安徽定远盐矿可培养嗜盐微生物多样性

【背景】嗜盐微生物多生活于高盐环境,具有独特的生理代谢特征,是一类重要的极端环境微生物资源。【目的】为更好地认识我国陆相盐矿的嗜盐微生物多样性组成,更好地开发利用嗜盐微生物资源积累丰富的微生物菌种。【方法】对安徽定远盐矿盐芯样品进行嗜盐微生物的纯培养分离,并对所分离菌株进行基于16SrRNA基因的测序和序列相似性分析,并对所分离菌株进行物种多样性分析。在此基础上,对代表菌株进行菌落形态和耐盐度及酶活测定。【结果】通过纯培养共分离获得了嗜盐微生物264株,其中嗜盐古菌150株,占56.8%;嗜盐细菌114株,占43.2%。嗜盐古菌物种分别来自于Halorubrum、 Halopenitus、 Haloterrigena、 Natrinema、 Natronoarchaeum和Natronomonas等6个属;嗜盐细菌物种分别来自于Pseudomonas、Aliifodinibius、Halobacillus、Halomonas和Halospina等5个属。通过代表菌株的酶活平板检测,发现产胞外蛋白酶菌株1株,酯酶1株,淀粉酶2株;能液化明胶菌株2株。在物种多样性组成方面,发现嗜盐古菌的物种多样性指数高于嗜盐细菌。【结论】本研究对我国安徽定远陆相盐矿的可培养嗜盐微生物多样性进行探究,积累了丰富的嗜盐微生物菌株资源。     

盐度对滨海土壤细菌多样性和群落构建过程的影响

滨海盐土是重要的农业土地后备资源。微生物是土壤中物质循环的关键动力,然而盐度对土壤微生物群落特征影响的研究还很缺乏。本研究采集滨海地区的土壤样品,研究非盐、轻盐和高盐3组不同盐度对土壤细菌数量、多样性和群落构建的影响。结果表明: 与非盐和轻盐土壤相比,高盐土壤的脱氢酶活性和细菌数量显著降低,而细菌α多样性没有变化,细菌群落结构发生分异。利用零模型反演群落构建过程,发现盐度是细菌群落构建过程的主控因子,盐度主导的高确定性过程控制了滨海盐土细菌的群落结构。说明在现有的盐度范围内,高盐土壤中同样含有丰富的微生物种质资源,具有盐土改良的生物学基础,然而由于高确定性的群落构建机制,外源物种很难定殖于滨海盐土。因此,在利用微生物技术改良滨海盐土时,应尽可能筛选耐盐的土著菌种,提高定殖效率。     

极端嗜盐多形性古菌病毒HRPV13的分离鉴定及生物学特性

【背景】与感染细菌和真核生物的病毒相比,目前发现的古菌病毒数量很少,但是却展现出形态多样性。因此,分离和鉴定新的古菌病毒具有重要意义。【目的】为了进一步了解古菌病毒的多样性,我们从青海省翡翠湖水样中分离到一株新的嗜盐古菌病毒株,研究其生物学特性并进行分类。【方法】首先通过挑取单菌落法分离嗜盐古菌,通过噬菌斑法获得嗜盐古菌病毒,PEG 6000两步沉淀法和CsCl密度梯度离心对病毒颗粒进行浓缩和纯化,用醋酸双氧铀对病毒负染染色,在透射电镜下观察病毒形态,提取病毒基因组后进行测序并进行生物信息学分析,以三氯乙酸法制备病毒蛋白样品并进行SDS-PAGE分析,分别用考马斯亮蓝和苏丹黑B染色并观察其蛋白和脂质条带。【结果】在以Halorubrum属极端嗜盐古菌K2菌株为敏感菌的双层平板上分离到了一株嗜盐古菌病毒,其噬菌斑为浊斑,透射电镜下呈多形性包膜病毒状,直径60 nm左右;含有9333bp大小的双链环状DNA基因组,与已报道的β多形包膜病毒属(Betapleolipovirus)的HRPV11、HRPV12和HRPV10具有约75%的一致性,是该属的一个病毒新种。根据形态及基因组特征,将其归...     

