荧光原位杂交技术对土壤石油降解菌的检测

微生物修复是石油污染土壤修复的主要方法之一，但是对修复过程中微生物的检测却是其主要限制因素，传统的检测方法存在很大的局限性，而荧光原位杂交技术结合了分子生物学的精确性和显微镜的可视化信息，可较为精确快速的对微生物进行定性和定量测定。 实验中利用辽河油田污染土壤中筛选纯化的高效石油降解菌，以原油为碳源，考察不同菌株处理后的去除率变化。发现有8株菌：B101、B381、B0502、B25 、B64、B2001、B2003、B18，在14 d内对石油的降解率都在20%以上，且族组分分析结果显示，被降解部分主要是烷烃和芳烃。经鉴定，此8株菌均为杆菌。 同时，对5株石油降解菌标准菌株（铜绿假单胞菌、巨大芽孢杆菌、枯草芽孢杆菌、苏云金芽孢杆菌和地衣芽孢杆菌）在基因库中比对，核苷酸同源性显示其相似性均在98%以上，因此针对其16S rRNA设计寡核苷酸探针。 将上述5种探针应用于荧光原位杂交实验，分别作用于8株降解菌和5株标准菌株，得出5种探针均符合特异性标准，仅地衣芽孢杆菌探针与B381之间有特异性荧光反应。添加B381（地衣芽孢杆菌）于泥浆反应器中降解石油烃。鉴于石油污染土壤的复杂组成，检测泥浆反应器中地衣芽孢杆菌的FISH条件优化结果为：样品固定时间17 h，杂交温度46 ℃，杂交时间3 h，杂交液中去离子甲酰胺浓度35%，冲洗缓冲液中与去离子甲酰胺对应的NaCl的浓度88 mmol∙L-1。运用上述FISH技术监测生物泥浆反应器中地衣芽孢杆菌量的变化，得出地衣芽孢杆菌的浓度在第5天达到最大，同时对泥浆反应器中去除率进行测定，前5天菌对原油的去除速率也最高，前8 d是石油烃被去除的主要作用阶段。微生物的数量变化和原油的去除率进行比较，两者的变化情况符合微生物降解石油的趋势，为监测含油污泥中微生物的变化提供了一种可行的技术。

