非豆科固氮树种沙棘与微生物联合共生体的纯培养研究

Symbiosis between Frankia,VA mycorrhizal fungi and Hippophae rhamnoides L.was studied in lab.The characteristic structures-arbuscules of VAM and nodules were confirmed in the root of H.rhamnoides L.,which was inoculated with VAMF and Frankia in pure artificial culture.Evaluated by the stimulation on the growth of the host plant,VAH is a better associated fungi and HR16 is a better Frankia for Hippophae rhamnoides L.

Soil microbial community analysis using terminal restriction fragment length polymorphisms

Terminal restriction fragment length polymorphism (T-RFLP) analysis is a polymerase chain reaction (PCR)-fingerprinting method that is commonly used for comparative microbial community analysis. The method can be used to analyze communities of bacteria, archaea, fungi, other phylogenetic groups or subgroups, as well as functional genes. The method is rapid, highly reproducible, and often yields a higher number of operational taxonomic units than other, commonly used PCR-fingerprinting methods. Sizing of terminal restriction fragments (T-RFs) can now be done using capillary sequencing technology allowing samples contained in 96- or 384-well plates to be sized in an overnight run. Many multivariate statistical approaches have been used to interpret and compare T-RFLP fingerprints derived from different communities. Detrended correspondence analysis and the additive main effects with multiplicative interaction model are particularly useful for revealing trends in T-RFLP data. Due to biases inherent in the method, linking the size of T-RFs derived from complex communities to existing sequence databases to infer their taxonomic position is not very robust. This approach has been used successfully, however, to identify and follow the dynamics of members within very simple or model communities. The T-RFLP approach has been used successfully to analyze the composition of microbial communities in soil, water, marine, and lacustrine sediments, biofilms, feces, in and on plant tissues, and in the digestive tracts of insects and mammals. The T-RFLP method is a user-friendly molecular approach to microbial community analysis that is adding significant information to studies of microbial populations in many environments.

Introduction to Molecular Analysis of Ectomycorrhizal Communities

A number of methods are available for those researchers considering the addition of molecular analyses of ectomycorrhizal (EcM) fungi to their research projects and weighing the various approaches they might take. Analyzing natural EcM fungal communities has traditionally been a highly skilled, time-consuming process relying heavily on exacting morphological characterization of EcM root tips. Increasingly powerful molecular methods for analyzing EcM communities make this area of research available to a much wider range of researchers. Ecologists can gain from the body of work characterizing EcM while avoiding the requirement for exceptional expertise by carefully combining elements of traditional methods with the more recent molecular approaches. A cursory morphological analysis can yield a traditional quantification of EcM fungi based on tip numbers, a unit with functional and historical significance. Ectomycorrhizal root DNA extracts may then be analyzed with molecular methods widely used for characterizing microbiota. These range from methods applicable only to the simple mixes resulting from careful morphotyping, to community-oriented methods that identify many types in mixed samples as well as provide an estimate of their relative abundances. Extramatrical hyphae in bulk soil can also be more effectively studied, extending characterization of EcM fungal communities beyond the rhizoplane. The trend toward techniques permitting larger sample sets without prohibitive labor and time requirements will also permit us to more frequently address the issues of spatial and temporal variability and better characterize the roles of EcM fungi at multiple scales.

石油污染土壤的生物降解研究

Biodeyradation has the characteristics of simple operation,low cost,and easy application.In this research,two fungus were chosen to degrade the oil in polluted soil in laboratory and fields.In the laboratory culture test with temperature controlled,the rate of oil degradation by native fungi (F) and foreign fungi (P) are 61.8% and 66.1% respectively after 50 days.In the field test the rate of oil degradation by native fungi (F) and foreign fungi (P) are 59.8% and 70.1% respectively after 150 days.

石油污染土壤生物修复过程中微生物生态研究

The analysis on microbiological ecology for four types of oil contaminates soils showed that the bacteria utilizing the oil as carbon sources increase,wheras the fugi become less .Zooloea and Bacillu are the dominant bacteria ; Mocor and Cunninghamella ,and Fursarium are the dominant fungi streptomyces take the superiority among the actinomyces.The anaiysis on esterase activity showed that the microbes above mentioned have abilies of degrading esters. The biodeg radationrates are 55.45%,56.74%,38.37% and 45.19%respectively,after 53 days,the biodegradation rate can be increased by 12.6% when the dominant microbes are added.

