Protoplast fusion in Bacillus species produces frequent,unbiased, genome-wide homologous recombination

In eukaryotes, fine-scale maps of meiotic recombination events have greatly advanced our understanding of the factors that affect genomic variation patterns and evolution of traits. However, in bacteria that lack natural systems for sexual reproduction, unbiased characterization of recombination landscapes has remained challenging due to variable rates of genetic exchange and influence of natural selection. Here, to overcome these limitations and to gain a genome-wide view on recombination, we crossed Bacillus strains with different genetic distances using protoplast fusion. The offspring displayed complex inheritance patterns with one of the parents consistently contributing the major part of the chromosome backbone and multiple unselected fragments originating from the second parent. Our results demonstrate that this bias was in part due to the action of restriction–modification systems, whereas genome features like GC content and local nucleotide identity did not affect distribution of recombination events around the chromosome. Furthermore, we found that recombination occurred uniformly across the genome without concentration into hotspots. Notably, our results show that species-level genetic distance did not affect genome-wide recombination. This study provides a new insight into the dynamics of recombination in bacteria and a platform for studying recombination patterns in diverse bacterial species.     

连续两个生长季大气CO2浓度升高对银杏希尔反应活力和叶绿体ATP酶活性的影响

近年来,随着温室气体浓度不断上升,有关CO2浓度升高对植物影响的研究已取得一定进展,但CO2浓度升高对植物光合作用的影响需要从生理生化水平上进一步深入的研究.以沈阳城市森林树种银杏(Ginkgo biloba L.)为研究对象,利用开顶式气室研究连续两个生长季大气CO2浓度升高对银杏光合特性的影响.结果表明,在大气CO2浓度为700μmol·mol-1条件下,与对照相比,第1个生长季CO2处理的银杏叶片净光合速率极显著增加(P<0.01),希尔反应活力极显著增大(P<0.01)、Ca2+/Mg2+-ATP酶活性显著(P<0.05)或极显著增强(P<0.01)、光合产物淀粉的含量极显著增多(P<0.01);第2生长季CO2处理的银杏叶片净光合速率显著增加(P<0.05),希尔反应活力在通气60d时极显著(P<0.01)增大,Ca2+/Mg2+-ATP酶活性在处理30d时显著降低(P<0.05),淀粉含量增多.与第1个生长季相比,第2个生长季CO2处理的银杏叶片净光合速率降低,希尔反应活力减小,Ca2+/Mg2+-ATP酶活性减弱,叶绿素含量增多,淀粉含量减少.试验中出现了光合适应现象.     

竹产业生态系统结构及演化规律——以贵州省赤水市为例

以"中国竹子之乡"命名的贵州省赤水市为例,研究了竹产业生态系统结构及演化规律。结果表明:(1)竹子各构件都有开发价值,包括生态服务、食药、竹材加工和文化产品等5方面,产业系统呈3级链环模式,第二、第三产业链环由近200家加工企业及6大系列近300个竹加工品种组成,包括一般加工和深加工产品,第一产业链环主要由占全市森林面积47.3%的竹林资源构成。第一产业链环是环境保障与原材料的基础,第二和第三产业链环是产业化能力,三者互相依存。(2)竹产业生态系统由环境资源、原生产业、外生产业、共生产业和产业创新5个子系统组成,子系统间相互作用、重要性不同,出现原生产业子系统为主(Ⅰ)、外生-原生产业子系统为主(Ⅱ)、原生-外生产业子系统为主(Ⅲ)和外生产业子系统为主(Ⅳ)4种状态,赤水市竹产业生态系统处于Ⅱ→Ⅲ级状态。竹外生产业子系统产值占全市GDP的40%以上、呈逐年上升的趋势,2014年竹产业生态系统中原生、外生和共生产业子系统产值分别为0.95×10~9、2.85×10~9元和2.30×10~9元,总产值6.10×10~9元。(3)依据各子系统间相互作用及生产要素转化过程,赤水市竹产业生态系统演化过程划分为4个阶段。原始发展阶段(约1950年以前)处于Ⅰ级状态,手工利用阶段(1951—1980年)整体处于Ⅰ级、毛竹林资源及加工利用部分有向Ⅱ级转化的趋势,工业利用阶段(1981—2000年)处于Ⅱ级状态,系统利用阶段(2001年至今)整体处于Ⅱ级、杂竹林资源及加工利用部分有向Ⅲ级转化的趋势。现时竹产业发展需提高原生产业子系统规模与生产力,依靠产业创新子系统引领竹产业生态系统走创新驱动之路。     

