Bacterial colonization of the phyllosphere of nineteen plant species and antimicrobial activity of their leaf secondary metabolites against leaf associated bacteria

Summary. The scope of this work was to examine whether leaf constitutive secondary metabolites play a role in determining bacterial colonization of the phyllosphere. To this aim, we surveyed nineteen native or cultivated plant species that share a common bacterial pool in a North Mediterranean area, and estimated the size of total and ice nucleation active (INA) bacterial populations on their leaves. Large differences in the colonization of their phyllosphere were found; the population size of epiphytic bacteria ranged from 7.5 × 10² to 1 × 10⁶ CFU/g fresh weight, in eucalypt and celery, respectively. Species native in Mediterranean-type climate areas, particularly those belonging to the group of aromatic plants, are characterized by scarce presence of INA bacteria. The antibacterial activity of essential oils, surface phenolics and leaf tissue extracts was also estimated against the INA strains P. syringae and E. herbicola, isolated from two of these plant species. E. herbicola proved more sensitive than P. syringae. Of the species examined, oregano [Origanum vulgare L. subsp. hirtum (Link.) Ietswaart], an aromatic plant, had the highest antimicrobial activity, whereas six species showed no activity at all. Further experiments were performed with oregano and bean (Phaseolus vulgaris L.) that represent two extremes in their secondary metabolite content. Both plants were inoculated with P. syringae. By the end of incubation, the bacterial population on bean plants was about 100 times higher than that on oregano leaves. Scanning electron micrographs showed that bacterial growth on oregano leaves was confined to sites away from glandular hairs. Results from the bacterial colonization survey together with those from the toxicity tests showed that all species rich in antibacterial secondary metabolites harbored low leaf bacterial populations. These results provide substantial evidence that leaf secondary metabolites function as constitutive defense chemicals against microbial invasions. However, the fact that species with non- or moderately active leaf secondary metabolites are not always highly colonized suggests mediation of other unknown factors, the contribution of which requires further investigation.     

蔬菜作物施用耐氨固氮菌的增产效果

研究在小区试验条件下,施用耐氨固氮型的催娩克氏菌和阴沟肠杆菌混合菌剂对蔬菜生长和产量的影响。结果表明，施用耐氨固氮菌可促进蔬菜生长，提高产量并改善品质，供试蔬菜的平均增产率达20％，增产效果明显。     

Ammonium-dependent regulation of aerobic methane-consuming bacteria in landfill cover soil by leachate irrigation

The impacts of landfill leachate irrigation on methane oxidation activities and methane-consuming bacteria populations were studied by incubation of landfill cover soils with leachate and (NH₄)₂SO₄ solution at different ammonium concentrations. The community structures and abundances of methane-oxidizing bacteria (MOB) and ammonia-oxidizing bacteria (AOB) were examined by PCRDGGE and real-time PCR. Compared with the pure (NH₄)₂SO₄ solution, leachate addition was found to have a positive effect on methane oxidation activity. In terms of the irrigation amount, ammonium in leachate was responsible for the actual inhibition of leachate. The extent of inhibitory effect mainly depended on its ammonium concentration. The suppression of the predominant methaneconsuming bacteria, type I MOB, was responsible for the decreased methane oxidation activity by ammonium inhibition. Methaneconsuming bacteria responded diversely in abundance to ammonium. The abundance of type I MOB decreased by fivefold; type II MOB showed stimulation response of fivefold magnification upon the first addition but lessened to be lower than the original level after the second addition; the amount of AOB was stimulated to increase for 20-30 times gradually. Accumulated nitrate from nitrification strengthened the ammonium inhibition on type I and type II MOB, as a result, repetitive irrigation was unfavorable for methane oxidation.     

聚磷生物膜的快速启动及微生物特性研究

在同步去除并富集磷的基础上,探究采用前期不排泥、后期排泥的挂膜方式对聚磷生物膜反应器的运行效能、微生物特征及群落结构的影响.结果表明,经过驯化运行,聚磷生物膜的蓄磷能力明显提升.在挂膜阶段,生物膜厚度及EPS含量出现一定程度的增长;PN/PS上升至3.12,PN/PS比值增加有利于微生物粘附在填料上.在好氧出水达标的情况下,富集液中磷酸盐浓度提升至89.5 mg·L~(-1),达到了鸟粪石回收标准.高通量测序结果表明,经过富集培养,微生物群落多样性呈下降趋势,群落组成变化明显.优势菌门为变形菌门(Proteobacteria),其丰度从33.6%增长至75.3%;反应器中的聚磷菌属丰度明显增加,从11.8%上升至23.2%,红环菌属(Rhodocyclaceae)、UKL 13-1为反应器中的优势聚磷菌.     

