高效产氢菌株Enterococcus sp. LG1的分离及产氢特性

采用Hungate厌氧培养技术分别从厌氧污泥、好氧污泥及河底泥中分离出12株厌氧产氢细菌,并对其中的Enterococcus sp.LG1(注册号:EU258743)进行了研究.结果表明,该株细菌为专性厌氧菌,经革兰氏染色结果为阴性.通过16S rDNA碱基测序和比对证实,该菌株是目前尚未报道过的1个新菌种,初步确定其细菌学上的分类地位.同时,以灭菌预处理的污泥为底物培养基,对该菌的产氢能力及污泥发酵过程中底物性质变化(SCOD、可溶性蛋白质、总糖和pH值等)进行了探讨.实验结果显示,产氢茵Enterococcus sp.LG1的发酵过程中只有H2和CO2产生,无CH4产生.产气量最高为36.48 mL/g TCOD,氢气含量高达73.5%,为已报道文献中以污泥为底物发酵制氢中之最高.根据污泥发酵产物分析得知,该菌的发酵类行为典型的丁酸型发酵.     

应用规模缩小方法的鸟苷发酵过程放大*

利用鸟苷生产菌株枯草芽孢杆菌(B.subtilis)754#,在50L发酵罐成功优化的基础上,分别在12M^3的中试规模和100M^3的生产规模进行了放大,产苷分别达到29．4g/L和21．4g/L;进而通过过程缩小(scale down)方法,从代谢流动态变化的角度研究了放大过程中存在的问题,发现DO是限制过程放大的另一重要因素,据此将50L规模克服代谢流迁移的优化工艺成功放大到生产规模,使产苦水平进一步提高了18％,达25．2g/L。     

mTORC1-independent translation control in mammalian cells by methionine adenosyltransferase 2A and S-adenosylmethionine

Methionine adenosyltransferase (MAT) catalyzes the synthesis of S-adenosylmethionine (SAM). As the sole methyl-donor for methylation of DNA, RNA, and proteins, SAM levels affect gene expression by changing methylation patterns. Expression of MAT2A, the catalytic subunit of isozyme MAT2, is positively correlated with proliferation of cancer cells; however, how MAT2A promotes cell proliferation is largely unknown. Given that the protein synthesis is induced in proliferating cells and that RNA and protein components of translation machinery are methylated, we tested here whether MAT2 and SAM are coupled with protein synthesis. By measuring ongoing protein translation via puromycin labeling, we revealed that MAT2A depletion or chemical inhibition reduced protein synthesis in HeLa and Hepa1 cells. Furthermore, overexpression of MAT2A enhanced protein synthesis, indicating that SAM is limiting under normal culture conditions. In addition, MAT2 inhibition did not accompany reduction in mechanistic target of rapamycin complex 1 activity but nevertheless reduced polysome formation. Polysome-bound RNA sequencing revealed that MAT2 inhibition decreased translation efficiency of some fraction of mRNAs. MAT2A was also found to interact with the proteins involved in rRNA processing and ribosome biogenesis; depletion or inhibition of MAT2 reduced 18S rRNA processing. Finally, quantitative mass spectrometry revealed that some translation factors were dynamically methylated in response to the activity of MAT2A. These observations suggest that cells possess an mTOR-independent regulatory mechanism that tunes translation in response to the levels of SAM. Such a system may acclimate cells for survival when SAM synthesis is reduced, whereas it may support proliferation when SAM is sufficient.     

Cross-seeding of WT amyloid-β with Arctic but not Italian familial mutants accelerates fibril formation in Alzheimer's disease

Alzheimer’s disease (AD) involves the neurotoxic self-assembly of a 40 and 42 residue peptide, Amyloid-β (Aβ). Inherited early-onset AD can be caused by single point mutations within the Aβ sequence, including Arctic (E22G) and Italian (E22K) familial mutants. These mutations are heterozygous, resulting in an equal proportion of the WT and mutant Aβ isoform expression. It is therefore important to understand how these mixtures of Aβ isoforms interact with each other and influence the kinetics and morphology of their assembly into oligomers and fibrils. Using small amounts of nucleating fibril seeds, here, we systematically monitored the kinetics of fibril formation, comparing self-seeding with cross-seeding behavior of a range of isoform mixtures of Aβ42 and Aβ40. We confirm that Aβ40(WT) does not readily cross-seed Aβ42(WT) fibril formation. In contrast, fibril formation of Aβ40(Arctic) is hugely accelerated by Aβ42(WT) fibrils, causing an eight-fold reduction in the lag-time to fibrillization. We propose that cross-seeding between the more abundant Aβ40(Arctic) and Aβ42(WT) may be important for driving early-onset AD and will propagate fibril morphology as indicated by fibril twist periodicity. This kinetic behavior is not emulated by the Italian mutant, where minimal cross-seeding is observed. In addition, we studied the cross-seeding behavior of a C-terminal-amidated Aβ42 analog to probe the coulombic charge interplay between Glu22/Asp23/Lys28 and the C-terminal carboxylate. Overall, these studies highlight the role of cross-seeding between WT and mutant Aβ40/42 isoforms, which can impact the rate and structure of fibril assembly.     

