泡囊假单胞菌的分离及鉴定

从一例心包积液患者的心包液中,分离到一株氧化代谢、氧化酶阳性,具单极毛运动的革兰氏阴性杆菌。经形态、生理生化特性鉴定,DNA的G+C mol％测定和Biolog自动系列的验证,确认为泡囊假单胞菌(Pseudomonas vesieularis)。指出该菌可使免疫力低下患者发生感染。对该菌分类地位的变迁进行了讨论。     

我国污染土壤生物修复技术的发展及现状

本文简要回顾了近30年来我国污染土壤生物修复技术的发展过程,主要包括生物修复技术在我国的发展阶段、生物修复的4大类型及其所适用对象与范围、生物修复用菌株的筛选与特性研究、活性菌株(菌剂)在典型污染土壤中的应用及其效果等,并针对土壤污染和生物修复技术的发展现状,简要讨论了未来生物修复技术的发展。     

凋落物分解对土壤生物的影响

凋落物分解是生态系统物质循环和能量流动的重要环节,而土壤生物是凋落物分解的主要完成者.大量研究分析了土壤生物在凋落物分解中的作用,然而有关凋落物分解对土壤生物影响的研究则相对较少,致使我们对凋落物分解和土壤生物的相互作用了解依然不够深入.本文综述了凋落物对土壤微生物和土壤动物的影响,并进一步探讨了其影响机制.凋落物对土壤微生物的影响与凋落物类型或组成、在土壤中的掩埋位置及其破碎化程度紧密相关;大多数研究表明凋落物对土壤动物群落结构有明显影响;凋落物对土壤生物的影响主要通过直接改变土壤生物的食源和生境.今后需要加强跨区域长期定位实验研究,注重整合研究凋落物分解和生态系统过程,深入研究凋落物分解与土壤生物的相互作用机制.     

接种促生菌对花生根际土壤微生物及营养元素的影响

植物根际促生菌是一类可促进植物生长的有益细菌,有效的根际促生菌剂可以减少化肥施用。以束村氏菌属(Tsukamurella sp.)P9、伯克霍尔德氏菌属(Burkholderia sp.)P10、以及P9和P10混合菌液作为接种菌株,研究促生菌对花生生长、植株及土壤营养、根际土壤微生物类群及功能的影响。30 d盆栽实验结果表明,接种组的花生鲜重、株高及根长均显著提高;根际土壤细菌总数、固氮菌和溶磷菌数均明显高于未接种组;氮循环功能菌群数量有不同程度提高,土壤蔗糖酶、脲酶及过氧化氢酶均高于对照;土壤碱解氮及速效钾显著提高,植株营养指标有所提升,尤以P10接种效果更优。本研究初步结论表明2株促生菌通过活化土壤微生物、提高植株的有效营养元素含量,促进了花生的生长。     

草本凋落物与尿素联合修复对油污土壤生物化学性质的影响

为研究植物残体配合施氮对石油污染土壤生物学和化学性质的综合修复能力,以紫花苜蓿(Medicago sativa)、铁杆蒿(Artemisia gmelinii)和小冠花(Coronilla varia)3种广泛分布于陕北石油污染区的草本植物凋落物为对象,分别在配合施氮调节土壤C∶N为25∶1和不配合施氮的条件下,将其与45.37 g/kg的重度石油污染土壤混合,在20—25℃、恒湿条件下进行为期180 d的室内修复试验,检测上述处理对油污土壤微生物数量、11种土壤水解酶和氧化还原酶活性以及速效N、P和K含量的影响。结果表明:(1)3种凋落物处理均可显著提高污染土壤中放线菌和真菌数量,蔗糖酶、淀粉酶、纤维素酶、木聚糖酶、脲酶、蛋白酶、磷酸酶、多酚氧化酶、过氧化氢酶和过氧化物酶活性以及速效养分含量,并显著降低土壤总石油烃含量。(2)配合施氮总体上显著强化了凋落物对污染土壤生化性质的修复作用,但对凋落物处理下木聚糖酶、脲酶、蛋白酶和脱氢酶活性和微生物数量的恢复可能产生不利影响。(3)单纯使用凋落物作为调理剂可以更为全面的修复油污土壤受损生化性质,具有高N和P含量、较低C/N、C/P比以及较低...     

