3种介导猪NR2F2基因转染PK15细胞方法的比较研究

试验旨在研究3种转染方法对猪肾上皮细胞（PK15）的转染效率，为以PK15细胞为模型研究外源基因的功能提供参考。本研究以PK15细胞为研究对象，用脂质体、电穿孔、慢病毒3种转染方法转染后，在荧光显微镜下观察绿色荧光蛋白的表达，采用实时荧光定量PCR检测EGFP和NR2F2基因的表达情况，采用CCK-8检测细胞存活率，进而比较3种方法的转染效果。结果显示，转染PK15细胞后，电穿孔和慢病毒方法转染效率极显著高于脂质体（P<0.01），但电穿孔方法和慢病毒方法之间差异不显著（P>0.05）；EGFP和NR2F2基因的实时荧光定量PCR结果显示慢病毒方法效果最好，脂质体方法较差，与细胞转染效率基本一致；CCK-8结果显示，电穿孔转染后细胞存活率最低，极显著低于对照组（P<0.01），脂质体方法显著低于对照组（P<0.05），慢病毒方法与对照组间差异不显著（P>0.05）。综合考虑转染效率及细胞活性，本研究认为慢病毒转染方法最适合转染PK15细胞，为今后高效转染PK15细胞提供了理想的方法。     

不同水平的青贮饲料对饲喂稻草秸秆奶牛瘤胃微生物数量和微生物蛋白合成的影响

本试验的目的是确定饲粮中青贮银合欢添加水平对奶牛瘤胃微生物种群、氮平衡和微生物蛋白合成的影响。试验采用4×4拉丁方设计,选择12头初始体重为163±16 kg、带有瘤胃瘘管的奶牛,随机分为4个组,每组3个重复,每个重复1头牛。基础日粮以100%的水稻秸秆为原料,处理组分别用30%、60%和100%的青贮银合欢替代水稻秸秆,奶牛自由采食水稻秸秆和青贮银合欢,每天按照体重的0.2%补充浓缩料。结果表明,用60%青贮银合欢替代水稻秸秆组奶牛瘤胃微生物群,尤其是纤维素分解菌、蛋白分解菌和厌氧真菌的数量显著增加(P<0.05),各组对淀粉分解菌群无显著影响(P>0.05)。原生动物种群数量随日粮青贮银合欢添加水平的增加呈线性下降(P<0.05)(P<0.05)。此外,氮平衡和微生物蛋白合成均随着日粮青贮银合欢添加水平的升高而升高(P<0.05),其中以60%青贮银合欢组最高。结论:在本试验基础上,用60%青贮银合欢替代水稻秸秆可以显著提高奶牛体内的微生物数量和微生物蛋白的合成。     

不同改良材料对灌漠土微生物量磷及生物有效性的影响

通过培养和盆栽试验，向灌漠土土壤中加入等量磷和不同添加比例的硫磺S（0.05%、0.15%、0.45%）、生物菌肥B（0.25%、0.50%、1.00%）、有机肥OM（0.50%、1.00%、2.00%）和小麦秸秆WS（1.00%、2.00%、4.00%），研究不同改良材料配施磷肥土壤微生物量磷的变化特征，及其与Olsen P、小麦吸磷量之间的关系。结果表明，添加不同改良材料处理的土壤微生物量磷含量均显著高于对照（不添加改良材料），且随着添加比例的增大而增加。在第16天时，各处理土壤微生物量磷含量达到最大值，OM₂2.00、B₂1.00、WS₂4.00和S₂0.45处理分别较对照显著增加了34.66%、34.52%、28.19%和23.89%；经过30 d的培养，硫磺、生物菌肥、有机肥和小麦秸秆处理的土壤微生物量磷含量较对照分别增加了19.51%、43.08%、47.92%和41.68%。在一定范围内（Olsen P约90 mg·kg^-1），土壤微生物量磷随土壤Olsen P提高而增加。在培养的第16天和第30天时，小麦植株吸磷量也与土壤微生物量磷存在极显著的相关关系。总体看来，有机肥或生物菌肥配合磷肥施用于灌漠土能促进土壤微生物量磷的增加，提高磷的生物有效性，对于灌漠土磷素高效利用具有重要意义。     

Long-term fertilization leads to specific PLFA finger-prints in Chinese Hapludults soil

Soil microbes play essential roles in the biogeochemical processes of organic carbon and nutrient cycling. Many studies have reported various short-term effects of fertilization on soil microbes. However, less is known about the effects of longterm fertilization regimes on the rhizosphere. Therefore, the objective of this study was to explore how the soil microbial communities in the rhizosphere respond to different long-term fertilization strategies. Based on a 21-year field treatment experiment in Guizhou, China, we extracted phospholipid fatty acids(PLFAs) to determine the microbial community structure in both the non-rhizosphere(NR) and rhizosphere(R). Six treatments were included: no fertilizer(CK), mineral nitrogen fertilizer(N), N with potassium(NK), phosphorus with K(PK), NPK, and NPK combined with manure(MNPK). The results showed that total PLFAs under unbalanced mineral fertilization(N, NK and PK) were decreased by 45% on average in the NR compared with CK, whereas MNPK increased fungi and G~– bacteria abundance significantly in both the NR(by 33 and 23%) and R(by 15 and 20%), respectively. In addition, all microbial groups in the R under these treatments(N, NK and PK) were significantly increased relative to those in the NR, except for the ratio of F/B and G~+/G~–, which might be due to the high nutrient availability in the R. Soil pH and SOC significantly regulated the soil microbial community and structure, explaining 51 and 20% of the variation in the NR, respectively. However, the rhizosphere microbial community structure was only significantly affected by soil pH(31%). We concluded that the soil microbial community in the NR was more strongly affected by long-term fertilization than that in the R due to the rhizosphere effect in the agricultural ecosystem. Rhizosphere nutrient conditions and buffering capacity could help microbial communities resist the change from the long-term fertilization.     

