冬青和女贞叶表面颗粒物微形态及叶际细菌群落结构

植物叶表面颗粒物和微生物的结构动态对近地表环境和生态具有重要意义.本研究以青岛市不同功能区冬青和女贞植物的新鲜叶片为研究对象,利用环境扫描电镜观察叶表面颗粒物与微生物微形态特征及季节变化规律;采用Illumina高通量测序技术分析植物叶际细菌群落结构.结果表明,冬青和女贞植物叶片颗粒物滞留量秋、冬季节高于春、夏两季,两种植物叶片更易滞留PM₁₀和PM_2.5,但冬青叶片的滞尘能力尤其是对粒径大于10 μm的颗粒物滞留效果高于女贞,城市主干道植物叶片滞尘能力高于文教区和城市公园植物,受地面扬尘和汽车尾气影响较大;另植物叶际微生物组成存在季节差异,叶片易滞留不同类型真菌、真菌孢子及菌丝体等,且两种植物因叶片革质更适宜菌丝体生长,此外女贞叶表皮还易镶嵌一种表面均匀褶皱的真菌孢子,微生物丰富度城市公园 > 城市主干道 > 文教区,湿度对微生物的生长繁殖呈现积极影响;高通量测序下不同季节的冬青和女贞植物叶际细菌群落差异显著,春季植物叶际细菌丰度最高,夏季最低,两种植物叶际细菌相对丰度最高的优势菌门和优势菌纲为变形菌门（Proteobacteria）和γ-变形菌纲（γ-Proteobacteria）,且同一功能区两种植物共用一个核心微生物群,另外受地面扬尘污染严重的城市主干道植物叶际细菌群落结构与其他两个功能区差异明显.本研究结果证明城市不同功能区典型绿化植物叶表面颗粒物与微生物群落结构之间存在显著相关性,为城市绿化建设提供参考.     

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.     

土壤和叶际微生物对啶虫脒的降解作用

深入研究了土壤、叶际原位和叶际可培养微生物对啶虫脒的降解作用及各条件下降解效果的差异.结果表明,不同环境下的微生物对啶虫脒有不同程度的降解效果.原位叶际微生物对啶虫脒降解影响的程度较小,在灭菌处理与对照叶面中啶虫脒的半衰期分别为8 1 d和6 6d.相对而言,土壤和叶际分离培养微生物对啶虫脒降解作用更加显著,在自然土壤和灭菌土壤中啶虫脒的降解半衰期分别为5.0 d和39.6d,相差为8倍;和原位叶际微生物相比,在油菜叶培养基中微生物的生物量大幅度提高,同时,对啶虫脒的降解效果也更加显著;在灭菌处理的豆叶培养基中(CK),啶虫脒42d的降解率为16.5%,相同时间内非灭菌处理的叶际分离微生物降解率高达95%,其降解速率与培养基中的微生物量呈正相关性,叶际和土壤中都存在能降解啶虫脒的微生物.     

青岛城市公园3种植物叶表面颗粒物形态特征及叶际细菌群落结构

探究植物叶表面颗粒物及微生物群落的分布特征,对深入了解植物叶片与环境互作机制及城市公园绿植选择具有重要意义。本研究选择青岛市中山公园冬青、女贞和樱花3种植物作为供试树种,利用环境扫描电镜观察植物叶片微观结构,采用16S rRNA高通量测序分析叶际细菌群落多样性与丰富度,通过组间差异对比分析不同季节不同植物的叶际细菌群落差异。结果显示：3种植物叶片对大气颗粒物的滞纳能力为冬青＞女贞＞樱花,总体趋势表现为灌木＞乔木,常绿植物＞落叶植物,且受季节变化、植物叶片生长情况与颗粒物累积等因素的影响,叶面颗粒物滞留量表现为春季显著低于其他季节;扫描电镜观察到植物叶片表面存在不同类型的真菌且存在明显季节变化,其中春夏季节植物叶表面存在孢子较多,而秋冬季节主要为丝状真菌,这与青岛秋冬季节出现灰霾天气导致空气气溶胶中真菌含量大相关;高通量测序发现3种植物的叶际微生物群落结构存在明显差异,叶际细菌丰富度为冬青＞女贞＞樱花,且夏季叶际细菌群落丰富度最低,但相对丰度最高的优势菌门均为Proteobacteria和Cyanobacteria,主要优势菌属为norank_f__norank_o__Chloroplast、Sphingomonas、Acinetobacter和Hymenobacter,预示环境因素对叶际微生物群落结构具有重要影响。本研究证明不同植物叶面颗粒物形态存在明显时空变化规律,冬青和女贞叶片滞尘能力强,可作为城市绿化植物的优选;植物类型是影响叶表面颗粒物及微生物群落分布特征的根本因素,而季节变化和位置是影响植物叶面微观特征的重要因素。     

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、pH、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.     