白腐菌选择性降解木质素的研究

对15株白腐真菌进行了以玉米秸秆为基质的初步筛选，从中获得一株选择性系数较高的菌株Y10，并对其降解玉米秸秆的情况进行了研究。结果表明，在30天的培养过程中菌株Y10对玉米秸秆降解的选择性系数都大于1，第15天选择性系数最高为3.88。对未经降解和降解过的玉米秸秆分别作了紫外光谱和红外光谱分析，结果表明，经该菌降解后玉米秸秆的化学成分发生了很大变化，且木质素的降解程度要大于纤维素的降解程度。对菌株Y10进行了ITS-5.8S rDNA序列鉴定，初步判定其为Cerrena sp.。 为了考查不同的外源添加物对菌株Y10降解玉米秸秆的影响，在以玉米秸秆为基质的固态发酵培养基中分别添加了7种金属离子、8种碳源、6种氮源。结果显示，这7种金属离子均能促进木质素的降解，并且一定浓度的某些离子明显抑制纤维素的降解；其中添加0.036%的MnSO4·H2O和0.36%的MgSO4·7H2O对纤维素降解的抑制作用比较强，降解率分别为0.96%和1.31%，木质素的选择性系数分别达到了34.40和20.17。8种碳源中除麦芽糖外都能促进木质素的降解，除微晶纤维素外都明显促进纤维素的降解。6种氮源中酒石酸铵、硫酸铵、草酸铵和氯化铵的添加都会使该菌生长变慢，而且氮源浓度越高菌丝生长越慢。外加碳源和金属离子对半纤维素降解和选择性系数的影响不大。 同时对菌株Y10在液态培养下产木质素降解酶的条件和培养基做了优化。结果表明，在初始产酶培养基中，菌株Y10的漆酶酶活在第10d达到最高，锰过氧化物酶酶活在第11d达到最高，基本上检测不到木质素过氧化物酶。菌株Y10产漆酶的最适温度为32℃，最适PH为6.0；产锰过氧化物酶的最适温度为32℃，最适PH为6.5。菌株Y10产漆酶的最佳碳源为甘露糖，最佳氮源为酒石酸铵，最适诱导剂VA浓度为3 mmol/L，最适表面活性剂TW-80浓度为1%。 利用响应面法对其产漆酶的培养基进行优化，优化后的培养基配方为葡萄糖10.00 g/L，酒石酸铵0.50 g/L，大量元素296.50 ml/L，微量元素100.00 ml/L，NTA 1.40 g/L，VA 5.00 mmol/L，吐温-80加入量为0.10%。进行了菌株Y10产漆酶的验证实验，实测酶活为5282.56 U/L，与预测酶活5162.73 U/L接近。在优化后培养基中，菌株Y10在第14 d达到生长的最高峰，第20 d时，漆酶酶活最高，为11325.00 U/L；第16 d时，锰过氧化物酶酶活最高，为30.77 U/L。 对菌株Y10的漆酶酶学性质做了初步的研究，结果显示，酶反应的最适温度为40℃-65℃，最适PH为3.0。在40℃，PH=3.0时，漆酶催化ABTS反应的米氏方程为 。 Fifteen white-rot fungi based on corn stalk were screened. One white-rot fungus Y10 with high selectivity value was obtained. The degradation of corn stalk was initially studied. The results indicated that the selectivity value was above 1 during the 30 day-cultivation and the highest was 3.88 after 15 days. The composition of untreated and treated stalk was analyzed through ultraviolet spectroscopy and infrared spectroscopy. It was found that the composition of treated stalk was greatly altered and the degree of the degradation of lignin is greater than the cellulose. Y10 was identified as Cerrena sp. by ITS -5.8S rDNA sequence analysis. The influence of metal ions, carbon sources and nitrogen sources on corn stalk degradation by white-rot fungus was studied. While all seven metal ions could promote lignin degradation, the cellulose degradation was best inhibited at certain ion concentrations. Notably, when 0.036% MnSO4·H2O and 0.36% MgSO4·7H2O were added into the medium, the cellulose degradation was restrained to the extents that the coefficients of lignin selectivity rose to 34.40 and 20.17 respectively. It was also found that all carbon sources except maltose can promote lignin degradation. The addition of carbon sources other than microcrystalline cellulose significantly promoted cellulose degradation. The addition of the nitrogen sources, ammonium tartrate, ammonium sulfate, oxalate, ammonium chloride, resulted in remarkable inhibition to mycelium growth; the larger the concentrations of nitrogen sources are, the slower the mycelium grew. The addition of carbon sources and metal ions had less impact on the degradation of hemicellulose and selectivity value. Meanwhile, we optimized the conditions and culture medium of the lignin-degrading enzyme production of strain Y10. The results showed that in the initial culture medium, the Lac activity was highest at the 10th day, the MnP activity was highest at the 11th day and the LiP could not be detected. The optimum condition of Lac was at temperature 32 and PH =6.0 and the optimum condition of MnP was at temperature 32 and PH =6.5. The optimum carbon source for Lac was seminose, the optimum nitrogen source was ammonium tartrate, the optimum content of VA was 3 mmol/L, the optimum content of TW-80 was 1%. PB and RSM were used to optimize the culture medium of laccase by white-rot fungus Y10. The optimum culture medium was consist of glucose 10.00 g/L, ammonium tartrate 0.50 g/L, macro elements 296.50 ml/L, trace elements 100.00 ml/L, NTA 1.40 g/L, VA 5.00 mmol/L, TW-80 0.10%. Under the optimal conditions, the activity of laccase was 5282.56 U/L and the experimental value agreed with the predicted value 5162.73 U/L. The biomass was highest at the 14th day, the Lac activity was highest at the 20th day, the MnP activity was highest at the 16th day. The results of the studies on the characteristics of Lac showed that the optimum temperature for Lac activity is 40℃-65℃ ; the optimum PH for Lac activity is 3.0 and under 40℃，PH=3.0, the Michaelis-menten equation of Lac catalized ABTS oxidation was .

QCD求和规则对S夸克质量的研究

QCD求和规则是强子物理中的一种非常有效的非微扰方法，它从流的算符乘积展开开始，引入算符乘积展开式的真空期望值，把微扰和非微扰效应分开处理：微扰效应包含在展开系数中，非微扰效应则由算符的真空凝聚值表示。然后利用色散关系，把算符乘积展开式的真空期望值与一个含有强子物理参数的色散积分联系起来，这样就能够计算有关强子的物理量。 本文首先系统介绍了QCD求和规则的基本原理、基本方法，然后结合Dominguez，Gend和Paver的工作[14]，展开式保留了的算符d=4凝聚值，采用新的参数化渐近自由阈以下谱函数的方法：即根据文献[25]，用实验上了解得比较清楚的两个共振态的贡献来参数化谱函数，计算了s夸克的质量，得到了在动量标度为1Gev时，s夸克的跑动质量为219MeV。在误差范围内，这是一个理论上可以接受的结果，本文计算得到的跑动质量的值能为考察动量转移为1GeV时的质量效应提供参考。 关键词：QCD求和规则 算符乘积展开 色散关系 Borel变换 夸克质

3,5-二甲基吡唑对尿素氮转化及NO_3~--N淋溶的影响

采用室内培养和田间试验相结合的方法,探讨了新型硝化抑制剂3,5-二甲基吡唑(DMP)对尿素氮转化及玉米田硝酸盐淋溶损失的影响.结果表明,DMP对尿素水解仅起短暂的抑制作用,但可在较长时间内显著抑制土壤铵的氧化,且随DMP用量的增加,抑制效应显著增强.培养第10 d时,DMP各处理(0.002 5、0.01及0.025 g/kg)的土壤NH4+-N累积量分别比CK提高了5.17、9.36和11.04倍,而NO3--N累积量于培养第14 d时差异最大,与CK相比分别降低了33.30%、61.19%和73.72%(p<0.01).土壤NO2--N只在尿素施用前期有少量累积,但第3 d DMP各处理土壤NO2--N含量降低幅度达95.77%~96.13%;土壤矿质氮总量于10 d以后,随DMP用量的增加,显著降低,而DMP1处理的土壤微生物量N在培养14~56 d期间显著提高.连续2 a的玉米田间试验原位取土测定结果表明,2004和2005年,DMP的施用使作物根系密集层以下(40~100 cm)土层的NO3--N累积总量分别比CK降低了28.77%和44.70%.因此,硝化抑制剂DMP与尿素配合施用是调控氮素转化、缓解氮肥污染的有效措施.

