Isolation and functional characterization of fatty acid delta5-elongase gene from the liverwort Marchantia polymorpha L

Bryophyte Marchantia polymorpha L. produces C22 very-long-chain polyunsaturated fatty acid (VLCPUFA). Thus far, no enzyme that mediates elongation of C20 VLCPUFAs has been identified in land plants. Here, we report the isolation and characterization of the gene MpELO2, which encodes an ELO-like fatty acid elongase in M. polymorpha. Heterologous expression in yeast demonstrated that MpELO2 encodes delta5-elongase, which mediates elongation of arachidonic (20:4) and eicosapentaenoic acids (20:5). Phylogenetic and gene structural analysis indicated that the MpELO2 gene is closely related to bryophyte Delta6-elongase genes for C18 fatty acid elongation and diverged from them by local gene duplication.     

病原微生物对抗菌肽抗性机制的研究进展

抗菌肽(antimicrobial peptides, AMPs)是生物先天免疫系统的重要组成部分,可帮助宿主有效应对病原细菌、真菌和病毒等微生物的胁迫,被认为是医疗、食品加工和农业领域最具前途和潜力的抗生素替代物。病原微生物在与抗菌肽的互作中进化出了多种有针对性的抗性机制,本文从病原微生物对AMPs的感应与基因调控、细胞壁/膜成份的修饰、分泌蛋白酶降解及利用外排泵排出等四个方面综述了国内外的研究进展,并对AMPs类制品的研究前景进行了讨论与展望。     

Autoradiographic reevaluation of the binding properties of 125I-[Leu31,Pro34]peptide YY and 125I-peptide YY3-36 to neuropeptide Y receptor subtypes in rat forebrain

125I-[Leu31,Pro34]peptide YY (PYY) and 125I-PYY3-36, initially described as selective neuropeptide Y Y1 and Y2 receptor ligands, respectively, were recently shown to label also Y4 and Y5 receptors. We used receptor autoradiography to assess whether these ligands can be reliably used to investigate the various neuropeptide Y receptors in rat forebrain. In most of the brain regions examined (in coronal sections at the level of dorsal hippocampus), specific 125I-[Leu31,Pro34]PYY binding was completely inhibited by 1 microM BIBP-3226, a selective Y1 receptor ligand, but unaffected by 10 nM rat pancreatic polypeptide, selectively inhibiting Y4 receptors, suggesting that Y4 receptors are present in negligible numbers compared with Y1 receptors in the areas examined. Significant numbers of BIBP-3226-insensitive 125I-[Leu31,Pro34]PYY binding sites were measured in the CA3 subfield of the hippocampus only, possibly representing Y5 receptors. 125I-PYY3-36 binding was unchanged by 1 microM BIBP-3226, whereas a population of 125I-PYY3-36 binding sites was sensitive to 100 nM [Leu31,Pro34]neuropeptide Y, likely representing Y5 receptors. The possibility of distinguishing between Y2 and Y5 receptors using 125I-PYY3-36 as radioligand was validated by their different regional distribution and their distinct changes 24 h after kainate seizures, i.e., binding to Y5 receptors was selectively decreased in the outer cortex, whereas binding to Y2 receptors was enhanced in the hippocampus. Thus, the use of selective unlabeled compounds is required for distinguishing the various receptor subtypes labeled by 125I-[Leu31,Pro34]PYY and 125I-PYY3-36 in rat brain tissue.     

Induced Resistance in Cucumbers against Fusarium oxysporum f. sp. cucumerinum and Rhizoctonia solani AG2–2 by Infection of the Cotyledons

Localized infection in cucumber cotyledons with Colletotrichum lagenarium induced resistance against infection after challenge inoculation with Rhizoctonia solani AG2–2 and Fusarium oxysporum f. sp. cucumerinum in the roots. The plants were unprotected in soil that was infested heavily with R. solani or in contact with the mycelium, and induced resistance was not observed. Wounding of the root also negated the effect of induced resistance to F. oxysporum .     