阿维菌素杀虫剂对甘蓝叶际微生物群落结构的影响

植物的叶际存在各种类型微生物包括不同种类的细菌、丝状真菌、酵母、藻类等,它们发挥着重要的生态功能.本研究首次利用磷脂脂肪酸分析（PLFA）和末端限制性片断长度多态性分析（T-RFLP）2种非培养方法评价2种不同剂量的阿维菌素杀虫剂喷施甘蓝后对叶际微生物群落的影响.结果表明,低剂量阿维菌素农药处理对甘蓝叶际微生物的生物量和群落结构影响不显著,而高剂量阿维菌素杀虫剂处理明显地改变了甘蓝叶际微生物的群落结构和组成,减少了总叶际微生物生物量、细菌生物量和革兰氏阳性菌与革兰氏阴性菌特征磷脂脂肪酸之间的比值,而真菌生物量变化不大.PLFA分析结果表明经高剂量阿维菌素农药处理后增加了不饱和脂肪酸16:1ω9t、 18:1ω7和环丙烷脂肪酸cy17:0、 cy19:0的含量,而减少了饱和脂肪酸i15:0、 a15:0、 i16:0 和 a17:0的含量.T-RFLP分析结果表明经高剂量阿维菌素农药处理后出现一些新的酶切片段（如58、 96、 236和420 bp）,这些片段所属的微生物对甘蓝叶片上阿维菌素的降解可能发挥重要的作用;而一些酶切片段消失（如51、 89、 99、 338、 66、 223和482 bp）,这些片段所属的微生物可能对阿维菌素类农药残留的指示具有重要意义.     

水位梯度对小叶章湿地土壤微生物活性的影响

通过3 a野外控制试验,研究了不同水位梯度(-10~30 cm)条件下,小叶章湿地植物地上生物量和0~15 cm以及15~50 cm土壤总有机碳(TOC)、微生物量碳(MBC)、基础呼吸(BR)、微生物商(Cmic/Corg)、代谢商(qCO2)的变化规律.结果表明,不同水位条件下植物地上生物量差异显著(p0.05),积水水位10 cm的植物地上生物量最高,且积水水位0~20 cm保持较高的生产力;积水水位变化对土壤TOC、MBC、BR、Cmic/Corg、qCO2影响显著(p0.05).土壤TOC和BR对积水水位变化的响应趋势一致,0~15 cm土壤TOC和BR在零积水水位最高;15~50 cm土壤TOC和BR随着积水水位的升高而降低,与土壤MBC和Cmic/Corg的变化趋势相同,其中MBC变化最为明显;而qCO2随着积水水位的增加而增大.随着积水水位的增加,土壤微生物群落发生改变和土壤微生物活性降低,其中积水水位30 cm的土壤微生物活性最低,对有机碳的累积与分解过程产生影响.     

某铀尾矿库土壤核素污染与优势植物累积特征

采用野外采样和电感耦合等离子体质谱仪分析法,测定某铀尾矿库内自然生长的14种优势植物及其根系土壤中核素的含量,并针对植物对核素的耐受性和富集性能进行分析. 结果表明:双穗雀稗(Paspalum distichum L.)、圆果雀稗(Paspalum orbiculare Forst.)、水莎草〔Juncellus serotinus (Rottb.) C. B. Clarke〕、水蜈蚣(Kyllinga brevifolia Rottb)和碎米莎草(Cyperus iria L.)的地上部分及金毛狗〔Cibotium barometz (Linn.) J. Sm.〕的地下部分对铀(U)和钍(Th)均有不同程度的累积. 莎草科的碎米莎草的地上部分对U的富集系数高达6.04,其余植物的富集系数均小于1;一部分植物样对U的转移系数小于1.莎草科的水蜈蚣对Th的转移系数为2.56,其他植物的转移系数均小于0.50.莎草科植物对U的吸收富集性能明显优于其他科植物,碎米莎草对U则表现出超耐受性和超富集性.      