嗜盐细菌噬菌体CJL-1的分离及其生物学特性研究

[目的]以嗜盐细菌为宿主,从运城盐湖水样中分离特异性噬菌体,并对其部分生物学特性进行研究。[方法]采用双层平板法分离获得噬菌体,并进行浓缩纯化。电镜观察噬菌体形态,以及限制性酶切分析、蛋白组成分析及特性研究。[结果]分离获得1株可感染Virgibacillus sp.GQ15的噬菌体CJL-1,其头部为正多面体结构,直径约110 nm,尾长约25 nm。其基因组为双链DNA,且衣壳中至少含有10种蛋白质,其中4种为主要结构蛋白:75 k Da、45 k Da、25 k Da和15 k Da。噬菌体CJL-1含有脂质包膜,且对低盐环境敏感。CJL-1最适MOI为1.0,最适保存温度为30~50℃,最适感染p H为7,且其宿主范围较广。[结论]噬菌体CJL-1属短尾噬菌体,为裂解性嗜盐噬菌体,研究结果可为运城盐湖极端微生物资源的开发利用提供参考。     

云南盐矿中病毒样颗粒的多样性

摘要:【目的】了解云南盐矿中病毒样颗粒的多样性及特征,为丰富嗜盐菌病毒资源及其生态功能研究奠定基础。【方法】采用电子显微镜直接观察、双层平板分离纯化对云南5 个盐矿样品病毒的分布和形态特征进行比较研究。【结果】电镜观察病毒富集液表明,其中含有3种不同形态的病毒样颗粒(头尾形、丝状、球状)。同时,分离获得3株嗜盐细菌病毒,其中2株感染盐单胞菌(Halomonas sp.),1株感染色盐杆菌(Chromoha lobacter sp.),分别命名为QHHSV-1、YPHSV-1 及YPCPV-1。QHHSV-1形成边缘清晰、透明的噬菌斑,出斑时间约为8 h,培养24 h,其噬菌斑直径可达3 mm,电镜观察显示,QHHSV-1呈头尾形,头部直径约为47 nm,其尾部极易断裂,长约75 nm;YPHSV-1形成边缘清晰、透明的噬菌斑,出斑时间约为12 h,培养24 h,其噬菌斑直径可达1 mm,电镜观察显示,YPHSV-1呈头尾形,头部直径约为50 nm,尾长约为140 nm;YPCPV-1形成边缘模糊、透光亦不易见的噬菌斑,出斑时间约为72 h,培养96 h,其噬菌斑直径可达2－4 mm,电镜观察显示,YPCPV-1呈球形,是表面有突起的多态形病毒,大小为20－50 nm。     

河南叶县岩盐可培养中度嗜盐菌的多样性

【背景】嗜盐微生物因为独特的生理和代谢特征而对高盐环境有着良好的适应能力,在环境污染治理、酶制剂等领域具有很高的应用和研究价值,是一类重要的极端环境微生物资源。【目的】为了更好地认识我国岩盐微生物的多样性,开发和利用嗜盐微生物资源,积累丰富的微生物菌种资源。【方法】在5%和10%的盐度下,使用Alkaline oligotrophic medium (AOM)、Neutral haloarchaeal medium (NHM)、Diluted modified marine agar (dmMA)和ISP3 medium (ISP3)四种培养基,分离和纯化河南叶县岩盐矿的卤水和盐土中的嗜盐菌,使用细菌通用引物27F和1492R扩增和测序纯化菌株的16SrRNA基因,使用Ez BioCloud和NCBI上的BLAST比对进行分子鉴定,使用MEGA5.0进行遗传进化分析。【结果】从河南叶县岩盐卤水和盐土中一共分离和纯化到78株细菌,菌株16S rRNA基因序列显示它们来自3个门:厚壁菌门(Firmicutes)的Bacillus 26株、Halobacillus 30株、Oceanobac...     

