滇中高原云南松林枯落物输入对土壤碳氮储量及其分布格局的影响

为了更好地理解土壤碳氮对枯落物输入变化的响应,通过枯落物添加与去除实验（DIRT）对滇中高原云南松林枯落物输入变化对土壤碳氮储量及其分布格局的影响进行了研究。2018年3月至2019年2月分别设置6种枯落物输处理,分别为对照（CO）、去除枯落物（NL）、双倍枯落物（DL）、去除根系（NR）、无输入（NI）以及去除有机层与A层（O/A-Less）,研究了不同处理条件下土壤剖面上碳氮储量的分布规律。研究结果表明：（1）不同处理全碳储量为134.49-170.92 t/hm²,全碳储量在不同处理间表现为：S_C（NL）=170.92 t/hm² > S_C（CO）=168.10 t/hm² > S_C（NR）=153.26 t/hm² > S_C（NI）=147.20 t/hm² > S_{C（O/A-Less）}=143.54 t/hm² > S_C（DL）=134.49 t/hm²,不同处理0-20 cm土层全碳储量占0-60 cm土层全碳储量的40.86%-53.56%;不同处理全氮储量表现为：S_N（CO）=11.83 t/hm² > S_N（NL）=9.70 t/hm² > S_N（DL）=8.70 t/hm² > S_N（NR）=8.35 t/hm² > S_{N（O/A-Less）}=8.21 t/hm² > S_N（NI）=8.09 t/hm²。不同处理0-20 cm土层的全氮储量占0-60 cm土层全氮储量的39.28%-46.04%。云南松林地枯落物添加去除实验发现去除枯落物短期内可以增加土壤碳储量,其他处理均在一定程度上减少了土壤碳氮储量。（2）地上枯落物输入对表层（0-20 cm）土壤碳氮影响显著,根系输入对深层（20-40 cm）土壤碳氮影响显著;（3）土壤C、N存在耦合关系,不同处理土壤全碳含量与全氮含量极显著正相关,并且土壤全碳含量与土壤各化学计量比均呈极显著正相关关系;土壤容重与土壤碳氮含量具有极显著负相关关系。     

螺旋藻POD、CAT和SOD同工酶的研究*

采用聚丙烯酰胺凝胶电泳法对毛乌素沙地碱湖的钝顶螺旋藻S3和鄂尔多斯螺旋藻S4与国外引进的钝顶螺旋藻S₁和极大螺旋藻S₂ 的POD、CAT和SOD同工酶进行了比较研究。结果表明 :4个样品的 3种酶同工酶带数目不同 ,依次是S₄ >S₃>S₁>S₂ ;S₃和S₄ 酶带数多 ,对环境适应性强 ,进化程度较高。螺旋藻不同种间的酶谱相似系数     

低温胁迫对螺旋藻脱氢酶活性的影响*

低温胁迫试验结果表明,处理的温度越低、天数越长,鄂尔多斯高原碱湖的钝顶螺旋藻S₁、引进的钝顶螺旋藻S₂和极大螺旋藻S₃细胞内脱氢酶活性越低。在每一种处理下,脱氢酶活性高低的排列顺序均为S₁>S₂>S₃。脱氢酶活性的Q₁₀值变化范围排列为S₁。     

Gc、Pi蛋白亚型的快速微量电泳分析

用快速微量等电聚焦技术对190名北京地区汉族健康人血清Gc蛋白亚型、Pi蛋白亚型进行分型鉴定和基因频率调查.上样量为1.5μl,电泳和染色各0.5h.Gc^1F=0.4891,Gc^1S=0.2432,Gc²=0.2678.观察值与期望值吻合良好.(∑X²=1.404,0.7<P<0.8).Pi^M₁=0.7542,Pi^M₂=0.1808,Pi^M₃=0.0650,观察值与期望值吻合也良好,(∑X²=1.1233,0.7<P<0.8).     

