Tissue expression and cellular localization of phospholipid hydroperoxide glutathione peroxidase (PHGPx) mRNA in male mice

Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is an ubiquitous antioxidant enzyme, but the exact expression pattern in mammalian tissues is still unknown. The expression and cellular localization of PHGPx mRNA were examined in male mice using real time-polymerase chain reaction and in situ hybridization techniques. The rank order of PHGPx mRNA expression across tissues exhibiting substantial levels of expression was:testes ≫ heart > cerebrum ≥ ileum > stomach = liver = jejunum ≥ epididymis. In testes, PHGPx mRNA was highly expressed in spermiogenic cells and Leydig cells. The signal was also expressed in the molecular layer, Purkinje cell layer, and white matter of cerebellum, the pituicytes of neurohypophysis, the parafollicular cells and follicular basement membrane of thyroid, the exocrine portion of pancreas, the tubular epithelium of kidney, the smooth muscle cells of arteries, and the red pulp of spleen. In the gastrointestinal tract, PHGPx mRNA expression was mainly observed in the keratinized surface epithelium of forestomach, the submucosal glands and serosa layers, and further the Paneth cells of intestines. PHGPx mRNA appeared to be ubiquitously expressed in the parenchyma of heart, liver, and lung. These results indicate that PHGPx exhibits a cell- and tissue-specific expression pattern in mice.     

萱草叶片响应低温胁迫的转录组分析

低温胁迫是萱草（Hemerocallis fulva）生长过程中经常会遭遇的一种非生物胁迫。比较了萱草叶片在低温处理（10、5、0 ℃）下转录组与对照（15 ℃）数据的差异,共筛选出差异表达基因2 457个,其中上调基因1 253个,下调基因1 204个。差异表达基因主要富集在细胞过程、代谢过程和催化活性等49个GO过程,代谢途径、次生代谢产物的生物合成、植物激素信号转导等42条KEGG代谢途径中。其中参与植物激素信号转导通路的差异表达基因发生了不同程度的变化,GH3.10基因上调至对照组的13.624倍,IAA1基因下调0.120倍;参与可溶性糖合成通路的差异基因发生了0.076~28.114倍不同程度的变化。随后对3个低温处理组共有的29个差异表达基因进行热图和网络调控分析,基于基因在网络调控中的位置,对ABCF5、OFPs和SWEETs等基因在冷应答的作用进行了分析。本研究结果为进一步挖掘萱草低温响应的关键基因及耐寒萱草种质开发、分子育种提供了一定的理论支撑。     

盐碱胁迫对染内生菌和不染菌苇状羊茅根系丛枝菌根真菌群落多样性和组成的影响

香柱菌属Epichloë内生真菌存在于宿主植物地上部组织,不仅能提高宿主植物对生物与非生物逆境的抗性,而且能对周围环境中的微生物产生影响。该研究以染内生菌(endophyte-infected,EI)和不染菌(endophyte-free,EF)苇状羊茅Festuca arundinacea为实验材料,探究内生真菌和不同水平盐碱胁迫处理对宿主根系丛枝菌根真菌(arbuscular mycorrhizal fungi,AMF)群落多样性和组成的影响。结果表明,内生真菌和盐碱胁迫处理对苇状羊茅根系AMF多样性影响存在交互作用。EF苇状羊茅根系AMF多样性随盐碱胁迫处理水平的增加而降低,内生真菌的存在缓解了这一效应,在200和400 mmol/L盐碱胁迫处理下,内生真菌感染增加了苇状羊茅根系AMF多样性;此外,内生真菌感染改变了苇状羊茅根系AMF群落组成,降低了优势属Funneliformis相对多度,增加了Claroideoglomus、Glomus和unclassified AMF相对多度。结构方程模型结果表明,内生真菌通过间接增加土壤总磷浓度对苇状羊茅根系AMF多样性产生影响。本研究为筛选盐碱污染区生态修复的植物-微生物共生体提供基础。     

基于地理探测器的宁夏贺兰山植被覆盖度时空分异及驱动因子

监测区域植被覆盖变化并分析其驱动因子,有利于实现生态环境的可持续发展。本研究以1989—2021年Landsat 5/8遥感影像为数据源,利用像元二分模型获取宁夏贺兰山植被覆盖度,基于地理探测器量化了环境和人为等10个因子对其时空分布的影响。结果表明: 1989—2021年间,宁夏贺兰山平均植被覆盖度为35.8%,时间尺度上总体呈增加趋势,平均增幅为0.043·(10 a)^-1,空间尺度上呈现从西南向东北递减的分布特征;研究区58.1%的区域植被覆盖度未来将持续性改善,但仍有30.7%的植被存在退化的潜在风险。降水是影响植被分布的主要环境因子,与单因子相比,环境因子和人为因子的交互作用对植被覆盖度的解释力更强,降水与其他因子的交互作用处于主导作用。     

