Nutrient-split feeding strategy for dialysis cultivation of Escherichia coli

Reduction in nutrient loss during dialysis cultivation of Escherichia coli on a glycerol medium was investigated. A dialysis reactor with an inner fermentation and an outer dialysis chamber was used. Aerobic condition was maintained by limiting the glycerol feed rate to an optimum value which was estimated from the oxygen requirements for glycerol oxidation and oxygen transfer capacity of the reactor. High reduction in nutrient loss was achieved by using water as the dialyzing fluid. However, osmotic movement of water from the dialysis to the fermentation chamber was observed, and the final cell concentration was low. With a nutrient-split feeding strategy (feeding glycerol directly to the fermentation chamber and dialyzing with salt solution), glycerol loss was small, there was no osmotic flux of water to the fermentation chamber, and the cell concentration was high. Both glycerol and salt loss could be avoided, and a cell concentration of 170 g/L was obtained when the dialysis process was substituted by addition of XAD adsorbents to the dialysis chamber. Application of this nutrient-split feeding strategy to cell cultivation in a stirred tank reactor, coupled with dialysis in external dialyzer modules, resulted in low cell concentrations. (c) 1993 Wiley & Sons, Inc.     

Antimicrobial activity of lipoprotein particles containing apolipoprotein Al

Human plasmain vitro inhibits the growth of coagulase negative staphylococci,S. epidermidis, which may be pathogenic in the immunocompromised host. To determine the antimicrobial components, serum was fractionated by column chromatography, which revealed that elution areas where lipoproteins can be yielded had high antimicrobial activity againstS. epidermidis. Therefore, lipoprotein fractions, including very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL), were separated by ultracentrifugation and incubated withS. epidermidis. All 3 lipoprotein fractions suppressed bacterial growth within the first 3 h but VLDL enhanced bacterial growth after 9 h of incubation compared with the control. HDL, however, inhibited bacterial growth throughout 21 h of incubation.To confirm these results, serum from healthy volunteers was separated by ion exchange column chromatography and again by HPLC to purify the antimicrobial fraction. In the protein analysis with gradient polyacrylamide-SDS gel, apolipoprotein Al (apo Al), which is a major apolipoprotein of HDL, was detected in the antimicrobial fraction. Therefore, this fraction was loaded onto an immunoaffinity column coupled with the anti-apo Al monoclonal antibody (Mab). Unbound fraction had no antimicrobial activity, but anti-S. epidermidis activity was recovered from the bound fraction which consisted mainly of apo Al, All and apo C in protein composition.These results indicated that the antimicrobial activity was associated with the apo Al-containing lipoprotein particles (HDL). This property of HDL may directly affect bacterial growth and promote the self-defense mechanisms of normal and immunocompromised individuals.     

A novel molecular fingerprint probe based on the endogenous avian retroviral element (EAV) of chickens

We have developed a novel molecular probe that is useful for DNA fingerprint analysis in chickens. The probe is based on the middle-repetitive, chicken endogenous retroviral (EAV) element. It consists of 1503 bp of the 3′ portion of the EAV element, extending from the downstream end of the envelope gene to the beginning of the downstream long terminal repeat (LTR). Unlike other probes that are currently in use for fingerprint analysis with chicken DNA, the EAV-based probe works well at normal levels of stringency, and with standard hybridization buffers. Digestion of chicken genomic DNA with a variety of restriction enzymes routinely yields up to 30 resolvable bands per bird in the 500 bp to 20kbp range. In order to test the efficacy of the EAV-based fingerprint probe, we have used it to estimate the degree of inbreeding in the inbred WG strain of White Leghorns. We find that the estimates derived with the EAV probe are very similar to those reported previously for the WG strain. These results suggest that molecular probes based on endogenous retroviruses and other middle-repetitive DNA elements should be useful for fingerprint analysis in chickens, and in vertebrates in general.     

