Isolation and identification of fungi tolerant to polycyclic aromatic hydrocarbons and coal tar from different habitats in Lithuania

A group of 36 fungal strains, belonging to the Lithuanian mycobiota, was collected and isolated from different locations, habitats, and matrices, including creosote-treated wood in storage yards for crosstie wastes. The eight most perspective strains selected according to preliminary assessment of tolerance to coal tar were subsequently identified combining taxonomical evaluation and molecular techniques. The tolerance of the eight identified fungal species (five basidiomycetes and three ascomycetes) to the presence of various concentrations of coal tar, and for the four most perspective fungal strains (Pleurotus sp., Schizophyllum sp., Irpex lacteus, Bjerkandera adusta) to polycyclic aromatic hydrocarbons was evaluated. The ligninolytic enzymatic activity assay of the isolated strains resulted in a good correspondence between the tolerance to pollutants and the capability to produce ligninolytic enzymes indicating that this group of white-rot fungi is perspective for further investigation and eventual usage for mycoremediation of polycyclic aromatic hydrocarbons polluted substrates.     

Community assembly in epiphytic lichens in early stages of colonization

Colonization studies may function as natural experiments and have the potential of addressing important questions about community assembly. We studied colonization for a guild of epiphytic lichens in a former treeless heathland area of 170 km2 in southwest Norway. We investigated if epiphytic lichen species richness and composition on aspen (Populus tremula) trees corresponded to a random draw of lichen individuals from the regional species pool. We compared lichen communities of isolated young (55-120 yr) and old (140-200 yr) forest patches in the heathland area to those of aspen forest in an adjacent reference area that has been forested for a long time. All thalli (lichen bodies) of 32 selected lichen species on trunks of aspen were recorded in 35 aspen sites. When data for each site category (young, old, and reference) were pooled, we found the species richness by rarefaction to be similar for reference sites and old sites, but significantly lower for young sites. The depauperated species richness of young sites was accompanied by a skew in species composition and absence of several species that were common in the reference sites. In contrast, genetic variation screened with neutral microsatellite markers in the lichen species Lobaria pulmonaria showed no significant differences between site categories. Our null hypothesis of a neutral species assembly in young sites corresponding to a random draw from the regional species pool was rejected, whereas an alternative hypothesis based on differences in colonization capacity among species was supported. The results indicate that for the habitat configuration in the heathland area (isolated patches constituting < 0.4% of the area) lichen communities may need a colonization time of 100-150 yr for species richness to level off, but given enough time, isolation will not affect species richness. We suggest that this contradiction to expectations from classical island equilibrium theory results from low extinction rates.     

4株多环芳烃降解菌的分离及鉴定

以多环芳烃(PAHs)为筛选培养基,从石油污染土壤和石油废水中筛选到4株能够降解PAHs的高效微生物菌株,分别定名为3-28、NF、EW和EY;并对其进行了形态学观察、生理生化指标测定及分子生物学鉴定.16S rDNA序列分析显示,这4株细菌分别有581、582、1209和1230个碱基,与Microbacterium、Cellulosimicrobium、Chelatococcus和Sphingopyxis等4个属有较高的序列同源性,分别为100%、97.8%、98.2%和99.0%.结合表型特征和16S rDNA序列分析,用DNAMAN构建系统发育树,并用Bootstraping法对其评价,初步将3-28、NF、EW和EY这4株PAHs降解菌分别归属于Microbacterium sp.、Cellulosimicrobium sp.、Chelatococcus sp.和Sphingopyxis sp..图3表1参22     

Microbial pathogens of public health significance in waste dumps and common sites

Microbial pathogens of public health significance found in waste and common sites were collected from four different dumping sites and assessed for pathogenic agents. The modified methods employed were based on the classical methods and basic principles of the reactions followed by biochemical enzymatic standards described for gram negative non fermenting bacteria. The results have shown presence of bacterial species including Pseudomonas, Mirococcus, Actinomyces, Neisseria, Bacillus and Klebsiella. These pathogens can infect wounds and cause sepsis and mortality and can even occur with such organisms to cause secondary infection. These groups of organisms are almost impossible to control since they are ubitiquous. Public health may be ensured from pathogenic agents at waste sites by prompt removal of waste and proper management (mechanical sorting and excavating) methods.     

