真菌-氧化氮还原酶细胞色素P-450nor 2 cDNA序列的测定

在真菌的反硝化作用中,一种细胞色素P-450起着一氧化氮还原酶的作用,被称为细胞色素P-450nor^[1]。最近的研究发现：真菌细胞色素P-450nor有三种类型。除了缣孢菌(Fusarium oxysporum)P+450nor(即F．P-450nor)外。还有两种存在于柱孢菌(Cylindrocarpon tonkinense)．即C. P-450norl和2^[2]。 F．P-450nox和C．P-450norl能以NADH为直接的电子供体,使NO还原生成N₂O。C. P-450nor2不仅能直接利用NADH。而且能直接利用NADPH．还原NO生成N₂O。F. P-450nor基因已被克隆和测序^[3-4]。本文测定了C．P-450nor2的eDNA编码区全序列,3’非编码区部分序列和5’引导序列。     

用Blue Sepharose CL-6B快速纯化天花粉蛋白

差光谱显示染料cibacron blue F3GA与天花粉蛋白(TCS)有特异性结合,复合物在可见光部分的最大吸收波长在690 nm,摩尔消光系数ε=2.6×10^-3(mol/L)^-1·cm^-1,解离常数K_d=1.8 μmol/L,0.5 mol/L NaCl可使复合物解离.根据这一特点,用Blue-Sepharose CL-6B凝胶从栝篓块茎中亲和纯化了TCS.此法快速、简便、高效,易于大量制备.     

外源H2O2对盐胁迫下小白菜种子萌发和幼苗生理特性的影响

以小白菜"甜脆青"为试材,研究不同浓度（5、10、25、50和100 mmol/L）过氧化氢（H₂O₂）浸种处理对100 mmol/L NaCl胁迫下小白菜（Brassica chinensis L.）种子萌发、幼苗生长及生理特性的影响。结果表明：100 mmol/L NaCl胁迫明显抑制小白菜种子的萌发状况和幼苗生长,发芽势、发芽指数、活力指数及幼苗根和芽长度和鲜重均明显降低,根和芽中CAT的活性及K⁺含量明显受到抑制,渗透调节物质、活性氧和MDA含量显著增加。不同浓度H₂O₂浸种处理提高了NaCl胁迫下小白菜种子发芽势、发芽指数和活力指数,促进小白菜根和芽的生长,增强了NaCl胁迫下根和芽中SOD、CAT和APX的活性及K⁺含量,降低O₂^-·产生速率及H₂O₂和MDA含量,进一步促进脯氨酸和可溶性糖含量的增加,降低体内Na⁺含量。其中以10 mmol/L H₂O₂处理缓解盐胁迫效果最好,明显缓解NaCl胁迫对小白菜种子萌发和幼苗生长的抑制。     

一种用分光光度计检测氧自由基的新方法

报告检测过硫酸铵／N, N,N′,N′-四甲基乙二胺体系所产生的氧自由基的新方法.O-·2与羟胺溶液反应生成NO-2,NO-2经对氨基苯磺酸和α-萘胺显色在波长530 nm处有专一吸收峰,其颜色深浅与产生的O-·2呈量效关系.氧自由基清除剂抗坏血酸对O-·2的清除作用也呈明显的量效关系.     

γ-氨基丁酸(GABA)种子引发缓解辣椒盐胁迫的效果及生理生化的变化

种子引发是提高作物生长期耐盐性的有效方法,而γ-氨基丁酸(GABA)种子引发对辣椒(Capsicum annuum)耐盐性的效果和作用机制尚不清楚。该研究以‘茂蔬360''朝天椒为材料,分析了不同浓度γ-氨基丁酸(0、1.0、2.0、4.0、6.0、8.0 μmol·L^-1)种子引发对4～6叶期100 mmol·L^-1 NaCl胁迫下的植株生物量、渗透调节物质、抗氧化能力、光合作用系统及钾钠离子吸收的影响。结果表明:(1)种子引发能显著增加盐胁迫下辣椒植株的生物量,以6.0 μmol·L^-1 GABA引发处理的效果最佳。(2)种子引发处理增加了盐胁迫下植株的可溶性糖、可溶性蛋白及脯氨酸的含量,同时,O₂^-和MDA的含量下降,抗氧化酶(SOD、POD、CAT和APX)活性增强,叶片叶绿素含量增加,叶片叶绿素荧光参数受胁迫影响程度低,叶绿素荧光指标包括F_v ''/F_m ''、qP_Lss、QY_Lss、NPQ_Lss和Rfd均有所上升。根、茎中的K⁺含量和K⁺/Na⁺比值下降。(3)灰色关联度分析表明,GABA种子引发主要通过提高POD、CAT的活性和渗透调节物质含量来缓解盐胁迫对辣椒植株的伤害。综上所述,6.0 μmol·L^-1 GABA种子引发可有效提高辣椒苗期的耐盐性,其作用机制可能是提高了盐胁迫下辣椒植株的抗氧化能力和渗透调节能力。     

