Nature and bioactivities of endolichenic fungi in Pseudocyphellaria sp., Parmotrema sp. and Usnea sp. at Hakgala montane forest in Sri Lanka

Aim:  The aim of this study was to investigate the nature and bioactivities of endolichenic fungi in three abundant lichens, Pseudocyphellaria sp., Usnea sp. and Parmotrema sp. in the lower elevation of Hakgala montane forest in Sri Lanka.
Methods and Results:  Endolichenic fungal strains, fungi that live asymptomatically in the lichen thallus, much the same way as endophytic fungi live within healthy plant tissues, were isolated from three abundant lichen species, Pseudocyphellaria sp., Usnea sp. and Parmotrema sp., at Hakgala montane forest in Sri Lanka, using the surface sterilization method. Nine endolichenic fungal strains were isolated from Parmotrema sp. and Usnea sp. separately, while 11 endolichenic fungi were recovered from the lichen Pseudocyphellaria sp. Isolation of endolichenic fungus Chrysosporium sp. 2 was common to all three lichen species. Substrate utilization patterns and antifungal activities of eight endolichenic fungal species were evaluated and the results revealed that all the test fungi were able to produce at least one enzyme to utilize the test substrates. Nigrospora sp., Chrysosporium sp. 1 and 2 and Cladosporium sp. showed antifungal activities on growth of some selected plant pathogenic fungi.
Conclusions:  Endolichenic fungal strains (29) were isolated from the lichens Parmotrema sp., Usnea sp. and Pseudocyphellaria sp. in Sri Lanka. Chrysosporium sp. 2 was common in all three lichens. Some of these endolichenic fungal strains showed antifungal activities against common plant pathogenic fungi and they are capable of utilizing the substrates by producing specific enzymes.
Significance and Impact of the Study:  The diversity and prevalence of the endolichenic fungi have not been studied extensively and this is the first report of isolation and identification of endolichenic fungi in lichens available in Sri Lanka.     

具ACC脱氨酶活性的碱蓬内生细菌的分离、鉴定及其生物学特性

【目的】具有1-氨基环丙烷-1-羧酸(ACC)脱氨酶活性的盐生植物碱蓬内生细菌的分离及生物学特性的研究有助于探索内生细菌与宿主植物耐盐性的关系。【方法】采用研磨法从健康碱蓬植株的根、茎、叶中分离具有ACC脱氨酶活性的碱蓬内生细菌,根据形态特征、生理生化、API鉴定系统和16S rRNA对菌种进行鉴定,并分别测定了菌株产ACC脱氨酶、铁载体、吲哚乙酸、赤霉素、脱落酸、蛋白酶及溶磷、固氮和拮抗病原菌的特性。【结果】将分离得到的内生细菌LP11、SS12、TW1和TW2分别鉴定为栖稻假单胞菌(Pseudomonas oryzihabitans)、假单胞菌(Pseudomonas sp.)、成团泛菌(Pantoea agglomerans)和恶臭假单胞菌(Pseudomonas putida),4株菌除具有较高的ACC脱氨酶活力之外,还可不同程度地产生铁载体、吲哚乙酸、赤霉素和脱落酸,且均有溶磷作用,但无固氮能力及蛋白酶活力,唯有菌株SS12对萝卜枯萎病菌(Fusariumoxysporum f.sp.conglutinans)和黄瓜枯萎病菌(F.oxysporum f.sp.cucumerinum)具有拮抗作用。【结论】从盐生植物碱蓬中分离到的假单胞菌属和泛菌属内生细菌,具有丰富多样的生物学特性。     