锈革孔菌科(担子菌门,非褶菌目)中国二新记录种(英文)

<正>1 INTRODUCTION During the investigation on wood-inhabiting fungi in China, two species of the Hymenochaetaceae were found: Fomitiporia texana (Murrill) Nuss and Inonotus ochroporus (Van der Byl) Pegler. Because they were not recorded in the Chinese fungal flora (Dai, 1999; Dai et al., 2001; Dai & Niemel?, 2006; Nú?ez & Ryvarden, 2000; Ryvarden, 2005), and their illustrated descriptions are given according to our collections.

中国伏革菌科二新记录种(英文)

I<正>NTRODUCTION Changbaishan Nature Reserve is one of the most important forest reserves in China and it is very rich in wood-rotting fungi (Dai, 1997, 2000, 2003; Dai et al., 2003). Nearly 100 species of the Corticiaceae sensu lato

Facile preparation of amperometric laccase biosensor with multifunction based on the matrix of carbon nanotubes-chitosan composite

The carbon nanotubes-chitosan (CNTs-CS) composite provides a suitable biosensing matrix due to its good conductivity, high stability, and good biocompatibility. Enzymes can be firmly incorporated into the matrix without the aid of other cross-linking reagents. The composite is easy to form insoluble film in solution above pH 6.3. Based on this, a facilely fabricated amperometric biosensor by entrapping laccase into the CNTs-CS composite film has been developed. At pH 6.0, the fungi laccase incorporated into the composite film remains better catalytic activity than that dissolved in solution. The system is in favor of the accessibility of substrate to the active site of laccase, thus the affinity to substrates is improved greatly, such as 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) diammonium salt (ABTS), catechol, and 0, with K. values of 19.86 mu M, 9.43 mu M, and 3.22 mM, respectively. The major advantages of the as-prepared biosensor are: detecting different substrates (ABTS, catechol, and 02), possessing high affinity and sensitivity, durable long-term stability, and facile preparation procedure. On the other hand, the system can be applied in fabrication of biofuel cells as the cathodic catalysts based on its good electrocatalysis for oxygen reduction.

Characterization of Full-Biodegradable Starch-PVA-Polyester Film

IntroductionConventional polymers such as polyethyleneand polypropylene persistfor many years after landdisposal.Furthermore,plastics are often soiled byfood and other biological substances,making phys-ical recycling of those materials impractical andgenerally undesirable. In contrast,biodegradablepolymers disposed in bioactive environment are de-graded by the enzymatic action of microorganismssuch as bacteria,fungi,and algae.The worldwideconsumption of biodegradable polymers increasedfrom1.4×107kg in ...

Comparison of two marine sponge-associated Penicillium strains DQ25 and SC10: differences in secondary metabolites and their bioactivities

Two strains of Penicillium, DQ25 and SC10, isolated from marine sponge Haliclona angulata (Bowerbank) and Hymeniacidon sp. respectively, were subjected to stationary cultivation under GYP medium for 30 days. The fermentation extracts were undergone bioactivities assays against human pathogens, phytopathogenic fungi and brine shrimp (Artemia salina). Bioassays-guided compounds isolation was performed by Silica gel columns and Sephadex LH-20 chromatography. Spectroscopic methods were used to structures elucidation of the compounds. Results showed the activities of secondary metabolites of strain DQ25 were generally stronger than that of strain SC10. Major bioactive molecules isolated from strain DQ25 were a 1,4-naphthoquinone derivative and an unidentified alkaloid. The two components were not isolated from the extract of strain SC10. ITS sequences revealed that these two species have the greatest similarity with Penicillium vinaceum and Penicillium granulatum respectively.

High level expression, purification, and characterization of the shrimp antimicrobial peptide, Ch-penaeidin, in Pichia pastoris