The Effects of Root-knot Nematode Infection and Mi-mediated Nematode Resistance in Tomato on Plant Fitness

The Mi-1.2 resistance gene in tomato (Solanum lycopersicum) confers resistance against several species of root-knot nematodes (Meloidogyne spp.). This study examined the impact of M. javanica on the reproductive fitness of near-isogenic tomato cultivars with and without Mi-1.2 under field and greenhouse conditions. Surprisingly, neither nematode inoculation or host plant resistance impacted the yield of mature fruits in field microplots (inoculum=8,000 eggs/plant), or fruit or seed production in a follow-up greenhouse bioassay conducted with a higher inoculum level (20,000 eggs/plant). However, under heavy nematode pressure (200,000 eggs/plant), greenhouse-grown plants carrying Mi-1.2 had more than ten-fold greater fruit production than susceptible plants and nearly forty-fold greater estimated lifetime seed production, confirming prior reports of the benefits of Mi-1.2. In all cases Mi-mediated resistance significantly reduced nematode reproduction. These results indicated that tomato can utilize tolerance mechanisms to compensate for moderate levels of nematode infection, but that the Mi-1.2 resistance gene confers a dramatic fitness benefit under heavy nematode pressure. No significant cost of resistance was detected in the absence of nematode infection.     

翻译和非翻译马铃薯Y病毒外壳蛋白基因介导的抗病性比较

利用RT－PCR方法克隆获得马铃薯Y病毒烟草叶脉坏死株系（PVY^N）的可翻译和不可翻译外壳蛋白（CP）基因,并分别插入pROKⅡ质粒中获得重组双元表达载体。通过根癌农杆菌（Agrobacterium tumefaciens）介导的基因转化方法,将可翻译和不可翻译的PVY^N-CP基因分别导入烟草栽培品种NC89叶片组织中,得到抗卡那霉素的再生植株,对所得到的抗卡那霉素的再生苗进行PCR检测表明,被导入可翻译和不可翻译CP基因的植株分别占卡那霉素抗性植株的95％和98％。攻毒实验表明,两种类型的转基因烟草对PVY^N的抗病性具有相似性,其表现型为：免疫、抗病和感病。免疫型转基因植株的抗病性不受接种物类型及其剂量的影响。Southern印迹杂交结果显示,目的基因已经整合到烟草基因组中。Northern印迹杂交证明,两种类型的CP基因都已在RNA水平上得到了表达,但细胞内RNA的积累量与转基因植株的抗病强度成负相关。Western印迹杂交表明,在表达不可翻译PVY^N－CP基因的转基因植株内 以CP蛋白,而在导入可翻译的PVY^N-CP基因的植株内检测到了CP蛋白,且CP蛋白含量与抗病性不存在正相关。本研究结果证明,表达可翻译与不可翻译PVY^N-CP基因的转基因烟草对PVY^N的抗病性均为RNA介导的抗病性。     

Conservation of globin genes in the “living fossil” Latimeria chalumnae and reconstruction of the evolution of the vertebrate globin family

