Cherry trees (Cerasus pseudocerasus (Lindl.) Loudon) in Bai Lu Yuan, Shaanxi Province, China, are susceptible to a serious disease that causes substantial economic losses. The pathogens causing this disease, designated black spot disease, are currently unknown. To identify the relationship between microbial community structure and the status of plant health, we obtained samples from the rhizosphere and tissues of both healthy cherry trees and those with symptoms of black spot disease. We used Illumina sequencing of 16S rRNA gene and internal transcribed spacer (ITS) amplicons to examine the bacterial and fungal communities in these samples. An alpha diversity analysis showed that there were no significant differences between diseased and healthy soils, while fungal diversity in diseased tissue was significantly higher than in healthy tissue (P < 0.01), suggesting that an imbalance in fungal flora may be related to black spot disease. Taxonomic classification and linear discriminant analysis effect size (LEfSe) indicated an increase in the relative abundance of some bacterial genera (Pseudomonas, Burkholderia, Sphingobium, Erwinia, Sphingomonas and Bradyrhizobium) and fungal genera (Phoma, Penicillium, Fusarium, Cystobasidium, Aureobasidium and Nakazawaea) in the diseased tissues (p < 0.05). Among these, the increased relative abundance of the bacteria Pseudomonas and Erwinia and fungi Penicillium, Phoma and Fusarium may be associated with blackspot of cherry tree. The bacterial genera Burkholderia, Sphingobium, Sphingomomas and Bradyrhizobium are probably antagonists of the pathogen(s) of black spot. These findings are an initial step in the identification of potential candidates for the biological control of the disease.

To obtain efficient halotolerant biosurfactant-producing bacteria,a halotolerant biosurfactant-produ-cing bacteria K1 was isolated from oil-water mixture in high salinity oil field by enrichment cultivation and oil-spreading diameter of culture supernatant. According to morphological,physiochemical characteristics and 16SrDNA sequence,strain K1 was identified as Klebsiella pneumonia. The biosurfactant produced by K1 was identified as Glycolipid by thin layer chromatography and infrared spectroscopy. The emulsifying activity,tem-perature and pH adaptability of the biosurfactant in highly mineralized water was studied. Experimental results are as follows:The biosurfactant can maintain a good emulsifying activity,and the emulsifying rate of diesel oil by the biosurfactant was 59. 5％;it has a good temperature stability and can tolerate the high temperature of 90 ℃;its activity is stronger when the pH between 6. 0 ～ 11. 0,and peracid greatly reduces its activity. The results show that the biosurfactant producted by strian K1 improves oil recovery in high mineralization oil field and it has a promising potential in bioremediation of crude oil contaminated environments.

在畜禽传染病的免疫预防实践中,经常会遇到免疫效果不好,保护率不高的情况[4],给养殖业免疫带来极大困惑,造成较大经济损失.实际上,要想动物免疫好,除了疫苗质量达到有效、安全、稳定和疫苗毒株匹配,疫苗运输保存科学规范,免疫程序(时间、剂量、频率)和免疫操作(严谨、认真、规范)这些基本条件外,还有更重要的是被免动物是否具有应答能力和感染是否处于临界发病状态以下.西安泰乐星生物科技有限公司生产的"增益素",是从T33#α-溶血性链球菌(泰乐星公司独家研发,荣获6项国家发明专利)代谢产物中提取的糖肽类物质,为一种新型高效免疫调节剂.为验证"增益素"的免疫调节效果,我们进行了"增益素"对猪伪狂犬疫苗免疫增效的试验,现报告如下.

The methods of traditional microbiology, molecular biology and biochemistry were combined to study the differences of rhizosphere soil physical and chemical properties, soil enzyme activities and the culturable microorganism diversities between healthy and diseased cherry trees in the White Deer Prairie Shaanxi for the prevention and control of 'Black tumor' disease. The results showed that the contents of total phosphorus, available potassium, available phosphorus and available nitrogen of healthy rhizosphere soil were significantly higher than those of diseased one, organic carbon content of healthy rhizosphere soil was significantly lower than that of diseased one. Total nitrogen, total potassium contents and pH had no significant difference between healthy and diseased rhizosphere soils. The activities of protease, urease, invertase and PPO enzymes of healthy rhizosphere soil were much higher than those of diseased one, but the activities of DHA and CAT enzymes of healthy rhizosphere soil were lower. The population of culturable bacteria and actinomyces of healthy rhizosphere soil were significantly more than diseased one, particularly of dominant bacteria, the mean population of streptomyces of healthy rhizosphere soil were lower than those of diseased one but without significant difference. The decrease of the number of culturable bacteria and actinomycetes, the imbalance proportion of dominant bacteria in the rhizosphere soil, the decrease of available fertilizer and enzyme activities may be the causes of Black tumor disease.

