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[目的] 探究镉吸附细菌是否能够高效固定土壤有效镉(Cd),为土壤有效Cd的微生物固定提供理论依据。[方法] 利用含Cd2+牛肉膏蛋白胨液体培养基对细菌进行Cd的耐受性测试筛选出镉抗性强的菌株;通过16S rRNA基因相似性及系统进化分析鉴定耐镉细菌,将菌细胞加入含CdCl2溶液中进行Cd2+吸附效率测定;通过土培模拟实验,测定土壤pH、碱解氮、有效磷、速效钾、有机质、CEC、有效Cd及微生物数量来分析镉吸附细菌对镉污染土壤的影响。[结果] 从德阳鱼腥草根际土壤中分离获得的57株细菌对Cd2+表现出不同程度的抗性,并从中筛选出3株耐Cd优势细菌普罗威登斯菌属(Providencia)DY8、芽孢杆菌属(Bacillus)DY3和芽孢杆菌属(Bacillus)DY1-4。其对溶液中的Cd2+表现出较好的吸附作用,吸附效率随着Cd2+浓度升高而降低。DY8、DY3、DY1-4能使镉污染土壤中有效Cd含量分别降低72.11%、68.55%、62.32%,同时显著提高镉污染土壤中碱解氮、有效磷的含量。[结论] Cd污染农田土壤中含有丰富的耐Cd微生物资源,Cd吸附细菌能降低土壤中有效Cd的含量,且能有效改善土壤养分条件。 相似文献
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对两种多孔菌科大型真菌槐栓菌(Trametes robiniophila)和木蹄层孔菌(Fomes fomentarius)子实体生物吸附Cd2+的影响因素(包括吸附剂用量、初始pH、吸附时间、初始Cd2+浓度)和吸附特性进行分析。结果表明,槐栓菌和木蹄层孔菌对低浓度的Cd2+(10 mg/L)吸附的最适pH为6;Cd2+的去除率随吸附剂用量和吸附时间的增加而增大,槐栓菌和木蹄层孔菌均在吸附剂用量为2g/L时达到吸附平衡,槐栓菌在吸附时间为30 min时达到吸附平衡,而木蹄层孔菌在吸附时间为60 min时达到吸附平衡;槐栓菌和木蹄层孔菌对10 mg/L Cd2+的最大去除率分别为98%和94%。Langmuir等温吸附平衡模型比Freundlich等温吸附平衡模型能更好的拟合两种大型真菌对Cd2+的吸附过程;槐栓菌和木蹄层孔菌对10 mg/L Cd2+的最大吸附量分别为17.40 mg/g和8.91 mg/g。对实验数据进行动力学模型拟合可知,两种大型真菌对Cd2+的生物吸附过程均符合准二阶动力学模型。槐栓菌和木蹄层孔菌生物吸附低浓度Cd2+的化学反应机理可能为离子交换。 相似文献
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【背景】目前对水库水体污染原因的研究往往专注于水体的富营养化、pH值、溶解氧、氨氮、菌落总数指标的变化,而重金属含量与环境因子的季节性变化相关性分析研究较少,同时对于典型季节原位微生物种群的多样性差异研究尚未见报道。【目的】研究浙江省台州市长潭水库底部水中正二价金属离子(二价锰离子Mn2+;二价铁离子Fe2+)浓度与不同环境因子的季节性变化规律,并对其相关性进行分析;富集平水期(2月)和丰水期(8月)水库底部水体中功能微生物菌群,分析其种类和丰度的差异。【方法】分别检测12个月的水库水Mn2+和Fe2+浓度及多种环境因子(水体溶解氧浓度、pH值、总磷浓度、浊度、水库环境温度及降水量),过滤并富集培养水库底部水体的功能微生物菌群,对其16S rRNA基因V3-V4区测序并分析其菌群结构。【结果】长潭水库水中Mn2+和Fe2+浓度呈季节性变化,每年的春夏交替季节水体中铁锰含量从零开始慢慢升高,至夏秋高温季节水体中Mn2+和Fe2+浓度达到最高值,然后慢慢降低,至秋末冬初检测不到含量。在检测的多种环境因子中,水体溶解氧浓度、水库环境温度及降水量呈明显的季节性规律变化。Mn2+和Fe2+浓度与温度、降水量和浊度有正相关性,与溶解氧浓度、pH值和总磷浓度有负相关性,其中在正负相关性分析中两种金属离子的浓度与溶解氧浓度的相关性最强,其次是环境温度及降水量。丰水期和平水期中富集获得的功能微生物菌群的种类和丰度差异很大,从属水平上分类,丰水期时菌群只包含不动杆菌(Acinetobacter)和鲑色沉积物杆状菌(Sediminibacterium)这2个属的菌株,含量各占约50%;平水期时菌群则主要由杆菌属(Bacillariophyta) (47.62%)和Limnohabitans(9.52%)等9个属的菌株构成。富集获得的平水期和丰水期的两个可培养的菌群均具有去除水库水中Mn2+的功能,去除率分别约为35.9%和11.4%。【结论】长潭水库底部水体中Mn2+和Fe2+浓度与不同环境因子均呈季节性规律变化,它们之间呈现不同的正负相关性,丰水期和平水期的功能微生物菌群结构差异很大。本研究为利用微生物进行重金属污染水体的治理储备了微生物资源,为实现国家“美丽乡村”的建设目标提供一定的参考价值。 相似文献
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【目的】嗜酸氧化亚铁硫杆菌(Acidithiobacillus ferrooxidans)作为湿法冶金的主要功能菌,其在重金属Ni2+胁迫下的亚铁氧化应激机制还不清楚。【方法】在两步法培养体系中,设计由低到高Ni2+胁迫浓度,利用分光光度法、电化学法和实时荧光定量PCR法等从宏观到微观探究Ni2+胁迫下A.ferrooxidans菌亚铁氧化应激效应。【结果】两步培养法体系下A.ferrooxidans菌能够耐受较高浓度的Ni2+毒性(≤30 g/L Ni2+)。当Ni2+胁迫浓度增加至40 g/L时,与对照组(不添加Ni2+胁迫)相比,A.ferrooxidans亚铁氧化率和速率显著降低(24 h的亚铁氧化抑制率约为59.4%),亚铁氧化代谢相关的rus操纵子各功能基因的表达量也显著下调,尤其是基因cyc1(最高下调了16倍)和coxBACD(4个不同亚基最高下调2.7–7.4倍);同时Fe2+氧化相关的胞外电子传递能力也降低,对照组(0 g/L Ni2+,29.0µA)最大还原峰电流值高于实验组(40 g/L Ni2+,24.5µA)。【结论】超高浓度Ni2+胁迫环境对A.ferrooxidans菌有毒性,导致该菌rus操纵子上各功能基因表达量下调,同时造成胞外电子传递速率降低,最终致使A.ferrooxidans菌亚铁氧化能力降低。研究结果将为含Ni固废高效生物浸出提供理论支持。 相似文献
