黄河三角洲刺槐根际与非根际细菌结构及多样性

为精确分析黄河三角洲刺槐根际与非根际土壤细菌群落定殖情况,本研究采用高通量测序方法对刺槐根际与非根际土壤细菌结构及多样性进行了研究。研究表明,根际土壤细菌共有36门214属,非根际土壤细菌共有33门153属。变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、酸杆菌门(Acidobacteria)细菌丰度超过15%,是根际与非根际土壤中的优势菌落。根际与非根际土壤中酸杆菌门、硝化螺旋菌门(Nitrospirae)丰度差异显著。根际土壤中红游动菌属(Rhodoplanes)、溶杆菌属(Lysobacter)、热单胞菌属(Thermomonas)、链霉菌属(Streptomyces)及非根际土壤中红游动菌属、溶杆菌属、链霉菌属、Kaistobacter细菌丰度超过4%。根际土壤中固氮菌丰度显著高于非根际土壤,解磷、解钾细菌丰度差异不显著。根际与非根际土壤细菌Chao丰富度分别为2 054、2 376,差异显著。根际与非根际土壤细菌之间的权重(Weighted Unifrac)距离在0.12~0.25之间。综上所述,黄河三角洲刺槐根际与非根际土壤细菌结构具有一定差异,多样性差异显著。     

马铃薯与玉米复合种植对土壤化感物质及土壤细菌群落结构的影响

为探究马铃薯与玉米复合种植对化感物质积累与细菌群落结构的影响,分析轮作、间作缓解连作障碍的机制,本研究以马铃薯连作、玉米连作、马铃薯||玉米间作、马铃薯-玉米轮作第8年的土壤为对象,利用GC-MS测定土壤中化感物质含量,并采用Illumina Miseq高通量测序技术对土壤细菌16Sr DNA V4-V5区域进行测序,分析土壤中细菌多样性和群落结构的变化,并对化感物质和优势菌属进行相关性分析。结果表明:玉米连作和马铃薯连作会导致化感物质的积累,玉米连作土壤积累了更多的油酸、亚油酸、花生酸、木焦油酸等脂肪酸,马铃薯连作土壤积累了更多的硬脂醇、二十烷醇等脂肪醇类物质。轮作降低了大部分化感物质的积累,间作降低的化感物质种类相对轮作较少。不同种植方式下土壤细菌群落结构发生了显著变化,相对于连作,间作和轮作Ace指数和Chao指数显著升高。在门水平上,轮作土壤放线菌丰度显著高于马铃薯连作土壤,间作土壤拟杆菌门丰度显著低于玉米连作土壤,两种连作土壤中酸杆菌门丰度都较轮作显著升高。在属水平上,一些有益细菌如节杆菌属、溶杆菌属等在复合种植土壤中相对丰度更高。通过相关性分析发现土微菌属、小梨形菌属与脂肪醇类物质呈显著正相关,黄杆菌属、溶杆菌属、微杆菌属等与脂肪酸类物质呈显著负相关。马铃薯与玉米复合种植降低了化感物质在土壤中的积累,从而抑制了土壤细菌丰度的降低,提高了有益菌属丰度,消减了连作障碍。     

PGPR菌剂对辣椒的促生效应及根际土壤细菌的响应研究

研究田间条件下PGPR菌剂灌根对辣椒根际土壤细菌数量及群落结构的影响,为深入探究PGPR菌剂的田间促生机制提供理论依据。以4株PGPR菌株制成PGPR菌剂,对辣椒进行灌根,并采用Illumina-MiSeq平台对辣椒根际土壤细菌V3-V4区进行Paired-end测序,结合细菌及功能菌群数量、辣椒产量指标,比较分析菌剂灌根配施全量化肥(PGPR-CF1)、菌剂灌根配施80%化肥(PGPR-CF2)和全量化肥(CK)处理后对辣椒根际土壤细菌数量及群落结构特征的影响。结果表明:PGPR菌剂灌根后辣椒根际土壤细菌及包含固氮、解磷及解钾等功能菌群的数量显著提高,且物种丰度和多样性均高于对照,细菌优势度则表现为PGPR-CF1和CK组高于PGPR-CF2。所有土样共得到有效序列573 896条,在相似水平为97%下聚类分析得到OTUs数分别为2 153个(CK)、2 337个(PGPR-CF1)、2 358个(PGPR-CF2);各处理辣椒根际土壤中的优势门为酸杆菌门、放线菌门、拟杆菌门、绿弯菌门、浮霉菌门和变形菌门,相对丰度总占比分别为83.7%(PGPR-CF1)、79.55%(PGPR-CF2)和79.77%(CK)。PGPR菌剂灌根后增加了伯克霍尔德氏菌目、芽孢杆菌目、鞘脂单胞菌目等的相对丰度,降低了芽单胞菌目、酸杆菌目的相对丰度。Pearson指数表明,辣椒产量和单果重与根际土壤细菌多样性和丰度正相关。研究显示,PGPR菌剂处理,可以提高根际土壤细菌和功能菌群的数量,同时影响了物种多样性和群落结构,并促进了辣椒增产。     