一株强裂解性大肠杆菌T1样噬菌体新成员的分离与鉴定

【目的】自然界中噬菌体种类繁多,其裂菌功能在针对细菌耐药方面具有潜在应用价值。不同噬菌体也呈现出显著的基因多样性及宿主特异性。从上海某猪场仔猪肠内容物样品中分离、纯化大肠杆菌的裂解性噬菌体,分析其生物学特性和病毒学特征,为探索应用噬菌体治疗细菌性感染提供研究材料。【方法】采用双层琼脂平板法分离、纯化噬菌体,观察噬菌斑特征,通过电镜观察噬菌体形态特征,测定其裂菌谱、最佳感染复数、一步生长曲线和生物学特性,进行噬菌体全基因组测序和遗传进化分析。【结果】分离、纯化获得一株能高效裂解大肠杆菌K-12菌株的噬菌体,命名为v B_Eco S_SH2(SH2),噬菌斑呈圆形、大而透明、边缘整齐。电镜观察SH2的头部呈二十面体立体对称,尾部较长。噬菌体的潜伏期为10 min,暴发期为60 min,裂解量高达121 PFU/感染细胞,其最佳感染复数为0.1。基因组测序和比对结果表明,SH2的核酸类型为ds DNA,基因组全长为49 088 bp,G+C%含量为45%,Gen Bank登录号为KY985004,结合电镜观察及BLASTp分析,确定其属于有尾噬菌体目长尾噬菌体科成员。同源性及进化分析表明,该噬菌体为大肠杆菌T1样噬菌体的新成员。【结论】分离鉴定了一株裂解效率极高的大肠杆菌T1样噬菌体,并确认其为T1样噬菌体新成员,为研究大肠杆菌噬菌体及其抗菌应用提供了新的实验材料。     

新疆罗布泊地区可培养嗜盐古菌多样性及其功能酶筛选

【目的】探索新疆罗布泊地区高盐环境可培养嗜盐古菌的多样性及其功能酶应用潜力。【方法】采集罗布泊地区13份土样,用纯培养并结合基于16S rRNA基因系统发育分析的方法来研究样品中嗜盐古菌的多样性。按系统进化树的聚类关系,挑选出一些菌株进行盐度耐受及淀粉酶、蛋白酶、酯酶的酶活检测。【结果】从13份土样中共分离到56株嗜盐古菌,经16S rRNA基因克隆测序,通过MEGA 4.0构建N-J树分析,56株菌分布于嗜盐古菌的10个生效发表属和5个潜在新属。运用Shannon-Wiener方法计算其多样性指数为1.820。挑选17株嗜盐古菌所测试盐浓度实验结果表明这一批嗜盐古菌的大部分生长范围在10%-35%之间,最适盐浓度在20%-25%之间。不同酶活检测结果为:淀粉酶酶活率为70.6%,蛋白酶酶活率为35.3%,酯酶酶活率为82.4%。【结论】新疆罗布泊周边地区由于气候及地理位置的独特性,蕴藏丰富的嗜盐古菌资源。本实验所设计的分离方法对嗜盐古菌的分离是极其有效的,为进一步研究新疆罗布泊及周边地区嗜盐古菌资源提供了技术基础。盐度耐受实验结果验证在低盐环境中分离嗜盐古菌新物种的可行性。同时,嗜盐古菌的酶活比率较高且活性较强为进一步开发利用嗜盐古菌资源提供了理论依据。     

嗜盐古菌噬菌体研究进展

嗜盐古菌噬菌体是噬菌体的一个重要分支,在群体生态学以及生命的起源与进化历程中扮演重要的角色.综述了嗜盐古菌噬菌体的形态多样性、研究方法、起源与进化几方面的研究报道,指出目前研究中存在的问题,对未来研究进行了展望.     

塔里木盆地荒漠盐碱生境嗜盐碱细菌的初步研究

为了探索塔里木盆地荒漠盐碱生境嗜（耐）盐碱细菌的分离方法,采用纯培养技术探讨了不同土壤预处理方法、盐度及不同分离培养基对不同盐度土壤中嗜（耐）盐碱细菌分离效果的影响。结果表明：高盐土壤嗜（耐）盐碱细菌的多样性高于中度盐分和低度盐分的土壤,而总菌落数则相反;半量的Horikoshi I（NaCl 10％～15％）为3种土样最佳的分离培养基,碱性复合培养基和高盐碱培养基A次之;分离嗜（耐）盐碱细菌以获得资源为主要目的时,富集培养法最佳。以反映土壤嗜（耐）盐碱细菌生态分布而言,用土壤悬液法;塔里木盆地嗜（耐）盐碱细菌生长盐浓度及pH值范围较宽,最适生长盐浓度为10％左右,pH值多为8—10左右。分离到的120株嗜（耐）盐碱细菌中,有33株为嗜盐碱细菌,占分离菌株的27．5％。     

