利用超氧化物歧化酶增加棉铃虫病毒的感染性

Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV) has been used as a commercial pesticide for the control of cotton bollworm. However, some kinds of phenolase (particularly peroxidase) secreted by cotton glands, which generate free radicals, diminish the infectivity of baculoviruses significantly on cotton leaves. Superoxide dismutase (SOD) acts on the polyphenolic substance and eliminates free radicals. Results in this research indicated that SOD-additive HaSNPV has a significantly higher efficacy to kill the bollworm on cotton leaves than HaSNPV alone. Therefore,SOD can be a promising enhancer for the HaSNPV pesticide.     

Extraction of Magnetosome from Acidthiobacillus ferrooxidans

In physiological property and growing environment aspects, there are some similarities betweenAcidthiobacillus ferrooxidans（ATF） and magnetotactic bacteria. ATF shows magnetotacxis under the microscope. Themagnetosome can be extracted from ATF by using the extracting method of magnetosome from magnetotactic bacteria.The membrane of ATF was crushed by ultrasonic wave and the magnetosomes in the ATF which mainly contains Fethrough chemical detection were attracted by using magnetite. After purifing with sucrose density gradient centrifugationand washing by PBS, the magnetosomes can be seen clearly through a transmission electron microscope. The resultsindicate there are a small amount of intracellular magnetosomes which made the ATF be magnetotactic under the applied magnetic field. It is the first time to find that ATF was magnetotactic. The magnetotaxis of ATF can be utilizedand isolated by the magnetotactic properties to gain the high - performance ATF strains of mineral leaching, whichhave high activity and different magnetism.     

Antibacterial, antifungal and cytotoxic activity of terrestrial cyanobacterial strains from Serbia

Cyanobacteria are known to be a rich source of biologically active compounds some of which can have pharmaceutical importance. In this work we present the screening results of cyanobacterial strains for their antibacterial, antifungal, and cytotoxic activity. Cyanobacterial strains were isolated from various soil types in province of Vojvodina and Central Serbia, Republic of Serbia. The screening included 9 strains of Anabaena and 9 strains of Nostoc. Both, extracellular products (from the culture liquid) and cellular crude lipophilic extracts were tested against 13 bacterial strains and 8 fungal strains. Cytotoxic activity was tested against three human cell lines. Methanol extracts were prepared according to ?stensvik. Antibacterial and antifungal activities were determined measuring inhibition zone, 48 h after inoculation. The cytotoxic activity was determined by sulforhodamine B (SRB) colorimetric assay. Of all cyanobacterial strains tested, 52% showed some antifungal and 41% antibacterial activity. Two out of six tested strains possessed cytotoxic activity. The cytotoxic activity of Anabaena strain S12 was found both in culture liquid and crude cell extract. It occurred specifically between the 21st and 42nd day of cultivation against HeLa and MCF7 cells, but had no activity against cell line derived from a healthy tissue. A high percentage of the active strains among the tested strains justify the effort of screening cyanobacteria that are isolated from terrestrial environments. The most promising strains for the fur- ther study are Anabaena strain S12 which showed strong cytotoxic and antibacterial activity and Ana- baena strain S20 which produces a potent antifungal compound. The future work, besides further screening and chemical identification of the active compounds, should also include the development of culture techniques that would lead to more efficient production of biologically active compounds.     

Efficient derivation of extended pluripotent stem cells from NOD-scid II2rg-/-mice