不同茬口和施氮水平对南瓜根际细菌碳分解潜力的影响

为探明根际细菌群落多样性及有机碳分解功能基因对耕作措施的响应特征,以收获小麦和西瓜后种植的南瓜根际土壤为研究对象,以两种施氮水平(纯氮施入量分别为1.0、2.0 g/kg)为处理,结合高通量测序和PICRUSt功能预测解析不同茬口和施氮水平下南瓜根际细菌群落结构和参与有机碳分解代谢细菌的关键功能基因丰度的差异。结果表明:各处理南瓜根际土壤细菌Shannon多样性指数和细菌群落结构无差异;所有处理中变形菌门、拟杆菌门、放线菌门、蓝藻细菌门相对丰度之和达到90%。冗余分析发现,小麦茬土壤中硝态氮的含量、西瓜茬土壤中铵态氮和硝态氮的含量对南瓜根际细菌群落结构的作用高于其他理化指标。高氮水平下,与西瓜茬相比,小麦茬南瓜根际土壤变形菌门的相对丰度降低,但拟杆菌门和放线菌门的相对丰度升高(P<0.05)。无论施氮水平高低,与西瓜茬相比,小麦茬南瓜根际土壤分解半纤维素和淀粉类物质诸如α-淀粉酶和β-半乳糖苷酶等产酶基因丰度提高。因此,施氮水平并未影响不同茬口土壤南瓜根际土壤微环境和细菌种群结构和多样性的稳定性;但高氮处理下,茬口类型影响参与有机碳分解的关键细菌门类相对丰度。与西瓜茬土壤相比,小麦茬土壤可能更能促进南瓜根际土壤有机碳的分解。     

轮作与连作对烟田土壤微生物区系及多样性的影响

为探讨轮作与连作对烟田根际土壤微生物群落的影响,采用实验室微生物培养联合微生物高通量测序技术,分析了玉米-烤烟轮作和烟叶连作对烟株根际土壤微生物数量、群落结构及其多样性的影响。研究结果表明,轮作烟田根际土壤中可培养细菌和放线菌数量均高于连作烟田根际土壤,但真菌数量低于连作烟田。土壤高通量测序结果表明,轮作烟田根际土壤真菌和细菌群落多样性均高于连作烟田;在烟株的生长过程中,轮作和连作烟田在旺长期和成熟期微生物群落相似度降低。相较于黑胫病/根腐病/根结线虫发病率较高(36. 67%)的连作烟田来说,轮作烟田发病率较低(0%),进一步分析真菌和细菌群落结构发现,轮作烟田根际土壤中拮抗菌如Sphingomonas (鞘脂单胞菌属)、Pseudomonas (假单胞菌属)、Aspergillus (曲霉属)等相对丰度均高于连作烟田,而连作烟田根腐病病菌Pythium (腐霉属)丰度较高。     