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【背景】角蛋白酶是一类特异性降解角蛋白的水解酶,在动物饲料、生物肥料、医学、洗涤、制革及环境治理等方面具有重要的应用潜力。【目的】对前期从海洋环境筛选出的一株铜绿假单胞菌Gxun-7的角蛋白酶基因进行克隆、表达,并探究重组酶酶学性质,为角蛋白酶在工业生产中的应用奠定基础。【方法】以铜绿假单胞菌Gxun-7基因组推定的角蛋白酶基因为基础,设计引物克隆获得角蛋白酶基因kp2,构建重组表达质粒pET22b-kp2,并转化到E. coliRosettagamiB (DE3)中进行诱导表达,同时对重组表达菌株的表达条件进行优化。利用镍柱分离纯化重组角蛋白酶并研究其酶学性质。【结果】重组角蛋白酶的分子量约为33 kDa,最适温度和pH值分别为40 ℃和8.0,在温度30-60 ℃和pH 6.5-8.0具有较好的稳定性。金属离子Co2+、Cu2+和化学试剂十二烷基磺酸钠(sodium dodecyl sulfonate,SDS)、乙二胺四乙酸(ethylenediaminetetraacetic acid,EDTA)、苯甲基磺酰氟(phenylmethylsulfonyl fluoride,PMSF)对酶活力有抑制作用,而Mg2+、K+、巯基乙醇和二硫苏糖醇(dithiothreitol,DTT)对酶活力有促进作用。重组角蛋白酶具有良好的耐盐性,在12.5%的NaCl作用下相对酶活为87.55%。以酪蛋白为底物时,酶的Km值为60.92 mg/mL、Vmax值为9.70 U/mL。【结论】海洋来源铜绿假单胞菌Gxun-7的重组角蛋白酶具有良好的温度、碱、盐稳定性,可应用于工业生产中。 相似文献
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为了探讨重金属Cd2+和Cu2+胁迫对泥蚶消化酶活性的影响,运用酶学分析的方法,按《渔业水质标准》(GB 11607)规定的Cd2+、Cu2+最高限量值的1、2、5、10倍设置重金属离子Cd2+、Cu2+浓度及其组合,研究了在重金属Cd2+、Cu2+胁迫下,30d内泥蚶3种消化酶活性的变化规律。结果表明:与空白对照组相比,在重金属Cd2+、Cu2+或其组合的胁迫下,较低浓度组泥蚶的淀粉酶活性实验前期增强(即被诱导),实验后期减弱(即被抑制),较高浓度组泥蚶的淀粉酶活性从实验一开始就减弱,并保持在较低水平,毒性比较,同一重金属高浓度 > 低浓度,不同重金属及其组合Cu2+ > (Cd2++Cu2+)组合 > Cd2+;泥蚶脂肪酶的活性实验前期增强,实验后期转为微减弱或减弱,毒性比较,同一重金属高浓度 > 低浓度,不同重金属及其组合(Cd2++Cu2+)组合 > Cu2+ > Cd2+;泥蚶胃蛋白酶的活性实验前期增强,且活性呈现升高-降低-再升高-再降低的变化,实验后期分别表现微增强、微减弱和减弱,毒性比较,同一重金属高浓度 > 低浓度,不同重金属及其组合(Cd2++Cu2+)组合 > Cu2+ > Cd2+。可见:环境中的Cd2+和Cu2+对泥蚶的消化酶活性起着明显的影响作用。 相似文献
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基因工程菌大肠杆菌JM109富集废水中镍离子的研究 总被引:4,自引:2,他引:4
利用通过基因工程技术所构建的在细胞内同时表达出高特异性镍转运蛋白和金属硫蛋白的基因工程菌富集水体中的镍离子。菌体细胞对Ni2+的富集速率很快,富集过程满足Langmuir等温线模型。与原始宿主菌相比,经基因改造的基因工程菌不仅最大镍富集容量增加了5倍多,而且对pH值、离子强度的变化及其它共存重金属离子的影响都呈现出更强的适应性。相比而言,Na+、Ca2+、Cd2+、Pb2+的影响较小,但Mg2+、Hg2+和Cu2+所引起的负面效应较大。进一步的实验表明基因工程菌对Ni2+的富集行为不需要外加营养物质。 相似文献
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NRAMP蛋白(natural resistance-associated macrophage proteins)家族在植物响应重金属胁迫时具有重要作用,能够转运Fe2+、Mn2+、Zn2+和Cd2+等重金属离子。为探究甘蔗ScNRAMP基因家族的特征,该文基于甘蔗割手密基因组鉴定了ScNRAMP基因家族,并进行了理化特性、基因结构、顺式作用元件、保守基序、结构域和进化关系等分析。结果表明:甘蔗ScNRAMP基因家族含有29个成员,不均匀分布在19条染色体上; 编码蛋白均为不稳定蛋白,无信号肽,亚细胞均定位在质膜上; 各成员保守基序有6~10个不等,跨膜数有6~12个不等,二级结构主要构成元件为α-螺旋和无规则卷曲; 顺式作用元件分析表明甘蔗ScNRAMP基因家族可能通过激素代谢参与逆境胁迫和生长发育等生物过程; 利用割手密的RNA-seq转录组表达数据进行的组织特异性分析发现,ScNRAMP在甘蔗不同发育阶段的叶和茎中具有时空表达特性; 进化树分析将甘蔗ScNRAMP家族成员分为3个亚家族(I、Ⅱ和Ⅲ)。该研究在全基因组水平上系统地鉴定了现代栽培甘蔗祖先种之一割手密NRAMP基因家族,既为进一步了解甘蔗NRAMP基因家族提供了基础,也为后续甘蔗重金属研究提供了重要候选基因。 相似文献
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细菌吸附Pd2+的研究 总被引:13,自引:0,他引:13
从不同来源的细菌菌株中筛选获得一株吸附Pd2+能力较强的菌株R08,经鉴定为地衣芽孢杆菌(\%Bacillus licheniformis)\%R08。R08死菌体吸附Pd2+的最适pH值为3.5,其吸附作用是一种快速而非依赖温度的过程。吸附作用受菌体浓度和Pd2+浓度影响。在起始Pd2+浓度200mg/L\,菌浓度0.4g/L\,pH35和30℃条件下,吸附45min,吸附量为2248mg/g。透射电镜观察显示,R08死菌体能够还原Pd2+成Pd0颗粒。红外光谱分析表明,细胞壁上的COO-和HPO42-基团可能与Pd2+的生物吸附有关。 相似文献
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A hyperbolic equation was fitted to the time dependence of Thiobacillus ferrooxidans adsorption on sulfur. This procedure allowed the determination of an asymptote value which represented a free cell count at equilibrium. The data thus obtained were used to calculate the numbers of adsorbed bacteria without attainment of equilibrium. The maximum number of adsorbed bacteria based on the Langmuir isotherm was 8.3 × 109 per g of sulfur. 相似文献