荒漠植物梭梭稳定碳同位素组成与环境因子的关系

以广泛分布于新疆荒漠地区的建群种植物——梭梭(Haloxylon ammodendron)为研究对象,通过对23个样地101份梭梭同化枝样品δ~(13)C值的测定,分析了梭梭稳定碳同位素组成的变化特征及其与环境因子(海拔、日照时数、潜在蒸散量、年平均降水量和年平均温度)的关系,并讨论了不同生境下梭梭同化枝δ~(13)C值的变化特征。研究结果显示:(1)梭梭同化枝δ~(13)C平均值为-14.15‰,其在95%置信区间的变化范围为-13.14‰—-15.38‰,表明梭梭是C_4光合途径的植物。(2)梭梭同化枝δ~(13)C值与年平均降水量和年平均温度呈显著负相关关系,而与日照时数、潜在蒸散量和海拔呈显著正相关关系。我们推测梭梭同化枝δ~(13)C值对各环境因子响应趋势的不同,可能是由气孔限制因素造成的,它是梭梭适应干旱荒漠环境的一种策略。(3)在不同生境下,梭梭同化枝的碳同位素组成存在显著差异。当梭梭群落中的主要伴生种为白刺、红砂时,其δ~(13)C值最高,当主要伴生种为沙拐枣和假木贼时,其δ~(13)C值最低。在灰漠土与灰棕漠土样地中的梭梭δ~(13)C值高于棕钙土、风沙土、石质土样地;盆地中梭梭同化枝δ~(13)C值低于平原、山地、丘陵地形条件下的样地。以上结果表明:梭梭水分利用效率在不同环境梯度和生境中,存在着显著不同,表现出显著的适应策略差异。     

Hemin with Peroxidase Activity Can Inhibit the Oxidative Damage Induced by Ultraviolet A

Excessive reactive oxygen species (ROS), a highly reactive substance that contains oxygen, induced by ultraviolet A (UVA) cause oxidative damage to skin. We confirmed that hemin can catalyze the reaction of tyrosine (Tyr) and hydrogen peroxide (H₂O₂). Catalysis was found to effectively reduce or eliminate oxidative damage to cells induced by H₂O₂ or UVA. The scavenging effects of hemin for other free-radical ROS were also evaluated through pyrogallol autoxidation, 1,1-diphenyl-2-picrylhydrazyl radical (DPPH·)-scavenging assays, and phenanthroline–Fe²⁺ assays. The results show that a mixture of hemin and tyrosine exhibits strong scavenging activities for H₂O₂, superoxide anion (O₂⁻·), DPPH·, and the hydroxyl radical (·OH). Furthermore, the inhibition of oxidative damage to human skin keratinocyte (HaCaT) cells induced by H₂O₂ or UVA was evaluated. The results show that catalysis can significantly reduce the ratio of cell apoptosis and death and inhibit the release of lactate dehydrogenase (LDH), as well as accumulation of malondialdehyde (MDA). Furthermore, the resistance to apoptosis was found to be enhanced. These results show that the mixture of hemin and tyrosine has a significantly protective effect against oxidative damage to HaCaT cells caused by UVA, suggesting it as a protective agent for combating UVA damage.     

干旱胁迫对银杏叶片光合系统Ⅱ荧光特性的影响

为了探讨银杏叶片在长期干旱胁迫下光合性能的变化,采用盆栽控水法对银杏幼苗进行干旱处理,并分别在处理后第0、20、30、40、50天,对荧光动力学曲线和荧光参数进行了测定和JIP-test分析.结果表明:随着干旱的加剧,叶片叶绿素含量逐渐递减.由于干旱导致银杏叶片的光合系统Ⅱ(PSⅡ)结构破坏和稳固性减弱,其荧光动力学曲线出现了K相和L相的升高,在干旱处理40 d后升高加剧.PSⅡ结构的变化导致了一系列荧光参数的变化,初始荧光F0逐渐升高,最大荧光Fm逐渐降低;代表单位反应中心活性的参数ABS/RC、ET0/RC、TR0/RC逐渐升高;单位面积捕获的光能TR0/CS0高于对照,单位面积用于电子传递的光能ET0/ CS0后期下降;表示受体侧电子传递活性的参数Sm、Ψ0、(φ)E0逐渐下降,M0、VJ、VI逐渐升高;表示热耗散的参数DI0/RC、DI0/CS0随干旱时间延长而急剧升高;活性反应中心数目RC/CS0减少;代表PSⅡ光合效率的参数Fv/Fm、PIabs逐渐下降;代表PSⅡ供体侧活性的参数Wk随干旱时间延长而逐渐升高,OEC逐渐减少.干旱胁迫导致银杏叶片PSⅡ反应中心失活,能流分配失衡,电子传递受阻,PSⅡ放氧复合体失活和PSⅡ稳固性减弱,从而破坏PSⅡ光合功能.而反应中心失活和受体侧电子QA-的累积是造成PSⅡ电子传递活性减弱的主要原因.干旱条件下,银杏PSⅡ的PIabs比Fv/Fm对干旱的反应更灵敏,可作为银杏叶片受到伤害的指标.     