有机无机肥料配施对植烟土壤养分及细菌群落结构的影响

为研究有机无机肥料配施对植烟土壤细菌群落结构的影响,采用Illumina MiSeq高通量测序技术,分析了连续3年施用化肥(CF)、烟草秸秆生物有机肥配施化肥(TBF)和高碳基土壤修复肥配施化肥(HCF)对土壤细菌多样性和群落结构的影响,同时结合土壤理化性质探究不同施肥条件下细菌群落变化的重要影响因素。与CF处理相比,HCF处理土壤碱解氮含量显著降低了34.09%。多样性指数分析显示有机无机肥料配施有提高土壤细菌多样性的趋势,HCF处理土壤细菌多样性最高。有机无机肥料配施提高了土壤放线菌门(Actinobacteria)、变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、芽单胞菌门(Gemmatimonadetes)、拟杆菌门(Bacteroidetes)和浮霉菌门(Planctomycetes)细菌丰度,降低了土壤绿湾菌门(Chloroflexi)、Saccharibacteria、厚壁菌门(Firmicutes)、WD272和蓝细菌门(Cyanobacteria)细菌丰度。有机无机肥料配施显著增加了土壤Patulibacter、短芽孢杆菌属(Brevibacillus)、农研丝杆菌属(Niastella)等有益细菌属丰度。Spearman相关性分析和典范对应分析(CCA)表明土壤pH值、碱解氮和速效钾是影响土壤细菌群落结构的重要因子。有机无机肥料配施对植烟土壤细菌多样性和群落结构产生了深刻影响,土壤pH值、碱解氮和速效钾是调控细菌群落结构的重要影响因素,在土壤培肥中应引起充分的重视。     

不同种植年限宁夏枸杞根际微生物多样性变化

为了解长期人工种植枸杞根际土壤微生物的种群结构变化特征,利用Illumina Mi Seq测序平台分别对种植5 a、10 a和15 a宁夏枸杞根际土壤微生物基因组总DNA中16S和18S r DNA基因的部分区域进行测序,经UPARSE pipeline和RDP classifier软件进行聚类分析和物种注释。结果表明,长期种植宁夏枸杞不会改变其根际土壤p H,但会导致根际土壤全磷、有效磷、全盐含量和电导率升高。测序结果表明,不同种植年限枸杞根际土壤细菌群落α多样性无显著变化,但真菌群落多样性在种植10 a枸杞中较种植5 a时显著降低(p0.05),表明根际细菌群落多样性受种植年限影响较小。从门的分类水平看,酸杆菌门、放线菌门、拟杆菌门、厚壁菌门、绿弯菌门、泉古菌门、蓝菌门、芽单胞菌门、变形菌门以及真菌子囊菌门、担子菌门、接合菌门的比例在不同种植年限的枸杞根际土壤中显著改变(p0.05)。属水平的分析也表明,共有27个细菌属和16个真菌属的比例发生改变(p0.05),这些结果表明枸杞根际土壤微生物群落组成受种植年限的影响更大。相关性分析结果表明,种植年限、土壤全磷及有效磷含量是影响枸杞根际微生物群落结构的主要因子。     

马铃薯连作栽培对土壤微生物多样性的影响

马铃薯是宁夏回族自治区南部山区重要的粮菜兼用作物,随栽培面积持续扩大,轮作倒茬困难,由此带来的连作障碍问题日渐突出。为探明马铃薯连作障碍机制,分别采集马铃薯正茬、连作1年、6年和10年的根际土壤样本,利用末端标记限制性片段长度多态性(terminal restriction fragment length polymorphism,T-RFLP)技术研究连作栽培对根际土壤微生物多样性的影响,以期揭示马铃薯根际土壤主要菌群动态变化规律及连作障碍的可能原因。结果表明:连作栽培后马铃薯土壤细菌和真菌DNA仍具有较高的T-RFLP多态性,但不同连作年限根际土壤中优势T-RFs片段发生变化,多年连作使某些T-RFs消失;随连作年限增加,根际土细菌Shannon-Wienen指数、Simpson指数和Sorenson指数下降,而真菌Shannon-Wienen指数和Simpson指数上升。菌群分析发现,土壤细菌中以厚壁菌门(Firmicutes)芽孢杆菌纲(Bacilli)和梭菌纲(Clostridia)所占比例最大。连作使细菌和真菌菌群发生变化,芽孢杆菌纲、鞘氨醇纲(Sphingobacteria)等比例下降甚至消失,β-变形菌纲(Beta proteobacteria)和异常球菌纲(Deinococci)比例上升;土壤真菌中座囊菌纲(Dothideomycetes)煤炱目(Capnodiales)随连作年限增加比例下降,粪壳菌纲(Sordariomycetes)肉座菌目(Hypocreales)随连作年限增加比例上升。马铃薯连作使根际土壤中芽孢杆菌属(Bacillus)等有益菌属的细菌减少,罗尔斯通菌属(Ralstonia)等致病菌属的细菌增加。连作导致马铃薯根际土壤细菌多样性水平降低,真菌多样性水平升高,根际土壤微生物多样性存在明显差异,连作破坏了根际土壤微生物群落的平衡,使其根际土壤微生态环境恶化。连作障碍可能是多因素综合互作用的结果,持续的研究有利于做出较合理的解释。     