The (hemo-)globins are among the best-investigated proteins in biomedical sciences. These small heme-proteins play an important role in oxygen supply, but may also have other functions. In addition to well known hemoglobin and myoglobin, six other vertebrate globin types have been identified in recent years: neuroglobin, cytoglobin, globin E, globin X, globin Y, and androglobin. Analyses of the genome of the “living fossil” Latimeria chalumnae show that the coelacanth is the only known vertebrate that includes all eight globin types. Thus, Latimeria can also be considered as a “globin fossil”. Analyses of gene synteny and phylogenetic reconstructions allow us to trace the evolution and the functional changes of the vertebrate globin family. Neuroglobin and globin X diverged from the other globin types before the separation of Protostomia and Deuterostomia. The cytoglobins, which are unlikely to be involved in O₂ supply, form the earliest globin branch within the jawed vertebrates (Gnathostomata), but do not group with the agnathan hemoglobins, as it has been proposed before. There is strong evidence from phylogenetic reconstructions and gene synteny that the eye-specific globin E and muscle-specific myoglobin constitute a common clade, suggesting a similar role in intracellular O₂ supply. Latimeria possesses two α- and two β-hemoglobin chains, of which one α-chain emerged prior to the divergence of Actinopterygii and Sarcopterygii, but has been retained only in the coelacanth. Notably, the embryonic hemoglobin α-chains of Gnathostomata derive from a common ancestor, while the embryonic β-chains – with the exception of a more complex pattern in the coelacanth and amphibians – display a clade-specific evolution. Globin Y is associated with the hemoglobin gene cluster, but its phylogenetic position is not resolved. Our data show an early divergence of distinct globin types in the vertebrate evolution before the emergence of tetrapods. The subsequent loss of globins in certain taxa may be associated with changes in the oxygen-dependent metabolism. This article is part of a Special Issue entitled: Oxygen Binding and Sensing Proteins.     

高压电场对A549细胞中ABCG2和V-ATPase的表达及其耐药性的影响

目的:探讨高压电场对A549细胞中ABCG2和V-ATPase表达量的影响;探讨高压电场对A549细胞耐药性的影响。方法:MTT法测细胞生长曲线,明确能导致细胞可逆性电穿孔的最高电场强度。慢病毒构建ABCG2和V-ATPase低表达的A549细胞系,并用电场处理,用q-RT-PCR和Western-blot法检测处理前后ABCG2和V-ATPase的m RNA和蛋白表达量的变化。最适强度的高压电场处理各组细胞,在处理前后的细胞中分别加入阿霉素,用高效液相色谱法检测各组细胞中阿霉素浓度。结果:当电场强度为1500 V/cm时,肿瘤细胞增殖最慢;电场强度为1500 V/cm时,肿瘤细胞中ABCG2和V-ATPase的m RNA和蛋白的表达量分别降至对照组的58%和61%,具有统计学差异;1500 V/cm强度的电场可以提高肿瘤细胞内阿霉素的浓度3-4倍。结论:高压电场可以显著降低肿瘤细胞中V-ATPase和ABCG2的m RNA和蛋白的表达量并降低肿瘤细胞的耐药性。     

植物抗病基因结构、功能及其进化机制研究进展

植物与病原菌在长期的共进化和相互选择的过程中,逐渐形成了组织障碍、非寄主抗性和小种专化抗性等有效的防御机制。小种专化抗性(基因对基因抗性)主要是由植物抗病基因识别相应的病原菌无毒基因并激活植物体内抗病信号进而抵御病原菌的侵染。从目前已克隆的 70 多个抗病基因来看,它们在结构上具有高度保守性,主要包括核苷酸结合位点(NBS),亮氨酸重复结构(LRR), 蛋白激酶结构域(PK), 果蝇蛋白 Toll 和哺乳动物蛋白质白细胞介素 1 受体[interleukin(IL)-1 receptor]类似结构域(TIR), 双螺旋结构(CC)或亮氨酸拉链(LZ)和跨膜结构域(TM)等,其在抗病基因与病原菌无毒(效应)蛋白互作以及植物内部免疫信号传导中起着重要的作用。同时,抗病基因又通过基因复制、遗传重组等进化机制形成多基因家族,为植物抗病的专化性和多样性提供了重要的遗传基础。本文主要讨论了近来已克隆抗病基因的结构特征、功能以及抗病基因进化机制研究的进展。     