农林复合生态系统防护林斑块边缘效应对节肢动物的影响

于2008和2009年在黄淮海平原典型地区商丘民权林场(34°31'-34°52'N, 115°00'-115°28'E)进行。选择杨树林、槐树林和杨槐混交林3种人工林模式,调查节肢动物群落构成、生物多样性及边缘效应对节肢动物群落结构的影响。结果显示,节肢动物科级水平上不同年份间的类群数和个体数差异都不显著(P>0.05),防护林不同模式间个体数(df=2, F=59.48, P<0.001)和类群数(df=2, F=15.44, P<0.001)差异均显著。3种防护林中以杨树林物种丰富度指数较高,S_杨树林(9.65)>S_{杨槐混交林}(8.78)>S_槐树林(7.47);生物多样性指数H'_杨树林(1.78)>H'_{杨槐混交林}(1.65)>H'_槐树林(1.46),其中杨树与槐树林的S和H'差异显著(P<0.05)。杨树林和槐树林边缘效应对节肢动物群落的影响大于杨槐混交林,林地边缘节肢动物的类群数和个体数均较高。节肢动物类群组成表现为林地边缘的偶见种和农田种多于林内。杨树林和槐树林边缘节肢动物多样性指数较高,向林内40m显著下降;而杨槐混交林边缘效应对节肢动物多样性指数的影响较小。综合分析认为,杨树林和槐树林边缘效应对节肢动物群落结构和多样性的影响距离为40m,杨槐混交林距离为20m。单一树种农田防护林斑块不小于80m宽,混交林斑块不小于40m宽,可以保护林内物种生存,维持农田生态系统生物多样性。     

氧化铈纳米颗粒种子引发对盐胁迫下辣椒种子萌发及幼苗生长的影响

氧化铈纳米颗粒(CeO₂NP_S),因具有较强的自由基清除能力和抗氧化酶特性,已被证明可提高植物的耐盐性,但其对辣椒种子引发作用和机制尚不明确。为揭示CeO₂NP_S种子引发处理辣椒对盐胁迫下的萌发及幼苗生长的影响,以辣椒品种(Capsicum annuum)茂蔬360为试验材料,设置了7个CeO₂NP_S浓度(0、0.05、0.1、0.2、0.3、0.4、0.5 mmol·L^-1),以未引发处理组为对照,研究不同浓度CeO₂NP_S种子引发处理后对盐胁迫下辣椒种子萌发、幼苗生物量和生理生化指标的影响。结果表明:(1)0.5 mmol·L^-1 CeO₂NP_S种子引发处理后的种子,其可溶性蛋白质、脯氨酸含量和过氧化氢酶(CAT)活性、抗坏血酸(AsA)含量和AsA/DHA比值显著提高,超氧阴离子(O₂^-)含量显著降低; 盐胁迫下,该处理种子的发芽率、发芽势、发芽指数、活力指数最大。(2)0.4 mmol·L^-1 CeO₂NP_S种子引发处理的幼苗在盐胁迫下的鲜重、干重和根长最大,幼苗的可溶性蛋白质、AsA含量和AsA/DHA比值均显著提高。综上认为,CeO₂NP_S引发处理不仅可通过降低种子水势、促进贮藏物质代谢和提高抗氧化能力提高种子在盐胁迫下的发芽率,还可在苗期通过增强蛋白合成和抗坏血酸-谷胱甘肽循环(AsA-GSH)促进盐胁迫下幼苗的生长。     