Identification of Products from Oxidation of Uric Acid Induced by Hydroxyl Radicals

The aim of the present study was to separate and characterise products formed by oxidation of uric acid by hydroxyl radicals with a view to probing for these products in vivo in clinical contexts. Aerated solutions of 200 μM uric acid, or its oxidation products, allantoin or parabanic acid, were exposed to gamma radiolysis, (52.0 Gy/min), as a source of HO- radicals, at pH 3.4 and 7.4. Aliquots were taken every 5 minutes for 20 minutes and oxidation products were separated by HPLC and analysed with a diode array detector. Identities of oxidation products were confirmed on the basis of similarity of retention times and absorbance spectra and peak purity parameters of known standards. Hydroperoxides were measured by tri-iodide formation in the 20 minute sample. Exposure of uric acid to such HO fluxes produced a net loss of the parent compound with formation of a complex mixture of products with allantoin and parabanic acid being the predominant products at pH 3.4. The rate of uric acid degradation at physiological pH was slower and the distribution of oxidation products was different. A small but significant amount of uric acid hydroperoxide was detected at both pHs. A mechanism for uric acid oxidation under these conditions is presented.     

高砷含水层参与腐殖酸-铁矿物转化的关键微生物群落及其对砷迁移转化的影响

【目的】探究不同腐殖酸浓度下参与含砷水铁矿转化的微生物类群组成和丰度变化及对砷释放的影响,预测原位高砷含水层中功能微生物群参与有机质—含砷铁矿物转化过程对砷转化释放的作用。【方法】对河套平原高砷地下水和同深度高砷沉积物中的铁还原功能群落进行富集培养,构建室内厌氧微宇宙体系,将富集菌群分别加入到实验室条件下合成的不同浓度腐殖酸(0、1.5、7、14 mg C/L)-含砷水铁矿体系中,通过体系中砷、铁形态及浓度的变化分析,结合高通量测序技术、X-射线衍射(X-ray diffractometer, XRD),探究不同条件下砷的释放固定和群落的演替。【结果】高砷地下水组(G组)和沉积物组(S组)富集得到的铁还原功能群落具有明显差异,G组中以Aeromonadaceae为特殊优势菌群,而S组中以Shewanellaceae为特殊优势菌群。微宇宙实验结果显示,S组的铁还原量相对较高且速率较快;G组与S组中液相砷形态存在明显差异,整个培养期内G组均以As(Ⅴ)为主,而S组中前期以As(Ⅴ)为主,当反应到达20 d时液相As(Ⅲ)高达3.4μmol/L,推测此时具有砷还原功能的群落占优势地位。当反应...     

Diverse flowering responses subjecting to ambient high temperature in soybean under short-day conditions

Flowering time is one of important agronomic traits determining the crop yield and affected by high temperature. When facing high ambient temperature, plants often initiate early flowering as an adaptive strategy to escape the stress and ensure successful reproduction. However, here we find opposing ways in the short-day crop soybean to respond to different levels of high temperatures, in which flowering accelerates when temperature changes from 25 to 30 °C, but delays when temperature reaches 35 °C under short day. phyA-E1, possibly photoperiodic pathway, is crucial for 35 °C-mediated late flowering, however, does not contribute to promoting flowering at 30 °C. 30 °C-induced up-regulation of FT2a and FT5a leads to early flowering, independent of E1. Therefore, distinct responsive mechanisms are adopted by soybean when facing different levels of high temperatures for successful flowering and reproduction.     

High-temperature stress in crops: male sterility,yield loss and potential remedy approaches

Global food security is one of the utmost essential challenges in the 21st century in providing enough food for the growing population while coping with the already stressed environment. High temperature (HT) is one of the main factors affecting plant growth, development and reproduction and causes male sterility in plants. In male reproductive tissues, metabolic changes induced by HT involve carbohydrates, lipids, hormones, epigenetics and reactive oxygen species, leading to male sterility and ultimately reducing yield. Understanding the mechanism and genes involved in these pathways during the HT stress response will provide a new path to improve crops by using molecular breeding and biotechnological approaches. Moreover, this review provides insight into male sterility and integrates this with suggested strategies to enhance crop tolerance under HT stress conditions at the reproductive stage.     

