The Extracellular Xylan Degradative System in Clostridium cellulolyticum Cultivated on Xylan: Evidence for Cell-Free Cellulosome Production

In this study, we demonstrate that the cellulosome of Clostridium cellulolyticum grown on xylan is not associated with the bacterial cell. Indeed, the large majority of the activity (about 90%) is localized in the cell-free fraction when the bacterium is grown on xylan. Furthermore, about 70% of the detected xylanase activity is associated with cell-free high-molecular-weight complexes containing avicelase activity and the cellulosomal scaffolding protein CipC. The same repartition is observed with carboxymethyl cellulase activity. The cellulose adhesion of xylan-grown cells is sharply reduced in comparison with cellulose-grown cells. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that cellulosomes derived from xylan- and cellulose-grown cells have different compositions. In both cases, the scaffolding protein CipC is present, but the relative proportions of the other components is dramatically changed depending on the growth substrate. We propose that, depending on the growth substrate, C. cellulolyticum is able to regulate the cell association and cellulose adhesion of cellulosomes and regulate cellulosomal composition.     

Identification of suitable reference genes for real-time quantitative PCR analysis of hydrogen peroxide-treated human umbilical vein endothelial cells

Background

Oxidative stress can induce cell injury in vascular endothelial cells, which is the initial event in the development of atherosclerosis. Although quantitative real-time polymerase chain reaction (qRT-PCR) has been widely used in gene expression studies in oxidative stress injuries, using carefully validated reference genes has not received sufficient attention in related studies. The objective of this study, therefore, was to select a set of stably expressed reference genes for use in qRT-PCR normalization in oxidative stress injuries in human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H₂O₂).

Results

Using geNorm analysis, we found that five stably expressed reference genes were sufficient for normalization in qRT-PCR analysis in HUVECs treated with H₂O₂. Genes with the most stable expression according to geNorm were U6, TFRC, RPLP0, GAPDH, and ACTB, and according to NormFinder were ALAS1, TFRC, U6, GAPDH, and ACTB.

Conclusion

Taken together, our study demonstrated that the expression stability of reference genes may differ according to the statistical program used. U6, TFRC, RPLP0, GAPDH, and ACTB was the optimal set of reference genes for studies on gene expression performed by qRT-PCR assays in HUVECs under oxidative stress study.

     

基于1H-NMR及GC-MS的中国块菌及其共生云南松的代谢物分析

采用代谢组学技术研究中国块菌(云南黑松露)及其共生云南松的小分子活性代谢产物.运用核磁共振(1H-NMR)和气相质谱(GC-MS)两种技术,分析新鲜中国块菌成熟子实体及松根的代谢成分;将1H-NMR及GC-MS数据导入SMICA-P软件进行多元统计分析,比较二者的差异代谢物.在块菌子实体及松根中分别共检测到了苏氨酸、甘...     

川西亚高山森林土壤呼吸和微生物生物量碳氮对施氮的响应

随着全球大气氮沉降的明显增加,将有可能显著影响我国西部地区受氮限制的亚高山森林生态系统。土壤微生物是生态系统的重要组成部分,是土壤物质循环和能量流动的重要参与者。由于生态系统类型、土壤养分、氮沉降背景值等的差异,土壤呼吸和土壤生物量碳氮对施氮的响应存在许多不确定性。而施氮会不会促进亚高山森林生态系统中土壤呼吸和微生物对土壤碳氮的固定?基于此假设,选择了川西60年生的四川红杉(Larix mastersiana)亚高山针叶林为研究对象,通过4个水平的土壤施氮控制试验(CK:0 g m~(-2) a~(-1)、N1:2 g m~(-2)a~(-1)、N2:5 g m~(-2) a~(-1)、N3:10 g m~(-2)a~(-1)),监测了土壤呼吸及土壤微生物生物量碳氮在一个生长季的动态情况。结果表明:施氮对土壤呼吸各指标和土壤微生物碳氮都有极显著的影响,施氮能促进土壤全呼吸、自养呼吸、异养呼吸通量和土壤微生物生物量碳氮的增长,施氮使土壤呼吸通量提高了11%—15%,土壤微生物量碳提高了5%—9%,土壤微生物量氮提高了23%—34%。在中氮水平下(5 g m~(-2) a~(-1))对土壤呼吸的促进最显著。相关分析发现,土壤呼吸与微生物生物量碳氮和微生物代谢商极呈显著正相关,微生物量碳氮与土壤温度呈极显著的正相关,与土壤湿度呈极显著负相关。通过一般线性回归拟合土壤呼吸速率与土壤10 cm温湿度的关系,发现土壤呼吸速率与土壤温度呈极显著的正相关,与土壤湿度极显著负相关(P0.001),中氮水平下土壤温度敏感性系数Q_(10)值(7.10)明显高于对照(4.26)。     