Distribution of alkB genes within n-alkane-degrading bacteria

Fifty-four bacterial strains belonging to 37 species were tested for their ability to assimilate short chain and/or medium chain liquid n-alkanes. A gene probe derived from the alkB gene of Pseudomonas oleovorans ATCC 29347 was utilized in hybridization experiments. Results of Southern hybridization of PCR-amplificates were compared with those of colony hybridization and dot blot hybridization. Strongest signals were received only from Gram-negative bacteria growing solely with short n-alkanes (C10). Hybridization results with soil isolates growing with n-alkanes of different chain lengths suggested as well that alkB genes seem to be widespread only in solely short-chain n-alkane-degrading pseudomonads. PCR products of Rhodococcus sp., Nocardioides sp., Gordona sp. and Sphingomonas sp. growing additionally or solely with medium-chain n-alkane as hexadecane had only few sequence identity with alkB though hybridizing with the gene probe. The derived amino acid sequence of the alkB-amplificate of Pseudomonas aureofaciens showed high homology (95%) with AlkB from Ps. oleovorans. alkB gene disruptants were not able to grow with decane.     

Antimicrobial properties of moderately halotolerant bacteria from cenotes of the Yucatan peninsula

AIMS: Isolation and antimicrobial evaluation of aquatic bacterial strains from two cenotes. METHODS AND RESULTS: A total of 258 bacterial strains were isolated from the water and sediment of two cenotes in the Yucatan peninsula, all of which were screened against six pathogenic micro-organisms. Antimicrobial activity was detected in 46 of the isolated strains against at least one of the target strains tested. Antimicrobially active isolates were identified as: Aeromonas, Bacillus, Burkholderia, Photobacterium, Pseudomonas, Serratia, Shewanella, Stenotrophomonas genera, and 13 remained unidentified. All antimicrobially active strains were able to grow in salt medium at a concentration of 75 g l(-1), thus classifying as moderately halotolerant bacteria. Most of the antimicrobially active strains exhibited a broad action spectrum, where 61% was because of uncharacterized antimicrobial substances, 25% because of bacteriocins and 13% because of siderophores. Ten strains were able to biosynthesize biosurfactant metabolites. CONCLUSIONS: Native bacteria from the Yucatan peninsula showed an interesting antimicrobial activity, diverse mode of action and moderate halotolerance to salt. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on bacterial isolates from cenotes of the Yucatan peninsula and their antimicrobial characterization, with great potential for future biotechnological applications.     

先锋牧草-香根草联合固氮菌多样性研究

摘要：【目的】香根草（Vetiver zizanioides）是一种多年生禾本科草本植物,具有极强的生态适应性和抗逆能力,可作饲料和水土保持用。通过研究香根草联合固氮菌多样性,为进一步研究和应用打下基础。【方法】采用无氮培养基,首次从香根草中分离到47株联合固氮菌,分别应用SDS-PAGE全细胞蛋白质电泳、DNA指纹图谱、唯一碳源和16S rDNA全序列测定等方法,进行聚类和多样性分析。【结果】SDS-PAGE、IS-PCR和Bio-BIQA碳源利用的聚类结果基本一致,将供试菌株分为6个类群和4个单菌株;     

土霉素暴露对小麦根际抗生素抗性细菌及土壤酶活性的影响

通过培养的方法研究了土霉素暴露和小麦根际抗性细菌的数量、种类、分布特征及土壤酶活性之间的剂量效应关系。结果表明,土霉素暴露下小麦根际单一抗生素抗性细菌数量和抗土霉素—链霉素双重抗性细菌数都明显增加,且与暴露剂量呈正效应关系;同时,土壤磷酸酶、脱氢酶活性下降,但与土霉素的剂量效应关系不明显。从土霉素暴露的土壤中分离到50株抗性细菌,经形态观察、RFLP分组和16S rDNA序列测定与分析,将它们聚集在Actinobacteria、Bacilli、Alphaproteobacteria、Gammaproteobacteria 和Sphingobacteria类群。其中放线菌最多（15株）,占抗性菌总数的30 %;其次是Bacillus属细菌（9株）和Pseudomonas属细菌（8株）,分别占18 %和16 %。同时,具有抗性的人类机会致病菌Pseudomonas、Sphingomonas和Stenotrophomonas属细菌在土霉素暴露的样品中均被分离到,分别占抗性菌株总数的16 %、8 %和4 %。值得注意的是,随着土霉素暴露剂量的增加,小麦根际优势促生菌Bacillus属细菌的抗性检出率逐步降低;但具有抗生素抗性的人类机会致病菌Pseudomonas、Sphingomonas和Stenotrophomonas属细菌的检出率却明显增加,提示可能会进一步增大其机会致病性。     

