Genomic analyses of metal resistance genes in three plant growth promoting bacteria of legume plants in Northwest mine tailings,China

To better understand the diversity of metal resistance genetic determinant from microbes that survived at metal tailings in northwest of China, a highly elevated level of heavy metal containing region, genomic analyses was conducted using genome sequence of three native metal-resistant plant growth promoting bacteria(PGPB). It shows that: Mesorhizobium amorphae CCNWGS0123 contains metal transporters from P-type ATPase, CDF(Cation Diffusion Facilitator), Hup E/Ure J and CHR(chromate ion transporter) family involved in copper, zinc, nickel as well as chromate resistance and homeostasis. Meanwhile, the putative Cop A/Cue O system is expected to mediate copper resistance in Sinorhizobium meliloti CCNWSX0020 while Znt A transporter, assisted with putative Czc D, determines zinc tolerance in Agrobacterium tumefaciens CCNWGS0286. The greenhouse experiment provides the consistent evidence of the plant growth promoting effects of these microbes on their hosts by nitrogen fixation and/or indoleacetic acid(IAA) secretion,indicating a potential in-site phytoremediation usage in the mining tailing regions of China.     

铜胁迫对小麦根系微域微生物群落的影响

小麦是我国主要的粮食作物,土壤重金属污染会严重威胁作物生长和粮食安全.大量研究表明根际微生物在调控作物发育和抗逆性方面具有非常重要的作用.因此,本研究通过高通量测序技术研究铜胁迫条件下小麦根际微生物群落的变化,从微生物学角度揭示铜污染胁迫对小麦根际的影响作用.采用盆栽培养试验对铜污染处理中小麦非根际、根际以及根内微生物...     

攀枝花矿区糙野青茅根际耐铬放线菌筛选及促生能力评价

从攀枝花矿区糙野青茅根际土中分离放线菌,筛选获得对重金属铬具有强耐受性、抗菌促生和产胞外酶能力的放线菌,为微生物-植物联合修复重金属污染提供菌种资源.采用稀释涂布法进行放线菌的分离,通过放线菌在不同铬浓度高氏一号培养基上的生长状况判断其对重金属铬的耐受性,进一步采用原子吸收分光光度法分析菌株对铬耐受性强的菌株耐铬及去铬能力;通过对菌株产铁载体、溶磷、产吲哚乙酸(IAA)、产纤维素酶、产淀粉酶、产木糖酶、产酪蛋白酶能力及抗菌活性的筛选,综合评价菌株抗菌促生能力.结果显示,分离获得的27株根际菌株,光学显微形态及扫描电镜结果均显示为放线菌,其中有14株能在含铬的培养基上生长,具有耐铬能力.16S rRNA基因测序结果显示,14株放线菌分属于链霉菌属(streptomyces)、小单孢菌属(Micromonospora)和冢村氏菌属(Tsukamurella).菌株SCAU9002和SCAU9008在铬浓度为300mg·L~(-1)的处理下培养6 d,去铬率分别达到81.17%和78.41%;菌株SCAU9002、SCAU9006、SCAU9008、SCAU9009、SCAU9013同时具有3种促生功能,盆栽实验结果进一步表明,在重金属胁迫下,经菌株处理后的玉米植株更高,生物量更大,根系更发达,促生效果较好.此外,菌株SCAU9006对6种病原菌具有拮抗作用,5株菌同时具有3种及以上产胞外水解酶的能力,4株菌能在7%的NaCl浓度下生长,13株菌在pH5.0~8.0的范围内均可生长.结果表明,根际放线菌在促进矿区植物生长和铬尾矿污染土壤的生物修复研究中具有很大潜力.     

海州香薷对铜的蓄积及铜的毒性效应

研究了不同浓度铜对海州香薷的毒性及海州香薷对铜的蓄积结果表明0.5 mmol/kg的铜能促进海州香薷的生长,使其生物量增大.低浓度的Cu²⁺使叶绿素a含量上升,而高浓度Cu²⁺则使叶绿素a下降,但Cu²⁺对叶绿素b的影响并不明显.反映到a/b上则表现为低浓度下Cu²⁺处理会提高a/b值.海州香薷的蓄铜浓度随土壤中铜浓度的增加而升高,可达1500mg/kg,达到了超量积累植物的标准.铜对SOD、POD活性的影响也表现为低浓度的刺激效应和高浓度的抑制效应,最高值出现在0.5 mol/kg.     

