Biodiversity,Phylogeny, and Antifungal Functions of Endophytic Fungi Associated with Zanthoxylum bungeanum

This study investigated the biodiversity, phylogeny, and antifungal activity of endophytic fungi isolated from Zanthoxylum bungeanum. A total of 940 isolates obtained were grouped into 93 morphotypes, 43 species, and 23 genera, which were authenticated by molecular identification based on rDNA internal transcribed spacer (ITS) sequence analysis. A high diversity of endophytic fungi from Z. bungeanum are observed with high species richness S (43), Margalef index D′ (6.1351), Shannon–Wiener index H′ (3.2743), Simpson diversity index D_s (0.9476), PIE index (0.9486), and evenness Pielou index J (0.8705) but a low dominant index λ (0.0524). Significant tissue specificity of the endophytic fungi was observed in Z. bungeanum, and the highest species richness and diversity indexes were obtained in the stem. Phylogenetic analyses of the 93 endophytic isolates were carried out by the neighbor-joining (NJ) method to demonstrate their evolutionary processes. Antifungal activities of endophytic fungi were assayed and eight endophytic isolates showed strong and long-lasting inhibition against host pathogenic fungi Fusarium sambucinum and Pseudocercospora zanthoxyli. Here, for the first time, we systematically demonstrate the biodiversity, phylogeny, and antifungal activity of endophytic fungi associated with Z. bungeanum and reveal the value of sampling different tissues of a given plant to obtain the greatest endophyte species diversity, which might offer a framework for further investigation and utilization of endophytic fungi as aunique source of interesting and useful bioactive compounds.     

Biosynthesis of Trehangelin in Polymorphospora rubra K07–0510: Identification of Metabolic Pathway to Angelyl‐CoA

Trehangelins are trehalose angelates discovered from endophytic actinomycete Polymorphospora rubra K07‐0510. We identified the trehangelin biosynthetic gene cluster, including genes that encode a glycoside hydrolase‐like protein (thgC), α‐amylase (thgD), 3‐ketoacyl‐ACP synthase III (thgI), 3‐ketoacyl‐ACP reductase (thgK), enoyl‐CoA hydratase (thgH) and acyl transferase (thgJ). Heterologous expression of thgH, thgI, thgJ and thgK confirmed the importance of these genes in the biosynthesis of trehangelin A. Enzymatic activity studies showed that ThgI catalyses the condensation of acetyl‐CoA and methylmalonyl‐CoA to 2‐methylacetoacetyl‐CoA (MAA‐CoA), ThgK catalyses NADPH‐dependent reduction of MAA‐CoA to 3‐hydroxy‐2‐methylbutyryl‐CoA (HMB‐CoA) and ThgH catalyses the dehydration of HMB‐CoA to angelyl‐CoA (AN‐CoA). This is the first report on the elucidation of the enzymatic formation of AN‐CoA.     

Production of lepidimoide by an endophytic fungus from polysaccharide extracted from Abelmoschus sp.: identification of the product and the organism producing it

A highly potent allelopathic factor, lepidimoide, was initially extracted from mucilage of germinated cress seeds. Polysaccharide extracted from okra (Abelmoschus esculentum Moench) is considered to have a similar structure to lepidimoide as its repeating unit. We therefore initiated the screening of enzymes capable of degrading okra polysaccharide into lepidimoide from endophytes. We discovered an endophytic fungal strain AHU9748 isolated from Coleus galeatus, which produced an oligosaccharide having similar properties to lepidimoide on thin layer chromatography. The physico-chemical data from ESI-MS, NMR spectra and other analyses also showed the purified product to be identical to lepidimoide. The strain AHU9748 was identified as a fungus belonging to the coelomycetes, closely related to the genus Colletotrichum, based on morphological characteristics and sequence analysis of the 18S rDNA and ITS region.     

植物内生菌在环境污染修复中的应用研究进展

植物内生细菌作为微生物学同植物微生态学的重要交叉点,近年来备受关注。国内外学者针对植物内生菌这一宝贵的微生物资源库进行了多方面的研究和探索。本文主要介绍植物内生菌的概念和特点,并重点阐述了植物内生菌在环境污染的生物修复中的应用研究动态。旨在为植物内生菌的实际应用提供新的思路。     

Anti-TMV Activity of Malformin A1, a Cyclic Penta-Peptide Produced by an Endophytic Fungus Aspergillus tubingensis FJBJ11