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Non-living, freeze-dried material of the brown algaEctocarpus siliculosus (Phaeophyceae) demonstrated high equilibrium uptake of Cd from aqueous solutions (Fehrmann & Pohl, 1993). The alga was grown in 250-L photobioreactors under various growth conditions (light, salinity and nutrient concentrations) in order to obtain larger quantities of biomass and to improve its Cd adsorption capacity. To derive further knowledge on the biosorbant phenomenon different adsorption parameters such as pH for the sorption process and kinetics of Cd adsorption were tested. The maximum adsorption capacity of the freeze-dried biomass exceeded 41 mg Cd per g biomass. After repeated addition of low Cd concentrations the maximum adsorption capacity was lower (31.4 mg Cd per g biomass). In comparison with other adsorbing materials (activated carbon, silica gel, siliceous earth)E. siliculosus showed significantly higher adsorption capacity. Desorption of deposited Cd with 0.1 M HCl resulted in no changes of the adsorption capacity through five subsequent adsorption-/desorption-cycles. Hence, driedE. siliculosus appears to be an efficient material for the elimination of Cd from industrial waste water.Transmission electron microscopic investigations showed an electron dense area in the outer surface layers of the cell wall after Cd adsorption indicating the most likely location of Cd fixation.Author for correspondence 相似文献
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Recovery of mouse endostatin produced by Pichia pastoris using expanded bed adsorption 总被引:3,自引:0,他引:3
Endostatin, a 20 KDa fragment of collagen XVIII, was shown to have an inhibitory effect on angiogenesis and can potentially be used as a tumor growth suppressor. To obtain the amount needed for testing, the protein was successfully cloned and expressed in Pichia pastoris. At the end of the fermentation process, the concentration of the endostatin in the culture was 50 mg per liter, accompanied by 400 gr per liter (wet weight) of biomass. Before the protein can be captured and purified on a packed bed of heparin-Sepharose, the biomass must be removed. Because of the high biomass concentration, conventional biomass removal techniques like centrifugation or filtration are inefficient and cumbersome. Therefore, the expanded-bed adsorption technique was chosen as an alternative approach. An efficient procedure for the initial recovery and purification of the endostatin was developed. The process utilized a cation- exchanger resin instead of a heparin-based affinity resin, because its dynamic