Isolation and identification of lactic acid bacteria from soil using an enrichment procedure

AIMS: To survey, and identify and classify the ecological distribution of lactic acid bacteria from soil in Japan and Taiwan. METHODS AND RESULTS: Acid-producing bacteria were isolated from 68 soil samples, collected from Japan and Taiwan, in the rhizospheres of fruit trees, from the floor of a henhouse and around a horse farm. All isolates were identified by physiological and genetic tests. Thirty-two of the 54 isolates were identified as lactic acid bacteria (LAB), 16 as spore-forming lactic acid bacteria, five as Clostridium and one as Bacillus. These lactic acid bacteria represent five genera: Lactobacillus, Lactococcus, Enterococcus, Leuconostoc and Weissella. CONCLUSIONS: A high rate of isolating lactic acid bacteria was obtained from soil. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests that soil may be a common source for the isolation of lactic acid bacteria.     

先锋牧草-香根草联合固氮菌多样性

[目的]香根草(Vetiver zizanioides)是一种多年生禾本科草本植物,具有极强的生态适应性和抗逆能力,可作饲料和水土保持用.通过研究香根草联合固氮菌多样性,为进一步研究和应用打下基础.[方法]采用无氮培养基,首次从香根草中分离到47株联合固氮菌,分别应用SDS-PAGE全细胞蛋白质电泳、DNA指纹图谱、唯一碳源和16S rDNA全序列测定等方法,进行聚类和多样性分析.[结果]SDS-PAGE、IS-PCR和Bio-BIQA碳源利用的聚类结果基本一致,将供试菌株分为6个类群和4个单菌株;16S rDNA序列测定表明,从香根草中分离的菌株包括了佛莱辛草螺菌(Herbaspirillum frisingense)、中型假食酸菌(Pseudacidovorax intermedius)、恶臭假单胞菌(Pseudomonas putida)、荧光假单胞菌(Pseudomonas fluorescens)、越南伯克氏菌(Burkholderia vietnamiensis)、阴沟肠杆菌(Enterobacter cloacae)、路德维希肠杆菌(Enterobacter ludwigii)和松江壳聚糖降解菌(Mitsuaria chitosanitabida)等不同菌种.[结论]香根草联合固氮菌具有较大的资源多样性,对固氮菌资源的扩展和将来牧草上的应用具有重要意义.     

中国横断山的地衣研究 (6)

通过对灰梅属(Canoparmelia)表型与基因型相结合的研究,澄清了云南南部报道的珊瑚大叶梅(Parmotrema conformatum)是裂芽灰梅(Cecaperata)的错误鉴定。由于珊瑚大叶梅和裂芽灰梅在经典分类鉴别上容易混淆,通过rDNA ITS序列的系统发育分析,证明了裂芽灰梅与粉芽灰梅(Ctexana)及同心灰梅(Cconcrescens)有较近的亲缘关系;首次记录了粉芽灰梅在云南的分布范围;文中对两个新记录种进行了详细描述和讨论,并给出了中国已知灰梅属的检索表。     

Diversity and antimicrobial susceptibility of oxytetracycline-resistant isolates of Stenotrophomonas sp. and Serratia sp. associated with Costa Rican crops