Plant-associated microorganisms are known to produce a variety of metabolites with novel structures and interesting biological activities. An endophytic fungus FJBJ11, isolated from the plant tissue of Brucea javanica (L.) Merr. (Simaroubaceae), was proven to be significantly effective in producing metabolites with anti-Tobacco mosaic virus (TMV) activities. The isolate was identified as Aspergillus tubingensis FJBJ11 based on morphological characteristics and ITS sequence. Bioassay-guided isolation led to the identification of a cycli penta-peptide, malformin A₁, along with two cyclic dipeptides, cyclo (Gly-l-Pro) and cyclo (Ala-Leu). Malformin A₁ showed potent inhibitory effect against the infection and replication of TMV with IC₅₀ values of 19.7 and 45.4 μg·mL⁻¹, as tested using local lesion assay and leaf-disc method, respectively. The results indicated the potential use of malformin A₁ as a leading compound or a promising candidate of new viricide.     

小豆内生真菌的筛选鉴定及其产玉米赤霉烯酮的可能性探究

玉米赤霉烯酮(zearalenone,ZEN)是目前全球范围污染谷物最广泛的霉菌毒素之一,对人体和动物健康都会产生极大威胁,目前从市售小豆检测到ZEN。【目的】筛选暴露ZEN的小豆内生真菌,结合经典方法和分子生物学方法鉴定所筛选内生真菌,并探索菌株潜在的ZEN产生风险。【方法】以暴露ZEN的小豆为样本,将分离纯化后的内生真菌进行经典形态观察;同时扩增真菌核糖体保守序列ITS1、ITS4,镰刀菌特异基因序列FU1、FU2确定小豆中镰刀菌种属,同时通过ZEN玉米赤霉烯酮产毒控制基因pks13进行PCR扩增,分析菌株ZEN合成的潜在风险。【结果】鉴别出3株镰刀菌,其中2株初步鉴定为砖红镰刀菌(Fusarium lateritium)、1株为花腐镰刀菌(Fusarium napiforme),未检测出ZEN合成关键基因;1株玛利节菱孢霉(Arthrinium marii),分子检测具有产ZEN玉米赤霉烯酮毒素的可能性。将以上4株菌分别回接至ZEN的产毒培养基,LC-MS/MS法未检测出ZEN毒素。【结论】暴露ZEN的小豆具有内生镰刀菌,存在可能产ZEN的内生节菱孢霉;回接培养基未检测到ZEN,但不排除小豆产生ZEN的风险。     

ATP,oxygen uptake rate and INT-dehydrogenase activity of actinomycete foams

The purpose of this study was to establish the validity of the INT-dehydrogenase test for determining the biochemical activity of actinomycete foams. Parallel measurements of INT-dehydrogenase activity, adenosine triphosphate and oxygen uptake rate were made on foam samples collected from three different activated sludge plants over a period of several months. Activity levels of actinomycete foams measured in this study were found to be comparable to those of activated sludge as reported in the literature. Good-to-excellent correlations (r² > 0.87) were found between INT-dehydrogenase activity and the alternative parameters. Precision of all assays tested was comparable. The INT-dehydrogenase test can therefore be used in lieu of ATP or oxygen uptake rate assays without sacrifice in either the reliability or reproducibility of results obtained.     

产纤维素酶放线菌的研究进展

介绍了由微生物产生的纤维素酶的组成及作用机制,概述了产纤维素酶放线菌的特点和其所产纤维素酶系的组成,以及放线菌产生的纤维素酶在工业上的应用,并简述了国内外的研究水平和发展趋势。     

烟草黑胫病拮抗内生细菌的分离、鉴定及防效测定

采用常规分离方法从健康烟株的根、茎、叶中分离得到内生细菌269株,从中筛选出7株对烟草黑胫病病原菌具有明显拮抗作用的菌株,它们的抑菌半径达到6 mm以上。菌株05-4004、05-2501培养液滤液中存在具有明显抑菌作用的活性成分。致病性试验中,发现菌株701-1在人工接种条件下有致病能力或潜在致病性,选择余下6株拮抗细菌进行了温室盆栽试验,以05-4004、05-2002两株烟草内生细菌对烟草黑胫病的防效最好,分别为46.6%、45.2%。促生试验中,NB培养液和清水对照的种子发芽率分别为84.3%、82.7%,而经过05-2002和05-4004除菌滤液浸种后,发芽率分别提高到93.5%和93.7%。经过05-4004、05-2002灌根处理后,株高、叶长、叶宽、有效叶片数、叶面积明显增加。证明菌株05-2002、05-4004同时还具有一定的促生作用。以细菌16S rDNA同源性为基础的系统发育分析表明05-4004和Bacillus pumilus(EU366363)、Bacillus safensis(AY030327)处于同一分支,相似性为100%,05-4004鉴定为芽孢杆菌属。