南京市铅锌矿采矿场土壤重金属污染评价及优势植物重金属富集特征

为了筛选可用于修复土壤重金属污染的植物材料,测定了南京市某铅锌矿采矿场土壤及样地内14种优势植物中Cd、Cr、Cu、Mn、Pb、Zn的含量,并分析了采矿场土壤重金属的污染状况和14种植物对这6种重金属元素的富集和转运能力.结果表明,矿区土壤中Cd、Mn、Zn、Pb的污染最为严重,单因子污染指数分别达到了45.71、11.68、10.40和4.46;样地的内梅罗综合污染指数为33.45,也达到重度污染级别.尽管各种重金属的含量不同,14种优势植物均表现出了较强的重金属耐受能力,其中井栏边草(Pteris multifida)和络石(Trachelospermum jasminoides)对6种重金属均表现出很高的富集能力.所有植物体内Zn含量均超出了正常范围,但仅马唐(Digitaria sanguinalis)对Zn的生物富集系数大于1,其他植物的重金属生物富集系数均小于1.14种植物对重金属的生物转移系数较高,其中菊芋(Helianthus tuberosus)和野菊(Dendranthema indicum)对6种重金属的生物转移系数均大于1.根据植物对重金属的吸收机制,可将14种植物分为3类:富集型植物,如菊芋(H.tuberosus)、野菊(D.indicum)、垂序商陆(Phytolacca americana)、爵床(Justicia procumbens)、马唐(D.sanguinalis)、山莴苣(Sonchus brachyotus)、龙葵(Solanum nigrum)和狗尾草(Setaria viridis);根部囤积型植物,如井栏边草(P.multifida)和络石(T.jasminoides);规避型植物,如一枝黄花(Solidago decurrens)、蛇莓(Duchesnea indica)、青绿苔草(Carex breviculmis)和贯众(Cyrtomium fortunei).     

Biosorption of copper and zinc by immobilised and free algal biomass, and the effects of metal biosorption on the growth and cellular structure of Chlorella sp. and Chlamydomonas sp. isolated from rivers in Penang, Malaysia

In this study, the biosorption of copper and zinc ions by Chlorella sp. and Chlamydomonas sp. isolated from local environments in Malaysia was investigated in a batch system and by microscopic analyses. Under optimal biosorption conditions, the biosorption capacity of Chlorella sp. for copper and zinc ions was 33.4 and 28.5 mg/g, respectively, after 6 hr of biosorption in an immobilised system. Batch experiments showed that the biosorption capacity of algal biomass immobilised in the form of sodium alginate beads was higher than that of the free biomass. Scanning electron microscopy and energy-dispersive X-ray spectroscopy analyses revealed that copper and zinc were mainly sorbed at the cell surface during biosorption. Exposure to 5 mg/L of copper and zinc affected both the chlorophyll content and cell count of the algal cells after the first 12 hr of contact time.     

云南兰坪铅锌矿区优势植物重金属富集特性及应用潜力

为筛选适合云南高原地区重金属污染土壤修复治理以及矿区生态复垦的植物材料,通过实地野外调查,采集云南兰坪铅锌矿区18种优势植物样品及相应根系土壤,测定其Cd、 Cr、 Cu、 Ni、 Pb和Zn含量,计算植物对重金属的富集和转运系数,通过聚类分析方法,综合评估植物的应用潜力.结果表明, 18种优势植物分属18属13科,均为草本植物.植物体内多种重金属含量均高于植物正常含量,其中,延胡索(Corydalis yanhusuo)、旱芹菜(Carum bretschneideri)和毛连菜(Picris hieracioides)地上部Cd含量分别达到62.29、 76.49和85.09 mg·kg^-1,Cd富集系数分别为0.57、 0.58和0.66,转运系数分别为0.89、 0.45和1.48,有着较强的吸收和转移土壤重金属Cd能力,具有修复Cd污染土壤潜力.狼毒(Euphorbia fischeriana)和倒提壶(Cynoglossum amabile)地上部Cd含量分别为0.46 mg·kg^-1和0.23 mg·kg^-1     