小兴安岭7种典型林型林分生物量碳密度与固碳能力

森林生物碳储量作为森林生态系统碳库的重要组成部分, 在全球碳循环中发挥着重要作用。以小兴安岭7种典型林型为研究对象, 通过外业样地调查与室内实验分析相结合的方法, 从林分尺度对林分生物量与碳密度进行计量, 分析了林分生物碳储量的空间分配格局, 并对林分年固碳能力与碳汇潜力进行了探讨。结果表明: 小兴安岭不同林型从幼龄林到成熟林的乔木层碳密度增长速率为: 蒙古栎(Quercus mongolica)林>兴安落叶松(Larix gmelinii)林>云冷杉(Picea-Abies)林>樟子松(Pinus sylvestris var. mongolica)林>山杨(Populus davidiana)林>红松(Pinus koraiensis)林>白桦(Betula platyphylla)林。7种典型林型不同龄组(幼龄林、中龄林、近熟林和成熟林)林分生物量碳密度分别为: 红松林31.4、74.7、118.4和130.2 t·hm^-2; 兴安落叶松林28.9、44.3、74.2和113.3 t·hm^-2; 樟子松林22.8、52.0、71.1和92.6 t·hm^-2; 云冷杉林23.1、44.1、77.6和130.3 t·hm^-2; 白桦林18.8、35.3、66.6和88.5 t·hm^-2; 蒙古栎林25.0、20.0、47.5和68.9 t·hm^-2; 山杨林19.8、28.7、43.7和76.6 t·hm^-2。红松林、兴安落叶松林、樟子松林和蒙古栎林在幼龄林时林分年固碳量较高, 其他林型在成熟林时林分年固碳量较高。7种典型林型不同龄组的林分生物量碳密度均随林龄增长而增加, 但不同林型的碳汇功能存在差异, 同一林型不同林龄的生物量碳密度增幅差异也较大。林分年固碳量在0.4-2.8 t·hm^-2之间, 碳汇能力较强、碳汇潜力较大。尤其是小兴安岭目前林分质量较差, 幼龄林和中龄林所占的比重较大, 具有较大的碳汇潜力。研究结果可为森林经营管理及碳汇功能评价提供参考。     

区域资源环境承载力科学概念及其生态学基础的讨论

资源环境承载力是生态学领域的一个重要概念,其理论和实践研究已成为衡量区域可持续发展的重要依据.但是有关生态学基础与资源环境承载力的科学联系仍未明确,其中有哪些生态学理论对资源环境承载力的发展起到了支撑作用还没被厘清,这使得资源环境承载力的科学概念十分模糊.本文在讨论资源环境、生态系统承载力科学概念及其发展的基础上,系统...     

川西亚高山林区不同土地利用与土地覆盖的地被物及土壤持水特征

川西亚高山森林是我国西南亚高山水源涵养林的重要组成部分.随着20世纪中叶以来农业人口的增加和森林的开发利用,受干扰强度、频度和干扰时间的影响,原始暗针叶林退化为耕地、草地、灌丛、次生阔叶林或人工林.1998年后,该区相继启动天然林保护工程和退耕还林工程.为评价工程效益,确定长江上游水源涵养林的恢复与重建模式,需要进行不同土地利用类型之间生态水文效应的对比分析.通过野外调查与室内实验,对比分析了川西亚高山林区农田、草地、退耕还林地、灌丛、次生桦木林、人工云杉林和老龄针叶林的地被物(苔藓与枯落物)和土壤持水特征,结果表明:不同土地利用与覆盖类型间地被物和土壤持水性能差异显著.随着干扰程度的增加,苔藓、枯落物蓄积量及最大持水量下降,土壤容重增加,土壤持水性能下降.苔藓最大持水量排序是老龄针叶林>人工云杉林>天然次生林>灌丛.枯落物最大持水量排序是老龄针叶林>天然次生林>人工云杉林>灌丛>草地>退耕还林地.人工云杉林与天然次生林之间、草地与退耕还林地之间苔藓和枯落物最大持水量没有显著差异.土壤0～40cm最大持水量排序是天然次生林>老龄针叶林>人工云杉林>灌丛>农田>草地>退耕还林地,其中天然次生林显著高于人工云杉林,草地与退耕还林地之间没有显著差异.对于森林恢复途径和树种的选择,需要考虑未来林分的多种生态系统服务功能.     

