Changes in the activities of key enzymes and the abundance of functional genes involved in nitrogen transformation in rice rhizosphere soil under different aerated conditions

Soil microorganisms play important roles in nitrogen transformation.  The aim of this study was to characterize changes in the activity of nitrogen transformation enzymes and the abundance of nitrogen function genes in rhizosphere soil aerated using three different methods (continuous flooding (CF), continuous flooding and aeration (CFA), and alternate wetting and drying (AWD)).  The abundances of amoA ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB), nirS, nirK, and nifH genes, and the activities of urease, protease, ammonia oxidase, nitrate reductase, and nitrite reductase were measured at the tillering (S1), heading (S2), and ripening (S3) stages.  We analyzed the relationships of the aforementioned microbial activity indices, in addition to soil microbial biomass carbon (MBC) and soil microbial biomass nitrogen (MBN), with the concentration of soil nitrate and ammonium nitrogen.  The abundance of nitrogen function genes and the activities of nitrogen invertase in rice rhizosphere soil were higher at S2 compared with S1 and S3 in all treatments.  AWD and CFA increased the abundance of amoA and nifH genes, and the activities of urease, protease, and ammonia oxidase, and decreased the abundance of nirS and nirK genes and the activities of nitrate reductase and nitrite reductase, with the effect of AWD being particularly strong.  During the entire growth period, the mean abundances of the AOA amoA, AOB amoA, and nifH genes were 2.9, 5.8, and 3.0 higher in the AWD treatment than in the CF treatment, respectively, and the activities of urease, protease, and ammonia oxidase were 1.1, 0.5, and 0.7 higher in the AWD treatment than in the CF treatment, respectively.  The abundances of the nirS and nirK genes, and the activities of nitrate reductase and nitrite reductase were 73.6, 84.8, 10.3 and 36.5% lower in the AWD treatment than in the CF treatment, respectively.  The abundances of the AOA amoA, AOB amoA, and nifH genes were significantly and positively correlated with the activities of urease, protease, and ammonia oxidase, and the abundances of the nirS and nirK genes were significantly positively correlated with the activities of nitrate reductase.  All the above indicators were positively correlated with soil MBC and MBN.  In sum, microbial activity related to nitrogen transformation in rice rhizosphere soil was highest at S2.  Aeration can effectively increase the activity of most nitrogen-converting microorganisms and MBN, and thus promote soil nitrogen transformation.      

土壤活性氮动态变化及氮素可利用性对紫云英翻压量的响应

通过田间定位试验,研究了减量化肥紫云英不同翻压量下土壤活性氮的含量、动态变化及氮素可利用性,探讨了紫云英鲜草的适宜翻压量和土壤氮素利用效率,为双季稻合理施用氮肥提供理论依据。在稻-稻-紫云英轮作体系典型时期紫云英翻压前、早稻分蘖盛期、早稻成熟期、晚稻分蘖盛期、晚稻成熟期分别采集土壤样品,监测稻田土壤微生物量氮（MBN）、可溶性有机氮（DON）含量动态变化及氮素可利用性,并分析晚稻成熟期土壤铵态氮（NH₄⁺-N）和硝态氮（NO₃^--N）含量。结果表明：与对照（CK）处理相比,各施肥处理均提高了土壤全氮（TN）、NH₄⁺-N和NO₃^--N含量,增幅分别为10.4%~21.2%、10.3%~44.1%和14.7%~52.9%。在翻压紫云英15.0~22.5 t·hm^-2时,土壤TN、NH₄⁺-N和NO₃^--N含量均随紫云英还田量增多而提高,之后则随还田量的增多而降低。与常规施肥处理相比,化肥减施下紫云英各翻压量处理均提高了土壤MBN、DON及活性氮含量,增幅分别为7.0%~28.7%、8.5%~22.5%和5.8%~26.6%,且随紫云英翻压量的增加呈先增加后降低的变化趋势,MBN和活性氮含量均在翻压量22.5 t·hm^-2时最高,DON含量在翻压量30.0 t·hm^-2时最高。MBN/TN在翻压量22.5 t·hm^-2时最高,DON/TN在翻压量30.0 t·hm^-2时最高。各处理不同时期土壤MBN、DON含量及MBN/TN、DON/TN有明显波动,总体来看,土壤MBN含量及MBN/TN在早稻分蘖盛期明显降低,早稻成熟期有所回升,至晚稻成熟期又逐渐降低;土壤DON含量及DON/TN在早稻成熟期降至最低,至晚稻成熟期再次上升。研究表明,减施40%化肥条件下长期翻压紫云英不仅能增加土壤活性氮含量,同时有利于提高土壤氮素可利用性,紫云英翻压量22.5~30.0 t·hm^-2时效果最好。     

Bacteria and antibiotic resistance detection in fractures of wild birds from wildlife rehabilitation centres in Spain

