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。     

三种不同种植模式对土壤细菌群落多样性的影响

采用变性梯度凝胶电泳(DGGE )技术,研究了不同种植模式对土壤细菌群落多样性的影响。结果表明,不同种植模式下DGGE图谱条带的数量及亮度有较大区别,且有几条特征性条带发生了明显的变化。0—15 cm土层细菌群落的丰富度、多样性指数及均匀度指数均表现为:“菜稻菜模式”（RVCs） “休闲轮作模式”（FRCs） “蔬菜连作模式”（VCCs）;15—30 cm土层细菌群落的丰富度、多样性指数及均匀度指数均表现为:FRCs RVCs VCCs。UPGMC聚类分析表明,RVCs 处理的细菌群落结构相似性较低。主成分分析表明,对各主成分起分异作用的主要是 RVCs和FRCs;与主成分因子1 正相关程度较高的种植模式为:RVCs和VCCs,与主成分因子2 正相关程度较高的是 FRCs。可见,水稻土细菌群落多样性与种植模式密切相关,这些变化对土壤细菌群落有重要的调节作用。     

茄子与香蕉轮作配施生物有机肥对连作蕉园土壤微生物区系的影响

连作现象在香蕉生产上非常普遍,而长期连作会导致严重的连作障碍。本文针对香蕉连作障碍,选择连作香蕉13年的地块,采用常规方法结合变性梯度凝胶电泳(PCR-DGGE)技术,在田间条件下研究了轮作茄子配施生物有机肥对高发枯萎病连作蕉园土壤可培养微生物数量、土壤化学性状以及土壤细菌群落结构的影响。结果表明:与连作香蕉相比,轮作茄子处理可显著降低可培养尖孢镰刀菌数量,使其数量从种植初的10~4 cfu·g~(-1)(干土)下降到10~3 cfu·g~(-1)(干土),同时提高了土壤p H,增加了土壤有机质、速效钾、碱解氮含量。无论是轮作还是连作种植模式,与配施普通有机肥相比,配施生物有机肥对可培养尖孢镰刀菌、真菌和细菌数量影响均不显著;但在轮作模式下,施用生物有机肥处理的细菌数量与真菌数量比值(B/F,381.2)显著高于配施普通有机肥处理(270.3)。PCR-DGGE分析结果表明,轮作茄子配施生物有机肥显著改变了土壤细菌群落结构,增加了细菌丰度、稳定性和多样性,其中多样性指数(Shannon-Wiener指数,3.22)较连作香蕉配施普通有机肥处理(2.89)显著增加。以上结果表明,茄子与香蕉轮作有利于连作蕉园土壤的微生态环境,同时轮作配施生物有机肥效果更优。     

设施种植模式对土壤细菌多样性及群落结构的影响

为了研究有机和常规设施种植模式及轮作对土壤细菌多样性和群落结构的影响,本研究采用Illumina平台Hiseq 2500高通量测序技术,于2016年6月(作物处于收获期)对北京市顺义区不同设施种植模式(分别为有机设施种植模式和常规设施种植模式下的叶菜连作、茄果连作和叶茄轮作)下土壤细菌进行16S r RNA测序。测序质控后共获得17 278个操作分类单元(operational taxonomic units,OTUs),共计318 851条有效序列。比较不同种植模式和轮作下土壤细菌多样性、细菌群落结构组成、相对丰度及土壤理化性质与细菌群落多样性关系的差异性。结果表明:土壤微生物群落结构在有机和常规设施种植模式下差异明显,有机设施种植土壤细菌多样性高于常规设施种植;有机设施种植下轮作与连作土壤细菌群落结构表现出明显差异,而常规设施种植下,两者没有明显差异;有机种植模式下,轮作土壤细菌群落多样性高于连作土壤;设施种植土壤细菌群落主要属于鞘氨醇单胞菌属(Sphingomonas,5.05%)和芽孢杆菌属(Bacillus,4.84%),相对丰度大于0.5%的共有14个属。有机设施种植土壤含有较多促进植物生长、有机质分解的细菌,常规设施种植土壤中降解化学杀虫剂、防治土壤病害、促进硝化过程的细菌较多。RDA分析结果显示土壤细菌群落主要受全磷、速效磷、有机质的影响。Tumebacillus、Candidatus Solibacter和Acidothermus都是分解有机质、利用碳源的细菌属,与土壤有机质含量呈正相关关系。由此可见,设施条件下,有机和常规种植土壤微生物群落结构的差异性主要源于肥料使用、有害生物防治措施和管理方式的不同。有机设施种植模式下,轮作更有利于发挥其改良土壤营养循环和防治土壤病虫害的作用。上述结果为在微生物水平上研究设施条件下不同种植模式的土壤生态质量差异提供了参考。     

不同连作年限烤烟根际土壤真菌群落18S rDNA-PCR-DGGE分析

采用PCR-DGGE技术,研究轮作、连作3年、连作6年烤烟根际土壤不同生育期真菌群落结构变化趋势,以期为烟草连作障碍调控提供依据。结果表明:无论是轮作还是连作,烤烟生育中后期植烟土壤真菌群落最为丰富。连作明显改变了烤烟根际土壤的真菌群落结构,且随着连作年限的增加影响越大。DGGE条带回收序列分析和RDA物种因子结果显示长期连作导致土壤真菌菌群结构发生变化,致病菌增多,而且土壤中烟草靶斑病原、腐霉科、被孢霉科、链格孢属、链壶菌科、生赤壳科、盘菌科、镰刀菌属和炭角菌目均与土壤营养代谢密切相关。因此,不同连作年限对烤烟根际土壤真菌群落有较大影响,烤烟连作后真菌种群结构的变化可能是引发烤烟连作障碍的主要原因之一。     