The sexual dimorphism displayed by the roots of mulberry (Morus alba) saplings depends on the sex of the neighboring plants

桑树幼苗根系表现出的性别二态性取决于相邻植株的性别 雌雄异株植物通常呈现性别二态性,但其雌株和雄株的根系性状和养分吸收如何响应相邻植株 的性别变化尚不清楚。本研究以中国广泛栽培的雌雄异株植物桑树(Morus alba)为试验对象,在水培环境 下,采取雌-雌、雄-雄和雌-雄3种组合种植模式进行试验。植株生长3个月后,分别测定雌雄植株根系的解剖结构、形态、营养浓度和生物量。研究结果表明,在水培环境中,与同性相邻相比,当桑树雄株与异性相邻时,其根系木质部变小,根和径生物量减少,根系中N、P、K浓度增加。相比,当桑树雌株与异性相邻时,其根系木质部变大,细根长度、表面积和体积以及根系δ¹³C和N、P、K浓度增加。雌雄植株根系δ¹³C和N、P、K浓度的变化与根系性状的改变有关。这些结果表明,相邻植株的性别影响了雌雄桑树根系解剖结构和形态。同时,雌雄植株对相邻植株性别改变的响应呈现性别二态性,这种二态性响应影响了雌雄植株水分利用效率和营养吸收。这些发现对于理解森林和自然系统中其它雌雄异株物种的种群动态变化具有重要意义。     

Volatile organic compound emissions from Microcystis aeruginosa under different phosphorus sources and concentrations

In eutrophicated water, cyanobacteria massively grow and release an abundance of volatile organic compounds (VOCs) that contribute to the water odor. To uncover the effects of different phosphorus (P) nutrients on the formation of cyanobacteria VOCs and water odor, the cell growth and VOC emissions of Microcystis aeruginosa were investigated under different P nutrient conditions. Among K₂HPO₄, Na₄P₂O₇, and (NaPO₃)₆, K₂HPO₄ showed the largest increase in cell density, while a reduction in P concentration decreased the cell density. There were 26, 23 and 22 compounds in M. aeruginosa VOCs with K₂HPO₄, Na₄P₂O₇ and (NaPO₃)₆ as the sole P source, respectively, including sulfocompounds, terpenoids, benzenes, hydrocarbons, alcohols, aldehydes, and esters. Non‐P markedly promoted the VOC emission, and six additional compounds were observed: α‐pinene, 1‐phenyl‐1‐butanone, 1H‐1‐ethylidene‐indene, 2,6,10‐trimethyl‐tetradecane, 2‐ethyl‐hexanal, and acetic acid 2‐ethylhexyl ester. It can be deduced that cyanobacteria release different VOC blends using various P forms in eutrophicated waters, and the reduction of P amount promotes VOC emission and increases the water odor.     

Lipid profiling of rat peritoneal surface layers by online normal- and reversed-phase 2D LC QToF-MS

An online, two-dimensional (2D) liquid chromatography (LC) quadrupole time-of-flight mass spectrometry (QToF-MS) method was developed for lipid profiling of rat peritoneal surface layers, in which the lipid classes and species could be simultaneously separated in one injection with a significantly increased sensitivity. Different lipid classes were separated on a normal-phase column in the first dimension and lipid molecular species were separated on a reversed-phase column in the second dimension, so that the ion suppression effects were reduced while the detection sensitivity was improved. Identified were 721 endogenous lipid species from 12 lipid classes, in which 415 structures were confirmed using tandem mass spectra, and the other 306 lipid molecular species were identified by accurate masses. The linearity, limit of detection, and repeatability were all satisfactory. The method was applied to the investigation of the lipid changes in rat peritoneal surface layer after peritoneal dialysis, and 32 potential lipid biomarkers were identified, as their concentrations in the dosed group were 2.2–12.5 times of those in the control group. The results revealed that this 2D LC-MS system was a promising tool for lipid profiling of complex biological samples.