Anatomic adaptations make birds more prone to open fractures with exposed bone parts losing vascularization. As a result of this exposure, fractures are colonized by different microorganisms, including different types of bacteria, both aerobic and anaerobic, causing osteomyelitis in many cases. For this reason, antibiotic treatment is common. However, carrying out antibiotic treatment without carrying out a previous antibiogram may contribute to increased resistance against antibiotics, especially in migratory wild birds. In this paper, bacterial counts regarding fracture type, bacterial identification and antibiotic resistance have been analysed in wild birds from wildlife rehabilitation centres in Spain. The results obtained showed that open fractures had higher bacterial counts (CFU/mL) than closed ones. Bacteria in family Enterobacteriaceae, identified were Escherichia spp., Enterobacter spp., Shigella spp., Hafnia alvei, Proteus mirabilis, Leclercia adecarboxylata and Pantoea agglomerans. Other bacteria present in wild birds’ fractures were Aeromonas spp., Enterococcus spp. Bacillus wiedmannii and Staphylococcus sciuri. All species found presented resistance to at least one of the antibiotics used. Wild birds can be implicated in the introduction, maintenance and global spreading of antibiotic resistant bacteria and represent an emerging public health concern. Results obtained in this paper support the idea that it is necessary to take this fact into account before antibiotic administration to wild animals, since it could increase the number of bacteria resistant to antibiotics.     

Five-year warming does not change soil organic carbon stock but alters its chemical composition in an alpine peatland

Climate warming may promote soil organic carbon(SOC) decomposition and alter SOC stocks in terrestrial ecosystems, which would in turn affect climate warming. We manipulated a warming experiment using open-top chambers to investigate the effect of warming on SOC stock and chemical composition in an alpine peatland in Zoigê on the eastern Tibetan Plateau, China. Results showed that 5 years of warming soil temperatures enhanced ecosystem respiration during the growing season, promoted above-and be...     

秸秆生物反应堆与菌肥对温室番茄土壤微环境的影响

为研究秸秆生物反应堆、微生物菌肥及两者配套措施对土壤理化性质和微生物功能多样性,以及作物生长的长期影响,试验以传统种植方式为对照(CK,常规栽培),研究了菌肥(T1,微生物菌肥4 kg/667 m2)、内置式秸秆生物反应堆(T2,秸秆(4 t/667 m2)+发酵沟菌剂(8 kg/667 m2)+腐熟猪粪(600 kg/667 m2))及2种措施配套处理(T3,秸秆(4 t/667 m2)+发酵沟菌剂(8 kg/667 m2)+微生物菌肥(4 kg/667 m2)+腐熟猪粪(600 kg/667 m2))对土壤理化性质和微生物功能多样性的影响。结果表明:1)与CK相比,秸秆生物反应堆能够在一定时期内提高土壤含水率;而菌肥能够在一定时期内降低土壤含水率,秸秆生物反应堆能够显著降低土壤酸性和电导率(EC,electrical conductivity)值,缓冲土壤酸化和次生盐渍化;而单施菌肥对土壤酸碱性和EC值没有显著影响。2)秸秆生物反应堆(T2)增加了土壤中有机质的含量和土壤微生物量,降低土壤中速效磷、钾的含量;微生物菌肥(T1)降低了土壤中有机质含量和微生物量,而显著提升了土壤的速效磷、速效钾含量,两种措施配套处理效果则更明显。3)菌肥能够改善土壤微生物对多聚物、碳水化合物和氨基酸的利用效率,而秸秆生物反应堆能够促进土壤微生物对于一部分氨基酸、羧酸类、酚酸类和胺类物质的利用。而2种措施同时使用时,其促进和改善微生物碳代谢能力的作用则更加显著。4)各处理均能够在一定程度上增加各年度番茄产量。综合考虑,认为内置式秸秆生物反应堆和菌肥配套处理(T3)能够更好的改善和修复日光温室连作土壤,增加作物产量,是一种较为有效的农艺措施。     

长期不同施肥对农田土壤养分与微生物群落结构的影响

为了阐明长期不同施肥对土壤养分与微生物群落的影响及影响机理,以杭嘉湖平原典型稻麦轮作区长期定位施肥试验土壤为对象,采用磷脂脂肪酸(Phospholipid fatty acid, PLFA)分析方法,研究了施用化肥(NPK)、秸秆(Straw)、栏肥(Manure)、化肥与秸秆配施(S+NPK)、化肥与栏肥配施(M+NPK)5种不同施肥方式对土壤主要养分指标与微生物量及微生物群落结构的影响,试验以不施肥处理为对照。结果表明,所有施肥处理均提高了土壤有机碳,其中秸秆与化肥配施、栏肥无论是单独施用还是与化肥配施所提高的有机碳含量均达到显著水平;施用化肥、有机肥、秸秆+化肥及有机肥+化肥均可在一定程度增加土壤氮、磷、钾养分。施用秸秆和栏肥显著提高了土壤中总PLFA、细菌、真菌、放线菌等含量以及微生物群落Shannon指数(P＜0.05),但施用化肥可引起土壤微生物量和多样性指数的下降。相关分析结果表明,土壤微生物PLFA含量及Shannon指数与土壤碳氮比呈显著正相关(P＜0.05)。典范对应分析结果表明,土壤有机碳含量(F=2.18,P=0.027)是影响微生物群落结构的主要环境因子。可得出结论,长期秸秆还田或施用栏肥可显著提高土壤微生物量并改变其群落结构,土壤有机碳含量及碳氮比对于土壤质量和肥力的提高具有至关重要的作用。     