连续水旱轮作对水稻冷浸田土壤细菌群落结构的影响

利用高通量测序技术,采用单季稻-冬闲(CK)、油菜-水稻(R-R)、春玉米-水稻(C-R)、紫云英-水稻(M-R)和蚕豆-水稻(B-R)五个处理探究水旱轮作模式对水稻冷浸田土壤细菌群落结构的影响,同时分析了土壤理化因子与细菌群落结构的关系。结果表明:水旱轮作能够显著增加水稻产量18%~44%。与CK相比,绝大多数水旱轮作降低了土壤细菌的丰富度,而细菌多样性与CK处理无显著差异。变形菌门和酸杆菌门为土壤中的优势细菌类群,其中变形菌门的丰度最高。与CK相比,轮作后酸杆菌、放线菌、硝化螺旋菌、慢生根瘤菌和亚硝化螺菌等参与碳氮循环的菌群丰度明显增加。聚类分析表明CK和M-R轮作土壤细菌群落单独聚为一类,而B-R、C-R和R-R轮作土壤聚为另一类。冗余分析表明土壤全磷、有效磷和p H是影响水旱轮作土壤细菌群落结构的主要因子。     

基于高通量测序的两种植被恢复类型根际土壤细菌多样性研究

为研究库布齐沙地生态恢复过程中不同植被恢复类型土壤微生物细菌群落结构、多样性的变化特征,以流动沙地为对照,运用高通量测序技术,对自然恢复的油蒿群落、人工种植的中间锦鸡儿群落根际和非根际土壤细菌多样性进行了研究,并分析了土壤理化性质对其分布的影响。结果表明:(1)与流沙对照相比,两种植被恢复类型对细菌多样性产生了正效应,细菌群落丰度、多样性和均匀度明显增加。其中,自然恢复的油蒿群落土壤细菌丰度高于人工种植的中间锦鸡儿群落;(2)变形菌门、酸杆菌门和放线菌门为研究区土壤中的优势细菌类群,其中变形菌门在各样地丰度比例最高,变形菌的4个亚群变化趋势一致,α-变形菌相对含量在油蒿和中间锦鸡儿群落根际土壤中明显增加,尤其是自然恢复的油蒿群落根际土壤中α-变形菌得到了很好的恢复;(3)土壤有机质、全氮、速效氮、速效钾含量和土壤含水量是影响土壤细菌群落丰度和多样性的主要土壤因子,典型相关分析表明土壤有机质、全氮、全钾、速效钾、速效氮含量对于研究区土壤细菌群落遗传多样性的变化起着重要作用。     

连作对再植枸杞根际细菌群落多样性和群落结构的影响研究

受枸杞道地产区土地资源等因素限制,连作障碍已成为影响枸杞产业发展的重要原因之一,导致严重的经济损失.研究连作条件下枸杞农田土壤生态系统微生物群落的演替规律对枸杞产业的可持续发展具有重要的理论意义.以宁夏银川市南梁农场连作多年的枸杞地为研究对象,利用Illumina MiSeq测序技术分析了连作对再植枸杞根际/非根际细菌群落的影响.结果表明,连作地显著抑制再植枸杞苗地径的增加,且其土壤pH较对照样地显著降低(p<0.05).测序结果证实,与对照样地相比,连作地再植枸杞根际土壤细菌物种数显著降低(p<0.05),细菌群落α多样性下降(p>0.05).主坐标分析表明,连作和对照样地间枸杞非根际细菌群落结构无明显差异,但连作显著改变再植枸杞根际细菌的群落结构.对细菌群落丰度的统计分析发现,连作地枸杞根际浮霉菌门、非根际假单胞菌门的相对丰度较对照样地显著降低(p<0.05).此外,冗余分析结果表明:枸杞园土壤pH和有效磷含量是影响枸杞非根际土壤细菌群落结构变化的主要因素,分别解释了41.8%和35.4%的群落结构变化(p<0.05),其他土壤因子无统计学意义,但土壤理化因子对再植枸杞根际细菌群落结构变化的影响均未达显著水平.这些结果证实连作能够显著抑制再植枸杞生长、影响再植枸杞根际细菌群落结构和多样性,干扰枸杞与土壤细菌群落间的互作关系.这些研究结果将为解析枸杞连作障碍机制提供理论基础.     

