High-rate acidophilic ferrous iron oxidation in a biofilm airlift reactor and the role of the carrier material

In this study, the feasibility and engineering aspects of acidophilic ferrous iron oxidation in a continuous biofilm airlift reactor inoculated with a mixed culture of Acidithiobacillus ferrooxidans and Leptospirillum ferrooxidans bacteria were investigated. Specific attention was paid to biofilm formation, competition between both types of bacteria, ferrous iron oxidation rate, and gas liquid mass transfer limitations. The reactor was operated at a constant temperature of 30 degrees C and at pH values of 0-1.8. Startup of the reactor was performed with basalt carrier material. During the experiments the basalt was slowly removed and the ferric iron precipitates formed served as a biofilm carrier. These precipitates have highly suitable characteristics as a carrier material for the immobilization of ferrous iron-oxidizing bacteria and dense conglomerates were observed. Lowering the pH (0.6-1) resulted in dissolution of the ferric precipitates and induced granular sludge formation. The maximum ferrous iron oxidation rate achieved in this study was about 145 molFe(2+)/m(3).h at a hydraulic residence time of 0.25 h. Optimal treatment performance was obtained at a loading rate of 100 mol/m(3).h at a conversion efficiency as high as 98%. Fluorescent in situ hybridization (FISH) studies showed that when the reactor was operated at high ferrous iron conversion (>85%) for 1 month, the desirable L. ferrooxidans species could out-compete A. ferrooxidans due to the low Fe(2+) and high Fe(3+) concentrations.     

健康与感染青枯病烟株根际土壤与茎秆细菌群落结构与多样性

【目的】了解健康烟株与感染青枯病烟株在根际土壤、茎杆发病部位、茎杆病健交界部位以及未发病茎杆的细菌群落结构与多样性。【方法】分别对土壤与茎杆样品中细菌的16S rRNA基因V3-V4区进行扩增,采用Illumina MiSeq测序技术对扩增片段进行高通量测序,然后对健康烟株与感染青枯病烟株不同部位细菌群落结构与多样性进行分析。【结果】感染青枯病烟株发病茎杆及根际土壤的细菌群落多样性高于健康烟株茎杆及其根际土壤样品,病健交界茎杆样品细菌群落多样性低于健康烟株。变形菌门(Proteobacteria)在所有样品中均为优势菌门;所有烟株根际土壤的优势菌门为拟杆菌门(Bacteroidetes)、酸杆菌门(Acidobacteria)、放线菌门(Actinobacteria)和绿弯菌门(Chloroflexi);健康烟株茎杆部位的优势菌门为蓝细菌门(Cyanobacteria);感染青枯病烟株发病茎杆和病健交界茎杆部位的优势菌门为蓝细菌门(Cyanobacteria)和厚壁菌门(Firmicutes)。所有根际土壤样品的优势菌属为劳尔氏菌属(Ralstonia)、假单胞菌属(Pseudomonas)、鞘脂单胞菌属(Sphingomonas)、黄杆菌属(Flavobacterium)和代尔夫特菌属(Delftia),而感染青枯病烟株根际土壤的劳尔氏菌属(Ralstonia)和假单胞菌属(Pseudomonas)相对丰度显著高于健康烟株根际土壤,鞘脂单胞菌属相对丰度显著低于健康烟株根际土壤。烟株茎杆的优势菌属为劳尔氏菌属和假单胞菌属等。感染青枯病烟株病健交界茎杆中劳尔氏菌属、肠杆菌属(Enterobacter)和泛菌属(Pantoea)相对丰度显著低于健康烟株样品。【结论】健康与感染青枯病烟株茎杆样品细菌群落的丰富度和多样性明显低于相应的根际土壤样品。较健康烟株而言,感染青枯病烟株根际土壤和茎杆样品细菌群落丰富度和多样性均表现出不同程度地增加,且根际土壤细菌群落结构变化较茎杆样品明显,而病健交界茎杆样品细菌群落丰富度和多样性降低。烟草青枯病为典型土传病害,其病原茄科劳尔氏菌尽管能在烟株维管束中蔓延扩增,但主要还是分布于土壤中;它的存在似乎对土壤细菌群落的影响大于茎杆样品的。该研究结果提示对于青枯病的防治不能局限于烟株本身,田间土壤也应加大防治力度。     