遮阴和施氮对黄檗幼苗叶片光合特性和碳氮计量特征的影响

选取大田环境下3年生黄檗幼苗,采用人工控制双因素随机区组试验,在不同光照［全光照(S₀)、轻度遮光21.4%(S₁)和重度遮光8.7%(S₂)］和不同氮添加［无添加对照(F₀)、轻度添加(F₁)和重度添加(F₂)］条件下,测定黄檗幼苗叶片的相对叶绿素含量(SPAD值)、气体交换参数及碳氮化学计量特征,探讨黄檗幼苗对遮阴和施氮的响应机制。结果表明：(1)随着遮光程度增强,黄檗幼苗叶片的SPAD值、蒸腾速率(T_r)、气孔导度(G_s)、碳氮比(C∶N)和瞬时光合氮利用率(PNUE)均呈现先增后降的趋势,两种遮光条件下SPAD均显著高于全光照环境;黄檗幼苗叶片的净光合速率(P_n)、水分利用率(WUE)和气孔限制值(L_s)逐渐降低;而叶片氮(N)和碳(C)含量均呈先降后升的趋势。(2)随氮添加量增加,黄檗幼苗叶片的P_n、WUE、L_s、N和C含量均呈先增后降趋势,T_r、G_s和PNUE则逐渐下降,而C∶N逐渐增加。(3)黄檗幼苗叶片的P_n在各光氮组合处理间均无显著差异;SPAD含量以S₁F₀、S₂F₀和S₁F₂处理组合显著较高,而以全光照(S₀)处理组合显著最低;T_r和G_s以轻度遮光(S₁)处理组合明显较高,而以S₂F₁、S₀F₂、S₂F₂明显较低;WUE和L_s均以S₀F₂显著最高,S₂F₂处理组合显著最低。黄檗幼苗叶片N和C含量在重度遮光/轻度氮添加(S₂F₁)时具有较大值,而其C∶N和PNUE在轻度遮光/无氮添加(S₁F₀)时具有较大值。(4)隶属函数综合评价认为,黄檗幼苗对光氮复合作用总体属中等耐受型,轻度遮光时不添加氮肥(S₁F₀)和轻度氮添加(S₁F₁)及全光照时轻度氮添加(S₀F₁)为适于幼苗生长的光氮组合。研究发现,光环境是影响黄檗幼苗光合作用和更新的主导因子,但黄檗苗期能耐受一定的遮阴胁迫;光照不受限制时,适当增加氮肥有利于黄檗幼苗生长;光照受限(重度遮光)时,施氮则抑制叶片叶绿素合成,降低了幼苗光能利用率,不利于其生长。     

平茬对半干旱黄土丘陵区柠条林地土壤水分的影响

半干旱黄土丘陵区多年生柠条人工林地发生土壤旱化,研究柠条林平茬对土壤水分影响对于防治土壤旱化具有重要意义。采用中子仪测定土壤水分,对未平茬和平茬柠条林地土壤水分进行测定,分析了平茬对土壤水分的影响。结果表明:未平茬和平茬柠条林地降雨补给量(R₁,R₂)同降雨量(P)显著正相关(P<0.05)。定义降雨耗损量(林冠截留量和地表径流之和)占降雨量的百分比为降雨耗损率,未平茬林地降雨损耗率(L₁)和平茬柠条林地降雨损耗率(L₂)分别与其降雨前土壤表层(0-20 cm)含水量(S₁,S₂)呈明显指数关系(P<0.05):L₁=2.54exp(0.22S₁),L₂=2.40exp(0.27S₂),表层含水量相同时,平茬林地降雨损耗率明显高于未平茬林地。平茬后,林地降雨最大入渗深度减小,土壤水分利用深度减小;短时间内(2个月左右)林地20-160 cm含水量增加,之后平茬林地土壤含水量与未平茬林地土壤含水量接近;丰水年和丰水年后的第一年,平茬林地含水量低于未平茬林地,0-400 cm土壤储水量比未平茬林地最多低45.9 mm。平茬后200-400 cm土层土壤水分有少量增加,但是0-200 cm土层土壤含水量损失更严重。平茬3a后,平茬对柠条林地土壤水分的影响减弱。     

直接法测定低密度脂蛋白胆固醇

为了提高血脂检验技术,探讨直接法测定低密度脂蛋白胆固醇(LDL-C),选用市售的两种直接测定LDL-C的试剂盒进行方法学的实验评价,结果表明：直接法具有简便、快速、准确和适合自动分析等特点.两种试剂盒的测定结果都能满足建立分析方法要求的目标,不精密度＜４％,两法显著相关r=0.994, y=1.0572x－0.1052, S_y/x=0.1913,且各种方法学检验结果相近,使用者可结合实际选作常规测定之用.     