Global network of specific virus-host interactions in hypersaline environments

Hypersaline environments are dominated by archaea and bacteria and are almost entirely devoid of eukaryotic organisms. In addition, hypersaline environments contain considerable numbers of viruses. Currently, there is only a limited amount of information about these haloviruses. The ones described in detail mostly resemble head-tail bacteriophages, whereas observations based on direct microscopy of the hypersaline environmental samples highlight the abundance of non-tailed virus-like particles. Here we studied nine spatially distant hypersaline environments for the isolation of new halophilic archaea (61 isolates), halophilic bacteria (24 isolates) and their viruses (49 isolates) using a culture-dependent approach. The obtained virus isolates approximately double the number of currently described archaeal viruses. The new isolates could be divided into three tailed and two non-tailed virus morphotypes, suggesting that both types of viruses are widely distributed and characteristic for haloarchaeal viruses. We determined the sensitivity of the hosts against all isolated viruses. It appeared that the host ranges of numerous viruses extend to hosts in distant locations, supporting the idea that there is a global exchange of microbes and their viruses. It suggests that hypersaline environments worldwide function like a single habitat.     

Gammaproteobacterial Diversity and Carbon Utilization in Response to Salinity in the Lakes on the Qinghai–Tibetan Plateau

The Gammaproteobacteria are widely and abundantly distributed in various environments, and they play important roles in the geochemical cycles of biogenic elements (e.g., C, N and S) in the ecosystems. Previous studies showed that Gammaproteobacteria dominate in saline and hypersaline lakes. However, little is known on how salinity influences gammaproteobacterial community composition and their ecological functions (i.e., organic carbon degradation). In this study, we investigated the gammaproteobacterial diversity and carbon utilization and their response to salinity in six saline/hypersaline lakes and one freshwater lake on the Qinghai–Tibetan Plateau (QTP). The results indicated that the gammaproteobacterial community composition was mainly influenced by the salinity of the studied QTP lakes. Salinity was also a key environmental factor influencing the carbon utilization ability of Gammaproteobacteria: within one genus (e.g., Halomonas, Pseudoalteromonas, Vibrio) the strains retrieved from low-salinity environments had stronger carbon utilization ability than their counterparts from high-salinity environments; within one genus, the strains isolated from lakes with different salinity shared similar carbon utilization preference, indicating that species belonging to the same genus may execute similar ecological functions in the environments regardless of salinity.     

Euryhalinity of Palaeozoic articulate brachiopods

OMonotypic and very low diversity virgianid shell beds from the Upper Ordovician to Lower Silurian dolomites of North Greenland were formed in marginal marine quiet-water hypersaline environments. In the light of this evidence the salinity tolerances of other Palaeozoic articulate brachiopods is evaluated. There are only a small number of species apparently invading hypersaline or brackish environments, but it is significant considering that previously all articulate brachiopods were thought to be fully marine. Two types of occurrence are noted, those species specifically related to marginally marine environments, disappearing with the introduction of fully marine faunas, and the majority of species which extend their normal marine range into marginal conditions. No brachiopod species appears to have invaded very hypersaline or truly brackish conditions. No single group of articulate brachiopods specifically specialised in colonising marginal marine environments, apart from possibly the virgianid pentamerids. Palaeozoic, Upper Ordovician, Lower Silurian, Brachiopoda. Pentamerida, Virgianidae, Greenland, palaeoecology. hypersaline environments, brackish environments .     