紫色土硅酸盐细菌的表型特征及溶磷解钾能力

对40株分离自四川、重庆等地的代表笥紫色土中的硅酸盐细菌和1株参照菌株进行了52个表型性状的数值分析，探讨了硅酸盐细菌液体振荡培养后pH值、菌数变化与溶磷、解钾量之间的关系，结果表明：紫色土硅酸盐细菌绝大部分属于胶胨样芽孢杆菌（Bacillus mucilaginosus），但在个体和群体生长特征、碳氮源利用、硝酸盐还原能力、耐Ⅰ菌株在86％相似水平分为2个亚群，不同的硅酸盐细菌菌株的溶磷、解钾特性差异较大，具有溶磷解钾双重特性的菌株约占供试菌株（含参考菌株）的22％，只具溶磷特性的约占29％,其溶磷效果较显著（有效磷增加最高达40.64%），而解钾效果不明显（有效钾增加最多的只有7.9%）。供试紫色土硅酸盐细菌的溶磷、解钾特性与菌株生长过程中产酸有关，其溶磷、解钾量与培养液中pH值的降低呈正相关，图5表3参20。     

Use of ecoacoustics to determine biodiversity patterns across ecological gradients

The variety of local animal sounds characterizes a landscape. We used ecoacoustics to noninvasively assess the species richness of various biotopes typical of an ecofriendly forest plantation with diverse ecological gradients and both nonnative and indigenous vegetation. The reference area was an adjacent large World Heritage Site protected area (PA). All sites were in a global biodiversity hotspot. Our results showed how taxa segregated into various biotopes. We identified 65 singing species, including birds, frogs, crickets, and katydids. Large, natural, protected grassland sites in the PA had the highest mean acoustic diversity (14.1 species/site). Areas covered in nonnative timber or grass species were devoid of acoustic species. Sites grazed by native and domestic megaherbivores were fairly rich (5.1) in acoustic species but none were unique to this habitat type, where acoustic diversity was greater than in intensively managed grassland sites (0.04). Natural vegetation patches inside the plantation mosaic supported high mean acoustic diversity (indigenous forests 7.6, grasslands 8.0, wetlands 9.1), which increased as plant heterogeneity and patch size increased. Indigenous forest patches within the plantation mosaic contained a highly characteristic acoustic species assemblage, emphasizing their complementary contribution to local biodiversity. Overall, acoustic signals determined spatial biodiversity patterns and can be a useful tool for guiding conservation.     

蔬菜灰霉病生防菌的筛选与防效试验初报

针对灰霉病对大棚蔬菜生产造成的严重危害,从不同生态环境条件下筛选到４种对灰霉病菌（由Ｂｏｔｒｙｔｉｓｃｉｎｅｒｅａ引起）有拮抗作用的生防菌株,并进行了初步鉴定．室内和田间试验表明,４种生防菌株中尤以绿色木霉（Ｔｒｉｃｈｏｄｅｒｍａｖｉｒｉｄｅ）ＬＴＲ２的防治效果为最佳,达到了８０％以上     

A Mechanistic Approach to Evaluation of Umbrella Species as Conservation Surrogates

Abstract:  Although species with large area requirements are sometimes used as umbrella species, their general utility as conservation tools is uncertain. We surveyed the species diversity of birds, butterflies, carabids, and forest-floor plants in forest sites across an area (1600 km²) in which we delineated large breeding home ranges of Northern Goshawk ( Accipiter gentilis ). We tested whether protection of the home ranges could serve as an effective umbrella to protect sympatric species of the four taxa. We also used an empirical habitat model of occupancy of home range to examine mechanisms by which the Northern Goshawk acts as an umbrella species. Among species richness, abundance, and species composition of the four taxa, only abundance and species composition of birds differed between sites located inside and outside home ranges, which was due to greater abundance of bird species that were prey of Northern Goshawks inside the home ranges. Thus, although home range indicated areas with high abundance of certain bird prey species, it was not effective as an indicator of the species diversity of all four taxa. We also did not find any difference in species richness, abundance, and species composition between sites predicted as occupied and unoccupied using the habitat model. In contrast, when we selected sites on the basis of each habitat variable in the model, habitat variables that selected sites either in agricultural or forested landscapes encompassed sites with high species richness or particular species composition. This result suggests that the low performance of the Northern Goshawk as an umbrella species is due to this species' preference for habitat in both agricultural and forested landscapes. Species that can adjust to changes in habitat conditions may not act as effective umbrella species despite having large home ranges.     

