黄顶菊入侵对土壤氨氧化古菌群落多样性的影响

为了明确入侵植物黄顶菊对土壤氮循环关键过程硝化作用的影响机制,本研究通过对其入侵地和未入侵地、根围土和根际土氨氧化古菌(ammonia-oxidizing archaea,AOA)的群落多样性分析,探讨了AOA对黄顶菊入侵的响应规律。结果表明:黄顶菊入侵增加了入侵地根围土AOA的多样性,AOA的Shannon指数表现为:入侵地根围土入侵地根际土未入侵地土壤,且差异显著。铵态氮含量与p H的变化都会影响土壤AOA的群落结构。系统发育树分析表明,土壤AOA主要隶属于氨氧化古菌的Nitrososphaera cluster。黄顶菊入侵导致的AOA的多样性水平的提升主要是由于入侵地氨氧化菌群种类增加所致。     

模拟条件下土壤硝化作用及硝化微生物对不同水分梯度的响应

以江苏滨海县一植稻土壤为研究对象,在微宇宙培养条件下设置了不同水分处理(最大持水量的30%、60%、90%和淹水2 cm深),研究了硝化作用及硝化微生物对水分变化的响应特征。结果表明:淹水处理显著降低了土壤的氧化还原电位(Eh),但所有处理土壤Eh变化范围为330~500 m V,土壤整体处于氧化态。在每7天向土壤加入10 mg kg-1NH+4-N的连续培养过程中,各个水分处理均观察到明显的NH+4-N降低和NO-3-N累积的现象,60%WHC处理下土壤硝态氮累积最显著和迅速,90%WHC处理次之,随培养时间延长,30%WHC和淹水处理也观察到明显的硝化作用。淹水处理中氨氧化细菌(Ammonia-oxidizing bacteria,AOB)的数量显著高于非淹水处理,且淹水处理中AOB在DGGE图谱上的条带更加清晰明亮,而氨氧化古菌(Ammonia-oxidizing archaea,AOA)的群落组成和数量在不同水分处理间无明显变化。表明该土壤中AOB对水分条件变化响应灵敏,是该土壤的硝化作用、尤其是淹水条件下硝化作用发生的主要原因。     

氮肥形态对香蕉种植土壤中氨氧化细菌与古菌的影响

氨氧化微生物通过影响氮素在土壤中的转化而间接影响作物对不同氮素形态的吸收。因此,本实验采集了海南省种植香蕉的滨海土壤,通过施用硫酸铵和硝酸钙,研究了氨氧化细菌(AOB)和氨氧化古菌(AOA)的数量与群落的变化,并测定了香蕉的生长与氮营养状况。通过构建氨氧化细菌和古菌的功能基因amo A文库后发现:施用硫酸铵的土壤中AOB的amo A基因拷贝数显著高于施用硝酸钙的土壤和不施肥的土壤,而AOA的拷贝数低于硝酸钙与不施肥处理。但是氨氧化古菌AOA的绝对数量都高于AOB。施用硫酸铵的土壤中AOB的群落多样性和丰富度明显增加,而施用硝酸钙的土壤却没有发生显著变化。施用硫酸铵和硝酸钙的土壤中AOA群落结构相似,但丰富度和多样性均高于不施肥处理。从施肥效果看,相比硫酸铵,硝酸钙显著提高了香蕉植株的生物量和全氮含量。上述结果表明,在南方酸性土壤中AOA占优势,虽然施用铵态氮肥可以促进AOB的丰度与多样性,但是很可能会抑制AOA的数量。因此,在海南滨海土中施用铵态氮肥后,通过硝化作用以满足香蕉吸收硝态氮的需求达不到理想的效果,应直接补充硝态氮肥来促进香蕉生长。     

