ratios does not necessarily result in the “best”; explanation of variation in yield.

“Classical”; growth equations were fitted to corn yields, but were not as precise in predicting yields as the polynomials. A modified stagewise regression procedure was used for fitting one mathematical model, but results were less satisfactory than those obtained with usual least squares procedures.     

人工养殖西伯利亚狍肠道微生物组成的雌雄差异

肠道微生物是一个复杂的生态系统,与宿主的健康、疾病以及其他因素有着密切的关系。目前,关于肠道菌群结构和功能的研究较多,但是对西伯利亚狍肠道菌群的研究还未见报道。西伯利亚狍(Capreolus pygargus)属于反刍动物,有独特的消化特性和微生物群来帮助它们适应高纤维的食物。本研究利用高通量测序技术在Illumina MiSeq平台上对14只狍肠道微生物进行16S rRNA基因分析。结果表明:个体样本中门类组成的差异,雌性个体JY3和雄性个体JY9除Firmicutes,Bacteroidetes,Proteobacteria这3类菌群外,其他门类菌群所占比例都很低。在科水平上,雌性个体中Ruminococcaceae在JY7和JY13肠道内含量相对较少,Bacteroidaceae在JY7,JY10和JY13中含量较高。雄性个体中Ruminococcaceae在JY11和JY14肠道内含量较少,Prevotellaceae在JY4中含量较少。在属水平上,雌性个体特有的菌属为Roseburia,Alloprevotella,Lachnospiraceae_incertae_sedis和Coprococcus;雄性个体特有的菌属为Fusobacterium,Succiniclasticum,Streptococcus和Clostridium sensu stricto。Alpha多样性分析表明,Ace指数范围2128.93—772.98,Chao1指数范围2128.89—785.27,Shannon指数范围5.44—3.78,Simpson指数范围0.07—0.01。在不同物种分类水平上,雌雄狍肠道内的优势菌群有些不同。PCoA(principal co-ordinates analysis)分析显示,狍样本几乎混在一起,没有明显差异。