核酸去甲基酶AlkB在毕赤酵母中的表达纯化及在tRNA相关研究中的应用

[目的]核酸的甲基化修饰是一种常见的化学修饰形式,具有重要的生物学功能,却也在一定程度上给一些核酸研究过程带来了技术难度。tRNA上具有的大量甲基化修饰会阻碍逆转录进程,从而降低荧光定量PCR (real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)和高通量测序对其的检测效率。来自大肠杆菌(Escherichia coli)的AlkB蛋白是一种多功能的脱烷基化酶,可以去除DNA和RNA上多种甲基化为代表的修饰,有望解决以上问题。[方法]针对大肠杆菌来源的AlkB,分别尝试在大肠杆菌和毕赤酵母(Pichia pastoris)表达系统中进行诱导表达和纯化,对纯化获得的AlkB进行酶学性质测定。最后以tRNA_UAU^Ile等两种tRNA为代表,研究AlkB的处理对于荧光定量PCR法检测tRNA表达水平的影响。[结果]AlkB在大肠杆菌中表达时多以包涵体形式存在,但是在毕赤酵母中可以成功分泌表达。使用镍柱分离纯化后获得了纯度高于95%的AlkB蛋白,其酶学性质参数如...     

生物淋洗修复钒污染土壤的性能与机理研究

【目的】土壤重金属污染问题日益受到关注,其中钒污染逐渐成为研究热点。淋洗是土壤修复的重要手段,但存在污染大、成本高的缺点。生物淋洗技术因其经济高效且环保的特点能够应用于土壤的修复,但其对钒污染土壤的修复,认识仍非常有限。【方法】本研究采用嗜酸性氧化亚铁硫杆菌对钒污染土壤进行了生物淋洗试验,通过影响因素试验探究了钒的最佳浸出条件,并应用扫描电子显微镜-能量色散X射线谱分析了钒在淋洗过程中的变化,最后对代谢产物进行了解析。【结果】微生物次生代谢产物能促进土壤中钒的溶出。氧化亚铁硫杆菌对土壤钒的浸出效率较高,生物淋洗20 d后土壤中钒的浸出率达到27.4%,进一步的影响因素试验表明,在固体浓度为3%、接种体积为10%、初始pH值为1.8、初始Fe²⁺的浓度为3.0 g/L的条件下,土壤中钒的浸出效果最佳。SEM-EDS分析证实生物淋洗后土壤中钒含量减少,其中以非残渣态形式存在的钒更容易被浸出。代谢组学分析显示氧化亚铁硫杆菌在浸出过程中产生了大量代谢产物来应对重金属胁迫。【结论】生物淋洗技术能够有效地实现土壤钒污染的修复,本研究为钒污染土壤提供了一种环境友好的修复方式。     

内生真菌印度梨形孢诱导小麦抗根腐病作用研究

【目的】明确印度梨形孢(Piriformospora indica)诱导小麦对根腐病产生抗性的作用机制。【方法】用印度梨形孢悬液浸种,以无菌培养液为对照,用病原菌禾谷镰孢菌(Fusarium graminearum)侵染小麦,对其相关生理生化指标及转录组变化进行分析。【结果】禾谷镰孢菌能诱导小麦产生过氧化氢,降低细胞内水含量,破坏细胞膜的稳定性;根部定殖印度梨形孢的小麦细胞内抗氧化酶活性增强,活性氧自由基含量降低,胞内水含量提高,细胞膜稳定性增强;印度梨形孢定殖能改变由于病原菌引起的mRNA转录组变化,抗性相关基因的表达增强。综合表明印度梨形孢定殖能有效地提高小麦对禾谷镰孢菌的抗性。【结论】研究结果为深入理解植物与微生物互作、开发新型高效环保抗根腐病生物制剂提供理论和实验依据。     

Phytopathogenic bacteria utilize host glucose as a signal to stimulate virulence through LuxR homologues