Penaeidins, members of a new family of antimicrobial peptides constitutively produced and stored in the haemocytes of penaeid shrimp, display antimicrobial activity against bacteria, and fungi. Here, a DNA sequence encoding the mature Ch-penaeidin peptide was cloned into the pPIC9K vector and transformed into Pichia pastoris. The transformed cells were screened for multi-copy plasmids using increasing concentrations of G418. Positive colonies carrying chromosomal integrations of the Chp gene were identified by phenotype and PCR. When transformed cells were induced with methanol, SDS-PAGE and Western blotting revealed the production of a similar to6100 Da recombinant CHP (rCHP) expression product. Large scale expression revealed that rCHP was produced at 108 mg/L under optimal conditions in the highest Chp-producing P. pastoris clone. The antimicrobial activities of rCHP were studied by liquid phase analysis, which revealed that rCHP exhibited activities against some Gram-negative and Gram-positive bacteria, but had a relatively low activity against some fungi. Purification of rCHP by cation exchange chromatography and subsequent automated amino acid sequencing revealed the presence of four additional amino acids (YVEF) at the N-terminus that belonged to the cleaved fusion signal peptide; these residues may account for the observed decrease in antifungal activity. Together, these observations indicate that rCHP is an effective antimicrobial peptide that can be successfully produced at high levels in the yeast, and therefore may be a potential antimicrobial candidate for practical use. (C) 2004 Elsevier Inc. All rights reserved.

Ergosterimide, a new natural Diels-Alder adduct of a steroid and maleimide in the fungus Aspergillus niger

Ergosterimide (1), a natural Diels-Alder adduct of ergosteroid and maleimide, was characterized from the culture extract of Aspergillus niger EN-13, an endophytic fungus isolated from the marine brown alga Colpomenia sinuosa. In addition, four known steroids including (22E,24R)-ergosta-5,7,22-trien-3 beta-ol (2), (22E,24R)-ergosta-4,6,8(14),22-tetraen-3one (3), (22E,24R)-5 alpha,8 alpha-epidioxyergosta-6,22-dien-3 beta-ol (4), and (22E,24R)-ergosta-7,22dien-3 beta,5 alpha,6 beta-triol. (5) were also isolated and identified. The structures of these compounds were elucidated by extensive analysis of 1D and 2D NMR and IR spectra and MS data. The plausible biosynthetic pathway of 1 was also discussed. To the best of our knowledge, 1 is the first natural Diels-Alder adduct of steroid and maleimide reported so far. (C) 2007 Elsevier Inc. All rights reserved.

CfLGBP, a pattern recognition receptor in Chlamys farreri involved in the immune response against various bacteria

Lipopolysaccharide and beta-1, 3-glucan binding protein (LGBP) is a kind of pattern recognition receptor, which can recognize and bind LPS and beta-1, 3-glucan, and plays curial roles in the innate immune defense against Gram-negative bacteria and fungi. In this study, the functions of LGBP from Zhikong scallop Chlamys farreri performed in innate immunity were analyzed. Firstly, the mRNA expression of CfLGBP in hemocytes toward three typical PAMPS stimulation was examined by realtime PCR. It was up-regulated extremely (P < 0.01) post stimulation of LPS and beta-glucan, and also exhibited a moderate up-regulation (P < 0.01) after PGN injection. Further PAMPs binding assay with the polyclonal antibody specific for CfLGBP proved that the recombinant CfLGBP (designated as rCfLGBP) could bind not only LPS and beta-glucan, but also PGN in vitro. More importantly, rCfLGBP exhibited obvious agglutination activity towards Gram-negative bacteria Escherichia coil, Gram-positive bacteria Bacillus subtilis and fungi Pichia pastoris. Taking the results of immunofluorescence assay into account, which displayed CfLGBP was expressed specifically in the immune cells (hemocytes) and vulnerable organ (gill and mantle), we believed that LGBP in C farreri, serving as a multi-functional PRR, not only involved in the immune response against Gram-negative and fungi as LGBP in other invertebrates, but also played significant role in the event of anti-Gram-positive bacteria infection. As the first functional research of LGBP in mollusks, our study provided new implication into the innate immune defense mechanisms of C. farreri and mollusks. (C) 2010 Elsevier Ltd. All rights reserved.