AIMS: To ameliorate the identification, evaluate the diversity, and determine the antimicrobial sensitivity of 19 oxytetracycline-resistant isolates of Stenotrophomonas sp. and Serratia sp. associated with Costa Rican crops. METHODS AND RESULTS: Phenotypical, chemotaxonomical, and molecular data allocated most isolates to the species Sten. maltophilia and Ser. marcescens. The API profiles, antimicrobial resistance patterns (ATB system), and BOX-polymerase chain reaction (PCR) genomic fingerprints of isolates of Stenotrophomonas sp. exhibited a higher degree of heterogeneity than those obtained for the isolates of Serratia sp. The former group of bacteria exhibited multiresistance to antimicrobials. In contrast, isolates of Serratia sp. were sensitive to the majority of the drugs tested. Changes in the results of the antibiograms throughout incubation, which indicate an induction of tolerance, were observed for isolates of both the species. Minimum inhibitory concentration of oxytetracycline, determined using E-test stripes, were rather elevated. CONCLUSIONS: The occurrence of two species of opportunistic pathogens in crop-associated materials poses a risk to consumers in the community. SIGNIFICANCE AND IMPACT OF THE STUDY: The phenotypic and genotypic data presented could support epidemiologist and physicians dealing with infections caused by environmental strains of these taxa.     

The Potentiality of Free Gram-negative Bacteria for Removing Oil and Grease from Contaminated Industrial Effluents

Summary Eight bacterial species were isolated from vegetable oil and grease-contaminated industrial wastewater, only four of which were found to have the ability to degrade oil and grease in the contaminated wastewater. These isolates were identified according to morphological and biochemical profiles as, Pseudomonas sp. (L1), P. diminuta (L2), P. pseudoalcaligenes (L3), and Escherichia sp. (L5). The degradative capabilities of the identified bacterial isolates for Tween 20 (Tw20) were investigated under different pH levels (6.5, 7, 7.5, and 8), different temperatures (30 and 37 °C) and different concentrations of Tw20 (1, 1.5, and 2%). Results revealed differences in their optimum conditions for maximum degradation of vegetable oil. Bacterial isolates were tested individually or in combinations using synthetic aqueous medium supplemented with 1% palm oil, incubated at 30 °C, and agitated at 150 rev/min for 13 days. All the tested bacteria were able to degrade the palm oil completely and utilized the free fatty acids (FFA) as a carbon source. The combination M1 (Pseudomonas sp. and P. diminuta) produced the highest degradative activity, followed by M3 (Pseudomonas sp., P. diminuta and P. pseudoalcaligenes). Also M1 produced the highest activity in reducing COD (93%) and BOD₅ (100%).     

Characterization of xylanolytic bacteria present in the bract phyllosphere of the date palm Phoenix dactylifera

AIMS: Despite the interest of phyllosphere microbiology, no studies have addressed the bacteria present in bract phyllosphere, an ecosystem that has special characteristics in palm trees because the dry bracts remain on the plant until pruning and may contain polymer-degrading bacteria involved in plant degradation. Therefore, the aim of this work was to characterize xylanolytic bacteria isolated from palm bract phyllosphere. METHODS AND RESULTS: Twelve xylanolytic strains were isolated and characterized by phenotypic features and complete sequencing of 16S rRNA gene. The results showed that the isolates were phenotypically and genotypically diverse. Gram-positive isolates were classified into genus Paenibacillus some of them belonging to hitherto undescribed species of this genus. Gram-negative isolates were classified into genera Pseudomonas and Acinetobacter. CONCLUSIONS: The results of this work confirm the complexity of the bacterial populations present in phyllospheric ecosystems and suggest that bacteria involved in plant degradation are present at the early degradation steps of this process in dry palm tree bracts. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study on bract phyllospheric bacteria able to hydrolyse vegetal polymers and offers a new perspective in the search of unexplored sources of xylanase-producing strains.     

松材线虫对其携带的细菌繁殖的影响

1.引言松材线虫病（Bursaphelenchus xylophilus）是松树的一种毁灭性病害,在日本、中国、韩国和北美、尼日利亚和葡萄牙等国家蔓延,造成了巨大经济损失,其中以日本和中国受害最重.一直认为松材线虫是引起该病的唯一病原,但近十几年来的研究发现,细菌在致病过程中可能起着重要作用,相继从病木和松材线虫体上分离到能对黑松苗有致萎活性的细菌.赵博光等首次根据实验提出松材线虫病是线虫和细菌共同侵染引起的复合侵染病害的假说,并在以后的试验中得到了验证.关于松材线虫对其细菌繁殖的影响研究鲜有报道.本试验采用从感病松树上分离并鉴定了的细菌菌株中选取假单胞属7株、其它属的细菌菌株3株,     