古尔班通古特沙漠藻类结皮中微生物群落空间分异特征

古尔班通古特沙漠短命和旱生植物在沙漠东西走向上形成明显的种群变化梯度,那么沙漠藻类结皮土壤中微生物是否也具有类似地空间分布特征?为研究该科学问题,采用高通量测序技术,探究沙漠东西走向上藻类结皮土壤中细菌和真菌群落结构分布规律。在古尔班通古特沙漠西部、中部和东部区域分别采集藻类结皮土样,进行土壤理化因子分析及细菌和真菌扩增子测序,对比不同区域土壤理化性质和微生物群落空间分异特征,分析微生物多样性与环境因子相关性,最后根据微生物物种种类对其功能进行预测分析。结果表明：藻类结皮土壤氮素、全磷、全钾、pH以及土壤化学计量比碳磷比(C∶P)和氮磷比(N∶P)在沙漠不同区域存在显著性差异,其余指标空间尺度上变化梯度较小。微生物α多样性结果显示3个区域藻类结皮土壤中细菌群落丰富度和多样性存在显著性差异,东部与西部区域产生明显分化;而真菌α多样性指数无显著性差异,群落结构较为均匀,未产生分化。β多样性结果显示细菌群落在西部与东部藻类结皮间存在显著性差异,沙漠中部为沙漠西部和东部的过渡带。门水平细菌优势菌群为蓝藻门、变形菌门和拟杆菌门;真菌为子囊菌门和担子菌门。属水平上,丰度前30个属在沙漠不同区域表现...     

甜樱桃对高温胁迫的生理响应

为探讨甜樱桃对高温胁迫的响应机制,以1年生的甜樱桃嫁接盆栽苗为试材,研究了自然高温处理及胁迫后恢复对其叶片生理和超显微结构的影响。结果表明：经连续3d日最高气温均值达57.7℃胁迫后,甜樱桃叶片过氧化氢酶(CAT)和抗坏血酸过氧化物酶(APX)活性、超氧阴离子(O₂^·-)产生速率、非光化学淬灭系数(qN)显著增加,渗透调节物质可溶性蛋白、可溶性糖及脯氨酸含量显著提高,谷胱甘肽还原酶(GR)活性、叶绿素含量、PSII最大光化学效率(Fv/Fm)和电子传递速率(ETR)显著下降,超氧化物歧化酶(SOD)和过氧化物酶(POD)活性及光化学淬灭系数(qP)变化不显著。同时对叶片显超微结构观察发现,胁迫引起叶绿体变性,基质内囊体发生扭曲,部分基粒类囊体片层消失,出现大量巨型淀粉粒及嗜锇颗粒。恢复2d后SOD、POD、CAT、APX活性以及可溶性糖、可溶性蛋白、叶绿素总含量均显著下降,而GR活性和脯氨酸含量显著升高。以上表明,连续3d自然最高气温达57.7℃对甜樱桃叶片抗氧化系统和光合特性等产生了显著影响,叶绿体超显微结构受到明显破坏,对叶片造成了不可逆...     

Antibiotic production by strains of Gliocladium virens and its relation to the biocontrol of cotton seedling diseases

Strains of the fungal antagonist Gliocladium virens were separated into two distinct groups on the basis of secondary metabolite production in vitro. Strains of the ‘P’ group produced the antibiotics gliovirin and heptelidic acid but not the antibiotic gliotoxin and its companion, dimethylgliotoxin. Strains of the ‘Q’ group produced gliotoxin and dimethylgliotoxin but not gliovirin or heptelidic acid. Strains from both groups produced the antibiotic viridin and phytotoxin viridiol. Gliovirin was very inhibitory to Pythium ultimum but had no activity against Rhizoctonia solani, and strains that produce it were more effective seed treatment biocontrol agents of disease incited by P. ultimum. Conversely, gliotoxin was more active against R. solani than against P. ultimum, and strains that produced it were more effective seed treatments for controlling disease incited by R. solani. These results indicate that the antibiotic profiles of strains should be considered when screening strains for biocontrol efficacy, and that it may be necessary to treat seeds with a combination of strains in order to broaden the disease control spectrum.