萱草响应非生物胁迫的研究进展

萱草是萱草属(Hemerocallis)多年生宿根花卉,被誉为中华母亲花,具有重要的观赏和药用价值。非生物胁迫导致光合效率降低,渗透调节物质浓度改变,活性氧(ROS)含量上升,膜系统持续受损,诱导AP2/ERF (APETALA2/ethylene-responsive element binding factors)、WRKY等家族基因的表达。该文综述了干旱、涝渍、盐碱、极端温度和重金属胁迫非生物胁迫因子对萱草形态学、生理生化及分子水平的影响,统计了各胁迫下的萱草抗性品种资源,认为地域与胁迫对萱草药用成分代谢变化的影响、抗逆相关基因调控网络与多种胁迫复合分子育种为未来的重点研究方向,为萱草资源开发利用与抗逆品种育种提供理论参考。     

炭样小单孢菌产抗生素对水稻细菌性条斑病菌的抗菌作用及其机理

【背景】水稻细菌性条斑病菌为水稻细菌性条斑病的病原菌,土壤中分离到的一株具有广谱抗菌活性的炭样小单孢菌JXNU-1,其发酵产物(即抗生素JX)对植物病原菌具有较强的抑菌活性。【目的】研究抗生素JX对水稻细菌性条斑病菌的抗菌作用及其机理。【方法】采用杯碟法测定抑菌圈大小,二倍稀释法测定最低抑菌浓度、最低杀菌浓度,并且从菌体形态观察、电导率变化、培养液大分子漏出、蛋白质合成、核酸合成和膜电位变化6个方面探究其作用机理。【结果】抗生素JX对水稻细菌性条斑病菌的抑菌圈直径达18.84±0.28mm,最低抑菌浓度和最低杀菌浓度分别为1.39μg/m L和2.78μg/mL,且杀菌速度很快,作用12 h的杀菌率达100%。在抗生素JX作用下,水稻细菌性条斑病菌的细胞形态发生改变,培养液电导率、膜电位和大分子漏出量均随抗生素浓度增加而增大,但菌体蛋白质含量随着抗生素浓度增加而降低,同时,通过实时荧光定量PCR方法检测发现ef-p表达量下调。【结论】抗生素JX对水稻细菌性条斑病菌具有较强的抗菌作用,推测其抑菌机理是通过抑制菌体蛋白质的生物合成和影响细胞膜完整性而起作用。     

海藻糖酶基因TRE2-like和TRE2在异色瓢虫羽化过程中的表达与功能

【目的】异色瓢虫Harmonia axyridis是一种重要的捕食性天敌昆虫,海藻糖在异色瓢虫的变态发育、羽化等整个生命过程都起着重要的作用。本研究以前期获得的类似膜结合型海藻糖酶(TRE2-like)与膜结合型海藻糖酶(TRE2)基因为基础,探讨在异色瓢虫羽化阶段这两个海藻糖酶的潜在功能,为阐明异色瓢虫从蛹发育到成虫时海藻糖代谢机制提供参考。【方法】根据TRE2-like和TRE2基因序列设计双链RNA(dsRNA)区域片段并合成对应的dsRNA,通过RNAi将其注射到异色瓢虫2日龄蛹中。采用实时荧光定量PCR(RT-qPCR)检测RNAi处理后羽化第1天的异色瓢虫成虫糖代谢相关基因的表达;同时采用蒽酮比色法、酶标法等分别测定RNAi处理后羽化第1天的异色瓢虫成虫主要糖类物质含量及TRE活性变化,并观察异色瓢虫羽化后的表型变化。【结果】结果表明,与对照组(dsGFP注射组)相比,异色瓢虫2日龄蛹被注射TRE2-like或TRE2 dsRNA后,其新羽化成虫体内TRE2-like和TRE2表达量均极显著下调,且少数个体出现了蜕皮与翅形成困难等畸形表型。可溶性海藻糖酶活性在注射dsTRE2-like后显著降低,膜结合型海藻糖酶活性在注射dsTRE2后显著降低;注射dsTRE2后糖原含量显著下降,注射dsTRE2-like后糖原和海藻糖含量显著下降,注射dsTRE2-like+dsTRE2后糖原和葡萄糖含量显著下降,且海藻糖含量极显著下降。注射dsTRE2-like, dsTRE2和dsTRE2-like+dsTRE2后可溶性海藻糖酶基因TRE1-1和TRE1-2表达下降或显著下降,而TRE1-5表达上升或显著上升,海藻糖合成酶(trealose-6-phosphate synthase, TPS)、糖原磷酸化酶(glycogen phosphorylase, GP)、糖原合成酶(glycogen synthase, GS)基因的表达均显著下调。【结论】TRE2-like和TRE2基因表达被抑制后,异色瓢虫海藻糖等代谢受到影响。研究结果为探究异色瓢虫体内膜结合型海藻糖酶的潜在功能和调控机制奠定了基础。     