纤维蛋白对尿激酶原激活纤溶酶原反应动力学的影响

反应体系中存在的纤维蛋白（fibrin）对尿激酶（UK）、scu-PA以及组织型纤溶酶原激活剂（t-PA）激活纤溶酶原（plasminogen）的反应有不同的作用：UK、t-PA激活plasminogen的反应可被反应体系中存在的fibrin所加强;fibrin对scu-PA激活plasminogen反应的动力学常数无明显影响;但对小分子质量scu-PA与单链抗体的嵌合分子激活plasminogen的反应起明显的抑制作用.为确定反应体系中存在的fibrin对scu-PA的K区插入突变体-InB激活plasminogen反应的影响,测定了在反应体系中存在fibrin的情况下的InB激活plasminogen反应的K_m^fibrin以及k_cat^fibrin.K_m^fibrin＝4.2 μmol·L^-1,远远大于无fibrin时的K_m＝0.379 μmol·L^-1,说明有fibrin存在时突变体InB与天然底物plasminogen的亲和性降低了.k_cat^fibrin＝0.107 s^-1,也远远大于无fibrin时k_cat＝0.0165 s^-1,说明有fibrin存在时突变体InB对plasminogen的反应活性增强了.原因可能是：与fibrin结合的plasminogen的构象发生了有利于被纤溶酶原激活剂水解的变化.     

Metabolic oxygen regulation and conformity during submergence in the salamandersSiren lacertina,Amphiuma means,andAmphiuma tridactylum,and a comparison with other giant salamanders

Summary The giant salamanders of North America include 4 genera, all of which are aquatic. We have compared the efficacy of aquatic O₂ uptake among them by measuring theVO₂ while submerged and determining the responses to progressive hypoxia at 10–240 mmHg at 20° C. Both species ofAmphiuma were metabolic O₂ conformers over the entire range ofPO₂. About half ofSiren lacertina were conformers over this range, and half were regulators with an average critical O₂ tension of 92 mmHg. There were no short-term changes (days) in the response ofSiren to progressive hypoxia, but one animal switched from conformation to regulation after 4–5 months. Neither genus is considered to have an exceptionally low metabolic rate. The “whole-body O₂ conductance”, defined asΔVO₂/ΔPO₂(μl O₂ · g⁻¹ · h⁻¹ · mmHg⁻¹) in the range of metabolic O₂ conformity, was least in the species most dependent upon air-breathing and most likely to be found in hypoxic waters (e.g., 0.076 forAmphiuma), and greatest in those that airbreathe less frequently and/or are found in relatively normoxic waters (e.g., 0.429 forNecturus). These conductances are considered to be adaptive in terms of preventing O₂ loss through the skin, or in facilitating its uptake, as correlated with the O₂ tensions normally prevailing in the environment of each species.     

Kinetics of urea uptake by Melosira italica (Ehr.) Kütz at different luminosity conditions

The kinetic parameters K_m and V_max for urea uptake by Melosira italica were determined at 160 μeinsteins m⁻² s⁻¹ and in the dark. The transport systems showed an affinity for the substrate and a storing capacity in the dark (K_m = 65.07 μM; V_max = 2.18 nmoles 10⁵ cells ⁻¹ h⁻¹) greater than under 160 μE m⁻² s ⁻¹ (K_m = 111.2 μM; V_max = 1.11 nmoles 105 cells⁻¹ h⁻¹). Similarly, a reduction in consumption rate of urea under increasing photon flux densities was observed. The use of an inhibitor (potassium cyanide) indicated that the uptake process requires metabolic energy. That urea transport is more important in darkness, may constitute a survival strategy in which this compound is utilized by cells mainly during heterotrophic growth.     