甘蔗宿根矮化病感病与非感病株根际土壤生物学性状及细菌群落结构特征

[目的]比较甘蔗宿根矮化病(ratoon stunting disease,RSD)感病植株与非感病植株根际土壤的生物学性状及细菌群落结构特征,旨在为构建甘蔗健康的根际微环境,筛选高效RSD生防细菌提供参考。[方法]通过田间调查和实验室鉴定,以甘蔗RSD感病植株为试材,非感病植株为对照,采集甘蔗RSD感病植株和非感病植株的根际土壤,并基于传统和现代高通量测序技术,分析了甘蔗RSD感病植株和非感病植株根际土壤的生物学性状和细菌群落结构特征。[结果]与甘蔗RSD非感病植株相比,感病植株根际土壤中指示土壤肥力与健康状况的生物学性状指标β-葡糖苷酶、磷酸酶和氨肽酶活性,以及微生物生物量碳、氮、磷显著降低;同时,指示细菌丰富度的Chao1指数和指示细菌多样性的Shannon指数显著下降。门分类水平与非感病甘蔗植株相比,RSD感病植株根际土壤中Proteobacteria(变形杆菌门)、Actinobacteria(放线菌门)、Gemmatimonadetes(芽单胞菌门)和Nitrospirae(硝化螺旋菌门)等优势门类细菌占比呈倍级降低,但Chloroflexi(绿弯菌门)、Acidobacteria(酸杆菌门)、Firmicutes(厚壁菌门)、Cyanobacteria(蓝细菌门)、Planctomycetes(浮霉菌门)、Bacteroidetes(拟杆菌门)等优势门类细菌占比呈倍级增加;在属分类水平,与非感病甘蔗植株相比,RSD感病植株根际土壤中Xanthobacteraceae(黄色杆菌属)、Acidothermus、Gaiellales、Roseiflexus(玫瑰菌属)、Micromonosporaceae(小单孢菌属)和Nitrospira(硝化螺旋菌属)细菌占比呈倍级降低,但Acidobacteria(嗜酸菌属)细菌及部分未知菌属却呈倍级提高。[结论]甘蔗RSD感病植株根际微环境中指示土壤肥力的生物学指标显著降低,细菌丰富度和多样性显著下降,部分优势细菌门属占比发生剧变可能是导致甘蔗RSD发生的重要原因。     

盐地碱蓬根际土壤细菌群落结构及其功能

盐地碱蓬作为生物改良盐碱地的理想材料,其根际土壤微生物对土壤改良发挥着重要作用。为了深入探索环渤海滨海盐碱地碱蓬根际土壤细菌群落结构组成及其功能,采用Illumina Misep高通量测序平台对环渤海地区滨海盐碱地盐地碱蓬根际土壤和裸地土壤进行测序。从16个样本中获得有效序列734 792条, 4 285个OTUs,归属于41门、100纲、282目、400科、892属、1 577种。盐地碱蓬根际土壤细菌群落由变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、绿弯曲门(Chloroflexi)、拟杆菌门(Bacteroidetes)、芽单胞菌门(Gemmatimonadetes)、酸杆菌门(Acidobacteria)、厚壁菌门(Firmicutes)、蓝藻细菌门(Cyanobacteria)、髌骨细菌门(Patescibacteria、浮霉菌门(Planctomycetes)组成。Alpha多样性计算结果表明,盐地碱蓬根际土壤细菌群落结构多样性高并与裸地土壤间差异显著;LEfSe(LDAEffectSize)分析发现,盐地碱蓬与裸地差异指示种明显不同。PCoA与相关性Heatmap表明,盐地碱蓬、速效氮、速效钾、速效磷、电导率是影响土壤细菌目类水平群落组成的主要因子。PICRUSt(Phylogenetic InvestigationofCommunitiesbyReconstructionofUnobserved States)分析表明微生物群落在新陈代谢等40个功能方面盐地碱蓬根际土壤比裸地土壤高。本研究表明盐地碱蓬覆盖能够降低土壤盐分,增加土壤养分,对土壤细菌群落多样性及其功能有积极作用。     

Long-term effect of plastic film mulching and fertilization on bacterial communities in a brown soil revealed by high through-put sequencing