红树林植物木榄和一株海藻内生真菌的化学成分研究

本论文对民间药用红树植物木榄（Bruguiera gymnorrhiza）中的次生代谢产物进行了研究，同时还对分离自似瘤凹顶藻（Laurencia similis Namet Saito）的内生真菌EN21（Exophiala oligosperma）进行了化学成分研究。对分离得到的部分化合物进行了初步的DPPH自由基清除活性和抗菌活性筛选。 采用常规的硅胶柱层析、反相硅胶柱层析、制备薄层层析、凝胶Sephadex LH-20 柱层析、半制备型高效液相色谱（HPLC）以及重结晶等手段，从木榄（Bruguiera gymnorrhiza）中分离得到23个化合物；从EN21（Exophiala oligosperma）中分离得到23个化合物。利用一维、二维核磁、质谱、元素分析等各种现代波谱技术及化学反应方法，确定了木榄中20个化合物的结构，包括黄酮单体化合物5个，三萜类化合物11个，甾体化合物2个，长链烷烃2个，其中4个化合物为首次从木榄中报道的黄酮；确定了EN-21中20个化合物的结构，其中脑苷脂1个，甾体化合物7个，芳环化合物3个，三萜类化合物2个，包括1个新天然产物。本文系首次报道Exophiala oligosperma 的化学成分。 对从木榄中分离得到的部分化合物进行了DPPH自由基清除活性筛选， 发现黄酮类化合物BRG-1、BRG-2、BRG-3 、BRG-4显示了较强的DPPH自由基清除活性，而三萜类化合物则未表现出DPPH自由基清除活性。抗菌筛选实验中所有测试样品未显示出活性。 研究结果充实了红树林植物和海藻内生真菌代谢产物的内容，为其有效利用提供了一定的科学依据。

黄槿内生真菌赤散囊菌及红海榄的次生代谢产物及其生物活性研究

由于生长环境的特殊性，红树林及其内生真菌的代谢产物在化学类型和生物活性方面都具有多样性，因此对其代谢产物的研究引起了人们越来越多的关注。本论文以菌丝体生物量、代谢产物量等指标及薄层色谱分析、高效液相色谱分析、抗菌活性测试等筛选手段对来源于我国海南红树植物的九株内生真菌在四种不同液体培养基上的静置发酵产物进行了综合评价，并从中选择了来源于半红树植物黄槿（Hibiscus tiliaceus）的内生真菌G2——赤散囊菌（Eurotium rubrum）进行了30 L规模发酵（采用PDB培养基）和次生代谢产物的研究，对分离得到的部分化合物进行了初步的生物活性评价。此外，本论文还对海南真红树植物红海榄（Rhizophora stylosa Griff.）的化学成分进行了研究，并对分离得到的部分化合物进行了二苯代苦味酰自由基（DPPH）清除活性的评价。 采用常规的硅胶柱层析、Sephadex LH-20柱层析、反相硅胶柱层析、制备薄层层析（pTLC）、重结晶等分离手段，分离纯化得到单体化合物。综合运用现代波谱技术 (IR、UV、MS、1D-NMR 和 2D-NMR) 以及与标准品或文献比对鉴定单体化合物的结构。从G2菌丝体和发酵液的合并提取物中鉴定了45个化合物的结构，其中13个为新化合物，结构类型包括6个苯甲醛类化合物（ER1*~ER6*）、4个蒽醌类化合物（ER15*~ ER18*）和3个含吲哚的二酮哌嗪生物碱类化合物（ER27*~ER29*）。 对以上分离鉴定的部分单体化合物进行了DPPH自由基清除活性、拒食杀虫活性、抗细菌活性以及体外细胞毒活性的初步评价。新化合物ER15*和三个已知化合物ER20、ER39和ER40都表现很强的DPPH自由基清除活性。化合物ER15*还表现较好的拒食杀虫活性，而化合物ER5*和ER18*不但没有杀虫活性，反而能促进幼虫的生长。所测试的化合物只有ER15*和ER18*表现出微弱的抗金黄色葡萄球菌活性。所测试的化合物对A-549、HL-60和P-388细胞株均未表现出有意义的体外细胞毒活性。 从红海榄枝条的提取物中分离鉴定了29个化合物，其中2个为新化合物，包括1个三萜酯（RS1*）和1个黄烷醇类化合物（RS12*）；另有1个三萜酯（RS5）和1个黄烷醇类化合物（RS11）作为新的天然产物被分离鉴定。 对从红海榄提取物乙酸乙酯相和正丁醇相分离得到的部分单体化合物进行了DPPH自由基清除活性的研究。黄烷醇类化合物RS16和RS17显示最强的活性。另外，实验结果说明黄烷醇类化合物的DPPH自由基清除活性与其分子中所含的羟基数目有一定关系，而且若芳香环上有多个邻位酚羟基，则该化合物的活性将增强。本论文实验结果为海南红海榄植物资源的利用提供了化学成分及抗氧化活性方面的科学依据。