螯合剂对铜毒性的影响

研究了螯合剂EDTA和DTPA对海州香薷蓄铜能力和铜毒性的影响.结果显示,EDTA和DTPA能抑制植物的生长,使其生物量降低,叶绿素含量下降,同时降低了植物的根系活力.EDTA和DTPA都可引起植物SOD、POD活性上升,在与Cu²⁺共同作用下,EDTA对Cu²⁺毒性影响较小,表现在SOD、POD活性仍按Cu²⁺独立作用下的变化规律而变化,且数值上变化不大.而DTPA则降低了Cu²⁺作用下SOD、POD活性的变化.EDTA使海州香薷地上部分铜浓度增加,DTPA则明显降低了海州香薷的铜含量.     

The role of arbuscular mycorrhiza on change of heavy metal speciation in rhizosphere of maize in wastewater irrigated agriculture soil

To understand the roles of mycorrhiza in metal speciation in the rhizosphere and the impact on increasing host plant tolerance against excessive heavy metals in soil, maize ( Zea mays L. ) inoculated with arbuscular mycorrhizal fungus ( Glomus mosseae) was cultivated in heavy metal contaminated soil. Speciations of copper, zinc and lead in the soil were analyzed with the technique of sequential extraction. The results showed that, in comparison to the bolked soil, the exchangeable copper increased from 26% to 43% in non-infected and AM-infected rhizoshpere respectively; while other speciation (organic, carbonate and Fe-Mn oxide copper) remained constant and the organic bound zinc and lead also increased but the exchangeable zinc and lead were undetectable. The organic bound copper, zinc end lead were higher by 15%, 40% and 20%, respectively, in the rhizosphere of arbuscular mycorrhiza infected maize in comparison to the non-infected maize. The results might indicate that mycorrhiza could protect its host plants from the phytotoxicity of excessive copper, zinc and lead by changing the speciation from bio-available to the non-bio-available form. The fact that copper and zinc accumulation in the roots and shoots of mycorrhia infected plants were significantly lower than those in the non-infected plants might also suggest that mycorrhiza efficiently restricted excessive copper and zinc absorptions into the host plants. Compared to the non-infected seedlings, the lead content of infected seedlings was 60% higher in shoots. This might illustrate that mycorrhiza have a different mechanism for protecting its host from excessive lead phytotoxicity by chelating lead in the shoots.     

极端暴雨条件下广东北江重金属污染特征

以2012年“6.25”暴雨径流过程为对象,研究广东省北江上游高桥断面在极端暴雨条件下水体理化性质与ρ(Pb)、ρ(Cu)、ρ(Zn)、ρ(Cd)(均为溶解态,下同)的变化过程;基于相关分析法,分析北江水质对水文过程的响应关系及各指标间的内联特点. 结果表明:暴雨径流期间,北江水质呈“大流量、高浓度”的特征. 暴雨径流发生前后4个典型阶段中,阶段Ⅰ(局部产流期)ρ(Cd)、ρ(Pb)仅为(0.48±0.09)和(1.91±0.29)μg/L,ρ(Cu)与ρ(Zn)基本未检出;阶段Ⅱ(以地表径流为主的全流域产流期)ρ(Cd)、ρ(Pb)、ρ(Cu)与ρ(Zn)均迅速上升,最高分别达1.80、12.34、6.18、52.57 μg/L;阶段Ⅲ(以浅层地下水径流为主的全流域产流期)重金属质量浓度整体较洪水期有所下降,但ρ(Pb)仍高于阶段Ⅰ正常水平,ρ(Zn)、ρ(Cu)和ρ(Cd)则低于阶段Ⅰ;阶段Ⅳ(深层地下水产流期)ρ(Cu)与ρ(Zn)已回落至未检出水平,ρ(Cd)持续下降,低于阶段Ⅲ和阶段Ⅰ,ρ(Pb)较阶段Ⅲ有所下降,但仍高于阶段Ⅰ. 暴雨径流期间,溶解态重金属质量浓度与流量及浊度间呈显著正相关(P<0.01),ρ(Zn)与ρ(Pb)、ρ(Cd)的相关系数均在0.85以上(P<0.01).      

