Exploration of swapping enzymatic function between two proteins: A simulation study of chorismate mutase and isochorismate pyruvate lyase

The enzyme chorismate mutase EcCM from Escherichia coli catalyzes one of the few pericyclic reactions in biology, the transformation of chorismate to prephenate. The isochorismate pyruvate lyase PchB from Pseudomonas aeroginosa catalyzes another pericyclic reaction, the isochorismate to salicylate transformation. Interestingly, PchB possesses weak chorismate mutase activity as well thus being able to catalyze two distinct pericyclic reactions in a single active site. EcCM and PchB possess very similar folds, despite their low sequence identity. Using molecular dynamics simulations of four combinations of the two enzymes (EcCM and PchB) with the two substrates (chorismate and isochorismate) we show that the electrostatic field due to EcCM at atoms of chorismate favors the chorismate to prephenate transition and that, analogously, the electrostatic field due to PchB at atoms of isochorismate favors the isochorismate to salicylate transition. The largest differences between EcCM and PchB in electrostatic field strengths at atoms of the substrates are found to be due to residue side chains at distances between 0.6 and 0.8 nm from particular substrate atoms. Both enzymes tend to bring their non‐native substrate in the same conformation as their native substrate. EcCM and to a lower extent PchB fail in influencing the forces on and conformations of the substrate such as to favor the other chemical reaction (isochorismate pyruvate lyase activity for EcCM and chorismate mutase activity for PchB). These observations might explain the difficulty of engineering isochorismate pyruvate lyase activity in EcCM by solely mutating active site residues.     

芦丁的资源、药理及主要剂型研究进展

芦丁广泛存在于多种天然植物如槐米、苦荞麦、尤曼桉中,在医药、日化、食品等领域有广泛的应用,近年来已成为研究热点.芦丁具有多种生理功能,对人体各个器官均有保护作用,如清除自由基、促进成骨细胞的生长、降低血糖、抑制结肠癌和前列腺癌等.本文对芦丁的资源来源、药理作用和剂型进行了简述.     

低分子肝素的绿色荧光蛋白荧光标记及其活性检测

增强型绿色荧光蛋白（EGFP）是生物领域常用的标记物. 本实验首先利用高碘酸法活化低分子肝素（LMWH）,与EGFP连接得到LMWH EGFP.然后用Sephacryl S-200 HR对其进行初步分离,再用Sephadex G-10 HR进行脱盐纯化. 采用Sephacryl S 200 HR检测LMWH EGFP的纯度,为单一对称峰. 经检测,LMWH-EGFP具有良好的热稳定性和耐碱性. 通过荧光分光检测器检测LMWH-EGFP的λEx为488 nm,λEm为509 nm. 通过抗凝实验发现LMWH EGFP仍具有抗凝活性. 本实验建立了LWMH荧光标记的方法,为多糖的荧光标记提供了新的选择.     

High genetic diversity is not essential for successful introduction

Some introduced populations thrive and evolve despite the presumed loss of diversity at introduction. We aimed to quantify the amount of genetic diversity retained at introduction in species that have shown evidence of adaptation to their introduced environments. Samples were taken from native and introduced ranges of Arctotheca populifolia and Petrorhagia nanteuilii. Using microsatellite data, we identified the source for each introduction, estimated genetic diversity in native and introduced populations, and calculated the amount of diversity retained in introduced populations. These values were compared to those from a literature review of diversity in native, confamilial populations and to estimates of genetic diversity retained at introduction. Gene diversity in the native range of both species was significantly lower than for confamilials. We found that, on average, introduced populations showing evidence of adaptation to their new environments retained 81% of the genetic diversity from the native range. Introduced populations of P. nanteuilii had higher genetic diversity than found in the native source populations, whereas introduced populations of A. populifolia retained only 14% of its native diversity in one introduction and 1% in another. Our literature review has shown that most introductions demonstrating adaptive ability have lost diversity upon introduction. The two species studied here had exceptionally low native range genetic diversity. Further, the two introductions of A. populifolia represent the largest percentage loss of genetic diversity in a species showing evidence of substantial morphological change in the introduced range. While high genetic diversity may increase the likelihood of invasion success, the species examined here adapted to their new environments with very little neutral genetic diversity. This finding suggests that even introductions founded by small numbers of individuals have the potential to become invasive.     