Dispersal Can Limit Local Plant Distribution

The ability of species to establish new populations at unoccupied sites is a critical feature in the maintenance of biological diversity, and it has taken on new importance as a result of global climate change and expected changes in species distribution. To examine the dispersal potential of plant species, seeds of four annual plant species were experimentally dispersed 40 to 600 m from existing populations in Massachusetts (U.S.A.) to 34 nearby unoccupied but apparently suitable sites. At three of these sites new populations were established that persisted for four generations and expanded slowly in area. At seven sites, a small initial population eventually died out. At the 24 other sites, new populations did not become established, indicating that the sites were in some way unsuitable, that not enough seeds arrived, or that conditions suitable for seed germination do not occur every year. These results suggest that some species may be unable to disperse naturally out of their existing ranges in response to global climate change, particularly if habitat fragmentation creates barriers to dispersal. These species may have to be assisted to reach suitable sites nearby to prevent their extinction in the wild.     

Screening for plants and rhizospheral fungi with bioremediation potency of petroleum-polluted soils in a Tehran oil refinery area

The contamination of soils by toxic and/or hazardous organic pollutants, especially with crude oil, is a widespread problem. This study was conducted in a petroleum-contaminated area in a Tehran oil refinery to find petroleum-resistant plants and their rhizospheral fungal strains with bioremediation potency. The plants growing in the oil-polluted area were collected and determined taxonomically. Root samples of the plant species were collected from a polluted area and fungal strains determined by laboratory methods and taxonomical keys. The growth ability of the isolated fungal strains was studied in media containing 1%–15% crude oil. Results showed that seven plant species were of the highest density in the contaminated area: Alhagi persarum, Hordeum marinum, Peganum harmala, Phragmites australis, Prosopis farcta, Salsola kali, and Senecio glaucus. The root-associated fungi were isolated and showed that the fungal variation in the oil-polluted area is higher than that in a non-polluted area. The growth assay of isolated fungal strains showed that all studied fungal strains were able to form colonies at the applied concentrations but Alternaria sp. and Rhizopus sp. were the most resistant ones. Some plants were resistant to oil pollution, which also had positive effects on the fungal strains.     

铜陵新桥矿区土壤中耐Cu微生物的筛选研究

选择安徽铜陵新桥矿区富Cu的污染土壤,充分利用微生物受自然环境重金属胁迫而产生耐性这一特点,进行土壤中耐Cu微生物的筛选研究。实验过程中分别配制三种不同的培养基,细菌、真菌和放线菌培养基。在水浴恒温振荡器中对土壤中耐受重金属的微生物进行驯化,将得到的对Cu2+耐受性最高的液体培养基作为菌源,在琼脂平板培养基上进行划线分离,并将得到的纯菌株在光学显微镜下进行形态观察,菌种经试管斜面富集培养后保存在4℃冰箱中以便后续使用。研究发现,土样中细菌和真菌对Cu的耐受性低,最高耐受质量浓度分别只达到500mg·L-1和1100mg·L-1。而放线菌表现出Cu高耐受性,分离得到的放线菌耐受Cu的质量浓度最高达到10000mg·L-1,初步鉴定该放线菌株为链霉菌属。该放线菌菌种可能同时对Cu有吸附降解特性,具有成为污染土壤生物治理的高效耐受吸附菌种的潜力巨大。     

Regional Turnover and Fluctuation in Populations of Five Plants Confined to Serpentine Seeps

Abstract: Spatial patterns in population turnover were analyzed for five plant species confined to a patchy and isolated habitat, serpentine seeps: annuals Helianthus exilis and Mimulus nudatus and perennials Senecio clevelandii , Astragalus clevelandii , and Delphinium uliginosum . A 3-year survey showed that, over a preceding 15-year period, 32 of 132 known populations in a 4200-ha area disappeared, and 65 new populations appeared. Populations that disappeared were significantly more isolated from conspecific populations than were surviving populations. Populations that disappeared were also marginally significantly closer to human-caused disturbances than were populations that survived. Vacant sites in which new populations appeared were significantly less isolated from conspecific populations than sites remaining vacant. Spatial patterns in population density were analyzed for three of the species for 3 years at 50 sites in a 20 × 40 km region. From 1997 to 1998, population densities of each species changed synchronously throughout the region. From 1998 to 1999, one species (   Mimulus nudatus ) again showed synchrony, and populations of the other two species (   Helianthus exilis , Delphinium uliginosum ) remained asynchronous and spatially uncorrelated. The five species we studied all appear to be persisting well at the regional scale, despite recent disturbance to the study region. These results suggest that for rare plants in isolated habitats, the spatial configuration of populations may have an important influence on local population persistence.     