铜尾矿白羊草重金属含量对叶际和根际真菌群落的影响

植物的叶际和根际微生物与宿主植物之间存在复杂的相互关系,能促进植物的生长,增强宿主植物对逆境胁迫的耐受性.本实验选择铜矿区主要恢复植被白羊草为研究对象,采用高通量测序的方法,分析白羊草叶际和根际真菌群落结构与多样性差异,探究白羊草叶片和根部富集的重金属含量对叶际和根际真菌群落特征的影响.结果表明,在白羊草的根际和叶际区域,子囊菌门（Ascomycota）和担子菌门（Basidiomycota）均为优势菌门.白羊草根际真菌群落的多样性和丰富度显著高于叶际真菌.白羊草植株叶际和根际真菌多样性受到不同重金属的影响,白羊草叶际多样性主要受到叶片Zn和Cu含量的影响,而根Pb含量是影响根际真菌群落多样性的关键因子.此外,叶际子囊菌门的孢菌科与叶片Cd含量有极显著正相关关系,根际丛赤壳科与根Zn含量显著正相关,可作为重金属污染区域生态恢复的指示菌种.本研究为铜尾矿生态恢复过程中发掘和利用叶际或根际真菌资源提供科学依据,也为筛选重金属污染区生态修复的植物-微生物共生体提供基础.     

Spatial characterisation of multi-level in-use copper and zinc stocks in Australia

A methodology has been developed to characterise the in-use stocks of copper and zinc at a variety of spatial levels. The approach employs representative concentrations of copper and zinc in their main in-use reservoirs (which account for virtually all the metal put into service) together with geographic information system (GIS) data sets of the spatial locations and densities of these reservoirs. The authors have applied this methodology to Australia at four spatial levels: central city, urban region, states/territories, and country, to produce what is believed to be the first multi-level spatial characterisations of the in-use stocks of technological materials. The results are presented quantitatively and as a series of stock density maps for Inner Sydney, Sydney Metro, all Australian states/territories, and Australia itself. The total stocks in Australia are estimated at about 4.3 Tg Cu (4.3 thousand million kg) and 3.8 Tg Zn (3.8 thousand million kg), or about 240 kg Cu/capita and 205 kg Zn/capita. A statistical analysis of the data shows that the metal stock density at a given spatial level is largely determined by a small number of high-density components at the next lower level. The spatial analysis of the in-use stocks indicates that 50% of all copper and zinc stock resides in just 10% of Australia's local government areas. The largest stocks occur in large urban regions, which can contain copper and zinc densities more than a hundred times higher than rural areas. These regions are expected to be major Australian “metal mines” in the future.     

铜污染对土壤细菌群落结构及重金属抗性基因的影响

重金属Cu在土壤中长期积累会对微生物产生毒害作用,为了探究Cu污染对土壤细菌群落及重金属抗性基因的影响,本研究建立了不同Cu浓度(50、100、200、300、500、750和1000 mg·kg-1)的土壤微宇宙,基于高通量测序和荧光定量PCR技术分析比较不同Cu浓度下培养14d后土壤细菌群落及重金属抗性基因的变化....     