The objective of this study was to access the effect of long-term fertilization and film mulching application on soil properties and bacterial community structure. We used 16S rRNA gene to investigate soil bacterial community composition by high through-put sequencing. The results demonstrated that predominant groups in the bacterial community were: Proteobacteria, Bacteroidetes, Actinobacteria, Acidobacteria, Firmicutes, Planctomycetes, Gemmatimonadetes, Verrucomicrobia, Chloroflexi and Cyanobacteria. Long-term fertilization of combined manure and nitrogen application caused significant decrease in soil bacterial diversity and richness compared to non-fertilization control, though manure fertilization alone played a significant role in restoring bacterial diversity. Film mulching and manure fertilization significantly increased the relative abundances of soil bacterial groups mentioned above. Furthermore, film mulching played significant role in shaping the bacterial community structure regardless of fertilization. Redundancy analysis (RDA) showed that soil moisture content, pH, total N and soil organic C had significant (P < 0.05) influence on dominant bacterial groups. Altogether, plastic film mulching and manure application prevented loss of soil bacterial diversity and abundance during long-term fertilization. These findings showed the detrimental use of combined manure and nitrogen fertilization to soil microbes and the useful application of manure fertilization coupled with film mulching to soil biodiversity in long-term fertilization experiments.     

寡糖对土壤微生物多样性及群落结构的调节作用

土壤微生物种群类型,对土壤质量和作物生长具有重要影响,研究寡糖对土壤微生物种群的影响特征,有助于正确、高效及安全使用寡糖。本研究利用人工气候室进行土壤培养,土壤施加50mg·L−1的壳寡糖（CSOS）和纤维寡糖（COS）溶液,以清水（CK）为对照处理,培养6d后取样,利用高通量测序技术,分析土壤微生物群落结构组成及多样性分布特征。结果表明：壳寡糖（CSOS）和纤维寡糖（COS）处理均显著改变细菌、真菌的群落结构,提高细菌的物种观测数。变形菌门（Proteobacteria）、酸杆菌门（Acidobacteria）、放线菌门（Actinobacteria）、绿弯菌门（Chloroflexi）、芽单胞菌门（Gemmatimonadetes）和拟杆菌门（Bacteroidetes）为优势细菌门,子囊菌门（Ascomycota）、担子菌门（Basidiomycota）和被孢霉门（Mortierellomycota）为优势真菌门。通过组间群落组成比较分析可知,壳寡糖（CSOS）和纤维寡糖（COS）处理均不同程度降低酸杆菌门（Acidobacteria）的相对丰度,增加变形菌门（Proteobacteria）、放线菌门（Actinobacteria）、芽单胞菌门（Gemmatimonadetes）、壶菌门（Chytridiomycota）以及有益菌属溶杆菌属（Lysobacter）、硝化螺旋菌属（Nitrospira）、Haliangium、芽球菌属（Blastococcus）和链霉菌属（Streptomyces）的相对丰度,但与纤维寡糖（COS）相比,壳寡糖（CSOS）处理微生物群落组成的变化幅度更大。此外,壳寡糖（CSOS）和纤维寡糖（COS）处理在调节土壤微生物群落结构上存在一定差异。其中,壳寡糖（CSOS）处理有益菌属Talaromyces的相对丰度增加195%,纤维寡糖（COS）处理有益菌属假单胞菌属（Pseudomonas）的相对丰度增加215%。综上,壳寡糖和纤维寡糖处理均能优化土壤微生物群落的结构组成,其调控差异性有助于理解不同寡糖的调控机制,推动寡糖的应用与推广。     

Members of soil bacterial communities sensitive to tillage and crop rotation

Pyrosequencing was used to study the effect of rotation and tillage on total bacterial communities. We designed primers to the bacterial 16s rDNA and amplified DNA from soil samples from a long-term tillage/rotation trial in Kansas for two seasons. The 2 × 2 factorial trial had two rotation treatments (wheat-wheat and wheat-soybean) and two tillage treatments (conventional and no-till). A total of 20,180 16s rDNA sequences were generated and 2337 operational taxonomic units (OTUs) were assembled using a 97% similarity cut-off. The phylum Proteobacteria represented 38% of 299 identified taxa. The second most abundant phylum was Acidobacteria, making up 20% of the sequences, the majority of which were Acidobacteria Group 1. The phyla Actinobacteria and Gemmatimonadetes comprised 12% and 3.5% of the sequences. Other groups detected included TM7, Nitrospira, Verrucomicrobia, and Bacteroidetes. Some clusters of Acidobacteria Group 1 were more frequent in continuous wheat versus wheat-soybean rotation, some Acidobacteria Group 2 were more frequent in no-till, and some Acidobacteria Group 4 were more frequent in wheat-soybean rotation. These results were validated by quantitative real-time PCR. Pyrosequencing provided taxonomic information about the overall bacterial community, and detected community shifts resulting from different cropping practices.     

