翻耕补播措施对宁夏荒漠草原土壤碳氮储量的影响

以3种翻耕补播方式(深翻耕补播、浅翻耕补播和免耕补播,播量30kg/hm²)及深翻补播不同播量(7.5kg/hm²、15kg/hm²和22.5kg/hm²)为研究对象,以封育未补播为对照,研究了不同翻耕补播措施对宁夏荒漠草原土壤碳氮储量的影响。结果表明:封育未补播处理可提高0～10cm土层土壤有机碳和全氮含量;深翻耕补播可显著提高10～20cm和20～40cm土层土壤有机碳含量,深翻耕补播和深翻-播量15.0kg/hm²可分别提高土壤10～20cm和20～40cm土层土壤全氮含量。深翻耕补播、深翻-播量15.0kg/hm²、7.5kg/hm²和封育未补播措施均可提高浅层(0～10cm)土壤碳氮比;浅翻耕补播和深翻-播量7.5kg/hm²可提高10～20cm土层土壤碳氮比。深翻耕且播量为30.0kg/hm²的人工补播措施较其他措施更有利于土壤有机碳和全氮的固存,但从操作性和成本的角度考虑,短期...     

鸡软骨细胞体外分离培养鉴定以及过表达miR-15a对软骨细胞的影响

为分析miR-15a在肉鸡不同组织中的表达情况,并探究过表达miR-15a对体外培养鸡软骨细胞的影响。本研究首先通过倒置显微镜观察、PCR、凝胶电泳和甲苯胺蓝染色对体外分离培养的软骨细胞进行鉴定。通过实时荧光定量PCR检测miR-15a在肢体内外翻畸形(valgus-varus deformity, VVD)组和健康组肉鸡中(各3只)各组织的表达量。之后通过CCK8和EDU方法分析软骨细胞过表达miR-15a后细胞增殖情况。软骨细胞转染miR-15a mimics后,通过qPCR检测软骨细胞的标志基因Collagen-2、Aggrecan、Collagen-10,成熟分化基因Runx2、Sox9、VEGF、MMP9,炎性因子IL-1β、IL-6、IL-8、IL-10、TNF-α、TGF-β3以及凋亡基因Fas、FasL、Bcl-2的表达量。并构建FKBP5 3'UTR的野生型载体和突变型载体,通过双荧光素酶检测报告检测miR-15a与FKBP5的靶向关系。结果表明,本研究所用的胰蛋白酶、Ⅱ型胶原酶和透明质酸酶联合消化法成功分离得到状态良好的软骨细胞。荧光定量结果显示,miR-15a在各组织中均有表达,与健康组相比,miR-15a在VVD组的肝(P < 0.01)、脾(P < 0.05)、胸腺(P < 0.01)中的表达量显著升高,在心和胸肌组织中的表达量显著降低(P < 0.01)。CCK8与EDU分析结果显示,与NC组相比,过表达miR-15a组软骨细胞增殖速度显著降低(P < 0.01),增殖细胞数量显著减少(P < 0.01)。qPCR结果显示,与mimics NC组相比,miR-15a mimics组的软骨细胞标志基因Aggrecan、成熟分化基因Sox9、Runx2表达量显著降低(P < 0.05),Fas基因表达量极显著上升(P < 0.01),FasL基因和抗凋亡基因Bcl-2极显著下降(P < 0.01)。成功构建了FKBP5 3'UTR野生型和突变型载体,双荧光素酶检测报告结果显示预测的靶基因FKBP5与miR-15a没有靶向关系。本研究成功分离并鉴定了鸡软骨细胞,过表达miR-15a抑制鸡软骨细胞增殖、成熟和分化并促进细胞凋亡。     