假单胞菌碱性木聚糖酶的纯化及性质

假单胞菌(Pseudomonas)G62可产生两种胞外木聚糖酶,即XynA和XynB。经过硫酸铵沉淀、阴离子和阳离子交换层析、分子筛色谱,最终得到 两种电泳纯酶。XynA的分子量及等电点分别为42kD和91,XynB的分子量和等电点分别是 20kD和88。经薄层色谱分析证明,两酶以不同的方式水解木聚糖,但都不产生木糖,即 两酶都为内切酶,它们的最适作用温度均为50℃。XynA的最适作用pH为7.0～9.8,而XynB的为7.0～7.5。在65℃时的半寿期XynA为6 min,XynB为140 min。XynA的Km和Vmax分别是5.56 mg·ml-1和543μmol·min-1·mg-1,XynB的Km和Vmax分别是7.72 mg·ml-1和819μmol·min-1·mg-1。两酶受Cu2+、Fe3+、Pb2+、Zn2+和Hg2+强烈抑制。化学修饰的初步结果表明,两酶的活性位点氨基酸均含有色氨酸和羧基氨基酸。     

沼泽红假单胞菌对健康小鼠肠道微生物的影响

探讨不同剂量沼泽红假单胞菌对健康小鼠生理功能及肠道微生物结构的影响。

将48只C57/BL6小鼠随机分为对照组(无菌生理盐水)以及沼泽红假单胞菌低剂量组(5×10⁸ CFU/mL)、中剂量组(5×10⁹ CFU/mL)、高剂量组(5×10¹⁰ CFU/mL), 每组12只; 不同剂量灌胃4周后, 测定小鼠胃泌素(Gas)、生长抑素(SS)含量, 观察肠道形态, 并应用Illumina-MiSeq测序平台进行16S rRNA基因V3-V4区测序, 分析其肠道微生物群落结构、多样性及组间差异。

与对照组相比, 低剂量组对促进小鼠Gas分泌差异具有统计学意义(t=6.426, P < 0.01), 高剂量组对抑制小鼠SS分泌差异具有统计学意义(t=3.711, P < 0.05);对照组小鼠回肠肠绒毛高度差异无统计学意义(P > 0.05), 且无肠道炎症刺激反应。沼泽红假单胞菌干预后细菌丰富度和多样性增加, 在门水平上, 与对照组相比, 实验组小鼠肠道菌群拟杆菌门丰度显著降低, 厚壁菌门丰度显著增加(均P < 0.05);在属水平上, 与对照组相比, 实验组小鼠肠道菌群Lachnospiraceae NK4A136 group、乳杆菌属、拟杆菌属、理研菌属和Prevotellaceae UCG-001相对丰度显著增加(均P < 0.05)。

沼泽红假单胞菌能够维持胃肠激素稳态和肠道形态结构, 且对健康小鼠肠道菌群的丰富度和组成产生一定的影响, 维持肠道平衡与稳定。

     

胞外和胞内菌胞外囊泡的功能及应用

细胞外囊泡(Extracellular Vesicles,EVs)是从细胞膜上脱落或者分泌的双层膜结构的囊泡状小体.真核生物、细菌、古细菌和支原体等具有细胞结构的生物均能够释放EVs.细菌分泌的EVs含有DNA、RNA及蛋白质等多种成分,其在细菌毒力保持、免疫逃逸、细菌间物质运输、宿主细胞免疫调节、宿主转录基因调节、耐...     