Refolding and purification of interferon-gamma in industry by hydrophobic interaction chromatography

A new technology for renaturation with simultaneous purification of the recombinant human interferon-gamma (rhIFN-gamma) in downstream of biotechnology is presented. The strategies to develop the new technology in industry scale were suggested. Based on chemical equilibrium and molecular interactions, the principle of rhIFN-gamma refolding by HPHIC was described. The kind of stationary and mobile phases were evaluated and found the former to contribute to the rhIFN-gamma refolding more than the latter. The extract containing the rhIFN-gamma in gram scale in 7.0 mol L(-1) guanidine hydrochloride solution of 700 mL was directly pumped into a unit of simultaneous renaturation and purification of proteins (USRPP, 10 x 300 mm i.d.) packed by small particle packings of hydrophobic interaction chromatography and a satisfactory recovery of bioactivity and mass of the rhIFN-gamma was obtained. With flow rate 100 mL min(-1) and a gradient elution by only one step in 4h, the purity and specific bioactivity approach to 95% and 8.7 x 10(7) IU(-1) mg, respectively. To evaluate the goodness of the presented new technology in this study, a usual method with the renaturation by dilution method firstly and then purification with a series of LC in literature was employed to compare with each other. The obtained result in terms of purity, recoveries of mass and bioactivity, cost time as well as expenses, the former is much better than the latter. Comparing the total bioactivity of rhIFN-gamma in the extract before to that after the renaturation by the USRPP, the total bioactivity of rhIFN-gamma increased 62-fold.     

酱油中黄曲霉毒素B1的风险评估

黄曲霉毒素B1(AFB1)是已知毒性最强的天然物质,在我国的酱油产品中普遍存在,对人体的危害及其严重,其安全性受到消费者的密切关注。为了解我国酱油中黄曲霉毒素B1的安全现状,建立我国酱油中黄曲霉毒素B1的应急对策,本文采用酶联免疫法(ELISA)对我国203个酱油样品中的AFB1含量进行了检测,并就现有的污染状况,从危害识别、危害描述、暴露评估和风险特征描述等方面对酱油中的AFB1进行了风险评估。     

Determination of cefazedone in human plasma by high performance liquid chromatography–tandem mass spectrometry: Application to a pharmacokinetic study on Chinese volunteers

A rapid, sensitive and simple high performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) method was developed for determination of cefazedone in human plasma using metronidazole as internal standard (IS). The chromatographic separation was achieved on an Ultimate XB-CN column (2.1 mm × 150 mm, 5 μm) with an isocratic mobile phase of acetonitrile and 20 mM ammonium acetate in 0.1% formic acid in water (15:85, v/v). Detection was performed using electrospray ionization in positive ion multiple reaction-monitoring mode (SRM), monitoring the transitions m/z 548.2 → 344.1 for cefazedone and m/z 172.2 → 128.1 for IS. Calibration curves were linear over a wide range of 0.20–401.12 μg/mL for cefazedone in plasma. The lower limit of quantification (LLOQ) was 0.20 μg/mL. The intra- and inter-day precisions were less than 7.2%. The average recovery of cefazedone was 90.8–91.0%. The validated method was successfully applied to the pharmacokinetic study of cefazedone in Chinese healthy volunteers following intravenous (IV) administration of 500, 1000 and 2000 mg cefazedone injection.     

石臼湖原生动物种群分布及其同质化

石臼湖地处长江中下游,是国内为数不多的通江淡水湖。为探讨湖泊与入湖支流不同生境中原生动物种群结构及其相似性,于2012年平水期和枯水期分别对石臼湖及其周边入湖支流进行原生动物调查,研究河流和湖泊区域原生动物的种类组成及其季节变化,同时与同一地区的相邻湖泊固城湖作对比,通过计算相似性指数,探讨原生动物对生境同质化的响应。结果表明:调查共采集到原生动物57种,平水期种类多于枯水期;石臼湖河流区各站点原生动物相似性指数在Ⅰ~Ⅲ级之间,为完全不同-轻度相似;湖区站点相似性指数在Ⅱ~Ⅳ之间,为极不相似-中度相似;河流区种类季节之间的相似度极低(0.050~0.267),而湖泊区种类季节之间处于中等相似水平(0.250~0.375),说明河流区原生动物种类的季节变化较湖泊区明显,生物组成的异质性也高于湖泊;原生动物分布对水质有很好的响应关系,氮磷元素在影响原生动物种类组成和分布中起了主要作用;通过石臼湖与固城湖及长江中下游其他湖泊的对比分析,表明在一定范围内,随着生境尺度的增加,生境的同质化会提高生物同质化水平,但超过景观尺度,原生动物地域性特征逐渐显现,即使生境同质,其生物也未必同质;且随着距离的增加,不同区域的生物相似性呈降低的趋势。