活性污泥微生物解壳聚糖松江菌中菌胶团形成相关大型基因簇的鉴定和分析

【背景】活性污泥法已广泛应用于城市污水和工业废水的处理,微生物菌胶团的形成在污泥通过重力沉淀实现泥水分离和污泥回用的过程中起着重要作用。从西安北石桥污水处理厂活性污泥中分离到一株菌胶团形成菌XHY-A6,经鉴定为解壳聚糖松江菌(Mitsuariachitosanitabida)。【目的】旨在揭示该株解壳聚糖松江菌菌胶团形成相关的基因及其菌胶团形成机制。【方法】结合分子遗传学,包括转座子插入突变技术和遗传互补分析以及基因组学方法分析与菌胶团形成相关的基因和基因簇。【结果】通过转座子插入突变技术获得了两株菌胶团形成缺陷的突变株,转座子插入位点在糖基转移酶(称为gt3)和多糖链长决定蛋白(wzz)基因内,且这两个基因位于一个与菌胶团形成相关的大型基因簇内,该基因簇内还包括与胞外多糖生物合成和分泌相关的基因、epsB2-prsK-psrR-prsT基因以及一个编码PEP-CTERM蛋白A的基因,遗传互补分析证明gt3基因、wzz基因及其下游wzc基因在菌胶团形成过程中是必需的。【结论】松江菌中菌胶团形成和调控机制极可能与活性污泥优势菌动胶菌(Zoogloea)非常相似,即由胞外多糖和PEP-CTERM家族胞外蛋白质共同介导。从武汉二郎庙、汤逊湖和深圳南山污水处理厂活性污泥中分离纯化出松江菌,这些松江菌属细菌可以用于富含几丁质和壳聚糖的市政污水和虾蟹类食品加工废水的净化和资源化利用。     

Mg2+ distribution in activated sludge and its effects on the nitrifying activity and the characteristics of extracellular polymeric substances and sludge flocs

Divalent cations act as bridges among extracellular polymeric substances (EPS) and form cross-linkage for the self-immobilization of microbial biomass. However, their effects on the nitrification performance during the biological nitrogen removal are still unclear. In the present study, the effects of Mg²⁺ on the nitrifying activity, EPS and floc characteristics were investigated using a lab-scale sequencing batch reactor. The distribution of Mg²⁺ was quantified at different level of sludge floc. The results indicated that the nitrification activity was significantly improved when influent Mg²⁺ was below 1.1 mmol/L, but suppressed at 3 mmol/L. The overall performance characterized by COD, NH₄⁺-N and TN, the particle size and sludge flocculation ability rapidly increased with the increase of Mg²⁺ concentration. Mg²⁺ was mainly distributed in the pellet and changed slightly in supernatant, LB-EPS and TB-EPS. The four fluorescence peaks detected by three-dimensional excitation-emission matrix spectra were attributed to PN-like substances and humic acid-like substances in the LB-EPS and TB-EPS. The results of XPS analysis demonstrated that LB-EPS and TB-EPS comprised similar elements. Therefore, the types of EPS functional groups was unchanged under varied Mg²⁺ concentrations, while their proportions changed and LB-EPS/EPS was key factor for the changes of bioflocculation.     

Electrical and chemical signals involved in short-term systemic photosynthetic responses of tobacco plants to local burning