Ecophysiological attributes of a Lactobacillus sp. and a Pseudomonas sp. on sterile beef fillets in relation to storage temperature and film permeability

AIMS: To determine the combined effect of packaging film and temperature on the rate and type of end-products caused by the growth of two main contrasting prevailing organisms in air and 100% CO2, Pseudomonas sp. and Lactobacillus sp., respectively. METHODS AND RESULTS: Pseudomonas sp. and Lactobacillus sp. were inoculated individually on sterile meat fillets. The samples were packed in air or 100% CO2, using a high and a low permeable film, and stored at 0 and 10 degrees C. Pseudomonas sp. grew aerobically and in 100% CO2 using high permeable film at both storage temperatures, while film permeability significantly affected the growth of Lactobacillus sp. only at 10 degrees C. Enzymatic kits and HPLC and GC analysis were used to determine the chemical changes of the samples throughout storage. Pseudomonas sp. presented a greater rate of consumption of glucose and lactate than Lactobacillus sp. in samples stored aerobically or with high permeable film. Propanol-1 and two unidentified organic acids were present only in samples inoculated with Pseudomonas sp., while acetaldehyde, ethanol, diacetyl and acetoin were detected in samples inoculated with Lactobacillus sp. CONCLUSION: Since different microbial species and introduction of new packaging methods affect spoilage reactions of meat either qualitatively or quantitatively, a combination of several chemical indicators should be thoroughly investigated. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study provides information on how and when such potential indicators can be exploited for the benefit of the industry and consumer.     

蜘蛛抱蛋属植物内生细菌的多样性及其抗菌活性筛选

【背景】药用植物中蕴含多样性丰富的内生菌资源,这些微生物产生的多种新型物质在制药领域表现出较好的应用前景。【目的】研究蜘蛛抱蛋属(Aspidistra Ker-Gawl.)植物内生细菌的多样性,探索药用植物内生细菌在药用活性产物方面的开发潜力,以期发现具有抗菌活性的次级代谢产物。【方法】对9种13株新鲜的蜘蛛抱蛋植物进行表面消毒,采用5种分离培养基分离内生细菌;根据菌落形态特征排除重复菌株,并测定其16S rRNA基因序列,构建系统进化树分析内生细菌多样性;将菌株分别用2种培养基发酵,使用耻垢分枝杆菌(Mycobacterium smegmatis ATCC 700044)、水稻白叶枯菌(Xanthomonas oryzae PXO99A)、白色念珠菌(Candidaalbicans ATCC 10231)、肺炎雷伯菌(Klebsiella pneumoniae ATCC 700603)和耐药粪肠球菌HH22(Enterococcus faecalis HH22)5种检定菌对分离菌株的发酵液进行抑菌活性筛选。【结果】从植物组织中分离得到了234株内生细菌,根据形态初步排重得到156株植物内生细菌;基于16S rRNA基因序列构建的系统进化树显示它们分属于3门10目22科29属,其中链霉菌属(Streptomyces)、芽孢杆菌属(Bacillus)、微杆菌属(Microbacterium)、类芽孢杆菌属(Paenibacillus)和根瘤菌属(Rhizobium)的菌株广泛地分布在不同种的蜘蛛抱蛋植株中,且占据一定优势;发现可能的潜在新分类单元6个;156株内生细菌中38株菌的发酵液具有抑菌活性,初筛阳性率为23.7%。【结论】蜘蛛抱蛋植物组织中含有种类多样的内生细菌,它们可能是抗菌生物活性次级代谢产物的有效来源。     

Isolation and characterization of lactic acid bacteria from dochi (fermented black beans), a traditional fermented food in Taiwan