Diversity,taxonomy, and evolution of archaeal viruses of the class Caudoviricetes

The archaeal tailed viruses (arTV), evolutionarily related to tailed double-stranded DNA (dsDNA) bacteriophages of the class Caudoviricetes, represent the most common isolates infecting halophilic archaea. Only a handful of these viruses have been genomically characterized, limiting our appreciation of their ecological impacts and evolution. Here, we present 37 new genomes of haloarchaeal tailed virus isolates, more than doubling the current number of sequenced arTVs. Analysis of all 63 available complete genomes of arTVs, which we propose to classify into 14 new families and 3 orders, suggests ancient divergence of archaeal and bacterial tailed viruses and points to an extensive sharing of genes involved in DNA metabolism and counterdefense mechanisms, illuminating common strategies of virus–host interactions with tailed bacteriophages. Coupling of the comparative genomics with the host range analysis on a broad panel of haloarchaeal species uncovered 4 distinct groups of viral tail fiber adhesins controlling the host range expansion. The survey of metagenomes using viral hallmark genes suggests that the global architecture of the arTV community is shaped through recurrent transfers between different biomes, including hypersaline, marine, and anoxic environments.

Comparative genomics and host range analysis reveals the remarkable diversity and evolution of tailed archaeal viruses of the order Caudoviricetes, which together with their bacterial relatives arguably represent the most abundant and widespread virus group on our planet.     

Characterization of protistan plankton diversity in ancient salt evaporation ponds located in a volcanic crater on the island Sal,Cape Verde

Salinity is an important factor when exploring the limits known for life. Therefore, hypersaline systems have attracted much attention in recent years. In this study, we investigated the protistan diversity and community composition in two natural salt evaporation ponds (27–30% salinity) located in an ancient volcanic crater on the Cape Verde island Sal using high-throughput DNA sequencing. Our study revealed a broad range of protistan taxa and a high taxonomic diversity within the Ciliophora, Dinophyceae, and Chlorophyta. We detected a total of 23 Dinophyceae families, although Dinophyceae were generally considered to be only this diverse in aquatic environments of less than 10% salinity. Moreover, we uncovered a high degree of genetic novelty in this habitat. The mean similarity of all detected OTUs to previously described sequences was only 93.6%. These findings strongly dispute the traditional view that extreme hypersaline environments generally maintain low protistan diversity. A meta-analysis covering our and previously published data from other inland and coastal salt ponds clearly showed that our samples clustered according to salinity and not biogeography. This result further supports the claim that salinity is a major transition boundary for protistan communities, regardless of their biogeographic origin.

     

SALINITY TOLERANCE OF DIATOMS FROM THALASSIC HYPERSALINE ENVIRONMENTS

Thirty-four benthic diatom strains were isolated from thalassic hypersaline marine environments and their salinity tolerance characterized in growth experiments conducted at salinities ranging from 0.5% to 17.5% (weight of total salts per volume, g·100 mL ^{− 1}). The results were compared with the patterns of diatom species distribution and abundance in hypersaline evaporation ponds and tidal channels of Guerrero Negro, Baja California Sur, Mexico. The isolated strains were representative of the diatom assemblages present in the saltern ponds but were less so of natural assemblages in tidal channels. In general, we found a clear decreasing trend of diatom diversity in the field and in the isolated strains with increasing salinity. With some exceptions, the upper limit of salinity tolerance in cultivated strains corresponded to their distribution in field samples. However, the relative abundance of species in the field was not correlated with growth rates achieved in culture for the same salinities. Most cultured strains exhibited extreme euryhalinity growing well from brackish to hypersaline conditions, but the particulars of salt tolerance were quite diverse among strains. The most halotolerant taxa, two Amphora species, Amphora cf. subacutiuscula Schoeman, Nitzschia fusiformis Grunow, and Entomoneis sp., grew well in salinities ranging from 0.5% to 15%. Three strains of Pleurosigma strigosum W. Smith that were unable to grow in salinities less than 5% total salts represent the only true halophilic diatoms ever reported. The fact that many strains displayed a remarkable halotolerance, with optimal or near-optimal growth rates at salinities as high as three times that of seawater, implies that diatoms from hypersaline environments are evolutionarily highly adapted to such environments.     