Short-term (up to 1 h) systemic responses of tobacco (Nicotiana tabacum cv. Samsun) plants to local burning of an upper leaf were studied by measuring the following variables in a distant leaf: extracellular electrical potentials (EEPs); gas exchange parameters; fast chlorophyll fluorescence induction; and endogenous concentrations of three putative chemical signaling compounds—abscisic (ABA), jasmonic (JA), and salicylic (SA) acids. The first detected response to local burning in the distant leaves was in EEP, which started to decline within 10–20 s of the beginning of the treatment, fell sharply for ca. 1–3 min, and then tended to recover within the following hour. The measured gasometric parameters (stomatal conductance and the rates of transpiration and CO₂ assimilation) started to decrease 5–7 min after local burning, suggesting that the electrical signals may induce stomatal closure. These changes were accompanied by systemic increases in the endogenous ABA concentration followed by huge systemic rises in endogenous JA levels started after ca. 15 min, providing the first evidence of short-term systemic accumulation of these plant hormones in responses to local burning. Furthermore, JA appears to have an inhibitory effect on CO₂ assimilation. The correlations between the kinetics of the systemic EEP, stomatal, photosynthetic, ABA, and JA responses suggest that (1) electrical signals (probably induced by a propagating hydraulic signal) may trigger chemical defense-related signaling pathways in tobacco plants; (2) both electrical and chemical signals are interactively involved in the induction of short-term systemic stomatal closure and subsequent reductions in the rate of transpiration and CO₂ assimilation after local burning events.     

Functional analysis of the essential bifunctional tobacco enzyme 3-dehydroquinate dehydratase/shikimate dehydrogenase in transgenic tobacco plants

In plants, the shikimate pathway occurs in the plastid and leads to the biosynthesis of aromatic amino acids. The bifunctional 3-dehydroquinate dehydratase/shikimate dehydrogenase (DHD/SHD) catalyses the conversion of dehydroquinate into shikimate. Expression of NtDHD/SHD was suppressed by RNAi in transgenic tobacco plants. Transgenic lines with <40% of wild-type activity displayed severe growth retardation and reduced content of aromatic amino acids and downstream products such as cholorogenic acid and lignin. Dehydroquinate, the substrate of the enzyme, accumulated. However, unexpectedly, so did the product, shikimate. To exclude that this finding is due to developmental differences between wild-type and transgenic plants, the RNAi approach was additionally carried out using a chemically inducible promoter. This approach revealed that the accumulation of shikimate was a direct effect of the reduced activity of NtDHD/SHD with a gradual accumulation of both dehydroquinate and shikimate following induction of gene silencing. As an explanation for these findings the existence of a parallel extra-plastidic shikimate pathway into which dehydroquinate is diverted is proposed. Consistent with this notion was the identification of a second DHD/SHD gene in tobacco (NtDHD/SHD-2) that lacked a plastidic targeting sequence. Expression of an NtDHD/SHD-2-GFP fusion revealed that the NtDHD/SHD-2 protein is exclusively cytosolic and is capable of shikimate biosynthesis. However, given the fact that this cytosolic shikimate synthesis cannot complement loss of the plastidial pathway it appears likely that the role of the cytosolic DHD/SHD in vivo is different from that of the plastidial enzyme. These data are discussed in the context of current models of plant intermediary metabolism.     

Bacterial community in the rhizosphere and rhizoplane of wild type and transgenic eucalyptus

The rhizosphere is a niche exploited by a wide variety of bacteria. The expression of heterologous genes by plants might become a factor affecting the structure of bacterial communities in the rhizosphere. In a greenhouse experiment, the bacterial community associated to transgenic eucalyptus, carrying the Lhcb1-2 genes from pea (responsible for a higher photosynthetic capacity), was evaluated. The culturable bacterial community associated to transgenic and wild type plants were not different in density, and the Amplified Ribosomal DNA Restriction Analysis (ARDRA) typing of 124 strains revealed dominant ribotypes representing the bacterial orders Burkholderiales, Rhizobiales, and Actinomycetales, the families Xanthomonadaceae, and Bacillaceae, and the genus Mycobacterium. Principal Component Analysis based on the fingerprints obtained by culture-independent Denaturing Gradient Gel Electrophoresis analysis revealed that Alphaproteobacteria, Betaproteobacteria and Actinobacteria communities responded differently to plant genotypes. Similar effects for the cultivation of transgenic eucalyptus to those observed when two genotype-distinct wild type plants are compared.     

Advances in aerobic granule formation and granule stability in the course of storage and reactor operation

Aerobic granulation is drawing increasing global interest in a quest for an efficient and innovative technology in wastewater treatment. Developed less than two decades ago, extensive research work on aerobic granulation has been reported. The instability of the granule, which is one of the main problems that hinder practical application of aerobic granulation technology, is still to be resolved. This paper presents a review of the literature in aerobic granulation focusing on factors that influence granule formation, granule development and their stability in the context of sludge granulation. The review attempts to shed light on the potential of developing granules with adequate structural stability for practical applications. The possibilities and perspective of using stored granule as inoculums for rapid startup, and as microbial supplement to enhance treatment of bioreactor systems are also discussed.     