capacity was higher, even though it was affected by the high linear flow on the expanded bed. After adjusting the conductivity, pH and biomass concentration, the complete broth was pumped directly on the expanded-bed matrix (Streamline SP XL). Though the yields of protein are similar, the expanded-bed approach is superior to the packed-bed method for several reasons. The expanded-bed process was shorter (only 8 hours compared to 16 hours for the packed bed), it is cheaper, and the product has higher specific activity (29% compared with 18%). Endostatin produced by the expanded-bed adsorption method showed the expected bioactivity and is currently being tested for its potential as a tumor suppressor. 相似文献
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Zamil SS Choi MH Song JH Park H Xu J Chi KW Yoon SC 《Applied microbiology and biotechnology》2008,80(3):531-544
The cells of psychrotrophic Pseudomonas fluorescens BM07 were found to secrete large amounts of exobiopolymer (EBP) composed of mainly hydrophobic (water insoluble) polypeptide(s) (as contain approximately 50 mol% hydrophobic amino acids, lacking cysteine residue) when grown on fructose containing limited M1 medium at the temperatures as low as 0-10 degrees C but trace amount at high (30 degrees C, optimum growth) temperature. Two types of nonliving BM07 cells (i.e., cells grown at 30 degrees C and 10 degrees C) as well as the freeze-dried EBP were compared for biosorption of mercury (Hg(II)) and cadmium (Cd(II)). The optimum adsorption pH was found 7 for Hg(II) but 6 for Cd(II), irrespective of the type of biomass. Equilibrium adsorption data well fitted the Langmuir adsorption model. The maximum adsorption (Q (max)) was 72.3, 97.4, and 286.2 mg Hg(II)/g dry biomass and 18.9, 27.0, and 61.5 mg Cd(II)/g dry biomass for cells grown at 30 degrees C and 10 degrees C and EBP, respectively, indicating major contribution of heavy metal adsorption by cold-induced EBP. Mercury(II) binding induced a significant shift of infrared (IR) amide I and II absorption of EBP whereas cadmium(II) binding showed only a very little shift. These IR shifts demonstrate that mercury(II) and cadmium(II) might have different binding sites in EBP, which was supported by X-ray diffraction and differential scanning calorimetric analysis and sorption results of chemically modified biomasses. This study implies that the psychrotrophs like BM07 strain may play an important role in the bioremediation of heavy metals in the temperate regions especially in the inactive cold season. 相似文献
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CK Bower J McGuire MA Daeschel 《Journal of industrial microbiology & biotechnology》1995,15(3):227-233