H9N2亚型禽流感病毒NS1基因真核表达载体的构建及其在293细胞瞬时表达

在293细胞中瞬时表达A/Chicken/Henan/1001/2010（H9N2）NS1基因蛋白,并对表达蛋白活性进行测定。试验利用PCR技术从NS1-T载体质粒上扩增NS1基因,将其克隆至pCAGGS载体,构建重组质粒NS1-pCAGGS;经酶切和测序鉴定正确后,重组质粒NS1-pCAGGS与脂质体按照一定比例混合后转染293细胞,用间接免疫荧光方法对瞬时表达细胞进行荧光信号反应。结果显示,禽流感NS1基因可以很好地在293细胞中瞬时表达,具有良好的反应原性。     

牛GDF9和BMP15基因遗传变异与双胎性状的关系研究

以生长分化因子9(Growth differentiation factor 9,GDF9)基因和骨形态发生蛋白15(Bone morphogenet-ic protein 15,BMP15)基因作为牛双胎性状的候选基因,研究了它们在鲁西牛、秦川牛、南阳牛和中国荷斯坦牛4个品种中的遗传变异,并在鲁西牛群体中研究了其多态位点与双胎性状的关系。结果表明:在鲁西牛中GDF9基因的3′UTR发现缺失突变,而其它3个品种中没有发现该突变。对鲁西牛群体中该多态位点与单、双胎性状之间进行卡方显著性检验表明,单胎牛群体与双胎牛群体基因型分布有极显著的差异(P=0.006),双胎牛群体的B等位基因频率明显大于单胎牛群体。通过生物信息学分析表明,突变体mRNA的二级结构与野生型相比总自由能值差异不大,但突变体mRNA翻译起始位点的二级结构稳定性明显大于野生型。在鲁西牛、南阳牛和秦川牛的BMP15基因中发现编码区第759~762位有GAAA 4个碱基存在缺失突变,但没有检测到突变纯合个体,中国荷斯坦牛中没有检测到该突变。卡方显著性检验表明单胎牛群体和双胎牛群体在该位点基因型组成差异不显著(P=0.947)。     

H1N1亚型猪流感病毒HA和NA基因双重RT-PCR定型检测方法的建立与应用

为了建立适用于临床诊断的H1N1亚型猪流感病毒快速检测方法,本研究根据GenBank已登录的H1N1亚型猪流感病毒HA和NA基因序列设计RT-PCR扩增引物,以H1N1亚型猪流感病毒、H3N2亚型猪流感病毒、猪瘟病毒和猪繁殖与呼吸综合征病毒为试验对照,通过优化RT-PCR反应条件和反应体系,建立了H1N1亚型猪流感病毒HA和NA基因双重RT-PCR定型检测方法。同时,运用H1N1亚型猪流感病毒血凝和血凝抑制试验方法和本研究建立的方法对165份猪病料样品进行了对比验证。结果表明,本研究建立的H1N1亚型猪流感病毒双重RT-PCR具有良好的特异性、敏感性、重复性,所扩增的目的基因片段大小分别为428 bp和678 bp左右,可检出最小基因组RNA浓度为2.9×10-5μg/μL。本研究建立的方法和H1N1亚型猪流感病毒血凝和血凝抑制试验方法均从同一份猪肺脏样品中检测出H1N1亚型猪流感病毒,其余样品中均未检出H1N1亚型猪流感病毒,两种方法符合率为100%。本研究建立的方法适用于H1N1亚型猪流感病毒双基因定型检测,可在H1N1亚型猪流感病毒流行病学调查和临床诊断中应用。     

我国西藏四种植物染色体数目观察

本文对西藏四种植物进行染色体观察，这四种植物的染色体数目均为国内外首次报道。     

稻草还田模式对稻田土壤速效氮、钾含量及晚稻生长的影响

设计了高桩翻耕抛栽、覆盖免耕抛栽和高桩免耕抛栽3种模式的田间小区试验。结果表明, 不同稻草还田模式对土壤NH₄⁺-N含量影响较大, 对土壤速效钾含量和土壤NO₃^--N含量影响不大; 高桩翻耕模式的土壤NH₄⁺-N、速效钾含量平均值比高桩免耕和覆盖免耕模式分别提高40.8%、7.5%和36.7%、2.0%; 高桩翻耕能促进水稻群体生长, 增加叶片叶绿素含量, 充分利用光能, 增强光合作用; 稻草还田以高桩翻耕模式的增产效果最好, 比高桩免耕、覆盖免耕分别增产5.8%和9.0%。以高桩翻耕稻草还田模式对土壤速效氮、钾含量及晚稻产量的影响最好。     