宁南山区典型植物根际与非根际土壤微生物功能多样性

选择宁南山区9种典型植物的根际与非根际土壤为研究对象,采用Biolog方法对土壤微生物功能多样性进行了研究。结果表明:9种不同植物根际土壤与非根际土壤的微生物活性(AWCD)、微生物多样性指数和微生物均匀度指数均存在明显差异;除冰草外,其他各种植物的根际土壤的微生物活性AWCD、微生物多样性指数和微生物均匀度指数均比非根际土壤的高;9种典型植物根际土壤微生物主要碳源利用类型是羧酸类和氨基酸类,非根际土壤微生物主要碳源利用类型是羧酸类、胺类、氨基酸类;微生物活性、微生物多样性指数和微生物均匀度指数两两之间均达到了极显著相关,与土壤化学性质各指标之间均未达到显著相关水平。     

药用植物根系土壤可培养粘细菌的分离鉴定

[目的]对药用植物根系土壤可培养粘细菌进行分离与鉴定,探讨药用植物根系土壤粘细菌资源多样性.[方法]采集华南植物园和南岭国家森林公园22种药用植物根系土,利用辅助菌诱导分离技术分离样品中可培养粘细菌;通过菌株显微形态和菌落形态观察,结合16S rDNA基因序列分析,确定分离粘细菌菌株的系统发育地位.[结果]分离获得50株粘细菌,分属于3个科,7个属,其中粘球菌属(Myxococcus)18株,珊瑚球菌属(Corallococcus)11株,孢囊杆菌属(Cystobacter)7株,原囊菌属(Archangium)8株,标桩菌属(Stigmatella)1株,软骨霉状菌属(Chondromyces)4株,匣状球菌属(Pyxidicoccus)1株.粘球菌属和珊瑚球菌属分布最为广泛.[结论]研究发现药用植物根系土壤可培养粘细菌与土壤pH和有机碳含量等环境因子有一定的相关性.粘细菌更适宜在有机质丰富、pH近中性的环境中生长;粘球菌属和珊瑚球菌属的菌株对pH适应性较强,而粘球菌属和孢囊杆菌属的菌株对土壤有机碳含量依赖性不强,在贫瘠土壤中仍有分布.本研究对后续粘细菌资源的开发和利用提供良好的实验材料和理论基础.     

A comparative study of Tam3 and Ac transposition in transgenic tobacco and petunia plants

Transposition of the Anthirrinum majus Tam3 element and the Zea mays Ac element has been monitored in petunia and tobacco plants. Plant vectors were constructed with the transposable elements cloned into the leader sequence of a marker gene. Agrobacterium tumefaciens-mediated leaf disc transformation was used to introduce the transposable element constructs into plant cells. In transgenic plants, excision of the transposable element restores gene expression and results in a clearly distinguishable phenotype. Based on restored expression of the hygromycin phosphotransferase II (HPTII) gene, we established that Tam3 excises in 30% of the transformed petunia plants and in 60% of the transformed tobacco plants. Ac excises from the HPTII gene with comparable frequencies (30%) in both plant species. When the -glucuronidase (GUS) gene was used to detect transposition of Tam3, a significantly lower excision frequency (13%) was found in both plant species. It could be shown that deletion of parts of the transposable elements Tam3 and Ac, removing either one of the terminal inverted repeats (TIR) or part of the presumptive transposase coding region, abolished the excision from the marker genes. This demonstrates that excision of the transposable element Tam3 in heterologous plant species, as documented for the autonomous element Ac, also depends on both properties. Southern blot hybridization shows the expected excision pattern and the reintegration of Tam3 and Ac elements into the genome of tobacco plants.     

