大鼠肝癌中α_2-巨球蛋白等几种分泌性蛋白基因表达的变化

用Northern blot方法对二乙基亚硝胺所诱发的大鼠肝癌中内源性蛋白酶抑制因子α_2-巨球蛋白(α_2-M)、非特异性免疫抑制剂α_1-酸性糖蛋白(α_1-AGP)及雄性激素正调控的α-2u球蛋白(α-2u)三种分泌性蛋白基因表达情况进行了分析。结果表明在大部分(14/16)肝癌样品中α_2-M RNA水平显著降低;而α_1-AGP RNA水平显著高于正常对照水平;α-2u RNA水平明显下降,但在某些雄性大鼠肝癌样品中该基因却有一定程度的表达。这些结果说明,一些肿瘤宿主血浆中α_2-M水平的显著下降及α_1-AGP水平的明显升高分别是由于基因表达活性的下降及升高所致。α-2u基因表达的异常提示,在癌变过程中机体的内分泌功能发生了某些变化。     

Accelerated organic matter decomposition in thermokarst lakes upon carbon and phosphorus inputs

Mineralization of dissolved organic matter (DOM) in thermokarst lakes plays a non-negligible role in the permafrost carbon (C) cycle, but remains poorly understood due to its complex interactions with external C and nutrient inputs (i.e., aquatic priming and nutrient effects). Based on large-scale lake sampling and laboratory incubations, in combination with ¹³C-stable-isotope labeling, optical spectroscopy, and high-throughput sequencing, we examined large-scale patterns and dominant drivers of priming and nutrient effects of DOM biodegradation across 30 thermokarst lakes along a 1100-km transect on the Tibetan Plateau. We observed that labile C and phosphorus (P) rather than nitrogen (N) inputs stimulated DOM biodegradation, with the priming and P effects being 172% and 451% over unamended control, respectively. We also detected significant interactive effects of labile C and nutrient supply on DOM biodegradation, with the combined labile C and nutrient additions inducing stronger microbial mineralization than C or nutrient treatment alone, illustrating that microbial activity in alpine thermokarst lakes is co-limited by both C and nutrients. We further found that the aquatic priming was mainly driven by DOM quality, with the priming intensity increasing with DOM recalcitrance, reflecting the limitation of external C as energy sources for microbial activity. Greater priming intensity was also associated with higher community-level ribosomal RNA gene operon (rrn) copy number and bacterial diversity as well as increased background soluble reactive P concentration. In contrast, the P effect decreased with DOM recalcitrance as well as with background soluble reactive P and ammonium concentrations, revealing the declining importance of P availability in mediating DOM biodegradation with enhanced C limitation but reduced nutrient limitation. Overall, the stimulation of external C and P inputs on DOM biodegradation in thermokarst lakes would amplify C-climate feedback in this alpine permafrost region.     

不同密度格木幼林的土壤理化与林下植被特征

为探索适合格木(Erythrophleum fordii)人工林在幼龄阶段的种植密度,在不同林分密度(2 m×1 m、2 m×2 m、2 m×3 m、3 m×3 m)的6 a生格木人工林下设置标准样地,采用土壤质量评价和灰色关联度等方法,探究不同密度下格木幼林的土壤理化与林下植被特征。结果表明,密度2 m×3 m下的林木胸径、树高最优,较最低水平高16.7%、27.9%;土壤总孔隙度最大,全N、硝态N、铵态N含量最高,灌木草本多样性最高。相关性分析表明土壤化学性质对灌木草本的多样性影响最大。不同林分密度下格木幼林土壤理化性质及林下植物多样性有显著差异,因此,选择合适的林分密度对人工林土壤肥力的可持续利用及林分的经营培育至关重要。     

粟穗螟滞育的形成和解除与环境条件的关系

粟穗螟Manpava bipunctella Ragonot在川南地区为二化性兼性滞育的昆虫。光周期是诱发滞育的主导因素,在中位温度下,滞育与否主要取决于幼虫发育期间的每日光照时数。在2s℃恒温下,临界光周期为14小时38分。幼虫对光照刺激反应的敏感期为低龄期。 温度和食料效应只发生在每天14小时以上的长光照下,低温有抵销长光照抑制滞育的作用,高温影响不显著;取食玉米的幼虫滞育率比高粱的高,并随寄主生育阶段的发展而增高。该虫滞育解除必需每天14-15小时的长光照;不利于滞育发育和解除,适宜温度为10一25℃。本文最后讨论了该虫滞育形成和解除的特点对发生规律的作用及在测报上的意义。     

酿酒酵母INO80染色质重塑复合物调控基因表达的研究进展

表观遗传调控是真核生物基因表达精细调控的重要组成部分,主要包括DNA甲基化、组蛋白修饰和染色质重塑。其中,染色质重塑因子可影响组蛋白修饰酶和转录因子与特定位点的结合,在基因表达调控中占有重要地位。INO80复合物是进化上保守的染色质重塑复合物,能利用ATP水解获得的能量促进核小体的滑动和驱逐。INO80复合物除了在DNA复制、修复中发挥重要功能外,还通过改变DNA可及性调控酿酒酵母的基因表达。本文综述了染色质重塑复合物的分类及组成,重点介绍了酿酒酵母多亚基复合物INO80在基因表达调控中的重要功能,包括驱逐RNA聚合酶Ⅱ、响应信号转导途径和改变基因表达水平等,并着重总结了其在酿酒酵母环境胁迫响应机理中的研究进展。深入研究INO80染色质重塑复合物的功能,可为理解真核生物精细代谢调控的机制,并进一步开发基于染色质重塑等表观调控水平的微生物代谢工程和合成生物学改造策略,提高菌株的环境胁迫耐受性和发酵性能提供基础。     