Chemical signal-mediated biological communication is common within bacteria and between bacteria and their hosts. Many plant-associated bacteria respond to unknown plant compounds to regulate bacterial gene expression. However, the nature of the plant compounds that mediate such interkingdom communication and the underlying mechanisms remain poorly characterized. Xanthomonas campestris pv. campestris (Xcc) causes black rot disease on brassica vegetables. Xcc contains an orphan LuxR regulator (XccR) which senses a plant signal that was validated to be glucose by HPLC-MS. The glucose concentration increases in apoplast fluid after Xcc infection, which is caused by the enhanced activity of plant sugar transporters translocating sugar and cell-wall invertases releasing glucose from sucrose. XccR recruits glucose, but not fructose, sucrose, glucose 6-phosphate, and UDP-glucose, to activate pip expression. Deletion of the bacterial glucose transporter gene sglT impaired pathogen virulence and pip expression. Structural prediction showed that the N-terminal domain of XccR forms an alternative pocket neighbouring the AHL-binding pocket for glucose docking. Substitution of three residues affecting structural stability abolished the ability of XccR to bind to the luxXc box in the pip promoter. Several other XccR homologues from plant-associated bacteria can also form stable complexes with glucose, indicating that glucose may function as a common signal molecule for pathogen–plant interactions. The conservation of a glucose/XccR/pip-like system in plant-associated bacteria suggests that some phytopathogens have evolved the ability to utilize host compounds as virulence signals, indicating that LuxRs mediate an interkingdom signalling circuit.     

Spatial scale impacts microbial community composition and distribution within and across stream ecosystems in North and Central America

A mechanistic understanding of factors that structure spatiotemporal community composition is a major challenge in microbial ecology. Our study of microbial communities in the headwaters of three freshwater stream networks showed significant community changes at the small spatial scale of benthic habitats when compared to changes at mid- and large-spatial scales associated with stream order and catchment. Catchment (which included temperate and tropical catchments) had the strongest influence on community composition followed by habitat type (epipsammon or epilithon) and stream orders. Alpha diversity of benthic microbiomes resulted from interactions between catchment, habitat, and canopy. Epilithon contained relatively more Cyanobacteria and algae while Acidobacteria and Actinobacteria proportions were higher in epipsammic habitats. Turnover from replacement created ~60%–95% of beta diversity differences among habitats, stream orders, and catchments. Turnover within a habitat type generally decreased downstream indicating longitudinal linkages in stream networks while between habitat turnover also shaped benthic microbial community assembly. Our study suggests that factors influencing microbial community composition shift in dominance across spatial scales, with habitat dominating locally and catchment dominating globally.     

Identification of tyrosines 154 and 307 in the extracellular domain and 653 and 766 in the intracellular domain as phosphorylation sites in the heparin-binding fibroblast growth factor receptor tyrosine kinase (flg).

Four tyrosine residues have been identified as phosphorylation sites in the tyrosine kinase isoform of the heparin-binding fibroblast growth factor receptor flg (FGF-R1). Baculoviral-insect cell-derived recombinant FGF-R1 was phosphorylated and fragmented with trypsin while immobilized on heparin-agarose beads. Phosphotyrosine peptides were purified by chromatography on immobilized anti-phosphotyrosine antibody and analyzed by Edman degradation and electrospray tandem mass spectrometry. Tyrosine residue 653, which is in a homologous spatial position to major autophosphorylation sites in the catalytic domain of the src and insulin receptor kinases, is the major intracellular FGF-R1 phosphorylation site. Residue 766 in the COOH-terminus outside the kinase domain is a secondary site. Tyrosine residues 154 and 307, which are in the extracellular domain of transmembrane receptor isoforms and are in an unusual sequence context for tyrosine phosphorylation, were also phosphorylated.     