假单胞菌WBC—3甲基对硫磷水解酶性质的初步研究

从最近分离到的有机磷农药降解菌Pseudomonas sp．WBC—3中获得了甲基对硫磷水解酶（Methyl parathion hydrolase,MPH,EC3．1．8．3)。该酶在48h的培养物中分布比例分别为：上清液2．1％,胞内86．2％和胞间质11．7％,说明MPH为胞内酶。经过CM—sepharose Fast Flow阳离子交换层析,获得电泳纯的酶。SDS—PAGE和凝胶过滤层析表明,该酶为单体蛋白,分子量约为34kD。动力学分析显示该酶为非特异性有机磷降解酶,但最适底物为甲基对硫磷。在pH9～12范围,酶表现较高活力水平,最高活力的反应温度为40℃。根据各类金属离子和鳌合剂对酶活的影响,推测MPH为金属酶。     

耐砷促生菌对超富集植物蜈蚣草砷吸收及根际微生物群落的调控作用

【目的】分析添加外源促植物生长微生物(plant growth-promoting bacteria,PGPB)对植物生长、砷富集和根际微生物的影响,为植物-微生物联合修复砷污染土壤提供参考。【方法】通过添加外源PGPB,研究蜈蚣草生物量和砷富集量与外源微生物促植物生长特性的关系,采用高通量测序技术分析蜈蚣草根际微生物群落在外源PGPB干预下的变化规律。【结果】2株根际菌(假单胞菌PG12、芽孢杆菌R19)和1株内生菌(恶臭假单胞菌S6)具备典型的促植物生长特性,对蜈蚣草的促生作用顺序为：PG12>S6>R19,与对照组相比,生物量分别提高了234%(P<0.01)、136%(P<0.01)和67%;添加外源PGPB后,蜈蚣草砷含量从对照的18.50 mg提高到了31.25-46.95 mg,增幅高达153%(PG12)和139%(S6),对应的蜈蚣草砷浓度从2616.34mg/kg降至1348.04-2 156.23 mg/kg,呈现出典型的砷“稀释效应”;α多样性指数Sobs、Chao和Ace显示,仅R19处理显著提高了根际微生物的群落多样性,而β多样性指数...     

Nickel localization and response to increasing Ni soil levels in leaves of the Ni hyperaccumulator Alyssum murale

We have previously developed phytoremediation and phytomining technologies employing Alyssum Ni hyperaccumulators to quantitatively extract Ni from soils. Implementation of these technologies requires knowledge of Ni localization patterns for the Alyssum species/ecotypes of interest under realistic growth conditions. We investigated Ni uptake and localization in mature Alyssum murale Kotodesh and AJ9ç leaves. Seedlings were grown in potting mix with an increasing series of NiSO₄ addition (0, 5, 10, 20, 40, 80 mmol Ni kg^–1), NiC₄H₆O₄ addition (0, 5, 10, 30, 60, 90 mmol Ni kg^–1), in Ni-contaminated soil from metal refining operations, and serpentine soil. Plants at Ni levels 0, 5, 10, 20 mmolkg^–1 and in native soils grew normally. Plants at 40 mmolkg^–1 exhibited the onset of phytotoxicity, and 60, 80, and 90 mmolkg^–1 were demonstrably phytotoxic, but symptoms of phytotoxicity abated within 6 months. Cryogenic complement fractures were made from frozen hydrated samples. High-resolution scanning electron microscope (SEM) images were taken of one half. The other half was freeze-dried and examined with SEM and semi-quantitative energy dispersive x-ray analysis. Ni was highly concentrated in epidermal cell vacuoles and Ni and S counts showed a positive correlation. Trichome pedicles and the epidermal tissue from which the trichome grows were primary Ni compartments, but Ni was not distributed throughout trichomes. Palisade and spongy mesophyll and guard/substomatal cells contained lesser Ni concentrations but palisade mesophyll was an increasingly important compartment as Ni soil levels increased. Ni was virtually excluded from vascular tissue and trichome rays.     

Salt (NaCl) tolerance in the Ni hyperaccumulator <Emphasis Type="Italic">Alyssum murale</Emphasis> and the Zn hyperaccumulator <Emphasis Type="Italic">Thlaspi caerulescens</Emphasis>

Many metal hyperaccumulating plants have to tolerate abiotic stresses in their native soils such as high metal concentrations, low nutrient status and drought. This paper tests the ability of the Ni-hyperaccumulator Alyssum murale and two races of the Zn-hyperaccumulator Thlaspi caerulescens (Prayon and Close House) to tolerate salinity. The plants were exposed to salt (NaCl) solutions ranging between 0 mM and 100 mM in conjunction with either high or low concentrations of Ni or Zn. Alyssum murale was most resistant to salt in terms of seedling emergence and survival of emerged seedlings. The two races of T. caerulescensand T. arvense were salt sensitive. High Ni or Zn concentrations did not have a clear effect on the salt tolerance of any of the plants tested. The implications of the findings are discussed for the development of metal phytoremediation/phytomining technologies on saline soils or where brackish water (e.g., mining wastewater) could be used to irrigate phytoremediation crops.     