体重和盐度对中国蛤蜊耗氧率和排氨率的影响

采用室内实验生态方法研究了不同软体部干重((1.022 ±0.821)、(0.557±0.266)、(0.303±0.277) g)和盐度(13、18、23、28、33)对中国蛤蜊(Mactra chinensis Philippi)耗氧率和排氨率的影响。结果表明:盐度、个体大小对中国蛤蜊耗氧率的影响极显著(P＜0.01),二者的交互作用对中国蛤蜊耗氧率影响显著(P＜0.05);中国蛤蜊单位体重耗氧率(R₀)与软体部干重(W)负相关,符合幂函数方程R₀=aW^-b, 其中a值的取值范围是0.695-1.762,平均值为1.449,b值的取值范围是0.446-0.587,平均值为0.542。盐度、个体大小对中国蛤蜊排氨率影响也极显著(P＜0.01);随着中国蛤蜊个体的增大,其单位体重排氨率逐渐降低;排氨率与其软体部干重呈负相关,它们之间可以用幂函数R_N=a₀W^-b₀表示。单位体重耗氧率和排氨率与盐度(S)、软体部干重(W)的二元线性回归方程分别为: R_O=2.111-1.817W+0.49S (R²=0.546, F=34.294, P＜0.001);R_N=168.186-120.589W+1.734S (R²=0.561, F=36.418, P＜0.001)。     

Molecular epizootiology and evolution of vesicular stomatitis virus New Jersey.

Vesicular stomatitis virus (VSV) has been shown previously to be capable of undergoing rapid mutational change during sequential experimental infections in various tissue culture cell systems (J. Holland, K. Spindler, F. Horodyski, E. Grabau, S. Nichol, and S. Vandepol, Science 215:1577-1585, 1982). The present study was undertaken to determine the degree of genetic diversity and evolution of the virus under natural infection conditions and to gain insight into the epizootiology of the disease. Between 1982 and 1985, numerous outbreaks of VSV of the New Jersey serotype were reported throughout regions of the United States and Mexico. A T1 RNase fingerprint analysis was performed on the RNA genomes of 43 virus isolates from areas of epizootic and enzootic virus activity. This indicates that virus populations were genetically relatively homogeneous within successive U.S. virus epizootics. The data included virus isolates from different epizootic stages, geographical locations, host animals, and host lesion sites. In contrast, only distant genome RNA T1 fingerprint similarities were observed among viruses of the different U.S. epizootics. However, Mexican viruses isolated before or concurrent with U.S. epizootics had very similar RNA genome fingerprints, suggesting that Mexico may have been the possible origin of virus initiating recent U.S. VSV New Jersey outbreaks. Comparison of T1 fingerprints of viruses with enzootic disease areas revealed a greater extent of virus genetic diversity in these areas relative to that observed in epizootic areas. The evolutionary significance of these findings and their relationship to experimental data on VSV evolution are discussed.     

Carbohydrate Structure of Sindbis Virus Glycoprotein E2 from Virus Grown in Hamster and Chicken Cells

Sindbis virus was used as a probe to examine glycosylation processes in two different species of cultured cells. Parallel studies were carried out analyzing the carbohydrate added to Sindbis glycoprotein E2 when the virus was grown in chicken embryo cells and BHK cells. The Pronase glycopeptides of Sindbis glycoprotein E2 were purified by a combination of ion-exchange and gel filtration chromatography. Four glycopeptides were resolved, ranging in molecular weight from 1,800 to 2,700. Structures are proposed for each of the four glycopeptides, based on data obtained by quantitative composition analyses, methylation analyses, and degradation of the glycopeptides using purified exo- and endoglycosidases. The largest three glycopeptides (S1, S2, and S3) have similar structures but differ in the extent of sialylation. All three contain N-acetylglucosamine, mannose, galactose, and fucose, in a structure similar to oligosaccharides found on other glycoproteins. Glycopeptide S1 has two residues of sialic acid, whereas glycopeptides S2 and S3 contain 1 and 0 residues of sialic acid, respectively. The smallest glycopeptide, S4, contains only N-acetyglucosamine and mannose, and is also similar to mannose-rich oligosaccharides found on other glycoproteins. Each of the complex glycopeptides (S1, S2, or S3) from virus grown in BHK cells is indistinguishable from the corresponding glycopeptides derived from virus grown in chicken cells. Glycopeptide S4 is also very similar in size, composition, and sugar linkages from virus derived from the two hosts. These results suggest that chicken cells and BHK cells have similar glycosylation mechanisms and glycosylate Sindbis glycoprotein E2 in nearly identical ways.     