Prevalence of the Crayfish Plague Pathogen Aphanomyces astaci in Invasive American Crayfishes in the Czech Republic

Abstract:  In Central Europe invasive North American crayfishes are carriers of the oomycete Aphanomyces astaci, which causes crayfish plague. This lethal disease currently represents one of the major threats to native European crayfishes. We used molecular methods—species-specific amplification and sequencing of the pathogen DNA—to investigate the prevalence of individuals latently infected with A. astaci in 28 populations of two invasive American crayfish species (6 of the signal crayfish [ Pacifastacus leniusculus ] and 22 of the spiny-cheek crayfish [ Orconectes limosus ]) in the Czech Republic. The pathogen occurred in 17 investigated populations. We recorded a high variation in positive reactions, ranging from 0% to 100%, in populations of O. limosus . In P. leniusculus, however, only one individual out of 124 tested positive for the pathogen. There was a clear relationship between the water body type and pathogen prevalence in O. limosus . Infection ratios in isolated standing waters were usually low, whereas in running waters, pathogen prevalence often exceeded 50%. Other evaluated characteristics of potential plague pathogen carriers (size, sex, and the presence of melanized spots in the cuticle) seemed to be unrelated to infection. Our data suggest that in contrast to other European countries, O. limosus seems to be the primary reservoir of crayfish plague in the Czech Republic. Although all populations of alien American crayfishes may be potential sources of infections and should be managed as such, knowledge on the prevalence of the plague pathogen at various localities may allow managers to focus conservation efforts on the most directly endangered populations of native crayfishes.     

应用全细胞蛋白SDS-PAGE分子标记技术验证含羞草根瘤菌的结瘤能力

对云南省热带及亚热带地区的含羞草根瘤菌进行了分离,选择其中40株菌为接种菌株,通过结瘤试验并采用全细胞蛋白SDS-PAGE分子标记方法研究了其结瘤能力.经过结瘤试验,发现除菌株SWF66075和SWF66093没有结瘤外,其它38株菌株均与含羞草植物结瘤.结瘤率为95%.从结瘤试验所获根瘤中,分离得到结瘤菌株,采用全细胞蛋白SDS-PAGE分子标记对结瘤菌株与接种菌株进行了比较研究.蛋白图谱及聚类分析显示,26株接种菌株与其结瘤菌株的全细胞蛋白分子图谱完全相同,在100%的相似水平上与其结瘤菌株聚在一起,说明宿主植物所形成的根瘤确系接种菌株侵入所致,因而可将这些菌株确认为根瘤菌菌株;而SWF66012、SWF66029、SWF66044和SWF66058等12株菌株的结瘤菌株与其各自接种菌株的全细胞蛋白图谱存在较大差异,推测这12株接种菌株与其结瘤菌株可能不是同一菌株,尚不能确定它们与含羞草植物的结瘤能力,这些菌株是否为根瘤菌菌株仍需进一步验证.研究结果表明,全细胞蛋向SDS-PAGE分子标记技术是一种快速、准确地验证根瘤菌结瘤能力的方法.该方法进一步完善了结瘤试验,并初步揭示了根瘤菌的竞争结瘤能力,适用于对大量根瘤内分离菌株进行根瘤菌的证实研究.图2参28     

The use of polymerase chain reaction for the identification of sciaenid eggs

Traditionally, sciaenid eggs have been identified based on morphological characteristics such as size, number of oil globules and/or pigmentation patterns. Identification of sciaenid eggs by these procedures is time consuming and often inaccurate due to considerable egg size overlap among species. The utilization of molecular techniques for the identification of economically important species has become a fundamental component in ecological studies involving fish eggs and larvae. This study reports the development of a series of both multiplex and individual polymerase chain reactions to identify the eggs of 11 sciaenid species commonly found in the Gulf of Mexico and estuaries near Port Aransas and Corpus Christi, TX, USA. Following method development, the discriminatory power of the assay was first determined with samples from adult fish collected from Aransas and Corpus Christi Bays, Galveston Bay and the lower Laguna Madre in northern Mexico. Most (97%) of these fishes were correctly identified to the level of species. To demonstrate the applicability of the assay, wild fish eggs were collected and analyzed from the Aransas Pass tidal inlet from September through December 2005. During this period, the eggs of four target species were positively identified which was in keeping with current knowledge regarding the spawning areas and seasons of these sciaenids based on the presence of mature females, eggs and/or larvae. Future use of this method can provide valuable information to better discriminate spawning sites and seasons of these species.     