Effect of arbuscular mycorrhizal fungal inoculation on heavy metal accumulation of maize grown in a naturally contaminated soil

A pot culture experiment was carried out to study heavy metal (HM) phytoaccumulation from soil contaminated with Cu, Zn, Pb, and Cd by maize (Zea mays L.) inoculated with arbuscular mycorrhizal (AM) fungi (AMF). Two AM fungal inocula--MI containing only one AM fungal strain (Glomus caledonium 90036) and MII consisting of Gigaspora margarita ZJ37, Gigaspora decipens ZJ38, Scutellospora gilmori ZJ39, Acaulospora spp., and Glomus spp.--were applied to the soil under unsterilized conditions. The control received no mycorrhizal inoculation. The maize plants were harvested after 10 wk of growth. MI-treated plants had higher mycorrhizal colonization than MII-treated plants. Both MI and MII increased P concentrations in roots, but not in shoots. Neither MI nor MII had significant effects on shoot or root dry weight (DW). Compared with the control, shoot Cu, Zn, Pb, and Cd concentrations were decreased by MI but increased by MII. Cu, Zn, Pb, and Cd uptake into shoots and roots all increased in MII-treated plants, while in MI-treated plants Cu, Zn, and Pb uptake into shoots and Cd uptake into roots decreased but Cu, Zn, and Pb uptake into roots and Cd into shoots increased. MII was more effective than MI in promoting HM extraction efficiencies. The results indicate that MII can benefit HMphytoextraction and, therefore, show potential in the phytoremediation of HM-contaminated soils.     

接种AM真菌对PAEs污染土壤中微生物和酶活性的影响

土壤灭菌条件下 ,添加 5 0mg·kg-1DEHP和 5 0mg·kg-1DBP ,在温室进行盆栽试验 ,观察土壤施加DEHP与DBP和接种AM真菌 (Acaulosporalavis,光壁无梗球囊霉 ,菌号 :34)后菌根际 (简称A)、菌丝际 (简称B)和常规土 (简称C)中土壤微生物和部分土壤酶活性的变化。结果表明 ,土壤施加DEHP和DBP后 ,A、B和C层土中土壤微生物数量和土壤酶含量下降 ;接种AM真菌后 ,受AM直接影响的A和B层土中细菌、放线菌和真菌数量比不接种低 ,而C层土中三菌数量比不接种高 ;A和B层土中中性磷酸酶和脲酶含量下降 ,脱氢酶含量在A、C层土中下降 ,在B层土中稍有增加 ,AM对土壤脱氢酶活性影响不大。接种AM真菌没有降低DEHP和DBP对土壤微生物生长和土壤酶活性不利影响的程度     

Relative contributions of soil chemistry, plant physiology and rhizosphere induced changes in speciation on Ni accumulation in plant shoots

The aim of this study is to rank the relative importance of soil properties, root uptake and root-to-shoot redistribution on the transfer of the trace element nickel from soil to the shoots of non hyperaccumulatings plants. Two contrasting soils and seven plant species have been studied using the radioactive isotope, ⁶³Ni. Shoots and roots were analysed separately and the specific activity of each plant has been measured. The isotopic exchange properties of rhizosphere soil where compared with control non rhizosphere soil. Possible changes in Ni speciation in the rhizosphere have been assessed by comparing the isotopic exchange properties of the rhizosphere and control soil and by comparing the specific activities of Ni in each plant. The capacity of soil to immobilise added radiotracer largely determines root uptake, leading to between a 4- and 40-fold difference between soils for a given species. The redistribution of nickel from roots to shoots was fairly constant for plants grown on the rendzina, but varied strongly between species for the acid soil. This variation enhanced the contrast between species of the soil-to-shoot transfer factor. Root action significantly enhanced immobilisation of added nickel in an acid soil due to the modification of speciation of initially non exchangeable soil nickel, but had little effect on a neutral rendzina. Changes in rhizosphere pH were similar on the two soils. In the acid soil, these pH changes were accompanied by changes in Ni speciation but a causative link has not been established. In the neutral soil pH changes may have modified root uptake properties.     

