Timing and abundance of sporangia production and zoospore release influences the recovery of different Phytophthora species by baiting

Analysis of soil samples using High Throughput Sequencing (HTS) frequently detects more Phytophthora species compared with traditional soil baiting methods. This study investigated whether differences between species in the timing and abundance of sporangial production and zoospore release could be a reason for the lower number of species isolated by baiting. Stems of Eucalyptus marginata were inoculated with ten Phytophthora species (P. nicotianae, P. multivora, P. pseudocryptogea, P. cinnamomi, P. thermophila, P. arenaria, P. heveae, P. constricta, P. gondwanensis and P. versiformis), and lesioned sections for each species were baited separately in water. There were significant differences between species in timing of sporangia production and zoospore release. P. nicotianae, P. pseudocryptogea, P. multivora and P. thermophila released zoospores within 8–12 h and could be isolated from lesioned baits within 1–2 days. In contrast, P. constricta did not produce zoospores for over 48 h and was only isolated 5–7 days after baiting. P. heveae and P. versiformis did not produce zoospores and were not recovered from the baits. When species were paired in the same baiting tub, those that produced zoospores in the shortest time were isolated most frequently, while species slow to produce zoospores, or which produced them in lower numbers, were isolated from few baits or not at all. Thus, species differences in the timing of sporangia production and zoospore release may contribute to the ease of isolation of some Phytophthora species when they are present together with other Phytophthora species in an environmental sample.     

The Cytotoxic and Proapoptotic Activities of Hypnophilin are Associated with Calcium Signaling in UACC‐62 Cells

Hypnophilin (HNP) is a sesquiterpene that is isolated from Lentinus cf. strigosus and has cytotoxic activities. Here, we studied the calcium signaling and cytotoxic effects of HNP in UACC‐62 cells, a human skin melanoma cell line. HNP was able to increase the intracellular calcium concentration in UACC‐62 cells, which was blocked in cells stimulated in Ca²⁺‐free media. HNP treatment with BAPTA‐AM, an intracellular Ca²⁺ chelator, caused an increase in calcium signals. HNP showed cytotoxicity against UACC‐62 cells in which it induced DNA fragmentation and morphological alterations, including changes in the nuclear chromatin profile and increased cytoplasmatic vacuolization, but it had no effect on the plasma membrane integrity. These data suggest that cytotoxicity in UACC‐62 cells, after treatment with HNP, is associated with Ca²⁺ influx. Together, these findings suggest that HNP is a relevant tool for the further investigation of new anticancer approaches. © 2013 Wiley Periodicals, Inc. J BiochemMol Toxicol 27:479‐485, 2013; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.21507     

New Natural Noncannabinoid Ligands for Cannabinoid Type-2 (CB2) Receptors

Since the discovery that Δ ⁹-tetrahydrocannabinol and related cannabinoids from Cannabis sativa L. act on specific physiological receptors in the human body and the subsequent elucidation of the mammalian endogenous cannabinoid system, no other natural product class has been reported to mimic the effects of cannabinoids. We recently found that N-alkyl amides from purple coneflower (Echinacea spp.) constitute a new class of cannabinomimetics, which specifically engage and activate the cannabinoid type-2 (CB₂) receptors. Cannabinoid type-1 (CB₁) and CB₂ receptors belong to the family of G protein-coupled receptors and are the primary targets of the endogenous cannabinoids N-arachidonoyl ethanolamine and 2-arachidonoyl glyerol. CB₂ receptors are believed to play an important role in distinct pathophysiological processes, including metabolic dysregulation, inflammation, pain, and bone loss. CB₂ receptors have, therefore, become of interest as new targets in drug discovery. This review focuses on N-alkyl amide secondary metabolites from plants and underscores that this group of compounds may provide novel lead structures for the development of CB₂-directed drugs.     

Concentration of Virus from Water by Electro-Osmosis and Forced-Flow Electrophoresis. II. Improvement of Methodology and Application to Tap Water

The ability of the Canalco Model CF-3 electro-osmosis (EO) apparatus to concentrate viruses from artificially seeded distilled water was improved. Modification of the physical arrangement of the equipment allowed for a 10–25 fold increase in concentration efficiency and a concomitant decrease in the process time. The major improvements involved modifications of the cell arrangement (which increased the membrane transport area), a change in the salt replenishing solution and the use of different membranes of higher flux. Viruses concentrated by E0 from seeded tap water resulted in lower recoveries when compared to distilled water. The lower yields were probably due to instability or aggregation of the agents in the menstruum and not directly related to the physical apparatus. Under the conditions used, one could detect virus at levels as low as 0.01 plaque forming units (PFU) per ml of initial input. The efficacy of a modification of the Canalco forced-flow electrophoretic (FFE) system was also evaluated. The maximum potential was applied with a constant value for pump rates. A 6-fold concentration of virus and a 12-fold decrease in water volume was obtained.     