Susceptibility of various ceil culture systems to pseudorabies virus

A comparative study was carried out to determine the susceptibility of five different cell lines to pseudorabies virus (PRV), a herpes virus of pigs. The cell systems tested were swine testicle (ST), mink lung (ML), equine dermal (ED), porcine kidney (PK₁₅), and bovine turbinate (BT) cells. Virus titers obtained were 10^4.88, 10^4.38, 10^3.75, 10^2.63, and 10^0.25 for ML, ST, PK₁₅, BT and ED cells, respectively indicating that ML, ST, and PK₁₅ are optimal cell lines for the growth of PRV whereas BT and ED are not very sensitive.     

Prokaryotic Expression and Immunogenicity Analysis of N Protein from Vesicular Stomatitis Virus

This study was aimed to clone and express the specific antigen nucleoprotein (N) of vesicular stomatitis virus (VSV), and then purify and analyze its immunogenicity. Based on published VSV genome N gene sequence in GenBank, two kinds of N genes of VSV with different serotypes were synthesized, respectively. After sequence analysis, one pair of specific primers was designed and synthesized, N gene fragment with about 1 300 bp length was amplified by PCR, and subcloned into pCold Ⅰ expression vector. The recombinant N protein was induced with IPTG and purified by Ni-NTA. The results of SDS-PAGE showed that the N gene was successfully expressed in E. coli and the molecular weight of protein was 50 ku; The results of Western blotting showed that this recombined protein specifically reacted with polyclonal antibody serum of VSV. The recombinant vector with VSV-IND and VSV-NJ were successfully constructed, and the N protein was solubly expressed in E. coli, and the purified protein demonstrated promising immunogenicity.     

Effects of straw treatment and nitrogen supplementation on digestibility,intake and physiological responses of water intake as well as urine and faecal characteristics

This study investigates the effects of feeding diet based on untreated (UT) or ensiled alkali‐treated (ET) barley straw with either urea or casein supplementation, on feed intake, digestibility, ruminal pH, water intake and faecal and urinary characteristics. Four sheep fitted with ruminal cannulas were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Barley straw was treated by the dry (spraying) method in pH adjusted of hydrogen peroxide (pH 11.5), ensiled for 6 weeks and included at 65% of the diet dry matter (DM). The results showed that straw pH reduced from 11.58 to 8.60 after 6 weeks of ensilage. The ET diet increased average DM digestibility and intake by 19% and 43% respectively. Total water intake was similar across treatments, while the water/DM intake ratio was 23% higher with the UT diet than with the ET one. Ruminal (6.73 vs. 6.84) and faecal (8.67 vs. 9.05) pH decreased but urinary pH (6.14 vs. 8.13) increased as a result of feeding animals on the ET diet compared with the UT diet. Compared with the UT diet, the ET one decreased faecal fibre (12%), moisture (32%) and water holding capacity, while it increased faecal ash (10%) and density (20%). The volume of urine excreted by the sheep fed with the ET diet increased by 67%, but their urine specific gravity (SG) decreased. No significant effects were observed for the dietary N supplementation and interactions between straw type × N supplementation with regard to any of the measured characteristics except for DM intake, which reduced due to the casein supplementation in the ET diet. These results indicate that the alkali treatment and ensilage of barley straw increased digestibility, intake, faecal consistency and urinary pH and dilution but decreased straw alkalinity as well as ruminal and faecal pH.