The efficacy of the antimicrobial peptide nisin was examined after adsorption to silica surfaces. Three protocols were used to evaluate nisin's activity against adhered cells ofListeria monocytogenes: bioassay usingPediococcus pentosaceous FBB 61-2 as the sensitive indicator strain; visualization and enumeration of cells by microscopic image analysis; and viability of adhered cells as determined by lodonitrotetrazolium violet uptake and crystallization. The activity of adsorbed nisin was highly dependent upon conditions of adsorption. The highest antimicrobial activity of adsorbed nisin occurred with high concentrations of nisin (1.0 mg ml–1) and brief contact times (1 h) on surfaces of low hydrophobicity. Sequential adsorption of a second protein (-lactoglobulin or bovine serum albumin) onto surfaces consistently resulted in decreased nisin activity. These data provide direction for the development of applications to limit microbial attachment on food contact surfaces through the use of adsorbed antimicrobial peptides. 相似文献
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An experimental malaria transmission blocking vaccine antigen, Pfs25H, expressed and secreted from Pichia pastoris was recovered and purified using a screenless expanded bed column equipped with a rotating fluid distribution system. This column was able to accommodate feed stock, containing 30% biomass, at a flow rate of 300–400 cm/h without affecting column stability. This capability is three times higher than the capability of the expanded bed column currently in use, which is equipped with a perforated plate fluid distribution system; this design could accommodate biomass concentrations of only up to 10%. The screen-less design did not affect the binding capacity, purification level or process yield and, therefore, shorten the process. Purified Pfs25H of 6.4 g were recovered from 37 l of Pichia pastoris culture in one step. 相似文献
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Competitive adsorption of Pb, Cd and Zn ions onto Eichhornia crassipes in binary and ternary systems
A batch sorption technique was used to study the biosorption of Pb2+, Cd2+ and Zn2+ ions onto the vastly abundant water hyacinth weed, Eichhornia crassipes biomass in binary and ternary systems at a temperature of 30 °C and pH 4.84. Mutual interference effects were probed using equilibrium adsorption capacity ratios, , where the prime indicates the presence of one or two other metal ions. The combined action of the metals was found to be antagonistic, and the metal sorption followed the order Pb2+ Cd2+ Zn2+. The behaviour of competitive biosorption for Pb–Cd and Pb–Zn combinations were successfully described by the Langmuir Competitive Model (CLM), whilst the model showed poor fitting to the Cd–Zn data. In conclusion, Pb2+ ions could still be effectively removed from aqueous solution in the presence of both Cd2+ and Zn2+ ions, but removal of the Cd2+ and Zn2+ ions would be suppressed in the presence of Pb2+. 相似文献
18.