Neogene origins and implied warmth tolerance of Amazon tree species

Tropical rain forest has been a persistent feature in South America for at least 55 million years. The future of the contemporary Amazon forest is uncertain, however, as the region is entering conditions with no past analogue, combining rapidly increasing air temperatures, high atmospheric carbon dioxide concentrations, possible extreme droughts, and extensive removal and modification by humans. Given the long‐term Cenozoic cooling trend, it is unknown whether Amazon forests can tolerate air temperature increases, with suggestions that lowland forests lack warm‐adapted taxa, leading to inevitable species losses. In response to this uncertainty, we posit a simple hypothesis: the older the age of a species prior to the Pleistocene, the warmer the climate it has previously survived, with Pliocene (2.6–5 Ma) and late‐Miocene (8–10 Ma) air temperature across Amazonia being similar to 2100 temperature projections under low and high carbon emission scenarios, respectively. Using comparative phylogeographic analyses, we show that 9 of 12 widespread Amazon tree species have Pliocene or earlier lineages (>2.6 Ma), with seven dating from the Miocene (>5.6 Ma) and three >8 Ma. The remarkably old age of these species suggest that Amazon forests passed through warmth similar to 2100 levels and that, in the absence of other major environmental changes, near‐term high temperature‐induced mass species extinction is unlikely.     

四种重楼属植物光合作用特征

Species of Paris (Trilliaceae) have often been used as medicinal plants. Because of excessive exploitation,in this regard the wild resource of Paris is almost exhausted. Some species of Paris were transplanted for use in photosynthesis research and for conservation purposes. In the present study, light and CO2 photosynthetic response curves were investigated in four Paris taxa: P.polyphylla var. yunnanensis and var. alba, P.mairei, and P.marmorata. Our results showed that P.marmorata had the highest maximum photosynthetic rate (Pmax; 8.6μmol·m-2·s-1), light saturation point (LSP; 827μmol·m-2·s-1), maximum electron transport rate (Jmax; 39.9mol·m-2·s-1), a relatively high maximum carboxylation rate (Vcmax; 28.9μmol·m-2·s-1) and carbon dioxide saturation point (Cisat; 726μmol·mol-1), but a lower light compensation point (LCP; 6.23μmol·m-2·s-1) and the lowest carbon dioxide compensation point (Г*; 20.7μmol·mol-1). This suggests that P.marmorata is well adapted to light and CO2; however it has a low ability to acclimate to environmental stress as indicated by low water use efficiency (WUE) in high light conditions. P.polyphylla var. yunnanensis had the highest light compensation point (LCP; 10.1μmol·m-2·s-1), carbon dioxide compensation point (Г*; 35.3μmol·mol-1), carbon dioxide saturation point (Cisat; 727μmol·mol-1), relatively high maximum photosynthetic rate (Pmax; 7.5μmol·m-2·s-1) and light saturation point (LSP; 728μmol·m-2·s-1), maximum light saturated electron transfer rate (Jmax; 37.7μmol·m-2·s-1), suggesting that it is suitable for conditions of higher light and CO2 concentration. This taxon can adapt to adverse conditions, as suggested by high WUE under increased CO2 concentration. In contrast, P.polyphylla var. alba exhibited a relatively lower apparent quantum yield (AQY; 0.037μmol·mol-1) and poorer growth performance than the other taxa. We suggest from our results that different light and water conditions are suitable for the growth of the different taxa. Photosynthesis assimilation efficiency and production can be increased by raising humidity for P.polyphylla var. yunnanensis and P.marmorata. To protect plants of P.polyphylla var. alba from strong sunshine, they should be shaded from March to mid June.     