利用Illumina高通量测序对复合重金属污染下根际和非根际稻田土壤细菌的研究

There is an increasing concern about rice (Oryza sativa L.) soil microbiomes under the influence of mixed heavy metal contamination.We used the high-throughput Illumina MiSeq sequencing approach to explore the bacterial diversity and community composition of soils in four paddy fields,exhibiting four degrees of mixed heavy metal (Cd,Pb and Zn) pollution,and examined the effects of these metals on the bacterial communities.Our results showed that up to 2 104 to 4 359 bacterial operational taxonomic units (OTUs) were found in the bulk and rhizosphere soils of the paddy fields,with the dominant bacterial phyla (greater than 1％ of the overall community) including Proteobacteria,Actinobacteria,Firmicutes,Acidobacteria,Gemmatimonadetes,Chloroflexi,Bacteroidetes and Nitrospirae.A number of rare and candidate bacterial groups were also detected,and Saprospirales,HOC36,SC-I-84 and Anaerospora were rarely detected in rice paddy soils.Venn diagram analysis showed that 174 bacterial OTUs were shared among the bulk soils with four pollution degrees.Rice rhizosphere soils displayed higher bacterial diversity indices (ACE and Chao 1) and more unique OTUs than bulk soils.Total Cd and Zn in the soils were significantly negatively correlated with ACE and Chao 1,respectively,and the Mantel test suggested that total Pb,total Zn,pH,total nitrogen and total phosphorus significantly affected the community structure.Overall,these results provided baseline data for the bacterial communities in bulk and rhizosphere soils of paddy fields contaminated with mixed heavy metals.     

Comparison of bacterial community structures at main habitats in paddy field ecosystem based on DGGE analysis

Bacterial communities at different habitats in a Japanese paddy field ecosystem were compared to understand the bacterial world in the ecosystem as a whole by analyzing data of the denaturing gradient gel electrophoresis (DGGE) band patterns and the sequenced DGGE bands. The habitats were floodwater, percolating water, microcrustacean inhabiting in floodwater, plow layer soil, rice roots, rice straw and rice straw compost incorporated in soil, rice straw placed on the soil surface, plant residues in paddy fields, and rice straw under composting process. Phylotype (band) richness, diversity, evenness, and stability of the bacterial communities at the respective habitats were evaluated based on the DGGE profile data. Phylotype richness was greater near plant residues, rice straw buried in soil and rice straw placed on soil surface, while it was smaller at microcrustacean and rice straw compost buried in soil. The samples from plow layer soil and rice straw compost buried in soil showed considerably higher index values for diversity, evenness, and stability, while those from rice straw placed on soil surface and microcrustacean had lower values of the indices than other habitats. Sequences of totally 250 DGGE bands were assigned to phyla or classes. Distribution of bacterial members to phylogenetic taxa was different among the respective habitats. Inhabitants in plow layer soil were most widely distributed among the groups (nine phyla: Proteobacteria, Chloroflexi, Chlorobi, Verrucomicrobia, Acidobacteria, Nitrospira, candidate division OP10, Cyanobacteria, and Actinobacteria), while those in floodwater and microcrustacean were restricted to only three phyla (Proteobacteria, Bacteroidetes, and Actinobacteria). Proteobacteria and Bacteroidetes were found at all the habitats and the habitats except for plow layer soil, respectively, whereas abundant members belonged to Chloroflexi and Actinobacteria in plow layer soil. “Comprehensive mapping” of DGGE fragments was conducted by principal component analysis based on evolutionary distances of the fragments to 202 reference bacterial strains to overview phylogenetic relationships of bacterial members among the respective habitats. The score plots with the first and second principal components distinctly characterized bacterial members at the respective habitats, and the similarity between the respective communities was clearly demonstrated. Overall, bacterial communities at the respective habitats were distinct and different in the diversity and stability to each other, which may have contributed to the diversity of overall bacterial communities in the paddy field ecosystem.