Determining the involvement of two aminopeptidase Ns in the resistance of Plutella xylostella to the Bt toxin Cry1Ac: cloning and study of in vitro function

The cloning, expression in vitro, and characterization of two aminopeptidase Ns (APN5s and APN2s) isolated from the midgut of Cry1Ac-resistant (R) and susceptible (S) strains of Plutella xylostella larvae are presented in this paper. The deduced amino acid sequences of APN5s included C-terminal GPI-modification sites, the gluzincin aminopeptidase motif GATEN, and three N-glycosylated sites; those of APN2s had no GPI-modification sites, had gluzincin aminopeptidase motif GAMEN, and had four N-glycosylated sites. O-glycosylated sites were not predicted for either APN. Because APN2R and APN2S cDNAs contained the same nucleotides, only full-length cDNAs encoding APN5R and APN5S were expressed in Trichoplusia ni cells. Far-Western blotting showed that the expressed receptor APN5 bound to the Cry1Ac toxin. An enzyme-specific activity experiment also showed that APN5 genes were expressed in T. ni cells. ELISA revealed no differences in the binding of expression proteins from the resistant and susceptible strain with Cry1Ac.     

泛素化修饰与植物免疫应答

植物暴露在细菌、病毒和真菌等病原微生物环境中,病虫害是限制农作物产量和品质的主要因素,而植物病虫害的防治依赖于对植物抗病机制的深入认识。近年来的研究表明,蛋白质泛素化广泛地参与植物防御调节。蛋白质泛素化是真核生物中重要的翻译后修饰方式之一,在植物中,泛素化修饰在多种信号传导途径中发挥作用,如激素、光、糖应答,发育调节和病原菌防御信号途径等。综述了蛋白质泛素化修饰在植物免疫应答中的调控作用。     

稻根象甲种群空间格局及其受耕法的影响

本文根据36块田的调查,用种群空间格局参数I、C_A、I_δ、m~*/m和α、β值进行分析,稻根象甲幼虫和(?)≥0.6头/样方(每样方为0.5m~2)的成虫种群,在稻田均属具公共k值的负二项分布;(?)<0.6头/样方的成虫种群则为随机分布。成虫聚块的大小和面积与平均密度有关。低密度时种群的聚集主要由于环境的异质性;高密度时种群的聚集则由本身的行为习性所致。随着耕作强度的增加,幼虫种群的聚集度和越冬死亡率增大。     

瓜子金化学成分的研究

自瓜子金(Palygala japonica)中分离出16个化合物,本文报道其中4个化台物的分离和结构鉴定。经化学和光谱解析,鉴定为β-谷甾醇(1)、β-胡萝卜甙(Ⅱ)、槲皮素(Ⅲ)和鼠李亭-3-O-葡萄吡喃糖甙(Ⅳ),均为首次从该植物中获得。     

Partitioning of triphenylalkylphosphonium homologues in gel bead-immobilized liposomes: chromatographic measurement of their membrane partition coefficients

Unilamellar liposomes of small or large size, SUVs and LUVs, respectively, were stably immobilized in the highly hydrophilic Sepharose 4B or Sephacryl S-1000 gel beads as a membrane stationary phase for immobilized liposome chromatography (ILC). Lipophilic cations of triphenylmethylphosphonium and tetraphenylphosphonium (TPP+) have been used as probes of the membrane potential of cells. Interaction of TPP+ and triphenylalkylphosphonium homologues with the immobilized liposomal membranes was shown by their elution profiles on both zonal and frontal ILC. Retardation of the lipophilic cations on the liposome gel bed was increased as the hydrophobicity of the cations increased, indicating the partitioning of lipophilic cations into the hydrocarbon region of the membranes. The cations did not retard on the Sepharose or Sephacryl gel bed without liposomes, confirming that the cations only interact with the immobilized liposomes. Effects of the solute concentration, flow rate, and gel-matrix substance on the ILC were studied. The stationary phase volume of the liposomal membranes was calculated from the volume of a phospholipid molecule and the amount of the immobilized phospholipid, which allowed us to determine the membrane partition coefficient (KLM) for the lipophilic cations distributed between the aqueous mobile and membrane stationary phases. The values of KLM were generally increased with the hydrophobicity of the solutes increased, and were higher for the SUVs than for the LUVs. The ILC method described here can be applied to measure membrane partition coefficients for other lipophilic solutes (e.g., drugs).     

烟粉虱内共生菌16S rDNA的变异与系统发生

对 5年连续饲养在不同种寄主植物上的B型烟粉虱北京种群的内共生菌 1 6SrDNA基因进行了PCR扩增和测序。结合已知序列 ,构建了不同寄主植物烟粉虱初生内共生菌约 1 0 0 0bp的 1 6SrDNA及次生内共生菌约1 2 5 0bp的 1 6SrDNA的分子系统树。结果表明 ,中国北京不同寄主植物的B型烟粉虱内共生菌及世界其它地区烟粉虱内共生菌可能是同一种的不同生态型 ,内共生菌在其宿主分化后进行了选择 ,之后与其宿主长期共同进化、共同适应 ,为宿主对不同生境的适应提供了一定的基础     

通过形成类胚体诱导人羊水多能干细胞向心肌细胞分化

由人羊水中分离羊水多能性干细胞,通过形成类胚体诱导其向心肌细胞分化.取人羊水标本进行体外培养,分离得到人羊水干细胞,已连续传代培养至42代,采用免疫细胞化学、RT-PCR和流式细胞仪技术对羊水干细胞的生物学特性进行检测.取10～15代羊水干细胞,悬浮培养使其形成类胚体,进而向心肌细胞诱导分化.培养的羊水干细胞呈成纤维样,表达部分胚胎干细胞特异标志基因,悬浮培养可形成类胚体.类胚体碱性磷酸酶(AP)检测呈阳性,表达三胚层特异标志基因fgf5、ζ-globin和α-fetoprotein.羊水干细胞形成类胚体后进行诱导,得到α-actin阳性细胞,表达心肌细胞特异标志基因Tbx5、Nkx2.5、GATA4和α-MHC.试验结果表明,从人羊水标本中可分离得到具有胚胎干细胞特性的细胞,经初步检测确定为羊水干细胞,并能通过形成类胚体诱导其向心肌细胞分化.     

Mechanisms of the acute ischemia-induced arrhythmogenesis--a simulation study

The underlying ionic mechanisms of ischemic-induced arrhythmia were studied by the computer simulation method. To approximate the real situation, ischemic cells were simulated by considering the three major component conditions of acute ischemia (elevated extracellular K(+) concentration, acidosis and anoxia) at the level of ionic currents and ionic concentrations, and a round ischemic zone was introduced into a homogeneous healthy sheet to avoid sharp angle of the ischemic tissue. The constructed models were solved using the operator splitting and adaptive time step methods, and the perturbation finite difference (PFD) scheme was first used to integrate the partial differential equations (PDEs) in the model. The numerical experiments showed that the action potential durations (APDs) of ischemic cells did not exhibited rate adaptation characteristic, resulting in flattening of the APD restitution curve. With reduction of sodium channel availability and long recovery of excitability, refractory period of the ischemic tissue was significantly prolonged, and could no longer be considered as same as APD. Slope of the conduction velocity (CV) restitution curve increased both in normal and ischemic region when pacing cycle length (PCL) was short, and refractory period dispersion increased with shortening of PCL as well. Therefore, dynamic changes of CV and dispersion of refractory period rather than APD were suggested to be the fundamental mechanisms of arrhythmia in regional ischemic myocardium.