以十红滩砂岩型铀矿床中不同矿带的岩石样为研究对象,利用传统微生物学研究方法对矿石样中可培养假单胞菌进行富集分离、鉴定,并结合变性梯度凝胶电泳(DGGE)技术,对各矿带中赋存的假单胞菌的类群分布特征进行探究,结果表明经生理生化实验及16S rDNA基因序列鉴定十红滩铀矿中分布的可培养的假单胞菌主要为Pseudomonas stutzeri,Pseudomonas putida.DGGE显示Pseudomonas stutzeri是该矿床中的优势假单胞菌.

从西洋参种植土壤中分离获得一株具有抗真菌活性的细菌YA-3,杯碟法测定表明,该拮抗菌对13种植物病原真菌均有不同程度的抑制作用,其中对番茄早疫病抑制率达82.2％;YA-3培养滤液可导致番茄早疫病病原菌菌丝畸变、枯萎或者断裂,阻碍菌丝生长,并能够抑制其孢子萌发,抑制率达92.3％以上;利福平抗性标记试验表明,YA-3在番茄叶片上有很强的定殖能力,第26 d仍能检测到该菌;经盆栽试验结果表明,YA-3发酵液可使番茄叶片发病率显著降低,防治效果可达到75.52％;根据其形态、生理生化特性及16S rDNA同源性分析结果,鉴定为枯草芽孢杆菌(Bacillus subtilis).

从污泥中分离获得了一株可直接以蔗糖为基质发酵合成聚合度高、韧性强的聚羟基烷酸酯(PHA)的细菌U-3,经鉴定其为产碱假单胞菌(Pseudomonas alcaligenes).U-3菌株以蔗糖为碳源,蛋白胨、酵母膏为基质,摇瓶发酵培养44 h细胞生长量达19.58 g/L,PHA聚合物产量为10.82 g/L.利用傅里叶变换红外光谱、核磁共振波谱、乌氏黏度法、差示扫描量热、电子万能材料试验机分析研究了U-3菌合成PHA聚合物的结构、热学性能及力学性能.结果表明,该聚合物为聚-β-羟基壬酸酯,其粘均相对分子质量为1.06×106;弹性模量2.1381MPa,断裂伸长率282.9061％,拉伸应力2.3594MPa,位移79.21371mm;熔点37.91℃,分解温度为45.43℃,结晶度为5.18％,该聚合物膜具有良好的力学性能和热塑性,是一种新型的PHA原料资源.

A newly bacterial strain w-14,efficiently transforming cornstalk to hydrogen,had been isolated from uranium ore samples with strictly anaerobic culturing method.It was finally nomenclatured as Clostridium butyricum on the basis of series of standard identification methods.The influences of glycolysis temperature,glycolysis time,glycolysis pH,bran koji concentration of Trichoderma koningii and Aspergillus bran koji e.t.c on the the substrate saccharification efficiency and hydrogen production by the strain w-14 were investigated.Two periods of fermentation were used to discuss the effect of glycolysis (glycolysis temperature,glycolysis time,glycolysis pH,bran koji concentration of Trichoderma koningii and Aspergillus bran koji) on the substrate saccharification efficiency and hydrogen production by the strain w-14.At the optimum of variables combination,such as 20 g/L of bran koji,an initial pH of 5.0,incubating at 45 ℃ for 84 h by the Two stages of fermentation technology,the maximum reducing sugar content (402 mg/g-cs) and cumulative H2 yield of 99.7 mL/g-cs were obtained.Orthogonal design were used to optimize bio-hydrogen production from cornstalk glycolytic substrates by submerged fermentation.The maximum cumulative H2 yield of 140.24 mL/g-cs,increased by 41.38％,was achieved at the optimized hydrogen production conditions from the assistance of orthogonal design.Furthermore,it demonstrated that maximum cumulative H2 and the hydrogen content could reach 140.24 mL/g-cs,55.63％ respectively in 5 L batch fermentation tank.

Study different solvent extracts of the four medicinal lichens' antimicrobial activity. Seven kinds of intestinal pathogenic bacteria are used to evaluate four kinds of lichen extracts of different solvents' antimicrobial activity and identify their antimicrobial activity sites by LC-ESI-MS /MS. Methanol extracts' antimicrobial activity of Thamnolia subliformis is stronger in Bacillus cereus and pneumobacillus. Methanol extracts' antimicrobial activity of Lethariella cladonioides is stronger in Klebsiella pneumoniae and Bacillus coli. Methanol extracts' antimicrobial activity of Umbilicaria tornata is all stonger in seven tested microbe. Methanol extracts' antimicrobial activity of Umbilicaria hypococcinea is stronger in Waxy staphylococcus aureus and bacillus cereus. The four kinds of lichens' methanol extract are all usnic acid. The result showed that methanol extracts' antimicrobial activity of the four kinds of lichens is the strongest,and usnic acid plays a main role in antimicrobial activity. The study of lichens resource development from Tai Bai Mountain provides a variety of information.