Characterization of the catalytically active Mn(II)-loaded argE -encoded N -acetyl-l -ornithine deacetylase from Escherichia coli

The catalytically competent Mn(II)-loaded form of the argE-encoded N-acetyl-l-ornithine deacetylase from Escherichia coli (ArgE) was characterized by kinetic, thermodynamic, and spectroscopic methods. Maximum N-acetyl-l-ornithine (NAO) hydrolytic activity was observed in the presence of one Mn(II) ion with k _cat and K _m values of 550 s⁻¹ and 0.8 mM, respectively, providing a catalytic efficiency (k _cat/K _m) of 6.9 × 10⁵ M⁻¹ s⁻¹. The ArgE dissociation constant (K _d) for Mn(II) was determined to be 0.18 μM, correlating well with a value obtained by isothermal titration calorimetry of 0.30 μM for the first metal binding event and 5.3 μM for the second. An Arrhenius plot of the NAO hydrolysis for Mn(II)-loaded ArgE was linear from 15 to 55 °C, suggesting the rate-limiting step does not change as a function of temperature over this range. The activation energy, determined from the slope of this plot, was 50.3 kJ mol⁻¹. Other thermodynamic parameters were ΔG ^‡ = 58.1 kJ mol⁻¹, ΔH ^‡ = 47.7 kJ mol⁻¹, and ΔS ^‡ = –34.5 J mol⁻¹ K⁻¹. Similarly, plots of lnK _m versus 1/T were linear, suggesting substrate binding is controlled by a single step. The natural product, [(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]leucine (bestatin), was found to be a competitive inhibitor of ArgE with a K _i value of 67 μM. Electron paramagnetic resonance (EPR) data recorded for both [Mn(II)_(ArgE)] and [Mn(II)Mn(II)(ArgE)] indicate that the two Mn(II) ions form a dinuclear site. Moreover, the EPR spectrum of [Mn(II)Mn(II)(ArgE)] in the presence of bestatin indicates that bestatin binds to ArgE but does not form a μ-alkoxide bridge between the two metal ions.     

不同灌水上限和施肥量对温室番茄产量与品质的影响

以番茄品种‘英石大红’为材料,在滴灌施肥种植条件下进行日光温室栽培试验;灌水上限设3个水平,即土壤相对含水量分别为田间最大持水量的70%(W₁)、80%(W₂)、90%(W₃),灌水下限统一设定为田间最大持水量的50%;N、P、K施肥设3个水平,分别为低肥F₁ ［N(228 kg/hm²)、P₂O₅(132 kg/hm²)、K₂O(300 kg/hm²)］、中肥F₂ ［N(285 kg/hm²)、P₂O₅(165 kg/hm²)、K₂O(375 kg/hm²)］和高肥F₃ ［N(342 kg/hm²)、P₂O₅(198 kg/hm²)、K₂O(450 kg/hm₂)］,以大水漫灌(8165 m³/hm²)、当地推荐施肥量(342 kg/hm² N、198 kg/hm² P₂O₅、450 kg/hm² K₂O) 组成共同水肥对照(CK),探讨不同灌水上限和施肥量处理组合对番茄产量、品质和干物质量的影响,以筛选日光温室栽培番茄的最佳灌水上限和施肥量组合。结果显示：(1)在水肥一体化条件下,番茄植株的地上部干鲜重、地下部干鲜重、根冠比除F₃W₁处理外均显著高于传统水肥CK。(2)番茄果实产量在不同灌溉上限和施肥量处理下表现各异,其中在F₂W₂处理下番茄果实单果重、经济产量、生物产量达到最高,且与CK差异显著。(3)在同一灌水上限条件下,在一定范围内增加施肥量可以显著提高番茄果实的可溶性糖、维生素C、可溶性蛋白、番茄红素含量,而番茄果实有机酸和硝酸盐含量随施肥量的增加而不断增加;在同一施肥量条件下,番茄果实有机酸含量随着灌水上限的上调而不断减少,而番茄果实硝酸盐含量却随着灌水上限的上调而不断增加。研究发现,在滴灌施肥条件下合理控制温室番茄的灌水量与施肥量能够显著增加番茄产量以及番茄果实中可溶性糖、维生素C、可溶性蛋白、番茄红素、有机酸等含量,降低硝酸盐含量,增加干物质积累量;在灌水上限为田间最大持水量的80%、灌水量为3 686.691 m³/hm²,施肥量为285 kg/hm² N、165 kg/hm² P₂O₅、375 kg/hm² K₂O是最佳水肥处理组合,且番茄产量可以比传统水肥处理显著增加15.03%,果实品质同时得到有效改善。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.