含油污泥的堆肥处理对微生物群落结构的影响

采用堆肥方法处理含油污泥,评价堆肥处理对含油污泥中石油烃的去除效果,并采用Biolog方法和构建16SrRNA基因克隆文库的方法对处理过程中微生物碳源利用特征和微生物群落结构进行了研究。结果表明,含油污泥经过90d的堆肥处理,石油烃降解率达53.3%±9.5%,显著高于对照处理。堆肥处理可以显著促进石油烃降解,是一种处理含油污泥的有效措施。Biolog分析结果表明,堆肥处理的孔的平均颜色变化率(AWCD)显著高于对照处理,堆肥处理提高了土壤微生物代谢活性。主成分分析结果表明,对照处理和堆肥处理的微生物碳源利用特征明显不同,堆肥处理改变了含油污泥中微生物的代谢功能特征。对照处理和堆肥处理的16SrRNA基因克隆文库之间存在显著差异,对照处理的优势类群是γ-Proteobacteria,堆肥处理的优势类群是Bacteroidetes,堆肥处理显著改变了含油污泥中的微生物群落结构。Marinobacter和Alcanivorax是对照处理中的优势菌,可能与石油烃的自然降解过程有关,而Pusillimonas和Agrobacterium可能对堆肥处理中石油烃的降解起一定作用。     

日粮精粗比对湖羊瘤胃发酵和菌群结构的影响

为了研究日粮精粗比对湖羊瘤胃发酵和菌群结构的影响,试验选择40只体重相近、体况良好且健康的湖羊羔羊,随机分为4组,每组10只(公母各半),各试验组羊分别饲喂精粗比70:30(A)、60:40(B)、50:50(C)和40:60(D)的全混合日粮,试验期为70 d。结果表明:1)试验B组的氨态氮、丙酸、丁酸和戊酸含量均显著高于试验D组(P<0.05)。2)4组共产生875个OTU,其中共有OTU为725个,占总OTU数量的82.86%,试验A和D组分别含特有OTU为6和1个。试验A组的细菌门数量和香浓指数显著高于试验C和D组(P<0.05),而辛普森指数则显著低于试验B和C组(P<0.05)。试验C组细菌多样性与试验A和B组差异显著(P<0.05)。3)在门水平上,试验A组的放线菌门(Actinobacteria)和螺旋菌门(Spirochaetae)相对丰度显著高于试验C和D组(P<0.05)。在属水平上,试验A组毛螺菌科_ND3007(Lachnospiraceae_ND3007)、密螺旋体属_2(Treponema_2)、魏斯氏菌属(Weissella)、厌氧支原体属(Anaeroplasma)、短杆菌属(Brevibacterium)、假丁酸弧菌属(Pseudobutyrivibrio)、未培养瘤胃杆菌属(Uncultured_rumen_ bacterium)和罗斯氏菌属(Roseburia)的相对丰度显著高于试验D组(P<0.05),而毛螺菌科_XPB1014(Lachnospiraceae_XPB1014)和未培养拟杆菌目_BS11(Uncultured _Bacteroidales_BS11)的相对丰度则分别显著低于试验C和D组(P<0.05)。综上所述,高精粗比日粮能够提高湖羊瘤胃细菌丰富度和多样性,促进瘤胃的发酵。     

基于构建微生物传感器的甲基对硫磷降解菌的分离鉴定及其降解特性研究

有机磷农药是目前环境中残留量最多的农药之一,对其残留量的检测及降解机制的研究对于环境污染及生态修复具有重要意义。微生物传感器由生物学元件与换能器构成,因具有成本低廉、易于微型化及选择性高等特点而被广泛应用于各种生化物质的分析和检测。本文从长期受农药污染的土壤中分离出4株能以甲基对硫磷为碳源生长的菌株,根据形态特征和16S r RNA基因序列同源性分析,对4株降解菌进行鉴定,利用高效液相色谱测定降解率,选取降解率最高的1株菌进行降解机制研究,以期将其应用于测定环境中甲基对硫磷残留的电位型微生物传感器的构建。结果表明,在甲基对硫磷初始浓度50 mg·L-1、30℃、p H 7.0的培养条件下培养7 d,4株菌对甲基对硫磷的降解率均在78%以上,其中1株菌的降解效率可达100%。16S r RNA基因序列测定表明,该菌株属于克雷伯氏菌属,命名为Klebsiella sp.MP-6。利用液相色谱-质谱联用对其降解产物的研究表明,菌株MP-6水解甲基对硫磷主要产生二甲基硫代磷酸(dimethyl thiophosphoric acid,DMTP)和对硝基苯酚(p-nitrophenol,PNP),极少部分PNP通过产生4-硝基邻苯二酚(4-nitrocatechol,4-NC)和1,2,4-苯三酚(1,2,4-BT)进一步代谢。结果表明,基于测定中间产物对硝基苯酚(p-nitrophenol,PNP)的电位响应信号,该菌株适用于构建测定海水及土壤等环境中有机磷农药的微生物传感器。