A New Species of the Genus Leptolalax(Anura: Megophryidae) from Northern Thailand

A new species of the genus Leptolalax is described from Doi Saket, Chiang Mai Province, northern Thailand. The new species is distinguished from its congeners by the following combination of characters: 1) adult size large(male SVL 45.8-52.5 mm); 2) tympanum distinct; 3) skin of dorsal surface mostly smooth, with distinct tubercles on dorsal and lateral parts of the body; 4) ventrolateral glandular ridge distinct and complete; 5) lateral body and groin covered with dark brown or black spots; 6) throat, chest, and belly nearly immaculate white; and 7) bi-colored iris, upper onethird golden, and lower two-thirds gray. Molecular phylogenetic analysis of mitochondrial 16S rRNA further distinguishes it from its congeners for which sequences are available.     

Pseudomonas fluorescens,a potential bacterial antagonist to control plant diseases

Abstract

Fluorescent Pseudomonads belong to plant Growth Promoting Rhizobacteria (PGPR), the important group of bacteria that play a major role in the plant growth promotion, induced systemic resistance, biological control of pathogens etc. Many strains of Pseudomonas fluorescens are known to enhance plant growth promotion and reduce severity of various diseases. The efficacy of bacterial antagonists in controlling fungal diseases was often better as alone, and sometimes in combination with fungicides. The present review refers to occurrence, distribution, mechanism, growth requirements of P. fluorescens and diseases controlled by the bacterial antagonist in different agricultural and horticultural crops were discussed. The literature in this review helps in future research programmes that aim to promote P. fluorescens as a potential bio-pesticide for augmentative biological control of many diseases of agriculture and horticultural importance.     

Endophytic bacterium,Bacillus amyloliquefaciens,enhances ornamental hosta resistance to diseases and insect pests

A total of 84 bacterial endophytes were isolated from seeds of 6 cultivars of ornamental hostas, and they were identified to 5 species based on morphological characteristics and 16S rDNA sequence analysis. Among them, the strain ‘Blu-v2’, which was isolated from the seeds of cultivar ‘Blue Umbrella’ and identified to be Bacillus amyloliquefaciens, showed highest antifungal activity and capacity to deter feeding by Fall armyworms (Spodoptera fruigiperda). Lipopeptides in cultures of Blu-v2 were determined using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and its antifungal activities were verified. However, the lipopeptide preparation did not show toxicity to larvae of Fall armyworms. In a greenhouse experiment, Blu-v2 was inoculated into small plantlets of hosta (cultivar ‘Rainforest Sunrise’). The leaves of plants with bacteria (endophyte-infected?=?E+) and without bacteria (endophyte-free?=?E?) were used in seven-day feeding experiments employing fourth-instar larvae of Fall armyworms. We found that there was a significant decrease in the weights of larvae fed with E+ compared to E? plants; and the mortality rate of larvae fed with E? leaves was lower (3.33%) compared to that of larvae fed with E+ leaves (30%). Based on our studies, we suggest that endophytic B. amyloliquefaciens strain Blu-v2 has potential value as a biocontrol agent to reduce damage from fungal diseases and insect pests of hosta cultivars.     

Farm and management characteristics associated with boar taint

Pig farms in the Netherlands producing boars have different levels of boar taint prevalence, as assessed by sensory evaluation with the human nose at the slaughter line. With a questionnaire to 152 Dutch pig producers (response rate 59%), farm and management characteristics were identified that are potentially associated with farm-level boar taint prevalence. Lower farm-level boar taint prevalence was associated with a smaller group size, a smaller pen surface per boar, newer housing equipment, not practicing restricted feeding in the last period before delivery, a longer fasting period before slaughter, a higher stocking weight and a lower fraction of boars from purebred dam line sows or from Pietrain terminal boars. These characteristics can be used to develop farm-level intervention strategies to control boar taint. More research effort is needed to establish causal relationships.