NaCl胁迫下黑麦草种子萌发过程中DNA甲基化与基因表达分析

为了解黑麦草（Lolium perenne）种子在正常条件与NaCl胁迫下萌发过程中DNA甲基化动态变化以及重要盐胁迫相关基因的表达。运用甲基化敏感扩增多态性技术分析对照和150 mmol·L^-1 NaCl处理下黑麦草种子萌发过程（0,1,2,4,7 d）的DNA甲基化水平及动态变化,并通过实时荧光定量 PCR（QRT-PCR）检测8个重要的盐胁迫相关基因的表达情况。结果表明：黑麦草种子萌发过程中甲基化水平呈下降趋势,以双链甲基化方式为主,甲基化变化同时发生在编码序列和非编码序列中。NaCl处理下DNA甲基化程度高于CK,而去甲基化程度低于CK,最终导致NaCl处理下基因组净的甲基化位点数目增加。黑麦草种子耐盐萌发过程中代谢增强,8个盐胁迫相关基因表达量呈上升趋势,而NaCl胁迫延缓了种子萌发进程,在大多数时间点的基因表达低于对照。     

Experimental studies with foot-and-mouth disease virus type Asia-1, responsible for the 2005 epidemic in China

This study was carried out to investigate the biological characteristics of the foot-and-mouth disease (FMD) virus strain Asia-1 China/2005, which is responsible for the 2005 epidemic in China. The result showed that this strain is not host restricted, and could not only cause FMD in cattle and sheep but also in pigs by either inoculation or direct contact.     

Spatiotemporal heterogeneity of PPARγ expression in porcine uteroplacenta for regulating of placental angiogenesis through VEGF-mediated signalling

Non-infectious prenatal mortality severely affects the porcine industry, with pathological placentation as a likely key reason. Previous studies have demonstrated that peroxisome proliferator-activated receptor gamma (PPARγ) deficiency causes defects in the uteroplacental vasculature and induces embryonic losses in mice. However, its role in porcine placental angiogenesis remains unclear. In the present study, PPARγ expression was investigated in porcine uteroplacental tissues at gestational day (GD) 25, GD40 and GD70 via quantitative polymerase chain reaction (qPCR), Western blot and immunohistochemistry (IHC). Moreover, the roles of PPARγ in porcine placental angiogenesis were investigated using a cell model of porcine umbilical vein endothelial cells (PUVECs) to conduct proliferation, migration and tube formation assays in vitro and a mouse xenograft model to assess capillary formation in vivo. The results showed that PPARγ was mainly located in the glandular epithelium, trophoblast, amniotic chorion epithelium and vascular endothelium, as indicated by the higher expression levels at GD25 and GD40 than at GD70 in endometrium and by higher expression levels at GD40 and GD70 than at GD25 in placenta. Moreover, PPARγ expression was significantly downregulated in placenta with dead foetus. In PUVECs, knocking out PPARγ significantly inhibited proliferation, migration and tube formation in vitro and inhibited capillary formation in mouse xenografts in vivo by blocking S-phase, promoting apoptosis and downregulating the angiogenic factors of VEGF and its receptors. Overall, the spatiotemporal heterogeneity of PPARγ expression in porcine uteroplacental tissue suggests its vital role in endometrial remodelling and placental angiogenesis, and PPARγ regulates placental angiogenesis through VEGF-mediated signalling.     

鸡lncRNA-MSTRG.15568.9及其预测靶基因的表达

试验旨在了解在鸡睾丸中高表达的1个长链非编码RNA （long non-coding RNA,lncRNA）及其预测靶基因的时空表达规律,研究二者在鸡弱精子症中的调控作用。根据弱精子症和正常北京油鸡公鸡睾丸转录组测序筛选到的1个高表达的lncRNA （MSTRG.15568.9）,采用顺式（cis）作用模式预测其潜在靶基因SPAG4（sperm-associated antigen 4）,进一步采用实时荧光定量PCR方法进行表达量分析。分别选择3只0、5、20、30、45、60周龄正常北京油鸡公鸡,检测MSTRG.15568.9与SPAG4基因在不同周龄公鸡睾丸中的表达量差异;选择30周龄3只正常公鸡,采集睾丸、肝脏和脾脏等8个部位组织样品,检测MSTRG.15568.9与SPAG4基因在不同组织间的表达规律;选择45周龄弱精子症公鸡和正常公鸡各3只,对比MSTRG.15568.9与SPAG4基因在睾丸的表达量差异。结果显示,MSTRG.15568.9与SPAG4存在明显的时空表达差异,且二者表达趋势基本一致。在不同周龄的鸡睾丸组织中,MSTRG.15568.9和SPAG4的表达趋势相近,MSTRG.15568.9在20周龄的表达量显著高于0、5、30、45、60周龄（P<0.05）,0和5周龄表达量显著低于20、30、45和60周龄（P<0.05）;SPAG4在45周龄表达量最高,其次是20周龄（P<0.05）。MSTRG.15568.9和SPAG4在睾丸和肝脏中的表达量均显著高于脾脏、肾脏等组织（P<0.05）;在正常睾丸组织中的表达量均显著高于弱精子症睾丸组织（P<0.05）。综上所述,MSTRG.15568.9与SPAG4基因具有较明显的组织表达特异性,且MSTRG.15568.9可能调控SPAG4基因的表达,参与精子发生与精子活力调控;但其具体作用机制需要进一步探索。本研究可为鉴定与鸡弱精子症调节机制相关的功能基因提供参考。     