【目的】对所筛选的1株耐镉甲基营养芽胞杆菌NTGB29进行了环境抗逆性的研究,及影响菌株吸附镉离子效率的条件优化。【方法】以发酵液活菌数为指标,研究其对不同NaCl浓度、酸碱度、镉离子浓度的耐受情况;进一步通过单因素实验和响应面法优化影响菌株镉离子吸附效率的发酵条件;以有效镉离子含量为指标,验证菌株在镉污染土壤中的吸附效果。【结果】结果表明,菌株NTGB29对NaCl浓度、酸碱度、Cd~(2+)浓度的最大耐受值分别为10%、pH11.0、50mg/L;菌株在发酵液初始Cd~(2+)浓度10 mg/L、起始pH 6.4、培养温度37°C、NaCl浓度4.2%、装液量50 mL/250 mL、培养时间24 h时,对Cd~(2+)的吸附率达到79.70%;菌株能有效降低镉污染土壤中的有效镉离子含量,吸附率为29.65%。【结论】菌株NTGB29在较高浓度Cd~(2+)浓度、NaCl浓度及强碱环境条件下仍然能够生长,具有良好的环境抗逆性及Cd~(2+)吸附能力,在镉污染土壤调理剂及微生物功能菌剂的研制方面能够提供有价值的菌种资源。 相似文献
19.
Fungi are capable of accumulating metals and, in soil, such accumulation may influence metal speciation and transport. The interactions between a common soil fungus, Trichoderma harzianum, and IIb elements were studied in the present investigation. The accumulation of the metals zinc, cadmium and mercury by starved and non-starved mycelium at different pH was determined by a batch technique using radioactive tracers; uptake of the metals was found to be large, with respective distribution coefficients of about 103.5, 102.5 and 104.0 for zinc, cadmium and mercury, respectively. Metal accumulation by a starved system was largely independent of pH in the range 3–9, where in a non-starved system an increased accumulation of zinc (at 10–
m) was observed at low pH (3–5). Potentiometric titrations performed on the two systems revealed significant differences in acid capacities, i.e. values close to zero for the starved system and 500–800 meq kg– for the non-starved system. The maximum metal uptake was at least 50 mmol kg– at pH 6.5 (calculated from adsorption isotherms). The present findings suggests that in the non-starved system a metabolite is produced and then released when the pH is within a certain range. 相似文献
20.
The present study was undertaken to comparatively investigate the attachment capacities of Azospirillum brasilenseSp245 and its lipopolysaccharide-defective Omegon-Km mutants KM018 and KM252, as well as their activities with respect to the alteration of the morphology of wheat seedling root hairs. The adsorption dynamics of the parent Sp245 and mutant KM252 strains of azospirilla on the seedling roots of the soft spring wheat cv. Saratovskaya 29 were similar; however, the attachment capacity of the mutant KM252 was lower than that of the parent strain throughout the incubation period (15 min to 48 h). The mutation led to a considerable decrease in the hydrophobicity of the Azospirillumcell surface. The lipopolysaccharides extracted from the outer membrane of A. brasilenseSp245 and mutant cells with hot phenol and purified by chromatographic methods were found to induce the deformation of the wheat seedling root hairs, the lipopolysaccharide of the parent strain being the most active in this respect. The role of the carbohydrate moiety of lipopolysaccharides in the interaction of Azospirillumcells with plants is discussed. 相似文献