发酵全混合日粮对肉羊消化吸收功能和生长性能的影响研究

旨在探讨添加混合微生物菌剂的发酵全混合日粮(Fermented total mixed ration，FTMR)对肉羊营养物质消化吸收功能和生长性能的影响。通过发酵和肉羊饲喂试验，测定了FTMR的品质变化、营养物质表观消化率、肉羊瘤胃发酵情况及日增重。结果表明:1)全混合日粮发酵后pH和中性洗涤纤维含量显著降低，乳酸和乙酸含量显著增加(P<0.05)；2)试验组肉羊瘤胃液pH显著降低，氨态氮含量降低50%以上(P<0.05)；粗蛋白质、粗脂肪、中性洗涤纤维、酸性洗涤纤维和酸性洗涤木质素(ADL)的利用率均显著高于对照组(P<0.05)，尤其是ADL利用率提高了29.92%(P<0.05)。3)试验组平均日增重较对照组提高50%。综上，混合微生物菌剂加入全混合日粮发酵后，适口性增强，可改善肉羊瘤胃液环境，提高肉羊的消化吸收功能和生长性能，有利于提高秸秆饲料化利用效率。     

瑞香狼毒化学成分抑制PC-12细胞神经突起生长活性研究

对瑞香狼毒(Stellera chamaejasme L.)的化学成分进行研究,采用硅胶色谱、凝胶色谱、高效液相色谱等多种色谱手段对瑞香狼毒的乙酸乙酯提取物进行分离纯化,利用NMR、MS波谱分析鉴定3个木质素、1个二萜原酸酯、3个双黄酮、2个黄酮。枇杷素(5)、异落叶松树脂醇(7)和(-)-落叶松树脂醇(8)为首次从该植物中分离得到。PC-12细胞活性试验表明,双黄酮类化合物(狼毒色原酮(2),新狼毒素B(3),异新狼毒素A(4))对NGF依赖的PC-12细胞突起生长有较强的抑制活性(P0.05)。     

微生物发酵在菜籽饼粕饲用品质改良中的应用研究进展

随着畜牧业的快速发展、优质饲料原料的严重不足以及国际进口贸易形势的日益紧张,开发非常规饲料资源的需求愈发迫切。菜籽饼粕是我国第一大油料作物油菜的加工副产物,饲用价值不高严重制约了在饲料工业中的应用。微生物发酵技术具有改良菜籽饼粕品质和提高畜禽生产性能的优势。本文综述了微生物发酵改良菜籽饼粕饲用价值的机理、影响因素以及微生物发酵菜籽饼粕饲料在畜禽生产中的应用,讨论了目前微生物发酵菜籽饼粕生产中存在的问题,展望了未来微生物发酵菜籽饼粕饲料的发展方向。     

不同产量水平下枣园土壤微生物量碳、氮与土壤和植株养分的关系

【目的】 研究不同产量水平下枣园土壤微生物量碳、氮与不同时期枣园土壤和植株养分的关系,为南疆红枣高产提供土壤管理的理论依据。【方法】 选取南疆高中低3种不同产量水平的枣园为研究对象,采用田间试验,研究不同时期不同产量水平下枣园土壤微生物量碳、氮与不同土层和植株氮、磷、钾养分含量之间的关系。【结果】 土壤碱解氮、速效磷含量随着土壤深度的增加逐渐降低,而速效钾含量逐渐升高。土壤微生物量碳(MBC)和土壤微生物量氮(MBD)含量之间呈极显著正相关,相关系数为0.836 7;土壤微生物量碳与骏枣展叶期0~20 cm土层速效钾含量呈显著正相关,与果实膨大期20~40 cm土层速效钾含量呈显著正相关关系。【结论】 枣园土壤微生物量碳、氮及各养分含量均保持在良好水平,土壤养分及植株养分能对微生物量碳、氮产生显著影响。     

Air-drying changes the distribution of Hedley phosphorus pools in forest soils

Hedley labile phosphorus(P)pools in soil tend to be several times larger than annual forest requirements,even in highly weathered soils characterized by P limitation.The discrepancy between plant and soil P status could be partly attributable to the frequently adopted air-drying pretreatment that tends to increase soil P solubility.In this study,the effects of air-drying on the distribution of Hedley P fractions were examined using soils collected under 4 forest types at Gongga Mountain,southwestern China.The results showed that the microbial biomass P(Pmic)in the organic horizon decreased markedly after air-drying.The concentrations of Hedley labile P in the air-dried samples were 31%–73%more than those in the field-moist samples.Consequently,the air-drying-induced increments of Hedley labile P pools in the surface soil horizons were 0.8–3.8 times the annual plant P requirements.The organic horizon was more susceptible to the air-drying-induced increases in Hedley labile P than the mineral horizon,probably because of the stronger release of Pmicand disruption of soil organic matter.The quality of P,indexed by the ratio of Hedley labile P to slowly cycling P,shifted in favor of the Hedley labile fractions after air-drying,further revealing that air-drying changed the distribution of Hedley P pools in forest soils.These indicated that the effects of air-drying could not be ignored when interpreting the discrepancy between the P status of plants and the Hedley labile P pools in forest soils.To more efficiently evaluate the P status in forest soils via the Hedley fractionation procedure,the use of field-moist soils is recommended.