牛支原体和牛病毒性腹泻病毒二重二温式PCR检测方法的建立

为建立一种牛支原体(Mycoplasma bovis,MB)和牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)的快速鉴别诊断方法,针对MB的uvr C基因和BVDV的5'端非编码区(5'-UTR)保守基因序列,分别设计两对特异引物,并将三温式PCR扩增程序简化为二个温度梯度,建立了鉴别MB和BVDV的二重二温式PCR方法。该方法能同时扩增MB和BVDV,扩增产物大小分别为412和170 bp。特异性试验结果显示,该方法对参试的所有毒株只扩增MB和BVDV基因组,对其它牛病原体无扩增;敏感性试验结果显示,该方法最低能同时检测到104拷贝的两种目的核酸;干扰性试验结果显示,该方法能同时检测两个模板不同浓度的组合,试验结果不受模板影响。综上,本研究所建立的二重二温式PCR方法特异、敏感、快速、简便,可应用于MB和BVDV临床鉴别诊断和流行病学调查。     

Salmonella enteritidis clearance and immune responses in chickens following Salmonella vaccination and challenge

Our previous work showed that the cell-mediated immunity (CMI) was enhanced by live Salmonella vaccine (LV). The objective of this study was to evaluate the impact of live and killed Salmonella vaccines on Salmonella enteritidis (SE) clearance and to determine if the clearance was mediated by cell-mediated and/or humoral immunity. Chickens were first immunized at 2 weeks of age followed by a booster dose at 4 weeks, challenged with live SE 2 weeks later (6-week-old) and tested for CMI, antibody response and SE clearance 1-week post SE-challenge (7-week-old). Spleen cell proliferation induced by SE-flagella and Concanavalin A (Con A) were significantly higher and SE shedding was significantly lower in the LV group. The splenic CD3 population was significantly lower and B cells were higher in the control group compared to all the SE-challenged groups (with and without vaccination). Serum antibody to SE-flagella and envelope were significantly higher in the KV group compared to all the other groups. These results suggest that LV protects against SE infection, probably by enhancing the CMI.     

广西野生动物非法利用和走私的种类初步调查

广西独特的地理位置使得广西成为全国野生动物贸易的集散地之一。为了解广西野生动物的走私种类及其季节性变化,作者统计了广西2007～2008年已破获的非法利用和走私野生动物案件涉及的种类及其季节性变化。结果表明2007和2008年分别查获案件54起和61起。野生动物走私和非法案件中涉及种类共有71种,隶属15目33科,其中两栖类2种,占所有种数的2.8%;爬行类27种（蜥蜴类8种、龟鳖类5种、鳄类1种、蛇类13种）,占38.0%;鸟类25种,占35.2%;哺乳类17种,占23.9%。2007和2008年查获的非法利用和走私野生动物分别是56种和32种,非法利用或走私的种类主要是珍稀濒危的种类,数量大,种类多。广西的野生动物走私和非法利用需要引起关注。     

EM在养羊业中的应用

EM是一种新的微生物制剂,在动物生产中有着广泛的应用.该文主要综述了在羊只生产中EM制剂对羊只的环境、生长、产品质量、繁殖率、疾病防治等方面的作用效果,为实际应用提供了参考.     

Relationship between oocyte maturation and fertilization on zygotic diversity in swine

Two experiments were conducted to examine how oocyte maturation and fertilization influence zygotic diversity in swine. In the first experiment, the distribution of oocyte maturation was compared to that of zygotic development. Oocytes were aspirated from follicles of 31 gilts and classified into stages of meiosis. Zygotes were flushed from oviducts of 19 additional gilts and classified into stages of meiosis and fertilization. The second experiment examined whether the time from ovulation to fertilization was constant among all oocytes. To test this premise, four to six oocytes from follicles of 10 mated gilts were aspirated just before or during ovulation, stained and transferred back into the oviducts of these same gilts. Zygotes were recovered 10 h later to determine whether the first oocytes ovulated were the more developed zygotes and, conversely, whether the last oocytes to be ovulated represented the lesser developed contemporaries. The skewed (P less than .05) distribution of oocyte maturation was similar to that of zygotic development. Regression of the frequency distribution describing early oocyte maturation resulted in a line with a slope (.59) that was similar to the slope (.58) of the regressed distribution of zygotic development. Likewise, the order of ovulation and order of subsequent stages of zygotic development were similar. These data suggest that variation in zygotic development in swine was due to variability in oogenesis; the time from ovulation to fertilization appeared to be constant.     

抗病虫基因新资源:海洋绿藻孔石莼凝集素基因

植物凝集素(Lecin)具有抵抗植物病原菌和农业害虫的活性以及抗病毒活性,在植物保护上具有广阔的应用前景。从海洋绿藻孔石莼中分离到一种凝集素基因序列,为培育抗病虫作物新品种提供了新资源。