武夷山地衣表生和内生芽孢杆菌种群的多样性

摘要:【目的】分析武夷山地衣表生和内生芽孢杆菌种群的多样性。【方法】从武夷山自然保护区采集扁枝衣属(Evernia)、珊瑚枝属(Stereocaulon)、孔叶衣属(Menegazzia)等分属于7科9属的地衣样品9份,分离地衣表面附生(表生)和内生芽孢杆菌,并根据16S rRNA基因序列同源性对分离得到的芽孢杆菌进行种的鉴定。【结果】未从扁枝衣属、树花属(Ramalina)和茶渍属(Lecanora)的3个样品中分离出表生或内生芽孢杆菌,从另外6个样品中分离出芽孢杆菌34株,分别鉴定为芽孢杆菌属(Bacillus)、类芽孢杆菌属(Paenibacillus)、短芽孢杆菌属(Brevibacillus)、赖氨酸芽孢杆菌属(Lysinibacillus)和绿芽孢杆菌属(Viridiibacillus)的24个种。类芽孢杆菌属、芽孢杆菌属是这些地衣表生、内生芽孢杆菌的优势种群,分别占分离得到菌株的41.2%(14株)和35.3%(12株);短芽孢杆菌属、赖氨酸芽孢杆菌属和绿芽孢杆菌属为首次报道从地衣中分离获得。从不同的地衣样品分离的表生和内生芽孢杆菌的种类、数量存在差异。蜈蚣衣属(Physcia)表生的芽孢杆菌数量最多、可达3.85×106 cfu/g,而珊瑚枝属表面附生和内生芽孢杆菌的种类最多、共有12个种。分离到的芽孢杆菌大部分来自单独的一种地衣;台中类芽孢杆菌(Paenibacillus taichungensis)、土壤短芽孢杆菌(Brevibacillus agri)等4个种存在于2－3种地衣中;蕈状芽孢杆菌(Bacillus mycoides)分布最广,在蜈蚣衣属、珊瑚枝属等4种地衣中都有存在。【结论】武夷山地衣表生和内生芽孢杆菌存在种类和数量的多样性。     

Retrograde NGF Axonal Transport—Motor Coordination in the Unidirectional Motility Regime

We present a detailed motion analysis of retrograde nerve growth factor (NGF) endosomes in axons to show that mechanical tugs-of-war and intracellular motor regulation are complimentary features of the near-unidirectional endosome directionality. We used quantum dots to fluorescently label NGF and acquired trajectories of retrograde quantum-dot-NGF-endosomes with <20-nm accuracy at 32 Hz in microfluidic neuron cultures. Using a combination of transient motion analysis and Bayesian parsing, we partitioned the trajectories into sustained periods of retrograde (dynein-driven) motion, constrained pauses, and brief anterograde (kinesin-driven) reversals. The data shows many aspects of mechanical tugs-of-war and multiple-motor mechanics in NGF-endosome transport. However, we found that stochastic mechanical models based on in vitro parameters cannot simulate the experimental data, unless the microtubule-binding affinity of kinesins on the endosome is tuned down by 10 times. Specifically, the simulations suggest that the NGF-endosomes are driven on average by 5–6 active dyneins and 1–2 downregulated kinesins. This is also supported by the dynamics of endosomes detaching under load in axons, showcasing the cooperativity of multiple dyneins and the subdued activity of kinesins. We discuss the possible motor coordination mechanism consistent with motor regulation and tugs-of-war for future investigations.     

A green fluorescent protein-based whole-cell bioreporter for the detection of phenylacetic acid

Phenylacetic acid (PAA) is produced by many bacteria as an antifungal agent and also appears to be an environmentally toxic chemical. The object of this study was to detect PAA using Pseudomonas putida harboring a reporter plasmid that has a PAA-inducible promoter fused to a green fluorescent protein (GFP) gene. Pseudomonas putida KT2440 was used to construct a green fluorescent protein-based reporter fusion using the paaA promoter region to detect the presence of PAA. The reporter strain exhibited a high level of gfp expression in minimal medium containing PAA; however, the level of GFP expression diminished when glucose was added to the medium, whereas other carbon sources, such as succinate and pyruvate, showed no catabolic repression. Interestingly, overexpression of a paaF gene encoding PAACoA ligase minimized catabolic repression. The reporter strain could also successfully detect PAA produced by other PAA-producing bacteria. This GFP-based bioreporter provides a useful tool for detecting bacteria producing PAA.     

Molecular characterization of fprB (ferredoxin-NADP+ reductase) in Pseudomonas putida KT2440

The fpr gene, which encodes a ferredoxin-NADP+ reductase, is known to participate in the reversible redox reactions between NADP+/NADPH and electron carriers, such as ferredoxin or flavodoxin. The role of Fpr and its regulatory protein, FinR, in Pseudomonas putida KT2440 on the oxidative and osmotic stress responses has already been characterized [Lee at al. (2006). Biochem. Biophys. Res. Commun. 339, 1246-1254]. In the genome of P. putida KT2440, another Fpr homolog (FprB) has a 35.3% amino acid identity with Fpr. The fprB gene was cloned and expressed in Escherichia coli. The diaphorase activity assay was conducted using purified FprB to identify the function of FprB. In contrast to the fpr gene, the induction of fprB was not affected by oxidative stress agents, such as paraquat, menadione, H2O2 and t-butyl hydroperoxide. However, a higher level of fprB induction was observed under osmotic stress. Targeted disruption of fprB by homologous recombination resulted in a growth defect under high osmotic conditions. Recovery of oxidatively damaged aconitase activity was faster for the fprB mutant than for the fpr mutant, yet still slower than that for the wild type. Therefore, these data suggest that the catalytic function of FprB may have evolved to augment the function of Fpr in P. putida KT2440.     