AIMS: To isolate, characterize and identify lactic acid bacteria (LAB) in dochi (fermented black beans), a traditional fermented food in Taiwan. METHODS AND RESULTS: A total of 30 samples were collected from three different dochi producers and analysed after different periods of storage. Fifty-two cultures of LAB were isolated from dochi samples and the isolates were divided into classes by phenotype and then into groups by restriction fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Phenotypic and biochemical characteristics identified six different bacterial groups (A-F) and showed that the majority of the isolates were homofermentative LAB. Enterococcus faecium was the most abundant of the dochi-isolated LAB. All isolated LAB were able to grow in MRS broth containing 6% NaCl, but only Enterococcus, Pediococcus and Tetragenococcus species could grow in MRS broth containing 10% NaCl. Furthermore, antibacterial activities of isolates were determined, and four isolates showed inhibitory activities against the indicator strain Lactobacillus sakei JCM 1157(T). CONCLUSIONS: These results suggest that Ent. faecium is the main LAB present during the fermentation of dochi. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report describing the distribution and varieties of LAB that exist in the dochi fermentation process.     

Humus bacteria of Norway spruce stands: plant growth promoting properties and birch, red fescue and alder colonizing capacity

We studied the potential of the humus layer of the Norway spruce stands to supply beneficial rhizobacteria to birch (Betula pendula), alder (Alnus incana) and fescue grass (Festuca rubra), representatives of pioneer vegetation after clear-cutting of the coniferous forest. Axenically grown seedlings of these species were inoculated with the acid spruce humus, pH 3.7-5.3. Actinorhizal propagules, capable of nodulating alder, were present in high density (10(3) g(-1)) in humus of long-term limed plots, whereas plots with nitrogen fertilization contained almost none (相似文献   

Vanillin production from simple phenols by wine-associated lactic acid bacteria

AIMS: The ability of lactic acid bacteria (LAB) to metabolize certain phenolic precursors to vanillin was investigated. METHODS AND RESULTS: Gas chromatography-mass spectrometry (GC-MS) or HPLC was used to evaluate the biosynthesis of vanillin from simple phenolic precursors. LAB were not able to form vanillin from eugenol, isoeugenol or vanillic acid. However Oenococcus oeni or Lactobacillus sp. could convert ferulic acid to vanillin, but in low yield. Only Lactobacillus sp. or Pediococcus sp. strains were able to produce significant quantities of 4-vinylguaiacol from ferulic acid. Moreover, LAB reduced vanillin to the corresponding vanillyl alcohol. CONCLUSIONS: The transformation of phenolic compounds tested by LAB could not explain the concentrations of vanillin observed during LAB growth in contact with wood. SIGNIFICANCE AND IMPACT OF THE STUDY: Important details of the role of LAB in the conversion of phenolic compounds to vanillin have been elucidated. These findings contribute to the understanding of malolactic fermentation in the production of aroma compounds.     

Isolation and characterization of monochloroacetic acid-degrading bacteria

Five Burkholderia strains (CL-1, CL-2, CL-3, CL-4, and CL-5) capable of degrading monochloroacetic acid (MCA) were isolated from activated sludge or soil samples gathered from several parts of Japan. All five isolates were able to grow on MCA as the sole source of carbon and energy, and argentometry and gas chromatography-mass spectroscopy analyses showed that these five strains consumed MCA completely and released chloride ions stoichiometrically within 25 h. The five isolates also grew on monobromoacetic acid, monoiodoacetic acid, and L-2-monochloropropionic acid as sole sources of carbon and energy. In addition, the five isolates could not grow with DCA but dehalogenate single chlorine from DCA. Because PCR analyses revealed that all five isolates have an identical group II dehalogenase gene fragment and no group I deh gene, only strain CL-1 was analyzed further. The partial amino acid sequence of the group II dehalogenase of strain CL-1, named DehCL1, showed 74.6% and 65.2% identities to corresponding regions of the two MCA dehalogenases, DehCI from Pseudomonas sp. strain CBS-3 and Hdl IVa from Burkholderia cepacia strain MBA4, respectively. The secondary-structure motifs of the haloacid dehalogenase (HAD) superfamily and the amino acid residues involved in substrate binding, catalysis, and hydrophobic pocket formation were conserved in the partial amino acid sequence of DehCL1.