Use of Arabidopsis thaliana and Pseudomonas syringae in the Study of Plant Disease Resistance and Tolerance

The interaction between Arabidopsis thaliana and the bacterium Pseudomonas syringae is being developed as a model experimental system for plant pathology research. Race-specific ("gene-for-gene") resistance has been demonstrated for this interaction, and pathogen genes that determine avirulence have been isolated and characterized. Because certain lines of both Arabidopsis and soybean are resistant to bacteria carrying the avirulence genes avrRpt2 and avrB, extremely similar pathogen recognition mechanisms are apparently present in these two plant species. Isogenic bacterial strains that differ by the presence of single avirulence genes are being used to analyze plant resistance. Plant resistance genes have been identified in crosses between resistant and susceptible lines. The extensive map-based cloning tools available in Arabidopsis are being used to isolate these resistance genes. In a related project, ethylene-insensitive Arabidopsis mutants are being used to examine the role of ethylene in disease development. Ethylene apparently mediates symptom formation in susceptible plants and is not required for resistance, suggesting possible strategies for enhancement of disease tolerance in crops.     

大豆对4个大豆花叶病毒株系的抗性及其连锁遗传研究

利用7个抗大豆花叶病毒1—4个株系的品种(系)与1个丰产感病品种杂交,配制成10个组合。从其P_1、P_2、F_1、F_2、F_3和BC_1F_16个世代的鉴定结果表明:大豆对S(?)、S_c、S(?)和S_h株系的抗性分别受单个显性基因R(?)、R_c、R(?)和R_h控制。R(?)与R_c、R(?)与R_h间的重组率分别为25.4±7.7％和27.3±5.4％;R(?)与R_h、R_a与R_g、R_c与R_g间的重组率分别为23.5±5.2％、41.1±8.5％和46.5±7.5％。4个基因的排列项序为R_g—R_h—R_a—R_c,它们不属第一和第八连锁群。     

Nucleotide sequence of the genes coding for alpha, beta and gamma subunits of the proton-translocating ATPase of Escherichia coli

A nucleotide sequence of 2328 base pairs comprising a portion of the gene cluster for the proton-translocating ATPase of $\text{E. coli}$ was determined. The sequence covers most of the gene for α subunit, the entire gene for γ subunit and the amino terminal portion of the gene for β subunit, along with the flanking regions of these genes. The amino acid sequences of these subunits deduced from the DNA sequences indicate that the α and γ subunits have 513 and 287 amino acid residues, respectively. A possible secondary structure for each subunit was estimated from the inferred primary structure. The intercistronic regions between the genes for α and γ and between γ and β are 49 and 26 base pairs, respectively. The significance of codon usage in these genes is discussed in correlation with their expression.     

Single amino acid (482) variants of the ABCG2 multidrug transporter: major differences in transport capacity and substrate recognition

The human ABCG2 protein is an ATP binding cassette half-transporter, which protects our cells and tissues against various xenobiotics, while overexpression of ABCG2 in tumor cells confers multidrug resistance. It has been documented that single amino acid changes at position 482 resulted in altered drug resistance and transport capacity. In this study, we have generated nine Arg-482 mutants (G, I, M, S, T, D, N, K, Y) of ABCG2, and expressed them in insect cells. All ABCG2 variants showed cell surface expression and, in isolated membranes, an ABCG2-specific ATPase activity. When methotrexate accumulation was measured in inside-out membrane vesicles, this transport was supported only by the wild-type ABCG2. In intact cells, mitoxantrone was transported by all ABCG2 variants, except by R482K. Rhodamine 123 was extruded by most of the mutants, except by R482K, Y and by wild-type ABCG2. Hoechst 33342 was pumped out from cells expressing the wild-type and all Arg-482 variants, but not from those expressing R482K and Y. Our study demonstrates that the substrate specificity of the Arg (wild-type) form is unique and that amino acid replacements at position 482 induce major alterations in both the transport activity and substrate specificity of this protein.     