Exploring the community phylogenetic structure along the slope aspect of subalpine meadows in the eastern Qinghai–Tibetan Plateau,China

Exploring the community assembly has been important for explaining the maintenance mechanisms of biodiversity and species coexistence, in that it is a central issue in community ecology. Here, we examined patterns of the community phylogenetic structure of the subalpine meadow plant community along the slope gradient in the Qinghai–Tibetan Plateau of China. We surveyed all species and constructed the phylogenetic tree of the plant community based on data from the Angiosperm Phylogeny Group III. We selected the net relative index (NRI) and evaluated the community phylogenetic structure along the five slope plants communities. We found that the phylogenetic structure varied from phylogenetic clustering to phylogenetic overdispersion with the slope aspect from north to south. In the north slope, the community phylogenetically cluster indicated that the limiting similarity played a leading role in the community assembly and the maintenance of biodiversity. Community phylogenetic overdispersion in the east, southeast, and south slopes indicated that habitat filtration was the driving force for community assembly. The NRI index of the northeast slope was close to zero, implying random dispersion. But it may be driven by the neutral process or limiting similarity, in that the community assembly process was the result of a combination of several ecological factors and thus required further study.     

Molecular mechanism of tumour necrosis factor alpha regulates hypocretin (orexin) expression,sleep and behaviour

Hypocretin 1 and hypocretin 2 (orexin A and B) regulate sleep, wakefulness and emotion. Tumour necrosis factor alpha (TNF‐α) is an important neuroinflammation mediator. Here, we examined the effects of TNF‐α treatment on hypocretin expression in vivo and behaviour in mice. TNF‐α decreased hypocretin 1 and hypocretin 2 expression in a dose‐dependent manner in cultured hypothalamic neurons. TNF‐α decreased mRNA stability of prepro‐hypocretin, the single precursor of hypocretin 1 and hypocretin 2. Mice challenged with TNF‐α demonstrated decreased expression of prepro‐hypocretin, hypocretin 1 and hypocretin 2 in hypothalamus. In response to TNF‐α, prepro‐hypocretin mRNA decay was increased in hypothalamus. TNF‐α neutralizing antibody restored the expression of prepro‐hypocretin, hypocretin 1 and hypocretin 2 in vivo in TNF‐α challenged mice, supporting hypocretin system can be impaired by increased TNF‐α through decreasing hypocretin expression. Repeated TNF‐α challenge induced muscle activity during rapid eye movement sleep and sleep fragmentation, but decreased learning, cognition and memory in mice. TNF‐α neutralizing antibody blocked the effects of TNF‐α; in contrast, hypocretin receptor antagonist enhanced the effects of TNF‐α. The data support that TNF‐α is involved in the regulation of hypocretin expression, sleep and cognition. The findings shed some lights on the role of neuroinflammation in neurodegenerative diseases including Alzheimer's disease and Parkinson's disease.     

Conductive Copper Niobate: Superior Li+‐Storage Capability and Novel Li+‐Transport Mechanism

Niobates with shear ReO₃ crystal structures are remarkably promising anode materials for Li⁺ batteries due to their large capacities, inherent safety, and high cycling stability. However, they generally suffer from limited rate capabilities rooted in their insufficient electronic and Li⁺ conductivities. Here, micrometer‐sized copper niobate (Cu₂Nb₃₄O₈₇) bulk as a new anode material having a high electronic conductivity of 2.1 × 10^?5 S cm^?1 and an impressive average Li⁺ diffusion coefficient of ≈3.5 × 10^?13 cm² s^?1 is exploited, which synergistically leads to an excellent rate capability (184 mAh g^?1 at 10 C) while remaining a large reversible capacity and superior cycling stability. Moreover, the fast Li⁺ transport pathways of grain boundary (micrometer scale) → lattice deformation area (nanometer scale) → (010) crystallographic plane (angstrom scale) are demonstrated in Cu₂Nb₃₄O₈₇. Therefore, these results could pave the way for practical application of Cu₂Nb₃₄O₈₇ in high‐performance Li⁺ batteries.     

Microbial Poly‐3‐Hydroxybutyrate (PHB) as a Feed Additive for Fishes and Piglets

The large‐scale use of petrochemical‐based plastics is damaging our environment. Discarded plastics are harmful to both marine and land animals, sometimes causing death when ingested. Biodegradable plastics have gained attentions from the public and the academia to reduce environmental burdens. Poly‐3‐hydroxybutyrate (PHB), the simplest and the best‐studied bioplastic member of the polyhydroxyalkanoate (PHA) family synthesized by many bacteria, has been studied as a feed additive for large yellow croaker fish and weaned piglets. The fish grow faster and gain more weight when 1% and 2% PHB is added as a feed additive, accompanied by increased survival rates. Weaned piglets are found to grow normally and showed no significant change in average daily weight gains, average daily feed intakes, feed efficiency, and organ developments when 0.5% PHB is added to the feed. It can therefore be concluded that biodegradable and biocompatible PHB is not harmful as a feed additive for marine large yellow croakers and sensitive weaned piglets. PHB therefore holds great promise as a plastic that combines biodegradability and biocompatibility with good tolerability as a feed supplement for animals.