烤烟根际烟碱降解细菌的多样性及其促生特性分析

【背景】挖掘兼具烟碱降解和植物根际促生细菌(Plant Growth-Promoting Rhizobacteria,PGPR)功能的细菌资源,有助于保护土壤质量,实现绿色种植。【目的】分析烤烟根际细菌多样性,筛选可降解高浓度烟碱的PGPR。【方法】采用纯培养法在选择性培养基上分离烟碱降解细菌。通过BOXA1R-PCR分析技术、16SrRNA基因测序及系统发育树构建,对菌株的遗传多样性和分类学地位进行分析。进一步评价了菌株的吲哚乙酸(Indole-3-Acetic Acid,IAA)活性、溶磷能力、病原菌拮抗能力等PGPR指标,以筛选出高效PGPR,最后通过盆栽试验验证其促生效果。【结果】分离得到58株烟碱降解细菌,根据BOXA1R-PCR指纹图谱选取11株菌进行16S rRNA基因序列测定,结果表明,58株菌分别属于芽孢杆菌属(Bacillus)、假单胞菌属(Pseudomonas)、拉乌尔菌属(Raoultella)和短波单胞菌属(Brevundimonas)4个属,以芽孢杆菌属(Bacillus)为优势菌属。58株细菌中48.28%的菌株可产IAA,27.59%具备溶磷能力,37.93%具备纤维素降解能力,G2-13、G2-3及HT2-8因促生与抗病特性突出而被筛选为目标功能菌。盆栽试验结果表明,G2-13菌株对幼苗生长的促进作用明显,可使株高与地上部鲜重分别增加33.05%与53.32%。【结论】烤烟根际存在较为丰富多样的烟碱降解细菌,它们在种植业上具有潜在的应用价值。     

Bacterial inoculation of maize affects carbon allocation to roots and carbon turnover in the rhizosphere

To assess the influence of bacteria inoculation on carbon flow through maize plant and rhizosphere,¹⁴C allocation after¹⁴CO₂ application to shoots over a 5-day period was determined. Plants were grown on C- and N-free quartz sand in two-compartment pots, separating root and shoot space. While one treatment remained uninoculated, treatments two and three were inoculated withPantoea agglomerans (D5/23) andPseudomonas fluorescens (Ps I A12), respectively, five days after planting. Bacterial inoculation had profound impacts on carbon distribution within the system. Root/rhizosphere respiration was increased and more carbon was allocated to roots of plants being inoculated. After five days of¹⁴CO₂ application, more ethanol-soluble substances were found in roots of inoculated treatments and lower rhizodeposition indicated intensive C turnover in the rhizosphere. In both inoculated treatments the intensity of photosynthesis measured as net-CO₂-assimilation rates were increased when compared to the uninoculated plants. However, high C turnover in the rhizosphere reduced shoot growth of D5/23 inoculated plants, with no effect on shoot growth of Ps I A12 inoculated plants. A separation of labeled compounds in roots and rhizodeposition revealed that neutral substances (sugars) constituted the largest fraction. The relative fractions of sugars, amino acids and organic acids in roots and rhizodeposition suggest that amino acid exudation was particularly stimulated by bacterial inoculation and that turnover of this substance group is high in the rhizosphere.     

Effect of nitrogen forms on growth and chemical changes in the rhizosphere of strawberry plants

The experiment was set up to examine the influence of different nitrogen forms: (NH₄)₂SO₄, Ca(NO₃)₂ or NH₄NO₃ on growth response, root induced pH changes in the rhizosphere, root-borne acid phosphatase activity in strawberry plants cv. Senga Sengana. The plants grown on sandy mineral soil were fertilized with 3 forms of nitrogen, in concentrations of 46 mg N·kg⁻¹ soil. The plants were grown in rhizoboxes with removable plexiglass lids. To ensure the root growth along the plexiglass lids, the rhizoboxes were placed at an angle of about 50° with the lid on the lower side. In case of ammonium supply, the nitrification inhibitor DIDIN was added (10 mg·kg⁻¹ of moist soil) to prevent conversion of ammonium into nitrate. The growth response (roots and shoots) of strawberry plants were determined after 11 weeks of treatment with different N forms. The best development of the root system and shoots (root and shoot dry weight and root length) was obtained, when ammonium nitrate was supplied. It is suggested therefore, that NH₄NO₃ stimulates vegetative growth of strawberry plants cv. Senga Sengana. However, there were no statistical differences in a leaf and flower number of the plants grown under different forms of N-fertilization. Determination of rhizosphere pH, and acid phosphatase activity were executed using non-destructive techniques, which enabled weekly measurement of chemical changes in the rhizosphere. The results revealed that the form of nitrogen supplied had a predominant effect on chemical changes in the rhizosphere of strawberry plants. The highest pH values (average pH 6.8) were measured in the rhizosphere of individual plants supplied with Ca(NO₃)₂. Whereas the lowest pH values (average pH 5.8) were detected in the presence of (NH₄)₂SO₄. The curve of rhizosphere pH measured along individual roots of the plants treated with Ca(NO₃)₂ represents the highest pH values whereas the curve of rhizosphere pH under (NH₄)₂SO₄ treatment had the lowest pH values. The highest activity of acid phosphatase were observed in the rhizosphere of strawberry plants grown in the presence of (NH₄)₂SO₄, at pH 5.8.     