Seasonal changes in the oxygen storage capacity and aerobic dive limits of the muskrat (Ondatra zibethicus)

Summary The oxygen storage capacity and partitioning of body oxygen reserves were compared in summer-and winter-acclimatized muskrats (Ondatra zibethicus). Blood volume, blood oxygen capacity, and skeletal muscle myoglobin content were higher in December than in July (P<0.02). Total lung capacity increased only slightly in winter (P>0.05). The oxygen storage capacity of a diving muskrat was calculated at 25.2 ml O₂ STPD · kg^-1 in July, compared to 35.7 ml O₂ STPD · kg^-1 in December. Blood comprised the major storage compartment in both seasons, accounting for 57% and 65% of the total oxygen stores in summer and winter, respectively. Based on available oxygen stores and previous estimates of the cost of diving, the aerobic dive limit (ADL) increased from 40.9 s in July to 57.9 s in December. Concurrent behavioral studies suggested that most voluntary diving by muskrats is aerobic. However, the proportion of dives exceeding the calculated ADL of these animals was shown to vary with the context of the dive. Only 3.5% of all dives initiated by muskrats floating in the water exceeded their estimated ADL. Provision of a dry resting site and access to a submerged food source increased this proportion to 18–61%, depending on the underwater distance that foraging muskrats were required to swim. Serial dives exceeding the estimated ADL were not accompanied by extended postdive recovery periods.Abbreviations ADL acrobic dive limit - Hb hemoglobin - Hct hematocrit - Mb myoglobin - PaO₂ arterial O₂ tension - STPD standard temperature and pressure, dry     

黄土高原北洛河流域林地枯落物特征及水分吸持效应

明确黄土高原不同林分类型枯落物的时空变化特征,理解其水土保持功能,为流域土壤侵蚀模拟和预测提供基础数据和理论支撑。野外实测了北洛河流域内山杨、刺槐和沙棘3种人工林分以及乔木初期（白桦）、中期（辽东栎-油松混交林）、亚顶级（油松）和顶级（辽东栎）4个次生演替阶段林分枯落物盖度、厚度和蓄积量。用浸泡法对枯落物持水能力和过程进行了研究。流域各林分类型枯落物厚度介于4.55-1.38 cm,蓄积量介于17.24-4.99 t/hm²,24 h持水深和有效拦蓄深分别介于2.73-0.96 mm和2.45-0.81 mm,表现为辽东栎-油松混交林 > 油松/辽东栎 > 山杨 > 刺槐 > 沙棘 > 白桦。枯落物持水量与浸泡时间存在对数函数关系（R²≥0.85,P<0.01）,2 h以内迅速增加,8 h基本饱和。半分解层厚度、蓄积量、持水量和吸水速率均大于未分解层。流域内次生演替林分水分吸持能力和拦蓄作用总体上优于人工林分。植被演替过程中,辽东栎-油松混交林枯落物盖度、蓄积量和持水能力表现最优,建议在黄土高原造林及林地抚育管理过程中重视混交林的建设和保护。     

Adaptive modifications in membranes of halotolerant and halophilic microorganisms

Halotolerant and halophilic microorganisms can grow in (hyper)saline environments, but only halophiles specifically require salt. Genotypic and phenotypic adaptations are displayed by halophiles; the halotolerants adapt phenotypically, but it is not established whether they show genotypic adaptation. This paper reviews the various strategies of haloadaptation of membrane proteins and lipids by halotolerant and halophilic microorganisms. Moderate halophiles and halotolerants adapt their membrane lipid composition by increasing the proportion of anionic lipids, often phosphatidylglycerol and/or glycolipids, which in the moderately halophilic bacteriumVibrio costicola appears to be part of an osmoregulatory response to minimize membrane stress at high salinities. Extreme halophiles possess typical archaebacterial ether lipids, which are genotypically adapted by having additional substitutions with negatively-charged residues such as sulfate. In contrast to the lipids, it is less clear whether membrane proteins are haloadapted, although they may be more acidic; very few depend on salt for their activity.