Recently we have established a new culture condition enabling the derivation of extended pluripotent stem(EPS)cells,which,compared to conventional pluripotent stem cells,possess superior developmental potential and germline competence.However,it remains unclear whether this condition permits derivation of EPS cells from mouse strains that are refractory or non-permissive to pluripotent cell establishment.Here,we show that EPS cells can be robustly generated from non-permissive NOD-sc/d Il2rg 1 mice through de novo derivation from blastocysts.Furthermore,these cells can also be efficiently generated by chemical reprogramming from embryonic NOD-sc/d II2rg-/-fibroblasts.NOD-sc/d II2rg-/-EPS cells can be expanded for more than 20 passages with genomic stability and can be genetically modified through gene targeting.Notably,these cells contribute to both embryonic and extraembryonic lineages in vivo.More importantly,they can produce chimeras and integrate into the E13.5 genital ridge.Our study demonstrates the feasibility of generating EPS cells from refractory mouse strains,which could potentially be a general strategy for deriving mouse pluripotent cells.The generation of NOD-sc/d II2rg-/-Yaqin Du and Ting Wang contributed equally to this work.Electronic supplementary material The online version of this article(https://doi.org/10.1007/s13238-018-0558-z)contains supplementary material,which is available to authorized users.EPS cell lines permits sophisticated genetic modification in NOD-scid II2rg-/-mice,which may greatly advance the optimization of humanized mouse models for biomedical applications.     

Effects of Elevated Carbon Dioxide on the Growth and Foliar Chemistry of Transgenic Bt Cotton

A field study was carried out to quantify plant growth and the foliar chemistry of transgenic Bacillus thuringiensis （Bt） cotton （cv. GK-12） exposed to ambient CO2 and elevated （double-ambient） CO2 for different lengths of time （1, 2 and 3 months） in 2004 and 2005. The results indicated that CO2 levels significantly affected plant height, leaf area per plant and leaf chemistry of transgenic Bt cotton. Significantly, higher plant height and leaf area per plant were observed after cotton plants that were grown in elevated CO2 were compared with plants grown in ambient CO2 for 1, 2 and 3 months in the investigation. Simultaneously, significant interaction between CO2 level x investigating year was observed in leaf area per plant. Moreover, foliar total amino acids were increased by 14%, 13%, 11% and 12%, 14%, 10% in transgenic Bt cotton after exposed to elevated CO2 for 1, 2 or 3 months compared with ambient CO2 in 2004 and 2005, respectively. Condensed tannin occurrence increased by 17%, 11%, 9% in 2004 and 12%, 11%, 9% in 2005 in transgenic Bt cotton after being exposed to elevated CO2 for 1, 2 or 3 months compared with ambient CO2 for the same time. However, Bt toxin decreased by 3.0%, 2.9%, 3.1% and 2.4%, 2.5%, 2.9% in transgenic Bt cotton after exposed to elevated CO2 for 1, 2 or 3months compared with ambient CO2 for same time in 2004 and 2005, respectively. Furthermore, there was prominent interaction on the foliar total amino acids between the CO2 level and the time of cotton plant being exposed to elevated CO2. It is presumed that elevated CO2 can alter the plant growth and hence ultimately the phenotype allocation to foliar chemistical components of transgenic Bt cotton, which may in turn, affect the plant-herbivore interactions.     

Establishment of an efficient transformation protocol and its application in marine-derived Bacillus strain

Marine-derived Bacillus strains have been proved to be a very promising source for natural product leads.However,transformation of environmental strains is much more difficult than that of domesticated strains.Here,we report the development of an efficient and robust electroporation-based transformation system for marine-derived Bacillus marinus B-9987,which is a macrolactin antibiotics producer and a very promising biological control agent against fungal plant diseases.The transformation efficiency was greatly enhanced 103-fold by using unmethylated plasmid to bypass modification-restriction barrier,and using glycine betaine to protect cells from electrical damages during electroporation.Addition of HEPES and 2 mmol L?1MgCl2 further improved the efficiency by additional 2-fold,with a maximum value of 7.1×104 cfu/μg pHT3101.To demonstrate the feasibility and efficiency of the protocol,a green fluorescent protein reporter system was constructed;furthermore,phosphopantetheinyl transferase gene sfp,which is essential to the biosynthesis of polyketides and nonribosomal peptides,was overexpressed in B-9987,leading to increased production of macrolactin A by about 1.6-fold.In addition,this protocol is also applicable to marine-derived Bacillus licheniforms EI-34-6,indicating it could be a reference for other undomesticated Bacillus strains.To our knowledge,this is the first report regarding the transformation of marine-derived Bacillus strain.     