大庆龙凤湿地底泥氨氧化细菌与氨氧化古菌多样性分析

为研究湿地底泥中氨氧化细菌(Ammonia-Oxidizing Bacteria,AOB)与氨氧化古菌(Ammonia-Oxidizing Archaea,AOA)的群落多样性,构建AOB与AOA的amo A基因克隆文库,同时利用限制性片段长度多态性(RFLP)技术将克隆文库阳性克隆子进行归类并测序。结果发现,从湿地底泥中均得到3个AOB与AOA的可操作分类单元(OTU)。其中,所有的AOB均属于亚硝化单胞菌属(Nitrosomonas),而AOA属于泉古菌门(Crenarcharota),与河口沉积物等环境中获得的序列具有较高同源性。     

Response of ammonia oxidizing microbes to the stresses of arsenic and copper in two acidic alfisols

Soil pollution by elevated heavy metals exhibits adverse effects on soil microorganisms. Ammonia oxidizing bacteria and ammonia oxidizing archaea perform ammonia oxidative processes in acidic soils. However, influence of heavy metal stress on soil ammonia oxidizers distribution and diversity is inadequately addressed. This study investigated the responses of ammonia oxidizing bacteria and archaea to heavy metals, Cu and As during short-term laboratory experiment. Two different acidic alfisols named as Rayka and Hangzhou spiked with different concentrations of As, Cu and As + Cu were incubated for 10 weeks. Significant reduction in copy numbers of archaeal-16S rRNA, bacterial-16S rRNA and functional amoA genes was observed along elevated heavy metal concentrations. Ammonia oxidizing archaea was found to be more abundant than ammonia oxidizing bacteria in all the heavy metal treatments. The potential nitrification rate significantly decreased with increasing As and Cu concentrations in the two soils examined. Denaturing gradient gel electrophoresis analysis revealed no apparent community shift for ammonia oxidizing archaea even at higher concentrations of As and Cu. Phylogenetic analysis of archaeal amoA gene from 4 clone libraries indicated that all the archaeal amoA sequences were placed within 3 distinct clusters from soil and sediment group 1.1b of Thaumarchaeota. Our results could be useful for the better understanding of the ecological effects of heavy metals on the abundance and diversity of soil ammonia oxidizers.     

双价转基因抗虫棉花对土壤氨氧化细菌和氨氧化古菌群落结构及丰度的影响

采用末端片段多态性分析(T-RFLP)和实时定量PCR(Quantitative real-time PCR,qPCR)方法,研究了不同生长时期双价转基因抗虫棉花和亲本非转基因棉花(对照)土壤氨氧化细菌(Ammonia-oxidizing bacteria,AOB)和氨氧化古菌(Ammonia-oxidizing archaea,AOA)群落结构和丰度的变化,以探讨双价转基因抗虫棉花种植对土壤氨氧化微生物的影响。结果表明,双价转基因抗虫棉花土壤中AOB和AOA优势种群与对照相同,且随生长时期无显著变化,但在各自样品中所占比例不同。不同生长时期双价转基因抗虫棉花土壤AOB的Shannon指数和Evenness指数均与对照无显著差异,AOA的Shannon指数与对照也无显著差异,而苗期时Evenness指数显著低于对照(P0.05),其他3个生长时期差异均不显著。qPCR分析表明,除花铃期双价转基因抗虫棉花土壤AOB丰度高于对照外,其他3个生长时期均低于对照,而AOA丰度在4个生长时期均低于对照。吐絮期时,双价转基因抗虫棉花与对照土壤AOB和AOA丰度均差异显著(P0.05)。双价转基因抗虫棉花对土壤AOB和AOA群落结构组成无显著影响,但降低了AOB和AOA的数量,双价转基因抗虫棉花对土壤微生物的某些类群可能存在潜在影响。     

Community composition of ammonia-oxidizing bacteria in the rhizosphere of intercropped wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.), maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.), and faba bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.)