铜尾矿白羊草叶际和根际细菌群落特征

植物叶际、根际微生物及其生存环境共同形成复杂的生态系统,其中,根际微生物也是土壤物质循环的主要动力,可以为植物的生长发育提供基础.叶际和根际微生物也可作为生态学指标,在矿区生态稳定与生态恢复中具有显著作用.本研究选择铜尾矿优势禾草白羊草为研究对象,采用高通量测序的方法,研究白羊草叶际和根际细菌群落特征,探究影响白羊草叶际和根际微生物群落结构与多样性的关键生态因子.结果表明,白羊草叶际和根际细菌群落结构具有显著差异.其中,白羊草叶际细菌优势属包括假单胞菌属、肠杆菌属和鞘脂单胞菌属,根际细菌优势属为Solrubrobacter和酸杆菌属.白羊草根际细菌群落的香农指数、ACE指数和Chao1指数均显著高于叶际细菌.土壤水分含量、p H、土壤砷和锌含量,白羊草总氮和总硫,及叶片和根内的镉和铬是影响白羊草叶际和根际优势细菌属的主要生态因子.此外,根际细菌群落香农指数与根内铜含量显著负相关,辛普森指数与根的总氮含量显著正相关,叶际细菌群落ACE指数与叶片总硫含量显著正相关.本研究结果可为铜尾矿生态恢复中发掘和利用叶际或根际细菌资源及提高矿区生态修复效率提供科学依据.     

基于宏基因组技术分析MBR膜清洗后污泥中抗性基因

污水处理厂作为抗生素抗性基因（antibiotic resistance genes,ARGs）的重要储存库,是自然界ARGs的主要来源之一.膜生物反应器（membrane bioreactor,MBR）被认为是一种能够有效去除污水处理厂中ARGs的技术工艺.MBR膜截留的废水中胶体、颗粒物、悬浮物及微生物代谢物中存在着大量的病原菌与抗性基因,而目前关于膜清洗后污泥中抗性基因的分布特征和规律尚不明确.本文采用宏基因组技术对MBR膜清洗后污泥中抗性基因进行了分析.结果显示,膜清洗后污泥中共检测出39门,其中优势菌门为Proteobacteria、Nitrospirae和Actinobacteria,优势菌属为Nitrospira、Pseudomonas和Bradyrhizobium.污泥样品含有的病原菌属占所有菌属的10.54%,其中Pseudomonas属相对丰度最高,占到所有菌属的3.94%.样品中共注释出17类ARGs和16类金属抗性基因（metal resistance genes,MRGs,15类单金属抗性基因和1类多重金属抗性基因）.其中,多药类抗生素抗性基因相对丰度最高,占49.08%.金属抗性基因中多重金属类抗性基因相对丰度最高,占该污泥样品的34.58%,单金属抗性基因中对铜的抗性基因数量最多,占19.99%.该膜清洗后污泥中微生物群落最主要的功能通路为代谢相关,并存在大量与人类疾病相关的代谢通路相关基因,其中涉及细菌耐药和细菌传染疾病的基因数量最多,分别为占人类疾病相关的代谢通路已注释序列的34.50%和16.62%.由此可见,膜清洗后污泥中蕴藏着丰富的ARGs、MRGs以及病原菌属,具有潜在的环境健康风险,需要加强对膜清洗后污泥中ARGs、MRGs以及病原菌的管控.本文为选择合适的技术工艺有效去除膜清洗后污泥中ARGs、MRGs以及病原菌提供指导.     

田间条件下养分调控八宝景天Cd修复效率

在田间条件下,采用四因素三水平正交试验研究了N、 P、 K三因素不同水平对Cd富集植物八宝景天的生长和Cd吸收性的影响.结果表明,八宝景天生物量随施N水平的提高而增加,高N水平下(337.5 kg·hm^-2)八宝景天地上部生物量最大,为不施肥处理的0.86～2.00倍,但对其地上部Cd含量无显著影响.K肥对八宝景天Cd吸收具有一定的促进作用,高K水平下其地上部Cd含量最高达22.05 mg·kg^-1,较不施肥处理提高29.3%.综合分析,NPK肥料配施条件下,N肥和K肥分别是八宝景天生长和Cd吸收效率的主要影响因素,且对八宝景天Cd积累表现出明显的交互作用.肥料对八宝景天Cd提取量影响顺序为N>K>P,其中对八宝景天Cd修复效率的促进作用主要通过提高八宝景天生物量,高量N肥配施适量P、 K肥是提高八宝景天Cd修复效率的最佳施肥措施,其地上部Cd积累量较不施肥处理提高了0.9～2.2倍,合理增施N肥是提高八宝景天Cd修复效率的主要影响因子.     