嗜盐高效降酚菌株Halomonas sp.H17的筛选及降解苯酚特性

从巴丹吉林沙漠盐湖沉积物中分离获得1株在高盐环境下高效降解苯酚菌H17.分析了H17生理生化特性、16S rDNA基因序列、苯酚降解特性及动力学,结果表明,H17属于盐单胞菌属(Halomonas sp.),能在0～20%的盐度下有效降解苯酚,每升外加适量的碳源(葡萄糖浓度0.8 g)和复合氮源(KNO_3 1 g、NH_4Cl 5 g、酵母提取物0.2 g和胰蛋白胨0.2 g)能够促进H17的生长及降解苯酚能力.在温度为30℃、pH 7～8、盐度5～10%的条件下,H17均能高效降解苯酚,最高降解率可达到88.5%.该菌株降解苯酚动力学符合Halane模型,经拟合其生长参数为μ_(max)=0.31 h~(-1),K_S=191.63 mg·L~(-1),K_i=683.05 mg·L~(-1).研究显示H17具有在高盐环境下降解和耐受苯酚的能力,同时对环境有较强的适应性,体现出其在高盐含酚废水实际处理中具有良好的应用价值.     

共生细菌对盐生小球藻亚砷酸盐代谢的影响

为探讨共生细菌对盐生小球藻（Chlorella salina）亚砷酸盐（As（III））富集和形态转化的影响,从C.salina培养液中分离培养出一株共生细菌,并采用抗生素处理C.salina获得无菌藻,设置0,75,150,300,750 μg/LAs（III）溶液处理带菌和无菌的C.salina,7d后测定C.salina对As（III）的吸收、吸附和富集量,以及培养液和藻细胞内As的形态,计算带菌和无菌C.salina对As（III）的氧化率和去除率;使用不同浓度的As（III）处理共生细菌7d,并以300μg/L的As（III）处理共生细菌不同时间,计算两种情况下共生细菌对培养液中As（III）的氧化率和去除率.结果表明：C.salina的共生细菌为根癌农杆菌（Agrobacterium tumefaciens WB-1）;与无菌C.salina相比,带菌C.salina生长更快、对As（III）的耐性更强.带菌C.salina对As（III）的富集量为103.10~448.12mg/kg、氧化率为78.93%~96.88%、去除率为18.92%~55.21%,显著高于无菌C.salina的富集量（11.68~91.39mg/kg）、氧化率（14.46%~26.39%）和去除率（12.82%~29.15%）,也高于A.tumefaciens WB-1对As（III）的氧化率（4.51%~30.61%）和去除率（1.86%~16.19%）.此外,带菌C.salina胞内还检测到少量的As（III）和甲基砷,而在无菌C.salina胞内没有甲基砷存在.共生细菌促进了盐生小球藻对As（III）的富集和形态转化.     

CO2倍增和施氮对水稻不同生长期土壤反硝化细菌丰度的影响

随着全球气候变化的不断加剧,大气CO₂浓度呈明显增加趋势,这将间接影响土壤-植物-微生物系统的氮循环过程.为研究典型水稻土壤反硝化细菌对CO₂浓度升高的响应规律和机制,借助水稻密闭培养箱,运用实时荧光定量聚合酶链式反应（Real-Time qPCR）分子技术,设置不施氮（0 mg/kg）和常规施氮（100 mg/kg）2个处理,研究CO₂倍增对水稻不同生长期土壤关键反硝化功能细菌（narG、nirK和nirS型）丰度的影响.结果表明:①在2种施氮水平,CO₂倍增显著促进了水稻分蘖期、孕穗期、扬花期和成熟期水稻根系生长（增幅为2.96%~28.4%）、地上部生物量增加（增幅为7.1%~107.3%）以及成熟期籽粒干质量的增加（增幅为19.5%和38.0%）,具有显著的增产效应.②反硝化细菌丰度对CO₂倍增的响应与生育期及施氮水平有关,CO₂倍增在2个施氮水平均抑制分蘖期反硝化细菌的繁殖,显著增加孕穗期反硝化细菌数量;在水稻扬花期,CO₂倍增促进了施氮处理narG和nirS型反硝化细菌数量的增加,在成熟期抑制未施氮处理下narG、nirK和nirS型反硝化细菌的生长.另外,narG、nirK、nirS型反硝化细菌丰度整体表现为narG > nirS > nirK,且随水稻的生长,其在成熟期的丰度均呈降低趋势.nirK和nirS基因同属亚硝酸还原酶,但nirS基因丰度高于nirK,且对CO₂倍增和施氮的响应有所差异.研究显示,CO₂倍增可显著增加水稻生长和产量,不同施氮水平对稻田土壤反硝化细菌丰度的影响存在差异.      

