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1.
背主动脉注射是生产转基因鸡的经典方法,该方法不需换壳培养,但壳内注射技术难度大,并且无法实时观察后期胚胎发育情况。本实验对背主动脉注射法进行了改进,将增强型绿色荧光蛋白(EGFP)腺相关病毒(Adeno-associated Virus,AAV)壳外注射至150枚HH 14~16期(Hamburger-Hamilton Stage 14~16)鸡胚背主动脉中,再进行换壳培养,继续孵化至出壳,观察和分析改良背主动脉注射法与传统背主动脉注射法和胚盘下腔注射法对发育8、14、18、21 d鸡胚存活率、孵化率以及EGFP阳性检出率的影响。结果表明:改良背主动脉注射组鸡胚存活率均高于传统背主动脉注射组与胚盘下腔注射组(P<0.05或P<0.01);改良背主动脉注射组的鸡胚孵化率(37%)高于传统背主动脉注射组(16%)与胚盘下腔注射组(28%)(P<0.01);荧光蛋白手电筒检测显示,改良背主动脉注射组鸡胚EGFP阳性率(17%)明显高于传统背主动脉注射组(13%)与胚盘下腔注射组(12%)(P<0.01)。综上,背主动脉壳外注射结合换壳培养提高了转基因鸡胚胎孵化率和EGFP阳性检出率,对提高转基因鸡效率具有重要价值。 相似文献
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Litter- and ecosystem-driven decomposition under elevated CO2 and enhanced N deposition in a Sphagnum peatland 总被引：1，自引：0，他引：1
Andy Siegenthaler Alexandre Buttler Luca Bragazza Edwin van der Heijden Jean-Michel Gobat 《Soil biology & biochemistry》2010,42(6):968-977
Peatlands represent massive global C pools and sinks. Carbon accumulation depends on the ratio between net primary production and decomposition, both of which can change under projected increases of atmospheric CO2 and N deposition. The decomposition of litter is influenced by 1) the quality of the litter, and 2) the microenvironmental conditions in which the litter decomposes. This study aims at experimentally testing the effects of these two drivers in the context of global change. We studied the in situ litter decomposition from three common peatland species (Eriophorum vaginatum, Polytrichum strictum and Sphagnum fallax) collected after one year of litter production under pre-treatment conditions (elevated CO2: 560 ppm or enhanced N: 3 g m−2 y−1 NH4NO3) and decomposed the following year under treatment conditions (same as pre-treatment). By considering the cross-effects between pre-treatments and treatments, we distinguished between the effects on mass loss of 1) the pre-treatment-induced litter quality and 2) the treatment conditions under which the litters were decomposing. The combination between CO2 pre-treatment and CO2 treatment reduced Polytrichum decomposition by −24% and this can be explained by litter quality-driven decomposition changes brought by the pre-treatment. CO2 pre-treatment reduced Eriophorum litter quality, although this was not sufficient to predict decomposition. The N addition pre-treatment reduced the decomposition of Eriophorum, due to enhanced lignin and soluble phenols concentrations in the initial litter, and reduced litter-driven losses of starch and enhanced litter-driven losses of soluble phenols. While decomposition indices based on initial litter quality provide a broad explanation of quantitative and qualitative decomposition, they can only be taken as first approximations. Indeed, the microbial ATP activity, the litter N loss and resulting litter quality, were strongly altered irrespective of the compounds' initial concentration and by means of processes that occurred independently of the initial litter-qualitative changes. The experimental design was valuable to assess litter- and ecosystem-driven decomposition pathways simultaneously or independently. The ability to separate these two drivers makes it possible to attest the presence of litter-qualitative changes even without any litter biochemical determinations, and shows the screening potential of this approach for future experiments dealing with multiple plant species. 相似文献