Isolation and characterization of tyrosinase produced by marine actinobacteria and its application in the removal of phenol from aqueous environment

The present study was focused on screening and characterization of tyrosinase enzyme produced by marine actinobacteria and its application in phenolic compounds removal from aqueous solution. A total of 20 strains were isolated from marine sediment sample and screened for tyrosinase production by using skimmed milk agar medium. Among 20 isolates, two isolates LK-4 and LK-20 showed zone of hydrolysis and these were taken for secondary screening by using tyrosiue agar medium. Based on the result of secondary screening LK-4 was selected for further analysis, such as tyrosinase assay, protein content and specific activity of the enzyme. The tyrosinase enzyme was produced in a SS medium and was partially purified by ammonium sulfate precipitation, dialysis and SDS PAGE. The isolate （LK-4） was identified as Streptomyces espinosus using 16S rRNA gene sequencing and named as ＂Streptomyces espinosus strain LK4 （KF806735）＂. The tyrosinase enzyme was immobilized in sodium alginate which was applied to remove phenolic compounds from water. The enzyme efficiently removed the phenolic compounds from aqueous solution within few hours which indicated that tyrosinasc enzyme produced by Streptomyces espinosus strain LK-4 can be potently used for the removal of phenol and phenolic compounds from wastewater in industries.     

Biomimetics of Campaniform Sensilla： Measuring Strain from the Deformation of Holes

We present a bio-inspired strategy for designing embedded strain sensors in space structures. In insects, the campaniform sensillum is a hole extending through the cuticle arranged such that its shape changes in response to loads. The shape change is rotated through 90° by the suspension of a bell-shaped cap whose deflection is detected by a cell beneath the cuticle. It can be sensitive to displacements of the order of 1 nm. The essential morphology, a hole formed in a plate of fibrous composite mate- rial, was modelled by Skordos et al. who showed that global deformation of the plate （which can be flat, curved or a tube） induces higher local deformation of the hole due to its locally higher compliance. Further developments reported here show that this approach can be applied to groups of holes relative to their orientation. , The morphology of the sensillum in insects suggests that greater sensitivity can be achieved by arranging several holes in a regular pattern; that if the hole is oval it can be ＂aimed＂ to sense specific strain directions; and that either by controlling the shape of the hole or its relationship with other holes it can have a tuned response to dynamic strains. We investigate space applications in which novel bio-inspired strain sensors could successfully be used.     

Horizontal transfer of genetic determinants for degradation of phenol between the bacteria living in plant and its rhizosphere

Phenol and other monocyclic aromatic compounds (MACs) are highly water-soluble and volatile pollutants that plants are unable to completely degrade. Endophytic bacteria with MAC-degrading ability will facilitate phytoremediation, beneficial to plant survival in contaminated soil. Endophytic bacteria, strains FX1-FX3, and rhizosphere bacteria, strains FX0, FX4, and FX5, were isolated from the root tissue of a corn plant (Zea mays) and the corn rhizosphere near a chemical plant, respectively. The strains FX1-FX5 were able to grow on phenol and reduce phenol concentration, but the strain FX0 was unable to. The strains FX1, FX3, and FX4 were classified as Pseudomonas fluorescens and FX0, FX2, and FX5 as Burkholderia cepacia. The plasmids isolated from the strains FX1-FX5 were found to possess similar traits and to be loaded with a gene encoding the catechol 2, 3-dioxygenase (C23O), a key enzyme in the phenol degradation pathway. Alignment and phylogenetic analysis inferred that in situ horizontal transfer of the C23O gene might have occurred. The horizontal transfer of the C23O gene between endophytic and rhizosphere bacteria was proved by using conjugal matings experiment, in which the transconjugants were found to acquire the plasmid with the C23O gene, able to grow on phenol and degrade phenol.     