潮棕壤线虫群落对土地利用方式的响应

作者对潮棕壤不同土地利用方式(旱田、撂荒地和林地)下土壤线虫群落时空分布特征进行了研究, 结果表明, 不同土地利用方式能够影响线虫群落及其优势属的时空分布。线虫优势属对不同土地利用方式的响应不同, 撂荒地和林地处理中板唇属(Chiloplacus)线虫主要分布在5–30 cm土层, 而其他线虫优势属则主要分布在0–20 cm土层; 在旱田处理中, 短体属(Pratylenchus)线虫均匀分布在各个土层。由于土地利用方式的改变而引起的土壤环境因素的变化能够对土壤线虫产生影响, 研究发现土壤孔隙度、土壤有机碳、全氮和碳氮比与土壤线虫优势属的数量具有显著的正相关关系。线虫区系分析结果表明, 撂荒地和林地处理中土壤环境相对稳定, 土壤食物网向较成熟的阶段演替。线虫区系分析方法可用来揭示不同土地利用方式下土壤食物网的变化, 为进一步研究土壤生态过程对土地利用方式的响应提供了有效的工具。     

Amazonian Anthrosols Support Similar Microbial Communities that Differ Distinctly from Those Extant in Adjacent, Unmodified Soils of the Same Mineralogy

We compared the microbial community composition in soils from the Brazilian Amazon with two contrasting histories; anthrosols and their adjacent non-anthrosol soils of the same mineralogy. The anthrosols, also known as the Amazonian Dark Earths or terra preta, were managed by the indigenous pre-Colombian Indians between 500 and 8,700 years before present and are characterized by unusually high cation exchange capacity, phosphorus (P), and calcium (Ca) contents, and soil carbon pools that contain a high proportion of incompletely combusted biomass as biochar or black carbon (BC). We sampled paired anthrosol and unmodified soils from four locations in the Manaus, Brazil, region that differed in their current land use and soil type. Community DNA was extracted from sampled soils and characterized by use of denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism. DNA bands of interest from Bacteria and Archaea DGGE gels were cloned and sequenced. In cluster analyses of the DNA fingerprints, microbial communities from the anthrosols grouped together regardless of current land use or soil type and were distinct from those in their respective, paired adjacent soils. For the Archaea, the anthrosol communities diverged from the adjacent soils by over 90%. A greater overall richness was observed for Bacteria sequences as compared with those of the Archaea. Most of the sequences obtained were novel and matched those in databases at less than 98% similarity. Several sequences obtained only from the anthrosols grouped at 93% similarity with the Verrucomicrobia, a genus commonly found in rice paddies in the tropics. Sequences closely related to Proteobacteria and Cyanobacteria sp. were recovered only from adjacent soil samples. Sequences related to Pseudomonas, Acidobacteria, and Flexibacter sp. were recovered from both anthrosols and adjacent soils. The strong similarities among the microbial communities present in the anthrosols for both the Bacteria and Archaea suggests that the microbial community composition in these soils is controlled more strongly by their historical soil management than by soil type or current land use. The anthrosols had consistently higher concentrations of incompletely combusted organic black carbon material (BC), higher soil pH, and higher concentrations of P and Ca compared to their respective adjacent soils. Such characteristics may help to explain the longevity and distinctiveness of the anthrosols in the Amazonian landscape and guide us in recreating soils with sustained high fertility in otherwise nutrient-poor soils in modern times.     