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Rates of degradation of 2-propenyl isothiocyanate (PrITC), benzyl isothiocyanate (BeITC) and 2-phenylethyl isothiocyanate (2-PeITC) in a soil known to biodegrade methyl isothiocyanate (MITC) at an accelerated rate, but never previously exposed to the other ITCs, were higher (persistence in soil increased by 1150, 80 and 100%, respectively,) than in a similar non-degrading soil. The rate of degradation of the same three ITCs was significantly lower in sterilised (autoclaved) soils than in the degrading soil. These results indicate that the three ITCs are susceptible to enhanced cross-biodegradation in soils where enhanced biodegradation of MITC has been induced by use of metham sodium soil fumigant. When Brassica plant tissue containing sinigrin (2-propenyl glucosinolate) as the predominant glucosinolate (GSL) was added to the degrading soil, the amount of PrITC present after 24 h was significantly lower than in the non-degrading soil at the same amendment rates. The toxicity to an insect test organism of the PrITC produced from the biofumigant plant tissue was correlated with the concentration of PrITC measured in the two soils, with 67% more plant tissue required in the degrading soil to cause 100% mortality as in the non-degrading soil (3.0 vs 5.0 mg g−1). The effectiveness of biofumigation using ITC-producing Brassica plants may be diminished in soil suffering from enhanced biodegradation of MITC. 相似文献
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多氯联苯复合污染土壤的土著微生物修复强化措施研究 总被引：11，自引：1，他引：10
通过室内模拟试验,以不同C源、C/N比、水分及通透性为调控因子,对多氯联苯(PCBs)长期复合污染土壤的土著微生物强化修复进行了初步研究。结果表明,PCBs长期复合污染土壤中,在土壤水分含量为田间持水量的60%时,加入淀粉、葡萄糖和琥珀酸钠均在一定程度上增加了细菌和真菌数量,从而促进土壤中PCBs的土著微生物降解。不同种类的C源对PCBs污染土壤的土著微生物降解效果存在明显差异,且其降解效果与C源的施用剂量密切相关。当淀粉加入量为C1.0g/kg土时,土壤中PCBs的降解效果较好,而葡萄糖和琥珀酸钠加入量为C0.2g/kg土时,PCBs的降解效果明显。土壤C/N比为10:1的处理效果优于C/N比为25:1和40:1。土壤人为翻动有利于PCBs污染土壤中细菌和真菌的生长,提高土著微生物的代谢活性,从而促进土壤中PCBs的自然降解。这为进一步探讨加速土壤中PCBs降解的最适条件和研发POPs污染土壤的生物修复技术提供了科学依据。 相似文献
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增铵营养对番茄植株伤流液组分及含量的影响 总被引：1，自引：1，他引：0
在总氮(N)浓度相等的条件下,研究全硝营养(100% NO3-)和25% 增铵营养(NH4+∶NO3- = 25%∶75%)对开花期和幼果期番茄植株伤流液各组分含量的影响.结果表明,增铵营养显著增加幼果期伤流液中 K的含量,对Ca、Mg、P元素含量没有显著影响;增铵营养下伤流液中 NO3- 的含量下降、NH4+ 含量增加,氨基酸、苹果酸等的含量均显著增加,氨基酸/硝态氮含量之比显著提高,表明喜硝作物适当增铵不仅能够提高根系活力,显著促进K的吸收以供果实发育之需,而且提高了植株整体同化N素的能力. 相似文献
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[目的]研究增强UV-B辐射和Cd2＋复合胁迫下绿豆（Phaseolus radiatus L.）幼苗Cd积累和光合作用的气孔和非气孔限制因素。[方法]试验设1个对照（CK）和3个试验组,分别为增强UV-B辐射（UV-B）组、Cd2＋（Cd）组、增强UV-B辐射＋Cd2＋（UV-B＋Cd）组,研究0.35 W/m2的UV-B辐射、1μmol/L Cd2＋及其复合胁迫下绿豆幼苗对Cd的吸收、分配和光合作用的气孔、非气孔限制,分别利用原子吸收分光光度仪和光合测定仪测定Cd在绿豆幼苗各部位的含量及第1对真叶的光合指标。[结果]绿豆幼苗中Cd的积累是根〉胚轴〉叶,UV-B对Cd在幼苗中的积累和分配没有影响。此外,各胁迫下幼苗的净光合速率（Pn）、气孔导度（Gs）、光合能力（A0）和羧化效率（dPn/dCi）含量均明显降低。UV-B组的细胞间隙CO2浓度（Ci）升高,气孔限制值（Ls）降低,Pn的降低是非气孔因素。Cd和复合处理组的Ci降低,而Ls升高,Cd组Pn的降低主要是气孔因素。复合组前期以非气孔因素为主,后期主要是气孔因素。[结论]该研究揭示了UV-B辐射和Cd2＋复合胁迫下绿豆幼苗光合变化的主要原因,可为进一步研究提供科学依据。 相似文献
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为了探讨外源蛋白在柞蚕蛹-柞蚕核型多角体病毒(ApNPV)表达系统的表达水平和稳定性,将增强型绿色荧光蛋白(EGFP)基因克隆到柞蚕核型多角体病毒转移表达载体pApM748BE中,获得重组质粒DNA,与ApNPV DNA共转染樗蚕(Phi-losamia cynthiam)培养细胞Pc-01后,用末端稀释法筛选获得重组病毒ApNPV-Δph/egfp+。将该重组病毒感染柞蚕蛹,采用SDS-PAGE和Western blot方法检测EGFP在柞蚕蛹中的表达,结果显示:在柞蚕蛹感染重组病毒ApNPV-Δph/egfp+后6 d,EG-FP就有明显的表达;感染后12～18 d,蛹体液中的EGFP含量保持较高水平,以EGFP标准样品定量分析EGFP在柞蚕蛹体液中的表达水平高于1 mg/mL;感染后30～39 d仍可以检测到蛹体液中EGFP的表达,而且蛋白稳定。研究结果表明,利用柞蚕核型多角体病毒作表达载体,可在柞蚕蛹中高效稳定地表达EGFP,并且EGFP在雌雄蛹间的表达水平无明显差异。 相似文献
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本试验通过筛选抗原性较好的多杀性巴氏杆菌,经禽胚培养增殖细菌,灭活后加入免疫增强剂左旋咪唑和亚硒酸钠维生素E,制成油乳剂强化苗。试验表明,每头份含抗原量100亿,左旋咪唑和亚硒酸钠维生素E合用组免疫效果最佳。注射本苗后,鸡群第4天产生免疫应答,第7天即可产生坚强的免疫力,试管凝集抗体效价达到1:86,攻毒保护率达83.3％,第14天即可达100％,免疫期6个月。免疫期内攻毒保护率为97.6％,当试管凝集抗体效价为1:40时,80％的试验鸡能抵御C_(48-1)的攻击。临床上免疫接种4个鸡场12批商品蛋鸡共14.5万羽,有效地控制了该病的流行。 相似文献
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[目的]建立一种检测8-羟基喹啉铜的方法.[方法]采用自制合成的吖啶酮衍生物10-甲基-3-硝基-吖啶酮(MAT)作为增强型荧光信号探针,建立8-羟基喹啉铜的测定方法.[结果]在最佳条件下荧光增强值与8-羟基喹啉铜的浓度在5×10-9 ～5 ×10-5 mol/L范围内成良好的线性关系,检测限为6×10-10 mol/L.[结论]该方法灵敏度高、检测范围宽,结果令人满意,可用于实际样品中8-羟基喹啉铜含量的直接测定. 相似文献