Context: Podophyllotoxin, a pharmaceutically important bioactive compound of Podophyllum sps. (Berberidaceae), is in great demand worldwide as an anticancer and antivirus drug precursor. However, the source of podophyllotoxin is very limited due to the endangered status of the Podophyllum plant.Objective: The aim of this study was to isolate podophyllotoxin-producing endophytic fungi from Sinopodophyllum hexandrum (Royle) Ying (1979) (Berberidaceae) plants of the Taibai Mountains of China in order to obtain bioactive compounds.Materials and methods: The strains producing kaempferol and podophyllotoxin were screened by thin-layer chromatography (TLC) analysis. The presence of kaempferol and podophyllotoxin in extracts of these strains was further confirmed by high-performance liquid chromatography (HPLC) and nuclear magnetic resonance (NMR) analyses.Results: Among six endophytic fungi isolated from the rhizomes of S. hexandrum, one strain was able to produce kaempferol. Another strain, named TW5, was able to produce both kaempferol and podophyllotoxin simultaneously according to the TLC, HPLC, and NMR results. The podophyllotoxin yield of TW5 was calculated to be 49.3 mu g/g of mycelial dry weight after 7-d fermentation. Strain TW5 was identified morphologically and phylogenetically to be Mucor fragilis Fresen. (Mucoraceae). These results suggest that the podophyllotoxin-synthesizing ability is obtained by uptaking genes involved in the podophyllotoxin synthesis from the host plant into endophytic fungal genomes.Conclusion: Our results showed, for the first time, that the endophytic fungus M. fragilis is able to produce simultaneously the same two bioactive metabolites, podophyllotoxin and kaempferol, as its host plant. Furthermore, the relatively high podophyllotoxin yield obtained may improve the industrial production of podophyllotoxin, which may help protect this endangered plant.

Actinomycetes are one of the most important bio - control micro - organisms which produce antibiotics and enzymes and are advantageous in the bio - control of plant diseases. This study addresses the bio - control of actinomycetes against plant diseases in terms of actinomycetes＇ action on phytopathogen and on the plants＇ disease resistance under the soil environ- ment. The paper also gives a brief introduction to the applications of actinomycetes in the bio - control of plant diseases, research hotspot and development trend.

α-Pinene was converted into α-terpinene by Wagner-Meerwein rearrangement under catalysis of protonic acid,followed by Diels-Alder cycloaddtion reaction with maleic anhydride to obtain α-terpinene-maleic anhydride adduct(3).N-amino-α-terpinene-maleimide(4) was prepared by reaction of 3 with hydrazine hydrate.Eight novel α-terpinene-maleimide-based disulfamides(5a~5h) were synthesized by reaction of substituted benzene sulfonyl chlorides with 4 using DMAP as the catalyst.The structures were characterized by 1H NMR,13C NMR,IR and ESI-MS.The preliminary herbicidal activities of 4 and 5 were investigated.The results showed that most compounds exhibited a certain growth inhibition against rape root(Brassica campestris),in which 4 had inhibition rate of 84.6% at the concentration of 100 μg·mL-1.

A number of microbial species isolated from the natural environment have been cultured in the laboratory so as to obtain the extant morphology of their colonies.These were then compared with certain discoidal Ediacara fossils whose affinities remain highly controversial.Our results show that 1) Iron-oxidizing bacterium Sphaerococcus when isolated from sandstone-type Uranium ore,enriched and cultured solely on Winogradsky medium,can form discoidal colonies with concentric growth zones,which resemble closely certain Ediacara discoidal fossils in morphology and structure,such as Tirasiana concentralis and Cyclomedusa;2) Sphaerococcus colonies are not only stable in form,but also have a rigid consistency and therefore are resistant to mechanical compaction to some extent.Therefore,it is entirely possible for such colonies to be preserved in rocks as impressions.Our research implies that some of the Ediacara fossils are likely to be impressions of microbial colonies.

Beer waste yeast protein and ethylene glycol were mixed at different ratios and hot press-molded to produce beer waste yeast protein plastic. The effects of ethylene glycol on the structure and property of beer waste yeast protein plastic were studied by material testing machine, integrated thermal analyzer, Fourier transform infrared spectrometer and scanning electron microscopy. The results show that plastic with ethylene glycol mass fraction of 35% had a maximum elongation. The water absorption of the plastic increased with increase of ethylene glycol content. The hydrogen bonds between the ethylene glycol and the beer waste yeast protein wcakend the hydrogen bonds among the protein molecules and enlarged the elongation of plastic. The ethylene glycol reuld improve the microstructure and toughness of beer waste yeast protein plastic.