Arsenic trioxide induces G2/M growth arrest and apoptosis after caspase-3 activation and bcl-2 phosphorylation in promonocytic U937 cells

Arsenic trioxide has recently been shown to inhibit growth and induce apoptosis in acute promyelocytic leukemia (APL), but little is known about the molecular mechanisms mediating these effects. Here we demonstrate that treatment of promonocytic U937 cells with arsenic trioxide leads to G2/M arrest which was associated with a dramatic increase in the levels of cyclin B and cyclin B-dependent kinase and apoptosis. We further show that apoptosis occurs after bcl-2 phosphorylation and caspase-3 activation followed by cleavage of PARP and PLC-gamma1 degradation and DNA fragmentation. The arsenic trioxide-induced apoptosis could be blocked by the protein synthesis inhibitor cycloheximide. In addition, pretreatment of U937 cells with the DNA polymerase inhibitor aphidicolin also blocked apoptosis, but did not cause the arrest of cells in the G2/M phase. The findings suggest that arsenic trioxide exerts its growth-inhibitory effects by modulating expression and/or activity of several key G2/M regulatory proteins. Furthermore, arsenic trioxide-mediated G2/M arrest correlates with the onset of apoptosis.     

Manganese-containing superoxide dismutase and its gene from Candida albicans

Mitochondrial manganese-containing superoxide dismutase was purified around 112-fold with an overall yield of 1.1% to apparent electrophoretic homogeneity from the dimorphic pathogenic fungus, Candida albicans. The molecular mass of the native enzyme was 106 kDa and the enzyme was composed of four identical subunits with a molecular mass of 26 kDa. The enzyme was not sensitive to either cyanide or hydrogen peroxide. The N-terminal amino acid sequence alignments (up to the 18th residue) showed that the enzyme has high similarity to the other eukaryotic manganese-containing superoxide dismutases. The gene sod2 encoding manganese-containing superoxide dismutase has been cloned using a product obtained from polymerase chain reaction. Sequence analysis of the sod2 predicted a manganese-containing superoxide dismutase that contains 234 amino acid residues with a molecular mass of 26173 Da, and displayed 57% sequence identity to the homologue of Saccharomyces cerevisiae. The deduced N-terminal 34 amino acid residues may serve as a signal peptide for mitochondrial translocation. Several regulatory elements such as stress responsive element and haem activator protein 2/3/4/5 complex binding sites were identified in the promoter region of sod2. Northern analysis with a probe derived from the cloned sod2 revealed a 0.94-kb band, which corresponds approximately to the expected size of mRNA deduced from sod2.     

Expression, purification, characterization and crystallization of flap endonuclease-1 from Methanococcus jannaschii

A gene coding for a protein homologous to the flap endonuclease-1 (FEN-1) was cloned from Methanococcus jannaschii, overexpressed, purified and characterized. The gene product from M. jannaschii shows 5' endo-/exonuclease and 5' pseudo-Y-endonuclease activities as observed in the FEN-1 in eukaryotes. In addition, Methanococcus jannaschii FEN-1 functions effectively at high concentrations of salt, unlike eukaryotic FEN-1. We have crystallized Methanococcus jannaschii FEN-1 and analyzed its preliminary character. The crystal belongs to the space group of P2(1) with unit cell dimensions of a = 58.93 A, b = 42.53 A, c = 62.62 A and beta = 92.250. A complete data set has been collected at 2.0 A resolution using a frozen crystal.