过量Mg~(2+)对土壤中酶活性的影响

目的研究过量Mg2+对土壤中脲酶、磷酸酶和过氧化氢酶活性的影响.方法通过在清洁土壤中投加不同质量浓度的MgC l2试剂,经过28 d的培养,期间在第1、3、7、14、28 d取样,测定土壤中的脲酶、磷酸酶和过氧化氢酶的活性.结果数据分析表明Mg2+对脲酶有激活作用,当Mg2+的质量分数为0.6%时,土壤脲酶活性提高最多,为91.6%,脲酶活性增加的程度为:0.6%>0.8%>0.4%>0.2%>1.0%>0,并且第1 d是观测脲酶变化的最佳时间.Mg2+对磷酸酶活性有先激活再抑制的作用,其活性在第3 d有大幅度增加,而从第14 d开始到第28 d,5个含不同质量分数Mg2+的土样,其磷酸酶活性全都低于空白试验。过氧化氢酶活性在试验的所有土样中均受到长效抑制.结论脲酶、磷酸酶和过氧化氢酶对Mg2+反映敏感,在Mg2+质量分数为0.6%时可以短期提高土壤肥力,但是长期大量的Mg2+介入土壤会严重影响土壤肥力,因此,可以考虑以Mg2+作为镁污染土壤的生态毒理指标.

模拟高原低氧对新生大鼠肾上腺皮质功能发育的影响

本文探讨新生大鼠肾上腺皮质对高原低氧的应答及模拟高原低氧对其功能发育的影响。结果表明, 当不同日龄大鼠暴露于5 km 及7 km 海拔24 h, 7 d、14 d 龄大鼠肾上腺皮质无明显应答反应。21 d 及28 d 龄大鼠肾上腺皮质酮水平随海拔增高而增加, 血浆皮质酮表现为抑制作用。当1 d 龄新生大鼠在5 km 海拔高度发育3 d 和7 d,其肾上腺皮质功能无异于正常发育大鼠; 但发育14 d、21 d 及28 d, 其血液及肾上腺中皮质酮含量均明显低于对照组, 肾上腺皮质功能发育严重受抑。

Purification and cloning of a novel C-type lectin-like protein with platelet aggregation activity from Trimeresurus mucrosquamatus venom

TMVA, a novel C-type lectin-like protein that induces platelet aggregation in a dose-dependent manner, was purified from the venom of Trimeresurus mucrosquamatus. It consists of two subunits, alpha (15,536 Da) and beta (14,873 Da). The mature amino acid sequences of the a (135 amino acids) and beta subunits (123 amino acids) were deduced from cloned cDNAs. Both of the sequences show great similarity to C-type lectin-like venom proteins, including a carbohydrate recognition domain. The cysteine residues of TMVA are conserved at positions corresponding to those of flavocetin-A and convulxin, including the additional Cys135 in the alpha subunit and Cys3 in the beta subunit. SDS-PAGE, mass spectrometry analysis and amino acid sequence showed that native TMVA exists as two convertible multimers Of (alphabeta)(2) and (alphabeta)(4) with molecular weights of 63,680 and 128,518 Da, respectively. The (alphabeta)(2) complex is stabilized by an interchain disulfide bridge between the two alphabeta-heterodimers, whereas the stabilization of the (alphabeta)(4) complex seems to involve non-covalent interactions between the (alphabeta)(2) complexes. (C) 2002 Elsevier Science Ltd. All rights reserved.

~(14)C标记1,2,7,8-TCDD在鲤体内分布及代谢的初步研究

利用14C标记1,2,7,8-四氯代二苯并二噁英(1,2,7,8Tetrachloro[U—14C]dibenzodioxin,14C1,2,7,8TCDD)初步研究了其在鲤体内的分布和代谢规律。14C1,2,7,8-TCDD溶解于丙酮/植物油中,腹腔暴露。暴露1、2、4、8、12d后取样,肝脏、胆汁、腹腔脂肪等消化制样后用液闪仪测量放射性活度。肝脏和胆汁内的放射性活度同步变化,都是第8d达到峰值后下降。腹腔脂肪内1—2d放射性明显高于随后取样的样品。肝和腹腔脂肪的分布量之比呈现“S”型变化趋势。暴露4d

Effect of dead carbon on the C-14 dating of the speleothem

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洞穴次生碳酸盐中“死碳”对~(14)C法测年的影响

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