Cluster formation in liverwort-associated methylobacteria and its implications

Pink-pigmented methylotropic bacteria of the genus Methylobacterium inhabit the surfaces of plant organs. In bryophytes, these methylobacteria enhance cell growth, but the nature of this plant–microbe interaction is largely unknown. In this study, methylobacteria were isolated from the upper surface of the free-living thalli of the liverwort Marchantia polymorpha L. Identification of one strain by 16S ribosomal RNA (rRNA) gene-targeted polymerase chain reaction (PCR) and other data show that these microbes represent an undescribed species of the genus Methylobacterium (Methylobacterium sp.). The growth-promoting activity of these wild-type methylobacteria was tested and compared with that of the type strain Methylobacterium mesophilicum. Both types of methylobacteria stimulated surface expansion of isolated gemmae from Marchantia polymorpha by about 350%. When suspended in water, the liverwort-associated bacteria (Methylobacterium sp.) formed dense clusters of up to 600 cells. In liquid cultures of Methylobacterium mesophilicum, single cells were observed, but no clustering occurred. We suggest that the liverwort-associated methylobacteria are co-evolved symbionts of the plants: Cluster formation may be a behavior that enhances the survival of the epiphytic microbes during periods of drought of these desiccation-tolerant lower plants.     

华中地区水库沉积物重金属抗性基因的赋存特征及影响因素分析

环境中重金属抗性基因（Metal resistance genes,MRGs）的迁移转化作为环境及生态领域的重要研究内容近年来备受关注,但目前环境中重金属是否是MRGs的重要驱动因素则尚不明确.本研究采用宏基因组技术对华中地区水库沉积物MRGs进行了定量,并分析了不同形态重金属在MRGs迁移转化过程中起到的作用.结果表明,trcop （抗Cu基因）及arsM （抗砷基因）是水库环境中的主要MRGs,供水水库之间MRGs分布并无地域性差异.czrA （多抗基因）的潜在宿主菌较为广泛,trcop及arsM很可能位于同一种可移动遗传元件上进行水平转移.重金属总量与MRGs之间并无明显联系,而可交换态及碳酸盐结合态（B1）则是环境中MRGs的重要驱动因素.     

乐安江沉积物重金属污染及生态风险性评价

对１９８７－１９９５年期间所采集的乐安江表层沉积物中的重金属含量进行了统计分析和生态风险性评价。结果表明,德兴铜矿的矿山酸性废水和洎水河河水,使乐安江沉积物受到重金属的严重污染。     

Excessive Cu and Zn affecting on distribution of the metals and activities of glycolytic and nitrogen incorporating key enzymes in mycelia of ectomycorrhizal fungi Suillus bovinus

ＩｎｔｒｏｄｕｃｔｉｏｎＥｃｔｏ ｍｙｃｏｒｒｈｉｚａｌｆｕｎｇｉａｒｅｋｎｏｗｎｔｏａｍｅｌｉｏｒａｔｅｍｅｔａｌｔｏｘｉｃｉｔｙｔｏｔｈｅａｓｓｏｃｉａｔｅｄｈｉｇｈｅｒｐｌａｎｔｓ (Ｂｒａｄｌｅｙ ,1981;1982 ;Ｂｒｏｗｎ ,1985;Ｂｕｃｋｉｎｇ ,1995) .Ｔｈｅｉｍｐｒｏｖｅｍｅｎｔｏｆｈｅａｖｙｍｅｔａｌｔｏｌｅｒａｎｃｅｂｙｍｙｃｏｒｒｈｉｚａｌｉｎｆｅｃｔｉｏｎｉｓｒｅｌａｔｅｄｔｏａｒｅｄｕｃｔｉｏｎｏｆｓｈｏｏｔｔｉｓｓｕｅｃｏｎｃｅｎｔｒａｔｉｏｎ ,ｃｏｕｐｌｅｗｉｔｈａｎｉｎｃｒｅａ…