磷对土壤中砷吸附的影响

在黄壤、红壤和褐土中进行As的等温吸附实验表明,在不添加P的情况下,3种土壤对As的吸附能力随土壤pH的升高而降低,随土壤粘粒含量的降低而减弱,3种土壤吸附能力为黄壤>红壤>褐土,As在土壤中的吸附受P存在的影响,60mg·kg^-1的P可以降低黄壤和红壤对As最大吸附量．吸附质中P：As的比例不同,P对土壤吸附As的影响差异很大：吸附质中P：As=1：2时,土壤对As的吸附能力均强于P：As=1：1和P：As=2：1,后两者差异不显著,表明土壤中的吸附点位对As和P的亲和性具有一定的差异,Languir方程能很好地拟合所测得的土壤平衡溶液中As浓度。     

超累积植物与高生物量植物提取镉效率的比较

利用植物修复污染的土壤已受到广泛的关注.采用土壤盆栽试验,比较了超累积植物遏蓝菜与3种高生物量植物印度芥菜、烟草和向日葵对长期施用含镉有机、无机肥料污染的土壤(总Cd,2.87mg·kg-1)的提取效率.研究结果表明,遏蓝菜富集镉的能力明显高于其他3种植物,其地上部镉含量可达43.7mg·kg-1,分别是烟草、印度芥菜和向日葵(叶)的10、27和56倍;而地上部生物量最高的植物烟草,其生物量干重为24.8g· pot-1,分别是遏蓝菜、印度芥菜、向日葵的35倍、3倍、2倍.4种植物提取镉最多的是烟草,每盆可以提取117μg,遏蓝菜和印度芥菜提取镉量分别为35μg·pot-1和30μg·pot-1,向日葵提取量最少,每盆仅为10μg左右.植物对土壤中镉的提取效率分别为:烟草 1%,遏蓝菜0.6%,印度芥菜 0.5%,向日葵0.08%.4种植物种植后,土壤总镉和有效态镉含量没有显著的变化.     

Effect of thallium fractions in the soil and pollution origins on Tl uptake by hyperaccumulator plants: a key factor for the assessment of phytoextraction

Phytoremediation is often discussed as a means of extracting trace metals in excess in the soil, but to increase its efficiency a better understanding of the factors controlling plant uptake is required. The main objective of this study was to examine the effect of origin (anthropogenic vs. geogenic) and mobility of thallium (Tl) in the rhizosphere on Tl uptake. Two Tl-hyperaccumulating Brassicaceae species, kale (Brassica oleracea acephala L. cv. Winterbor F1) and candytuft (Iberis intermedia Guers.), were grown in a rhizobox system to investigate the dynamics of Tl in the rhizosphere soil. Four different soils were used. Two soils contained high Tl amounts due to anthropogenic sources (emissions from a cement plant and mining activities). High Tl content in the two other soils was due to a high rock content (geogenic origin). On completion of growth in the rhizoboxes, the depletion of Tl in seven different chemical fractions, determined by sequential extraction, was compared to the plant uptake. Most of the Tl taken up was derived from the so-called "easily accessible" fractions in both soils with geogenic Tl as well as in the soils polluted by mining activities. Due to the small amounts of easily accessible Tl in the geogenic soils, Tl uptake by Brassicaceae was low. On the other hand, for the air emission-polluted soil, a high depletion of Tl from "less accessible" fractions was observed in addition to depletion of the easily accessible fractions. Hence, the latter soil demonstrated the highest potential for effective soil decontamination by phytoextraction within an appropriate time frame.