Salt stress and plasma-membrane fluidity in selected extremophilic yeasts and yeast-like fungi

We have investigated changes in plasma-membrane fluidity in relation to NaCl concentrations in yeasts and yeast-like fungi that were isolated from either subglacial ice or hypersaline waters. In both of these natural environments, these organisms are exposed to low water activity, due to either high NaCl concentrations or low temperatures. Our data indicate that the fluidity of the plasma membrane can be used as an indicator of fitness for survival in extreme environments. Fungi that can survive in such extreme environments, such as Hortaea werneckii in the hypersaline waters of salterns, and Cryptococcus liquefaciens in subglacial environments, showed similar profiles of plasma-membrane fluidity in response to raised salinity. The same was seen for ubiquitous fungi, which are generally adapted for different types of stress, such as Aureobasidium pullulans and Rhodotorula mucilaginosa. Representatives of both of these groups modulated their plasma-membrane fluidity differently. When salinity exceeded their optimal range, the ubiquitous stress-tolerant species (A. pullulans, Rh. mucilaginosa) showed increased plasma-membrane fluidity, whereas in the dominant extremophiles (H. werneckii, Cr. liquefaciens), it decreased. On the other hand, the plasma membranes of the fungi with a narrow ecological amplitude (Arctic A. pullulans and Rhodosporium diobovatum) showed different responses.     

Biodegradation of organic pollutants by halophilic bacteria and archaea

Hypersaline environments are important for both surface extension and ecological significance. As all other ecosystems, they are impacted by pollution. However, little information is available on the biodegradation of organic pollutants by halophilic microorganisms in such environments. In addition, it is estimated that 5% of industrial effluents are saline and hypersaline. Conventional nonextremophilic microorganisms are unable to efficiently perform the removal of organic pollutants at high salt concentrations. Halophilic microorganisms are metabolically different and are adapted to extreme salinity; these microorganisms are good candidates for the bioremediation of hypersaline environments and treatment of saline effluents. This literature survey indicates that both the moderately halophilic bacteria and the extremely halophilic archaea have a broader catabolic versatility and capability than previously thought. A diversity of contaminating compounds is susceptible to be degraded by halotolerant and halophile bacteria. Nevertheless, significant research efforts are still necessary in order to estimate the true potential of these microorganisms to be applied in environmental processes and in the remediation of contaminated hypersaline ecosystems. This effort should be also focused on basic research to understand the overall degradation mechanism, to identify the enzymes involved in the degradation process and the metabolism regulation.     

High diversity and isolated distribution of aquatic heterotrophic protists in salars of the Atacama Desert at different salinities

The species richness of eukaryotes in the hypersaline environment is generally thought to be low. However, recent studies showed a high degree of phylogenetic novelty at these extreme conditions with variable chemical parameters. These findings call for a more thorough look into the species richness of hypersaline environments. In this study, various hypersaline lakes (salars, 1–348 PSU) as well as further aquatic ecosystems of northern Chile were investigated regarding diversity of heterotrophic protists by metabarcoding studies of surface water samples. Investigations of genotypes of 18S rRNA genes showed a unique community composition in nearly each salar and even among different microhabitats within one salar. The genotype distribution showed no clear connection to the composition of main ions at the sampling sites, but protist communities from similar salinity ranges (either hypersaline, hyposaline or mesosaline) clustered together regarding their OTU composition. Salars appeared to be fairly isolated systems with only little exchange of protist communities where evolutionary lineages could separately evolve.     

Salinity Regulation of the Interaction of Halovirus SNJ1 with Its Host and Alteration of the Halovirus Replication Strategy to Adapt to the Variable Ecosystem

Halovirus is a major force that affects the evolution of extreme halophiles and the biogeochemistry of hypersaline environments. However, until now, the systematic studies on the halovirus ecology and the effects of salt concentration on virus-host systems are lacking. To provide more valuable information for understanding ecological strategies of a virus-host system in the hypersaline ecosystem, we studied the interaction between halovirus SNJ1 and its host Natrinema sp.J7-2 under various NaCl concentrations. We found that the adsorption rate and lytic rate increased with salt concentration, demonstrating that a higher salt concentration promoted viral adsorption and proliferation. Contrary to the lytic rate, the lysogenic rate decreased as the salt concentration increased. Our results also demonstrated that cells incubated at a high salt concentration prior to infection increased the ability of the virus to adsorb and lyse its host cells; therefore, the physiological status of host cells also affected the virus-host interaction. In conclusion, SNJ1 acted as a predator, lysing host cells and releasing progeny viruses in hypersaline environments; in low salt environments, viruses lysogenized host cells to escape the damage from low salinity.