利用SpyTag/SpyCatcher构建胞内自组装多酶复合体实现高效生物合成

SpyTagr和SpyCatche可通过自发反应形成共价键,产生稳定的分子自组装体。酶分子自组装体因具有高效有序的催化特性在合成生物学和纳米技术领域具有重要的应用价值。为探索SpyTag/SpyCatcher在大肠杆菌胞内多酶复合体系形成有序自组装分子能力,将SpyTagr和SpyCatche分别与P450BM3m单加氧酶和葡萄糖脱氢酶GDH进行融合表达,以期产生具有辅酶再生循环系统、高效生物合成靛蓝分子的SpyTag/SpyCatcher双酶自组装复合体。首先,通过电泳及质谱对重组工程菌表达蛋白进行分析,证实SpyCatcher-P450BM3m与SpyTag-GDH在胞内成功形成了自组装多酶复合体;然后,系统分析不同培养条件下组装体合成靛蓝的能力。结果发现,经0.5mmol/L IPTG诱导后,菌体在16℃继续培养18h后,工程菌对吲哚(2mmol/L)与葡萄糖(4mmol/L)的全细胞催化能力最强,靛蓝产量最高达258mg/L,是未组装多酶系统的1.9倍,比P450BM3m单酶表达系统高约2.4倍;反应70min后达到反应平衡,转化率为52%。成功实现了SpyTag/SpyCatcher介导的多酶体系在大肠杆菌细胞中的自组装和高效转化体系,为胞内多酶复合物组装体的设计提供了新思路。     

用气象卫星对东亚季风区的生态过渡带的遥感监测研究

用极轨气象卫星的归一化植被指数(NDVI)对中国东亚季风区的生态过渡带(样带)进行遥感监测研究。研究结果表明,在过渡带内的各个生态区的NDVI与东亚季风进退的关系极为密切。在正常季风年NDVI有正常分布;在异常季风年,NDVI呈反常分布;把遥感过渡带内季风气候的变化简化为遥感生态区植被指数的变化。分析了在过渡带内主要生态区,即荒漠草原、干草原、草甸草原、农耕区、森林区ND-VI的变化规律,以及旬NDVI与旬降水量之间的关系。在过渡带内每年NDVI随时间变化的曲线形状可以作为每年季风气候变化特征的量度。     

Expression of baculovirus anti-apoptotic genes p35 and op-iap in cotton (Gossypium hirsutum L.) enhances tolerance to verticillium wilt

Background

Programmed cell death plays an important role in mediating plant adaptive responses to the environment such as the invasion of pathogens. Verticillium wilt, caused by the necrotrophic pathogen Verticillium dahliae, is a serious vascular disease responsible for great economic losses to cotton, but the molecular mechanisms of verticillium disease and effective, safe methods of resistance to verticillium wilt remain unexplored.

Methodology/Principal Findings

In this study, we introduced baculovirus apoptosis inhibitor genes p35 and op-iap into the genome of cotton via Agrobacterium-mediated transformation and analyzed the response of transgenic plants to verticillium wilt. Results showed that p35 and op-iap constructs were stably integrated into the cotton genome, expressed in the transgenic lines, and inherited through the T₃ generation. The transgenic lines had significantly increased tolerance to verticillium wilt throughout the developmental stages. The disease index of T₁–T₃ generation was lower than 19, significantly (P<0.05) better than the negative control line z99668. After treatment with 250 mg/L VD-toxins for 36 hours, DNA from negative control leaves was fragmented, whereas fragmentation in the transgenic leaf DNA did not occur. The percentage of cell death in transgenic lines increased by 7.11% after 60 mg/L VD-toxin treatment, which was less than that of the negative control lines''s 21.27%. This indicates that p35 and op-iap gene expression partially protects cells from VD-toxin induced programmed cell death (PCD).

Conclusion/Significance

Verticillium dahliae can trigger plant cells to die through induction of a PCD mechanism involved in pathogenesis. This paper provides a potential strategy for engineering broad-spectrum necrotrophic disease resistance in plants.     

单细胞RNA测序技术在病毒研究中的应用

单细胞RNA测序(single-cell RNA sequencing, scRNA-seq)技术已经成为不同领域中研究细胞异质性的有效工具。在病毒研究领域中,利用该技术分析病毒和细胞的转录组,可以在单细胞水平上检测病毒感染的动态变化,了解病毒与细胞间复杂的相互作用。本文简述了scRNA-seq技术,着重介绍病毒感染宿主细胞后scRNA-seq研究的最新进展,同时也描述了细胞周期、基因表达、细胞状态等细胞异质性对病毒感染过程的影响,以及病毒变异对其本身感染过程的影响。此外,本文还分析了scRNA-seq在研究病毒–宿主互作动态变化方面具有的独特优势,及其在病毒研究领域中广阔的应用前景,为揭示病毒的感染与致病机制、抗病毒靶标的开发等提供参考。