可改良棉粕蛋白的菌株选育及其发酵

棉粕蛋白质含量高达38％~50％,然而作为饲料其蛋白质品质较差,饲料利用率较低。为提高其蛋白质利用率,从自然界筛选出了能够高效降解棉粕大分子蛋白质的优良菌株4株,经生化与分子生物学试验鉴定为Bacillus subtilis(2株,编号为H1和H7),Bacillus cereus (1株,编号为H9)和Aspergillus niger (1株,编号为P1)。通过单菌及组合发酵实验比较发现:最适宜菌种组合为B. subtilis(H1)和A. niger (P1)。两菌株组合发酵后,棉粕TCA-NSI(三氯乙酸氮溶指数)提高了25.34%,小分子多肽含量提高了11%,游离必需氨基酸提高了24%,蛋白质的体外消化率由44.56%提高到了78.61%,明显高于其他菌种组合,研究结果表明其组合发酵可有效改良棉粕蛋白品质,显著提高棉粕蛋白的饲用价值。     

Effect of culture medium composition on peroxidase biosynthesis by the fungus Pleurotus ostreatus UzBI-I105

Production of extracellular peroxidase during submerged cultivation of the xylotrophic bazidiomycete Pleurotus ostreatus UZBI-I105 in nutrient media with lignocellulosic wastes, exhausted cottonseed oil cake, cotton stalks, rice husks, or ambary hemp was studied. The enzyme production increased threefold to fivefold in the presence of exhausted cottonseed oil cake extract in the nutrient medium. The dynamics of peroxidase production in various media was investigated.     

复合微生物菌剂发酵在棉籽饼脱毒中的应用

以棉籽饼为原料,采用黑曲霉(Aspergillus niger)、热带假丝酵母(Candida tropicalis)、短小芽胞杆菌(Bacillus pumilus)、保加利亚乳杆菌(Lactobacillus bulgaricus)4种微生物的复合菌剂作为棉籽饼固体发酵的主要菌种。通过测定发酵液中糖含量的变化,考察4种菌株两两之间的关系,测定棉籽饼脱毒率和发酵前后蛋白含量。结果显示:4种菌株可以互惠共生;当黑曲霉、热带假丝酵母、短小芽胞杆菌、保加利亚乳杆菌的接种比例为2:2:1:1,固体发酵后蛋白含量为37.71%,比发酵前提高了2.74%,是良好的蛋白饲料。棉籽饼脱毒率为84.63%,棉酚含量为116.20mg/kg,达到饲用标准。     

The Effect of Nutrient Medium Composition on Peroxidase Biosynthesis by Bazidiomycete Pleurotus ostreatus,strain UzBI-I105

Production of extracellular peroxidase was studied during the submerged cultivation of xylotrophic bazidiomycete Pleurotus ostreatus, strain UZBI-I105 in nutrient media with lignocellulosic wastes, exhausted cottonseed oil cake, cotton stalks, rice husks, or ambary hemp. Enzyme production was enhanced 3–5 times in the presence of exhausted cottonseed oil cake extract in the nutrient medium. The dynamics of peroxidase production in various media was investigated.     

Biodegradation of phenol by a filamentous fungi isolated from industrial effluents—identification and degradation potential

Thirty filamentous fungal strains were isolated from effluents of a stainless steel industry (Minas Gerais, Brazil) and tested for phenol tolerance. Fifteen strains of the genera Fusarium sp., Aspergillus sp., Penicillium sp. and Graphium sp. tolerants up to 10 mM of phenol were selected and tested for their ability to degrade phenol. Phenol degradation was a function of strain, time of incubation and initial phenol concentration. FIB4, LEA5 and AE2 strains of Graphium sp. and FE11 of Fusarium sp. presented the highest percentage phenol degradation, with 75% degradation of 10 mM phenol in 168 h for FIB4. A higher starting cell density of Graphium sp. FIB4 lead to a decrease in the time needed for full phenol degradation and increased the phenol degradation rate. All strains exhibited activity of catechol 1,2-dioxygenase and phenol hydroxylase in free cell extracts obtained from cells grown on phenol, suggesting that catechol was oxidized by the ortho type of ring fission. These data reported demonstrate the prospect after the application of filamentous fungal strains in protecting the environment from phenol pollution.     