Some characteristics of new tissue-binding proteins for metabolites of vitamin D other than 1,25-dihydroxyvitamin D

Protein(s) have been found in a wide range of tissues which have a high affinity for 25-hydroxycholecalciferol. Of the tissues examined only erythrocytes do not have this protein. The properties of the protein have been examined and it has been found that the association constants range from 2 · 10⁹ to 5 · 10⁹ M⁻¹ and the sedimentation constants between 5.0 and 6.0 S. It was not possible to distinguish the proteins from the different tissues by their S values, mobility on gel electrophoresis or behaviour on ion-exchange chromatography. These techniques were all used, however, to show that the tissue 25-hydroxycholecalciferol binding protein is distinct from the main plasma binding protein for this steroid and from the intestinal 1,25-dihydroxycholecalciferol-binding protein. A protein has been found in the plasma of rachitic animals but not of normals, which is apparently indistinguishable from this new tissue 25-hydroxycholecalciferol-binding protein. The steroid specificity of this new binding protein has been shown to be dependent upon a C-25 hydroxyl group, and an intact conjugated double bond system. Possible functions for this protein have been briefly discussed.     

A yeast assay for high throughput screening of natural anti-viral agents

Over the last decade the yeast Saccharomyces cerevisiae has become a popular organism for studying heterologous gene expression and in vivo protein-protein interactions. Many variations of these basic systems have originated over the years. Besides these vast and varied applications of the yeast expression system, S. cerevisiae has also been used extensively in fundamental research as a model simple eukaryote. We have used the S. cerevisiae system to design a high throughput screen for anti-viral agents from natural sources. The design of the assay rests on the ability of the L-A helper virus and the M(1) satellite virus to detect small variations in -1 ribosomal frameshifting. A minor change in frameshifting efficiencies can be detected and clearly shown phenotypically in terms of zones of clearing on an agar plate. Using such a process, we have initiated a high throughput screening process for natural anti-viral agents.     

Effects on protease inhibition by modifying of helicase residues in hepatitis C virus nonstructural protein 3

This study of the full-length bifunctional nonstructural protein 3 from hepatitis C virus (HCV) has revealed that residues in the helicase domain affect the inhibition of the protease. Two residues (Q526 and H528), apparently located in the interface between the S2 and S4 binding pockets of the substrate binding site of the protease, were selected for modification, and three enzyme variants (Q526A, H528A and H528S) were expressed, purified and characterized. The substitutions resulted in indistinguishable K(m) values and slightly lower k(cat) values compared to the wild-type. The K(i) values for a series of structurally diverse protease inhibitors were affected by the substitutions, with increases or decreases up to 10-fold. The inhibition profiles for H528A and H528S were different, confirming that not only did the removal of the imidazole side chain have an effect, but also that minor differences in the nature of the introduced side chain influenced the characteristics of the enzyme. These results indicate that residues in the helicase domain of nonstructural protein 3 can influence the protease, supporting our hypothesis that full-length hepatitis C virus nonstructural protein 3 should be used for protease inhibitor optimization and characterization. Furthermore, the data suggest that inhibitors can be designed to interact with residues in the helicase domain, potentially leading to more potent and selective compounds.     

Altered utilization of splice sites and 5'' termini in late RNAs produced by leader region mutants of simian virus 40.