不依赖天然外显子的蛋白质内含子定向进化筛选系统

蛋白质剪接技术为在蛋白质水平上直接对蛋白质进行修饰和加工提供了一种全新的解决方案,因而在蛋白质工程及相关领域具有非常广阔的应用前景。现阶段,大部分天然的蛋白质内含子在异源蛋白质中剪接活性非常低,极大限制了蛋白质内含子的开发和应用。为了开发一个可以同时对蛋白质内含子通用性和剪接活性进行筛选的系统,利用Bsa I限制性内切酶识别位点和切割不重合的特性,将Ter ThyX内含子(不含外显子序列)插入到卡那霉素抗性蛋白基因的多个位点。并且摒弃了以往需要结合天然外显子以实现剪接的方法,可以同时对蛋白质内含子的剪接活性和通用性进行筛选。Western blot结果和卡那霉素平板生长结果表明,通过卡那霉素筛选系统可以精确的将蛋白质内含子剪接反应与卡那霉素抗性结合起来,仅从卡那霉素平板上的菌落生长情况即可完成蛋白质内含子剪接活性阳性突变的筛选,是一个快速,稳定的定向进化筛选系统。     

锥栗自然居群遗传多样性的ISSR分析

采用20条ISSR分子标记对栗属中国特有种-锥栗(Castanea henryi)的16个自然居群进行了遗传多样性与遗传关系分析。在449份试材上共扩增得到379个位点,其中多态性位点378个,多态性位点百分率(PPL)达99.74%,平均期望杂合度(He)为0.2950,Shannon信息指数(I)为0.4522;居群水平遗传多样性为46.09%,且不同居群遗传多样性水平有较大差异,16个自然居群中以湘西居群的遗传多样性水平最高(PPL=53.30%,He=0.1861,I=0.2781),其次为靖州、庆元、昭通及都江堰居群,南平居群最低(PPL=36.94%,He=0.1202,I=0.1817)。Nei’s遗传多样性和AMOVA分析表明,居群间产生了较大的遗传分化(Gst=0.4466),锥栗自然居群内的遗传变异稍占优势(52.51%)。UPGMA聚类分析将16个锥栗居群分为2大类5亚类。湘西地区可能是锥栗的次生分布中心和现代遗传多样性分布中心,是锥栗研究的资源中心,也是最有价值的基因库,需要重点保护。     

微生物降解石油烃的功能基因研究进展

微生物对石油烃的降解在自然衰减去除土壤和地下水石油烃污染的过程中发挥了重要作用。微生物通过其产生的一系列酶来利用和降解这类有机污染物,其中,编码关键降解酶的基因称为功能基因。功能基因可作为生物标志物用于分析环境中石油烃降解基因的多样性。因此,研究石油降解功能基因是分析土著微生物群落多样性、评价自然衰减潜力与构建基因工程菌的重要基础。本文主要介绍了烷烃和芳香烃在有氧和无氧条件下的微生物降解途径,重点总结了烷烃和芳香烃降解的主要功能基因及其作用,包括参与羟化作用的单加氧酶和双加氧酶基因、延胡索酸加成反应的琥珀酸合酶基因以及中心中间产物的降解酶基因等。     

绿瘦蛇捕食鳄蜥的现象

鳄蜥(Shinisauruscrocodilurus)是濒危的孑遗物种,生存遭受多方威胁。除人为猎杀和生境受到破坏之外,天敌也是其致命的威胁之一,但目前关于其被天敌捕食的研究非常匮乏。本文报道了绿瘦蛇(Ahaetullaprasina)捕食鳄蜥的现象,以全事件记录法观察了这一行为过程。鳄蜥面对的被捕食风险可能较广。除蛇外,鳄蜥的天敌也可能是鸟类等动物。在饲养繁育中,要注意加强防范举措,以免鳄蜥被天敌侵害。     

Identification of a novel mutation in ZAP70 and prenatal diagnosis in a Turkish family with severe combined immunodeficiency disorder

Protein tyrosine kinases (PTKs) play an important role in T cell development and activation. In vitro and in vivo defects, resulting in variable deficiencies in thymic development and in T cell antigen receptor (TCR) signal transduction, in PTKs have been shown. ZAP70, one of those PTKs, is a 70-kDa tyrosine phosphoprotein and associates with the ζ chain and undergoes tyrosine phosphorylation following TCR stimulation. It is expressed in T and natural killer (NK) cells. Several mutations were shown to lead to an autosomal recessive form of severe combined immunodeficiency disease (SCID).