单增李斯特菌末端氧化酶亚基QoxB的生物学功能探究

【目的】本研究旨在构建单增李斯特菌末端细胞色素aa3氧化酶亚基qoxB基因缺失株,并探索其在细菌生长及感染过程中发挥的生物学功能。【方法】利用同源重组方法构建获得缺失株ΔqoxB后,对野生株EGD-e和缺失株ΔqoxB的生长能力、细菌运动能力和细胞内黏附、侵袭、增殖及胞内迁移能力进行比较,同时利用荧光定量PCR方法检测ΔqoxB中鞭毛相关基因转录水平的变化。【结果】缺失qoxB基因后细菌在体外培养过程中生长能力没有差异,细菌的鞭毛运动能力显著降低,在30℃培养24 h和48 h后ΔqoxB运动圈直径分别较EGD-e下降35.86%和34.20%,且22个鞭毛相关基因转录水平显著降低。通过细胞感染试验发现缺失qoxB基因后细胞黏附、侵袭、增殖及胞内迁移能力均显著下降。【结论】本研究首次证实末端氧化酶亚基QoxB能降低单增李斯特菌的运动能力和对细胞的感染能力,此研究为进一步阐明末端细胞色素氧化酶影响单增李斯特菌的致病机制提供重要依据。     

鸭疫里氏杆菌hemH基因功能初步鉴定及其缺失株的转录组学分析

血红素是绝大多数细菌生长繁殖所必需的一种营养物质,其参与了细菌多种重要的生理过程。细菌可以通过自身合成和从外界摄取2种方式获得血红素。然而过多的血红素对细菌是有毒性的,细菌则会利用外排、螯合和降解等多种方式减轻血红素毒性。鸭疫里氏杆菌(Riemerella anatipestifer, RA)是一种感染禽类的革兰氏阴性菌,前期研究表明该菌可通过转运的方式从外界环境摄取血红素。然而,是否该菌也可以自身合成血红素未知。基因组分析发现鸭疫里氏杆菌ATCC11845菌株中的基因RA0C＿RS08070编码铁螯合酶(ferrochelatase) HemH,是参与将铁插入卟啉中心,形成血红素的关键酶。hem H缺失后会导致铁离子和卟啉的积累,对细菌造成毒性。【目的】为鉴定Hem H在合成血红素中的功能及查找参与鸭疫里氏杆菌铁离子和卟啉解毒相关基因。【方法】本研究构建了RAATCC11845Δhem H缺失株,并检测亲本株和hem H缺失株在GCB以及GCB添加血红蛋白液体培养基中的生长情况;随后对亲本株和hem H缺失株进行转录组测序并进行比较分析。【结果】RAATCC11845Δhem H缺失...     

发酵乳杆菌CSC-19胞外粗多糖提取物抑制单核细胞增生李斯特菌生物被膜形成能力的研究

【目的】筛选能有效抑制单核细胞增生李斯特菌(Listeria monocytogenes,LM)形成生物被膜的乳酸菌,分析其活性成分并进行功能表征。【方法】采用结晶紫染色法筛选抑制LM形成生物被膜的不同乳酸菌提取物;通过酸中和、蛋白酶处理及热处理,推测抑制生物被膜活性物质以胞外多糖(extracellular crude polysaccharide,ECP)为主;乙醇沉淀法提取目标乳酸菌分离株胞外粗多糖,分析其抑制生物被膜形成活性和对LM生长的影响;运用激光共聚焦扫描显微镜(laser confocal scanning microscopy,LCSM)和扫描电子显微镜(scanning electron microscopy,SEM)观察胞外粗多糖对生物被膜细胞形态和结构的影响。【结果】发酵乳杆菌CSC-19发酵上清液对1516-2LM生物被膜的抑制率为81.7%;经热和蛋白酶处理后,发酵上清抑制生物被膜形成的活性未发生显著变化(P>0.05),表明发酵上清液中抑制生物被膜形成的物质可能为胞外多糖;在不抑制LM生长的条件下所提取的胞外粗多糖抑制生物被膜形成能力具有浓度依赖性。激光共聚焦扫描显微镜和扫描电子显微镜结果显示,胞外粗多糖显著抑制了生物被膜的形成能力,生物被膜三维、有组织的蜂窝状结构被破坏,仅有少量的粘附细胞分散于细胞爬片表面。【结论】发酵乳杆菌CSC-19胞外粗多糖能有效抑制LM生物被膜的形成,有望应用于高效防控该菌污染食品。     

Calcineurin B-like interacting protein kinase 31 confers resistance to sheath blight via modulation of ROS homeostasis in rice

Sheath blight (ShB) severely threatens rice cultivation and production; however, the molecular mechanism of rice defence against ShB remains unclear. Screening of transposon Ds insertion mutants identified that Calcineurin B-like protein-interacting protein kinase 31 (CIPK31) mutants were more susceptible to ShB, while CIPK31 overexpressors (OX) were less susceptible. Sequence analysis indicated two haplotypes of CIPK31: Hap_1, with significantly higher CIPK31 expression, was less sensitive to ShB than the Hap_2 lines. Further analyses showed that the NAF domain of CIPK31 interacted with the EF-hand motif of respiratory burst oxidase homologue (RBOHA) to inhibit RBOHA-induced H₂O₂ production, and RBOHA RNAi plants were more susceptible to ShB. These data suggested that the CIPK31-mediated increase in resistance is not associated with RBOHA. Interestingly, the study also found that CIPK31 interacted with catalase C (CatC); cipk31 mutants accumulated less H₂O₂ while CIPK31 OX accumulated more H₂O₂ compared to the wild-type control. Further analysis showed the interaction of the catalase domain of CatC with the NAF domain of CIPK31 by which CIPK31 inhibits CatC activity to accumulate more H₂O₂.