以陕西产地不同年限西洋参根际土壤微生物（包括解钾、解磷及自生固氮菌）为研究对象,通过对根际土壤微生物进行分离、培养、计数和鉴定,并对比栽参后的玉米轮作根际土壤微生物数量和类群的变化特征,研究了根际土壤微生物的动态变化对西洋参连续种植的影响。结果表明:与轮作根际土壤相比,随西洋参生长期的增加其根际土壤中细菌数量的变化不明显,但优势细菌种类减少;放线菌数量及种类均有不同程度的减少;引起西洋参病害的霉菌类群明显增加;解磷、解钾、自生固氮菌的比例明显失调。此种变化可引起土壤理化性质的改变,进而影响西洋参的连续种植情况。

The biological hydrogen (bio-H2) production from apple pomace (AP) by fermentation using natural mixed microorganisms in batch process was studied under various experimental conditions. The river sludge was used as a seed after being boiled for 15min. The results show that the optimal pretreatment for AP was to soak it in the ammonia liquor of 6% for 24h at room temperature. An optimal fermentation condition for bio-H2 production was proposed that the pretreated AP at 37°C, the initial pH of 7.0 and the fermentation concentration of 15g/l could produce a maximum cumulative H2 yield (CHYm) of 101.08ml/g total solid (TS) with an average H2 production rate (AHPR) of 8.08ml/gTS/h. During the conversion of AP into H2, acetic acid, ethanol, propionic acid and butyric acid were main liquid end-products.

An anaerobic fermentation process to produce hydrogen from cornstalk wastes was systematically investigated in this work. Batch experiments numbered series I, II and III were designed to investigate the effects of acid pretreatment, enzymatic hydrolysis (enzymatic temperature, enzymatic time and enzymatic pH) on hydrogen production by using the natural sludge as inoculant. A maximum cumulative H2 yield of 126.22mlg−1-CS (Cornstalk, or 146.94mlg−1-TS, Total Solid) and an average H2 production rate of 9.58mlg−1-CSh−1 were obtained from fermentation cornstalk with a concentration of 20g/L and an initial pH of 7.0 at 36°C through an optimal pretreatment process. The optimal process was that the substrate was soaked with an HCl concentration of 0.6wt% at 90°C for 2h, and subsequently enzymatic hydrolysis for 72h at 50°C and pH 4.8 before fermentation. The biogas consisted of only H2 and CO2. In addition, the fermentation system was the typical ethanol-type fermentation according to ethanol and acetate as the main liquid by-products.

[Objective]The aim was to study the best extration method and technology of protein from waste beer yeast.[Method]Four extraction methods were adopted in this research,which were ultrasonic extraction,freeze-thaw extraction,salt extraction and alkali extraction.[Result]Under the best extraction process and conditions,the protein extraction rate of ultrasonic extraction,freeze-thaw extraction,salt extraction and alkali extraction were 3.47%,4.15%,5.66% and 22.81% respectively.The best extraction technology of protein from beer waste yeast was alkali extraction,the best conditions of alkali extraction were 3∶10 of yeast to liquid ratio,1% of NaOH concentration,1 h of extraction time and 60 ℃ of extracting temperature.[Conclusion]It might provide the theoretic and experimental foundation for the extraction of protein from beer waste yeast in large scale.

To synthesize acetylsalicylic acid ethyl ester and evaluate its antibacterial activity.The target compound was synthesised under acetylsalicylic acid and ethanol in 1,4 -dimethyl aminopyridine catalyst. Drug-min paper aim to determine the antibacterial activity of compounds.The infrared chromatography mass spectrometry analysis and flight test results to determine the acetylsalicylic acid ethyl ester chemical structure.0.5g/L concentration of target compounds of the largest common intestinal pathogenic bacteria inhibition zone for 16.10mm; 1g/L concentration of target compounds under the common intestinal pathogenic bacteria inhibition zone for the maximum 23.20mm.Acetylsalicylic acid ethyl ester indeed has a clear antibacterial activity,it has the value of development and utilization.

It was to construct suicide vector of Xna gene of Xenorhabdus nematophila and establish the genetic transformation system.Firstly,the left and right homologous recombinant fragments of Xenorhabdus nematophila var.Pekingensis and kanamycin resistance gene of PEASY-T1 vector were amplified.Secondly,the three fragments were assembled together and cloned into suicide vector PJQ200.Finally,the homologous recombinant vector was introduced into donor bacteria by electroporation and then transformed into Xenorhabdus nematophila var.pekingensis by two parent′s conjunction.The homologous recombinant vector was successfully constructed and transformation system of Xenorhabdus nematophila var.pekingensis was also established.