4种草本植物对镉的富集特征

以4种草本入侵植物粗毛牛膝菊、洋野黍、莠狗尾草和欧黑麦草作为研究材料,利用盆栽实验研究了这4种植物在土壤镉（Cd）含量（mg·kg^-1）为0（T0）、5（T5）、25（T25）和50（T50）处理下的耐受和富集特征,以期筛选出对Cd污染土壤具有修复潜力的植物.结果表明,粗毛牛膝菊在T5~T50处理下,地上部分和根部生物量与对照（T0）相比均没有显著变化,而其他3种禾本科植物的地上部分和根部生物量在T25或T50处理下与对照相比显著降低,表明粗毛牛膝菊比其他3种植物具有更强的Cd耐受性.在不同土壤Cd含量处理下,4种植物地上部分和根中的Cd含量均随土壤Cd含量增加而显著升高,但4种植物地上部分富集系数随土壤Cd含量升高有显著降低的趋势.在不同土壤Cd含量处理下,粗毛牛膝菊和洋野黍的地上部分富集系数都大于1,莠狗尾草和欧黑麦草地上部分富集系数小于1.在T5、T25和T50处理下,粗毛牛膝菊的转移系数分别为0.93、0.73和1.04,分别显著高于同一Cd含量处理下的其他3种植物.此外,在相同的Cd处理下,粗毛牛膝菊积累的Cd总量和地上部分Cd含量均显著高于其它3种植物,并且粗毛牛膝菊能将90%以上的Cd转移到地上部分,显著高于其他3种植物.综合考虑对Cd的耐受和富集能力,本研究认为粗毛牛膝菊是一种潜在的Cd高富集植物,具有一定的植物修复潜力.     

人工湿地系统中氮、磷迁移转化规律试验研究

基于美人蕉垂直流人工湿地装置,研究废水中不同形态的氮和磷在人工湿地中的迁移转化规律,探讨人工湿地在脱氮除磷过程中填料、植株与微生物之间的关系和作用.结果表明,湿地装置1#出水口中各种污染物的去除效率均优于其他出水口,处理效果总体上沿程下降;装置中的基质对废水中氮、磷的吸附能力在湿地运行初期最强;美人蕉对氮、磷的累积量地上部分为1 407.68 mg/m2和260.52 mg/m2,地下部分为460.76mg/m2和62.01 mg/m2.表明植物对氮、磷污染物的吸收主要集中在地上部分.     

Effect of cypermethrin insecticide on the microbial community in cucumber phyllosphere

Cucumber (Cucumis sativus) is one of the most widely used vegetable in the world,and different pesticides have been extensively used for controlling the insects and disease pathogens of this plant.However,little is known about how the pesticides affect the microbial community in cucumber phyllosphere.This study was the first attempt to assess the impact of pyrethroid insecticide cyperemethrin on the microbial communities of cucumber phyllosphere using biochemical and genetic approaches.Phospholipid fatty ac...     

Film mulching reduces antibiotic resistance genes in the phyllosphere of lettuce

Phyllosphere is an important reservoir of antibiotic resistance genes(ARGs), but the transfer mechanism of ARGs from soil and air to phyllosphere remains unclear. This study demonstrated that soil-air-phyllosphere was the dominant ARG transfer pathway, and blocking it by film mulching can reduce typical phyllosphere ARGs in lettuce by 80.7%-98.7%(89.5% on average). To further eliminate phyllosphere ARGs in lettuce grown with film mulching, the internal soil-endosphere-phyllosphere transfer pathw...     

Leaf metabolic influence of glyphosate and nanotubes on the Arabidopsis thaliana phyllosphere

Chemical exposure can indirectly affect leaf microbiota communities, but the mechanism driving this phenomenon remains largely unknown. Results revealed that the co-exposure of glyphosate and multi-carbon nanotubes (CNTs) caused a synergistic inhibitory effect on the growth and metabolism of Arabidopsis thaliana shoots. However, only a slight inhibitory effect was induced by nanotubes or glyphosate alone at the tested concentrations. Several intermediate metabolites of nitrogen metabolism and fatty acid synthesis pathways were upregulated under the combined treatment, which increased the amount of energy required to alleviate the disruption caused by the combined treatment. Additionally, compared with the two individual treatments, the glyphosate/nanotube combination treatment induced greater fluctuations in the phyllosphere bacterial community members with low abundance (relative abundance (RA) <1%) at both the family and genus levels, and among these bacteria some plant growth promotion and nutrient supplement related bacteria were markable increased. Strikingly, strong correlations between phyllosphere bacterial diversity and metabolites suggested a potential role of leaf metabolism, particularly nitrogen and carbohydrate metabolism, in restricting the range of leaf microbial taxa. These correlations between phyllosphere bacterial diversity and leaf metabolism will improve our understanding of plant-microbe interactions and the extent of their drivers of variation and the underlying causes of variability in bacterial community composition.     