微生物镉解毒机制及微生物-植物互作修复研究进展

镉(cadmium,Cd)是引起粮食减产的主要金属之一,具有高溶解性及高迁移性,易被植物吸收和积累。微生物长期在镉胁迫的条件下进化出一系列的镉解毒机制。微生物对镉的解毒包括抑制Cd(Ⅱ)的进入、促进Cd(Ⅱ)的外排,以及将进入胞内的Cd(Ⅱ)进行“扣押”。微生物的Cd(Ⅱ)钝化是通过细胞吸附和胞外沉淀将游离态的Cd(Ⅱ)进行钝化,这类微生物具有较强的土壤镉污染治理潜力。本文主要介绍微生物的镉解毒机制、微生物-微生物互作、微生物-植物互作机制及其在镉污染生物修复中应用的最新研究进展。     

湖南省发现云开脊蛇兼记棕脊蛇新地模标本

云开脊蛇(Achalinus yunkaiensis)之前仅在广东和广西有分布报道。本文基于形态比较及线粒体COI基因序列分子系统关系分析结果,确定采集于湖南省新宁县的1号雌性脊蛇标本(CIB 119041)为云开脊蛇,为湖南省新记录种。该标本鼻间鳞沟约等于前额鳞沟,上颔齿24枚;背鳞通身23行,腹鳞150枚,尾下鳞55枚;尾长与体长之比为0.203。至此共有4种脊蛇分布于湖南省。此外,本文还报道了棕脊蛇(A. rufescens)1号雄性地模标本(CIB 119042),该标本鼻间鳞沟长于前额鳞沟;背鳞通身23行,腹鳞153枚,尾下鳞62枚;尾长与体长之比0.201。分子系统发育结果显示,棕脊蛇种组各支系的系统地位还需进一步研究厘定。     

辽宁省爬行动物名录及地理区划更新

基于此前辽宁省爬行动物调查资料和分类学研究,结合近年来野外调查进展,对辽宁省爬行动物名录和区系进行修订,并报道辽宁省蛇类新记录二种——乌梢蛇（Ptyasdhumnades）和黑头剑蛇（Sibynophischinensis）。根据从辽宁省凌源市野外获得的影像资料进一步确认黑眉锦蛇（Elaphe taeniura）在辽宁省的分布。另依据采集自葫芦岛市的1号王锦蛇（E. carinata）标本对该种在辽宁省的分布予以补充描述。截至2023年5月,辽宁省记录爬行动物2目11科21属37种,其中,龟鳖目3科4属5种,有鳞目蜥蜴亚目3科5属10种,蛇亚目5科12属22种。爬行动物中,国家一级重点保护野生动物3种,国家二级重点保护野生动物6种。此外,有15个物种被《中国生物多样性红色名录脊椎动物第三卷爬行动物》评估为受威胁物种。根据辽宁省爬行动物最新分布信息,将辽宁省划分为6个动物地理省,分别为辽东山地丘陵省、辽东半岛山地丘陵省、辽河平原省、努鲁儿虎山北麓丘陵台地及西辽河沙地省、辽西山地丘陵省和冀辽山地省,其中,冀辽山地省为本研究中新增动物地理省。扩大原辽东半岛丘陵省涵盖范围,并将其名称修订为“辽...     

乙醛脱氢酶基因ALDH2对雄性小鼠生育能力的影响

本研究使用乙醛脱氢酶2(ALDH2)基因全敲除的雄性C57BL/6(B6)小鼠（Musmusculus domesticus）,通过分析不同周龄小鼠的睾丸脏器系数、睾丸组织细胞形态、精子运动参数、配种后母鼠产仔数及子代雄雌比等生育指标,探讨ALDH2基因敲除对雄性小鼠生育能力的影响。结果表明,与野生型（WT）小鼠相比,5、7、10周龄ALDH2基因敲除型（KO）雄性小鼠睾丸脏器系数显著降低（P <0.05）;睾丸组织细胞间质变大,精子活率显著降低（P <0.05）;产仔数和雄雌比显著降低（P <0.05）。本研究为揭示乙醛脱氢酶ALDH2基因在雄性小鼠生殖中的作用提供了一定的基础。