温度和DO对MBBR系统硝化和反硝化的影响

在缺氧/好氧/好氧串联运行的移动床生物膜反应器（MBBR）系统中考察了温度和好氧反应器中溶解氧（DO）水平对生物膜硝化和反硝化过程氮素去除的影响,并通过高通量测序技术探究温度和DO的变化造成的MBBR系统中脱氮功能菌群结构的差异,从而在微观水平解释硝化和反硝化受温度和DO影响的生物学机理.结果表明,系统温度的升高可以同时强化生物膜硝化和反硝化过程,且好氧反应器中DO水平的提高对硝化过程有利,从而提高系统的脱氮效果.本研究中,在系统连续运行阶段,当系统温度和好氧O₁反应器的DO浓度为本研究范围内的最高水平时（即温度=20~22℃、DO=5~8mg O₂/L）,比硝化负荷可达1.60g NH₄⁺-N/（m²·d）以上,而相同温度范围内比反硝化负荷可高达2.84g NO₃^--N/（m²·d）,从而使MBBR系统在该工况条件下获得了最佳的NH₄⁺-N和TN去除率（分别达到了98.7%和85.7%）.温度和DO影响硝化和反硝化的根本原因是温度和DO变化引起了脱氮功能菌群数量和群落结构的改变：当好氧反应器的DO水平下降时,硝化功能细菌的OTUs比例显著降低,尤其是异养硝化细菌的生长受到了严重的抑制;而温度的变化对反硝化细菌的影响主要体现在群落结构的变化.     

FNA对短程硝化污泥菌群结构的影响

在SBR反应器增加游离亚硝酸（FNA）预处理单元,投加浓度为1.2mgHNO₂-N/L的FNA进行缺氧搅拌4.5h,连续处理3d,考察短程硝化污泥中FNA对氨氧化菌（AOB）,丝状菌和微生物菌群结构的影响.研究表明,FNA对AOB有短时抑制作用,并能够抑制优势丝状菌Candidatus_Microthrix（微丝菌属）和Cytophagaceae（噬纤维菌）的增殖,分别由5.1%和1.1%下降到0.78%和几乎不可见.SVI从281mL/g降低到100mL/g左右.NAR能够维持在90%左右,短程硝化不受到破坏.高通量结果显示,FNA处理后微生物菌群结构多样性与丰度出现下降,但Thauera（陶厄氏菌属）和Ottowia出现了增殖,分别增加到5.58%和7.82%,同步硝化反硝化（SND）作用明显,这使得即便只有短程硝化,总氮去除率依然能达到60%以上.     

模拟酸雨对福州平原稻田土壤细菌丰度及多样性的影响

为了探讨酸雨对稻田土壤细菌的影响,以福州平原某稻田为研究对象,测定和分析模拟不同酸度酸雨处理下早、晚稻土壤细菌群落组成及其丰度.结果表明：模拟酸雨处理提高了早稻土壤细菌的多样性,但却降低了晚稻土壤细菌的多样性;酸雨改变了稻田土壤细菌的丰度及群落结构,经模拟不同酸度的酸雨处理,稻田土壤细菌的优势菌属及其丰度并不一致;早稻pH3.5处理组与早稻对照组之间的物种多样性及群落结构差异最大,晚稻pH2.5处理组与晚稻对照组之间的物种多样性及群落结构差异最大;在酸雨作用下,溶杆菌属（Lysobacter）和产黄杆菌属（Rhodanobacter）的丰度明显升高,而马赛菌属（Massilia）的丰度则显著降低,说明溶杆菌属（Lysobacter）和产黄杆菌属（Rhodanobacter）为耐受酸雨的主要菌属,马赛菌属（Massilia）则为受酸雨影响最大的菌属;细菌属H16的相对丰度与土壤电导率（EC）呈极显著负相关（P<0.01）,Lysobacter的相对丰度与土壤pH值呈极显著负相关（P<0.01）,Arenimonas的相对丰度与土壤总有机碳（SOC）呈极显著正相关（P<0.01）.