氯代酰胺类除草剂降解菌的分离及降解性能

从生产乙草胺的农药厂废水生物处理池活性污泥中分离到一株氯代酰胺类除草剂降解细菌,命名为Y3B-1.根据表型特征、生理生化特性和16S rDNA序列系统发育分析,将其鉴定为副球菌属(Paracoccus sp.).研究了菌株Y3B-1在不同条件下对多种氯代酰胺类除草剂的降解性能.结果表明:菌株Y3B-1能以乙草胺为碳源生长,并能降解乙草胺、丁草胺和丙草胺,3 d对这3种氯代酰胺类除草剂的降解率分别达到86.7%、65.5%和69.1%,不能降解异丙甲草胺.该菌降解乙草胺的最适温度为30℃,最适pH为7.0,对乙草胺的降解效果与接种量成正相关,对较低浓度的乙草胺有很好的降解效果,过高的起始浓度抑制其对乙草胺的降解,外加营养如酵母膏和土壤悬液则显著促进其对乙草胺的降解.图7参23     

红树林沉积物Bt菌株的分离与cry基因型的鉴定（Isolation of Bacillus thuringiensis Strains from Mangrove Sediment Samples and Identification for cry Genes）

从收集自泉州洛江红树林保护区的400个沉积物土壤样品中,分离到18株苏云金芽孢杆菌(Bacillus thuringiensis,Bt).利用PCR-RFLP体系对Bt菌株杀虫晶体蛋白的基因型进行了鉴定,发现15株Bt含有cry1基因,4株含有cry2基因,有3株分别含有cry7、cry8、cry9基因.克隆了一个新型的cry2Ab基因,其编码蛋白与现有Cry2Ab型杀虫晶体蛋白的最高同源性为95%.     

Orientation and recruitment behavior in the ponerine ant Pachycondyla tesserinoda (Emery): laying of individual-specific trails during tandem running

Summary Tandem leaders of Pachycondyla tesserinoda mark their way during nest moving. While scouting and foraging for food in an unknown area, chemical orientation is used as well. The origin of the trail substance could not be identified. Secretions of gastral exocrine glands did not induce trail following behavior. Nor do these secretions elicit tandem-following reactions; rather the latter is released by the general body surface odor. The trail substance and the substance used for marking the nest entrance are colony-specific. Moreover, individual tandem leaders recognize and show a preference for their own trails. This extraordinary effect is independent of the age of the trail. P. tesserinoda workers search individually for food and new nest sites. Targets which are important for the colony are directly shown to nestmates by tandem running. Due to this type of foraging and scouting individual-specific trails may be advantageous for this ant.     

Impact of rhizobacteria on growth and chromium accumulation in Scirpus lacustris L. grown under chromium supplementation

Four chromate tolerant rhizobacterial strains viz., RZB-01, RZB-02, RZB-03 and RZB-04 were isolated from rhizosphere of Scirpus lacustris collected from Cr-contaminated area. These strains characterized at morphological and biochemical levels. The most efficient chromate tolerant strain RZB-03 was inoculated to fresh plant of S. lacustris and grown in 2 microg ml(-1) and 5 microg ml(-1) of Cr+6 supplemented nutrient solution under controlled laboratory condition. The effects of rhizobacterial inoculation on growth and chromium accumulation in S. lacustris were evaluated. The inoculation of rhizobacteria increased biomass by 59 and 104%, while total chlorophyll content by 1.76 and 15.3% and protein content increased by 23 and 138% under 2 microg ml(-1) and 5 microg ml(-1) concentrations of Cr+6, respectively after 14 d as compared to non-inoculated plant. Similarly, the Cr accumulation also increased by 97 and 75% in shoot and 114 and 68% in root of inoculated plants as compared to non inoculated plants at 2 microg ml(-1) and 5 microg ml(-1) Cr+6 concentrations, respectively after 14 d. The chromate tolerant rhizobacteria which play an important role in chromium uptake and growth promotion in plant may be useful in development of microbes assisted phytoremediation system for decontamination of chromium polluted sites.     

紫花苜蓿根际氢氧化细菌的分离与鉴定

利用气体循环培养体系从陕西乾县HUP-豆科植物紫花苜蓿(Medicago sativa)根际土壤中分离获得37株细菌.菌株氧化氢能力测定结果表明,8株菌氧化氢和自养生长能力较强,初步确定为氢氧化细菌类群;根据其形态特征、培养特征和生理生化特性,鉴定为7个不同属:假单胞菌属(Pseudomonas)、邻单胞菌属(Plesiomonas)、脂肪杆菌属(Pimelobacter)、黄色杆菌属(Xanthobacter)、勒米诺氏菌属(Leminorella)、地杆菌属(Terrabacter)和稀有杆菌属(Rarobacter);其中氧化氢能力最强的优势菌株WMQ-7 16S rDNA序列(GenBank登录号为EU807744)长度为1 451bp,GC含量为53.8%,其核苷酸序列与假单胞菌属同源性高于99%,在系统发育树上位于同一分支,将WMQ-7菌株鉴定为假单胞菌属(Pseudomonas).图2表5参16