Phenol degradation by Paenibacillus thiaminolyticus and Bacillus cereus in axenic and mixed conditions

In this study, seven aerobic bacterial strains were screened for phenol tolerance at different concentration of phenol. Bacterial strains were unable to utilize phenol in absence of glucose, indicated the phenomenon of co-metabolism. Among the seven isolated bacterial strains, only ITRC BK-4 and ITRC BK-7 found potential and identified as Paenibacillus thiaminolyticus (DQ435022) and Bacillus cereus (DQ435023), respectively. Phenol degradation was monitored routinely with spectrophotometer and further confirmed by HPLC analysis. ITRC BK-4, ITRC BK-7 and mixed culture degrade 700 ppm phenol up to 51.72, 70.00 and 84.57% respectively in mineral salt medium (MSM) at temperature 37 ± 1°C, pH 7.5 ± 0.2, 120 rpm in presence of 1% glucose (w/v) within 144 h incubation. The mix culture was found more potential for phenol degradation compared to axenic strains. Hence, the axenic and mixed strains of these bacteria would be useful for the removal/mineralization of phenol from industrial waste waters.     

Phenol degradation by immobilized cold-adapted yeast strains of Cryptococcus terreus and Rhodotorula creatinivora

Three strains were isolated from hydrocarbon-polluted alpine habitats and were representatives of Cryptococcus terreus (strain PB4) and Rhodotorula creatinivora (strains PB7, PB12). All three strains synthesized and accumulated glycogen (both acid- and alkali-soluble) and trehalose during growth in complex medium containing glucose as carbon source and in minimal salt medium (MSM) with phenol as sole carbon and energy source. C. terreus strain PB4 showed a lower total accumulation level of storage compounds and a lower extracellular polysaccharides (EPS) production than the two R. creatinivora strains, PB7 and PB12. Biofilm formation and phenol degradation by yeast strains attached to solid carriers of zeolite or filter sand were studied at 10°C. Phenol degradation by immobilized yeast strains was always higher on zeolite compared with filter sand under normal osmotic growth conditions. The transfer of cells immobilized on both solid supports to a high osmotic environment decreased phenol degradation activity by all strains. However, both R. creatinivora PB7 and PB12 strains maintained higher ability to degrade phenol compared with C. terreus strain PB4, which almost completely lost its phenol degradation activity. Moreover, R. creatinivora strain PB7 showed the highest ability to form biofilm on both carriers under high osmotic conditions of cultivation.     

苯酚的生物降解基因组成及其调控机制

苯酚的生物降解受多组分的降解基因控制,形成邻位和间位等不同的代谢途径.结合部分实验结果,介绍了邻位和间位代谢系统的基因组成及其作用机制等方面的研究进展,讨论了调节基因在苯酚降解菌中的作用和不同因素对苯酚降解的影响.     

高效棉酚降解枯草芽孢杆菌M-4菌株的常压室温等离子体诱变及发酵工艺优化

【背景】棉粕中游离棉酚的存在制约了棉粕作为饲料蛋白源的利用,棉酚的微生物降解问题成为研究热点。本实验室前期发现枯草芽孢杆菌(Bacillus subtilis) M-4菌株具有较强的降解棉酚能力,而且已经应用于棉粕脱毒工业。【目的】进一步提高枯草芽孢杆菌M-4菌株降解棉粕中棉酚的能力,扩大棉粕在养殖业的应用领域。【方法】采用常压室温等离子体诱变(atmospheric and room temperature plasma,ARTP)技术对菌株M-4进行诱变,以液体培养条件下的棉酚降解率为初筛指标,筛选获得正向突变株。以棉粕固体发酵条件下的棉酚降解率为复筛指标,测定初筛获得的正向突变株实际降解棉粕中游离棉酚的能力。以棉酚残存量和棉酚降解率为检测指标,采用单因素试验优化固体条件下棉粕发酵条件,获得突变株发酵棉粕的最适工艺参数。【结果】初筛获得正向突变株19株。复筛得到一株高效的突变株MY-4-17,在棉粕固体发酵条件下其棉酚降解率高达97.15%,比菌株M-4的棉酚降解率提高了2.55%。经过5次传代培养,突变株MY-4-17的遗传稳定性良好。突变株MY-4-17发酵棉粕的最适工艺参数为...