Cereal/cereal and cereal/legume intercropping systems are popular in the north, northwest, and southwest of China and often result in yield increases compared to monocropping. Rhizosphere interactions may play a significant role in the yield increases, particularly with respect to nutrient availability. The aim of this study was to investigate the effects of intercropping on N availability and community composition of ammonia-oxidizing bacteria in the rhizosphere of wheat, maize, and faba bean at different growth stages. Denaturing gradient gel electrophoresis (DGGE) based on 16S rRNA genes was used to analyze the community composition of bacterial ammonia oxidizers belonging to β-proteobacteria. The results showed that intercropping with faba bean significantly increased nitrate concentrations in the rhizosphere of wheat and maize at the second sampling time (20 June) compared to monocropping or intercropping between maize and wheat. Intercropping significantly affected the community composition of ammonia-oxidizing bacteria in the rhizosphere compared to monocropping, and the effects were most pronounced in the maize/faba bean and wheat/maize intercropping systems when faba bean and wheat were at anthesis and maize was in seedling stage. In wheat/faba bean intercropping, the effects of intercropping on community composition of ammonia-oxidizing bacteria were less pronounced at the seedling stage of the two species but were significant at anthesis.     

鸡粪菌渣好氧堆肥过程中氨氧化古菌及氨氧化细菌群落的动态变化

以amo A基因为标记,通过Real-Time PCR和限制性片段长度多态性(Restriction fragment length polymorphism,RFLP)法对鸡粪菌渣好氧堆肥过程中的氨氧化古菌(Ammonia-oxidizing archaea,AOA)和氨氧化细菌(Ammonia-oxidizing bacteria,AOB)进行了丰度及群落结构的分析。结果表明,在堆制初期、好氧发酵高温期及后熟期,AOB的amo A基因丰度均占主导优势,是AOA的38~992倍。进入好氧发酵高温期,AOA amo A基因丰度下降至发酵前的0.9%,AOB下降至17.6%,后熟期AOA与AOB的amo A基因丰度与好氧发酵高温期相当。在上述3个阶段AOA与AOB各自存在一个绝对优势菌群,分别为Cluster 3和Nitrosomonas europaea,其中Cluster 3克隆子数目分别占整个克隆文库的70.73%、54.28%、72.45%,Nitrosomonas europaea克隆子数目分别占整个克隆文库的78.44%、93.20%、94.27%。堆肥3个阶段AOA的多样性指数变化不大,Shannon-Wiener值维持在1.53~1.60,但群落结构发生明显演替,随着堆肥温度升高,堆肥前期的一些菌群(Cluster 4、Cluster 5、Cluster 6)逐渐消失,新的菌群Cluster 1出现并成为堆肥中后期的第二大优势菌群。AOB无论是多样性指数还是群落组成,都发生剧烈的变化。AOB在堆肥前期Shannon-Wiener指数值最大(1.47),种群数最多(6个基因簇,分别为Nitrosomonas europaea Cluster,Nitrosomonas halophila Cluster,Nitrosomonas communis Cluster,Nitrosomonas nitrosa Cluster,Nitrosospira briensis Cluster,Nitrosospira multiformis Cluster);进入高温发酵期,Shannon-Wiener下降至0.45,群落结构单一,只有Nitrosomonas europaea Cluster和Nitrosomonas halophila Cluster;进入后熟期,AOB多样性及种群数得到一定程度的回升。     

冰岛硫化叶菌xpb基因缺失突变体的构建及遗传学分析

为在体内研究古菌核苷酸切除修复(nucleotide excision repair,NER)机制,利用基因敲除的方法,对泉古菌核苷酸切除修复机制进行研究,构建了冰岛硫化叶菌编码真核生物NER解旋酶XPB同源蛋白的基因xpb1和xpb2单缺失突变体,从而在体内分析xpb1和xpb2基因的功能。结果表明:基因xpb1或xpb2不是冰岛硫化叶菌存活的必需基因。表型分析发现,xpb1和xpb2基因单缺失突变体相较野生型菌株REY15A,对DNA损伤试剂4-NQO 4-硝基喹啉-1-氧化物(4-nitroquinoline 1-oxide,4-NQO)分别表现出轻微敏感性和不敏感性,暗示NER解旋酶功能在冰岛硫化叶菌体内存在多重冗余。