We compared the 5' termini and splices of the late 16S and 19S RNAs synthesized by wild-type simian virus 40 and five mutants containing deletions in their late leader region. All mutants produced more unspliced 19S RNA than did wild-type virus, and in two mutants, unspliced 19S RNA constituted more than 60% of the total 19S species. The other three mutants each utilized predominantly a different one of the three spliced species of 19S mRNA. All mutants also produced decreased quantities of 16S mRNA, indicating that they may be defective for splicing both late RNAs. None of the 5' termini of the 16S and 19S RNAs made by the five mutants predominated as in those made by the wild type. Some of the mutant 5' termini were the same as those used by the wild type, whereas others were different. Although present, the major 5'-end positions used by the wild type were frequently not used as major sites by the mutants. In addition, mutants with very similar deletion endpoints synthesized RNAs with different 5' ends. Thus, downstream mutations have a pronounced effect on the location of 5' ends of the late RNAs, and there is no obvious involvement of a measuring function in the placement of 5' ends. For all mutants and wild-type virus, the 5' termini used for 16S and 19S RNAs showed major differences, with some degree of correlation found between the 5' ends and the internal splices of specific mRNA species. A model for the regulation of simian virus 40 late gene expression is presented to explain these findings.     

Shape of proteins S3 and S17 from the small subunit of Escherichia coli ribosome.

The shapes of proteins S3 and S17 purified from the 30 S subunit of Escherichia coli A19 were studied by hydrodynamic methods. The proteins have s020,w values of 2.1 +/- 0.1 S and 1.2 +/- 0.1 S and D020,w values of 7.4 +/- 0.5 . 10(-7) cm2/s and 11.4 +/- 0.6 . 10(-7) cm2/s. The respective molecular weights determined by sedimentation equilibrium are 25 800 +/- 500 and 9900 +/- 300. The intrinsic viscosity values for the two proteins are 5.8 +/- 0.3 ml/g and 4.2 +/- 0.2 ml/g. From these hydrodynamic parameters a slightly elongated shape for S3 and a globular shape for S17 have been concluded.     

Localization of the Single-Stranded RNA-Binding Domains of Bluetongue Virus Nonstructural Protein NS2

The S2 gene of bluetongue virus, serotype 17, has been cloned, and the nonstructural protein NS2 has been expressed. Synthetic peptides matching regions within the amino acid sequence of NS2 were used to map three single-stranded RNA (ssRNA)-binding regions within the protein. A prokaryotic expression system was then used to generate a series of deletion mutants with the ssRNA-binding domains of NS2 removed, singly and in different combinations. These truncated proteins were expressed on a large scale and purified to near homogeneity. The affinity of each truncated protein towards ssRNA was then assayed by electrophoretic mobility shift assays. As a result, the three ssRNA-binding domains of BTV nonstructural protein NS2 have been conclusively localized, and removal of these three domains completely abrogates the ability of NS2 to bind to ssRNA.     

Fusogenic properties of the Ectodomain of HCV E2 envelope protein

The steps leading from hepatitis C virus (HCV) attachment to the hepatocytes to the fusion of viral and cellular membranes remain uncharacterized. In this regard, we have studied the mechanism underlying the HCV fusion process using liposomes and a truncated form of E2 protein lacking the transmembrane region, E2₆₆₁ (amino acids 384–661). E2₆₆₁ has been previously obtained by using the baculovirus expression system and shown to behave as an independent folding domain (M. Rodriguez-Rodriguez, D. Tello, B. Yelamos, J. Gomez-Gutierrez, B. Pacheco, S. Ortega, A.G. Serrano, D.L. Peterson, F. Gavilanes, Structural properties of the ectodomain of hepatitis C virus E2 envelope protein, Virus Res. 139 (2009) 91–99). This form has been used in lipid-protein interaction studies with different model vesicles, at different pHs and by employing a variety of fluorescent assays. The obtained results indicate that E2₆₆₁ induces vesicle aggregation, lipid mixing and liposome leakage, reaching higher values in the presence of negatively charged phospholipids and cholesterol at acidic pH. Therefore, the results of these studies would be indicative of an HCV infection process through receptor mediated endocytosis. Accordingly, E2 might be important in the HCV initial infective steps, interacting with the target membranes and giving rise to their subsequent destabilization.