Assessing the impact of fungicide enostroburin application on bacterial community in wheat phyllosphere

Fungicides have been used extensively for controlling fungal pathogens of plants. However, little is known regarding the effects that fungicides upon the indigenous bacterial communities within the plant phyllosphere. The aims of this study were to assess the impact of fungicide enostroburin upon bacterial communities in wheat phyllosphere. Culture-independent methodologies of 16S rDNA clone library and 16S rDNA directed polymerase chain reaction with denaturing gradient gel electrophoresis (PCR-DGGE) were used for monitoring the change of bacterial community. The 16S rDNA clone library and PCR-DGGE analysis both confirmed the microbial community of wheat plant phyllosphere were predominantly of the γ-Proteobacteria phyla. Results from PCR-DGGE analysis indicated a significant change in bacterial community structure within the phyllosphere following fungicide enostroburin application. Bands sequenced within control cultures were predominantly of Pseudomonas genus, but those bands sequenced in the treated samples were predominantly strains of Pantoea genus and Pseudomonas genus. Of interest was the appearance of two DGGE bands following fungicide treatment, one of which had sequence similarities (98%) to Pantoea sp. which might be a competitor of plant pathogens. This study revealed the wheat phyllosphere bacterial community composition and a shift in the bacterial community following fungicide enostroburin application.     

Investigating the bacterial community and amoebae population in rural domestic wastewater reclamation for irrigation

Reclamation of domestic wastewater for agricultural irrigation is viewed as a sustainable option to create an alternative water source and address water scarcity. Free-living amoebae(FLA), which are amphizoic protozoa, are widely distributed in various environmental sources. The FLA could cause considerable environmental and health risks. However, little information is available on the risk of these protozoa. In this study, we evaluated the feasibility using rural domestic wastewater for agricultural irrigation, and analyzed dynamic changes of the microbial community structure and FLA populations in raw and treated wastewater, as well as the phyllosphere and rhizosphere of lettuce production sites that were irrigated with different water sources. The bacterial community dynamics were analyzed by terminal restriction fragment length polymorphism(T-RFLP). The bacterial community structures in the influent were similar to that in the effluent, while in some cases relative abundances varied significantly. The populations of Acanthamoeba spp. and Hartmannella vermiformis in the anaerobically treated wastewater were significantly higher than in the raw wastewater. The vegetables could harbor diverse amoebae, and the abundances of Acanthamoeba spp. and H. vermiformis in the rhizosphere were significantly higher than in the phyllosphere. Accordingly, our studies show insight into the distribution and dissemination of amoebae in wastewater treatment and irrigation practices.     

Profile of the culturable microbiome capable of producing acyl-homoserine lactone in the tobacco phyllosphere

Bacterial populations coexisting in the phyllosphere niche have important effects on plant health. Quorum sensing (QS) allows bacteria to communicate via diffusible signal molecules, but QS-dependent behaviors in phyllosphere bacterial populations are poorly understood. We investigate the dense and diverse N-acyl-homoserine lactone (AHL)-producing phyllosphere bacteria living on tobacco leaf surfaces via a culture-dependent method and 16S rRNA gene sequencing. Our results indicated that approximately 7.9%-11.7% of the culturable leaf-associated bacteria have the ability to produce AHL based on the assays using whole-cell biosensors. Sequencing of the 16S rRNA gene assigned the AHL-producing strains to two phylogenetic groups, with Gammaproteobacteria (93%) as the predominant group, followed by Alphaproteobacteria. All of the AHL-producing Alphaproteobacteria were affiliated with the genus Rhizobium, whereas the AHL-producing bacteria belonging to the Gammaproteobacteria mainly fell within the genera Pseudomonas, Acinetobacter, Citrobacter, Enterobacter, Pantoea and Serratia. The bioassays of supernatant extracts revealed that a portion of the strains have a remarkable AHL profile for AHL induction activity using the two different biosensors, and one compound in the active extract of a representative isolate, NTL223, corresponded to 3-oxo-hexanoyl-homoserine lactone. A large population size and diversity of bacteria capable of AHL-driven QS were found to cohabit on leaves, implying that cross-communication based AHL-type QS may be common in the phyllosphere. Furthermore, this study provides a general snapshot of a potential valuable application of AHL-producing bacteria inhabiting leaves for their presumable ecological roles in the phyllosphere.