厦门市七种药用植物根围AM真菌的侵染率和多样性

调查了福建省厦门市7种常见药用植物根围丛枝菌根真菌分布情况、侵染率及其多样性。结果表明:7种药用植物均能与AM真菌形成良好的共生关系,且不同药用植物形成菌根的能力差异明显,盐肤木根围的孢子密度最高,29.0个/g土,喜树的侵染率最高,100%。含笑根围的孢子密度最低,4.7个/g土,鱼腥草的总侵染率最低,4.5%。共分离鉴定AM真菌4属63种,其中球囊霉属(Glomus)39种、无梗囊霉属(Acaulospora)18种、巨孢囊霉属(Gigaspora)4种、盾巨孢囊霉属(Scutellospora)2种,其中Glomus、Acaulospora为优势属,黑球囊霉G.melanosporum为优势种。盐肤木根围AM真菌种类最丰富,Shannon-Weiner指数H达到1.29。侵染率与各土壤因子均无显著相关;孢子密度与pH值极显著负相关;种的丰度与pH值显著负相关、与电导率极显著负相关、与孢子密度极显著正相关;Shannon-Weiner指数H与有机质极显著负相关;均匀度与有机质、孢子密度极显著负相关。厦门地区AM真菌资源十分丰富,多样性程度高,宿主植物不同,土壤因子对其侵染率、孢子密度、种的丰度、Shannon-Weiner指数、均匀度的影响也不同。为实现AM真菌生物技术应用于中药材规范化种植提供宝贵种质资源和理论依据。     

Plant genotype, micronutrient fertilization and take-all infection influence bacterial populations in the rhizosphere of wheat

The relationship between micronutrient efficiency of four wheat (Triticum aestivum L.) genotypes, tolerance to take-all disease (caused by Gaeumannomyces graminis (Sacc.) Arx and Olivier var. tritici Walker), and bacterial populations in the rhizosphere was tested in soil fertilized differentially with Zn and Mn. Plant growth was reduced by Mn or Zn deficiency and also by take-all. There was an inverse relationship between micronutrient efficiency of wheat genotypes when grown in deficient soils and the length of take-all lesions on roots (efficient genotypes had shorter lesions than inefficient ones). In comparison to the rhizosphere of control plants of genotypes Aroona and C8MM receiving sufficient Mn and Zn, the total numbers of bacterial cfu (colony forming units) were greater in the rhizosphere of Zn-efficient genotype Aroona under Zn deficiency and in Mn-efficient genotype C8MM under Mn deficiency. These effects were not observed in other genotypes. Take-all decreased the number of bacterial cfu in the rhizosphere of fully-fertilized plants but not of those subjected to either Mn or Zn deficiency. In contrast, the Zn deficiency treatment acted synergistically with take-all to increase the number of fluorescent pseudomonads in the rhizosphere. Although numbers of Mn-oxidising and Mn-reducing bacteria were generally low, take-all disease increased the number of Mn reducers in the rhizosphere of Mn-efficient genotypes Aroona and C8MM. Under Mn-deficiency conditions, the number of Mn reducers in the rhizosphere increased in Aroona but not in C8MM wheat. The results suggest that bacterial microflora may play a role in the expression of Mn and Zn efficiency and tolerance to take-all in some wheat genotypes.