种植方式对玉米不同生长期土壤微生物群落功能多样性的影响

为探讨玉米不同种植方式下土壤微生物群落功能多样性的差异,进行田间定点试验,采用Biolog方法分别研究了4行轮作、4行连作、8行轮作和8行连作的种植方式对玉米种植前、拔节期、抽穗期和收获期土壤微生物功能多样性的影响。结果表明:4种种植方式的土壤微生物均在种植前代谢活性最弱、功能多样性最低,在玉米抽穗期土壤微生物代谢活性最强,功能多样性最高。在种植玉米前,轮作的土壤微生物代谢活性和功能多样性高于连作,8行轮作和4行轮作土壤微生物的物种多样性指数分别比相应的连作高22.93%和11.42%;4行轮作的土壤微生物物种多样性指数比8行轮作低3.17%,而4行连作比8行连作高6.83%。在玉米拔节期、抽穗期及收获期连作土壤微生物功能多样性略高于轮作,且有4行连作大于8行连作的趋势,但差异均未达显著水平。种植前,4种种植模式的土壤微生物对6大类碳源的利用程度整体上都较低,降解碳水化合物类、羧酸类和聚合物类碳源的微生物是种植方式影响的主要土壤微生物类群;随着玉米的生长,土壤微生物对6大类碳源的利用都逐渐增强,玉米拔节期、抽穗期和收获期之间土壤微生物特征碳源没有较大差异,4种种植方式的土壤微生物对聚合物类碳源利用程度差异都不显著。PLS-EDA分析结果表明种植方式对土壤微生物产生较大影响,种植前8行轮作和4行连作的土壤微生物碳源利用模式具有相似性;种植玉米后4种种植方式的土壤微生物对碳源的利用模式存在较大差异,其中4行连作的土壤微生物在玉米拔节期和收获期对碳源的利用模式与其他3种种植方式差异最大。试验说明作物长期连作栽培会影响土壤微生物群落功能,降低土壤微生物物种多样性,引起土壤微生物群落结构与功能的失调。     

亚热带四种主要植被类型土壤细菌群落结构分析

应用PCR-DGGE方法分别对天目山和玲珑山的四种典型森林土壤细菌群落结构进行了研究分析。结果表明,阔叶林、杉木林、马尾松林和毛竹林在天目山土壤中的DGGE条带数分别为49、45、51和51条,在玲珑山土壤中的DGGE条带数分别为42、4i多是奥前啊2、44和48条。不同森林植被土壤DGGE带谱差异不明显,天目山和玲珑山4种森林土壤分别有31条和23条共性条带,分别占总数的61%和48%以上,然而每种林分也有各自的特征条带。相对而言,两个样地杉木林与其他三种林分的土壤细菌群落结构相似度较低,相似值分别为0.72和0.68,其土壤细菌多样性指数也明显低于其他3种林分,而毛竹林和马尾松林多样性指数则没有显著差异。同时分析两个采样区的土壤细菌DGGE发现,土壤性质对细菌群落结构的影响大于植被。     

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。     

菇菜套作对土壤微生物群落的影响

套作是防治连作障碍的有效方法之一,但是蔬菜和可食用菌之间的套作机理研究鲜见报道,尤其是其土壤微生物学机制。本研究建立菇菜套作体系,利用实时荧光定量PCR和PCR-DGGE技术研究土壤细菌和真菌群落的变化。结果表明,菇菜套作显著提高了番茄生物量,且其番茄果实产量最高,硝酸盐含量最低。与对照相比,菇菜套作下土壤细菌和真菌基因拷贝数量均无显著变化;DGGE指纹图谱表明,不同处理下的细菌群落无明显差异,但是菇菜套作下真菌群落结构发生了分异,主要表现为尖孢镰刀菌(Fusarium oxysporum)和稻黑孢菌(Nigrospora oryzae)代表型条带的强度的下降。     

棉花连作对北疆土壤酶活性、致病菌及拮抗菌多样性的影响

本文通过测定土壤酶活性与微生物PCR-DGGE指纹图谱研究了北疆棉区5年棉花连作(CtN5)、10年棉花连作(CtN10)及15年棉花连作(CtN15)对土壤过氧化氢酶、蔗糖酶、芳基硫酸酯酶、脱氢酶和蛋白酶酶活性的影响,分析了土壤细菌、真菌、镰刀菌和枯草芽孢杆菌群落结构多样性对北疆棉田长期连作的响应。结果表明:过氧化氢酶、蔗糖酶、脱氢酶活性随棉花连作年限延长而下降。CtN15处理的过氧化氢酶、蔗糖酶和脱氢酶活性分别比CtN10处理下降15.0%、6.4%和12.0%,比CtN5处理下降16.8%、58.6%和49.5%(P0.05);芳基硫酸酯酶与蛋白酶活性随连作年限的增加呈先下降后升高的特点。土壤细菌、真菌多样性指数随连作年限的增加明显下降。CtN15的细菌条带数比CtN10下降7.41%,Ct N10比CtN5降低1.72%。CtN15真菌条带数和Shannon-Wiener多样性指数分别为78和3.22,比CtN5处理低17.02%和5.29%。土壤镰刀菌和枯草芽孢杆菌的条带数、多样性指数均表现为先下降后升高。CtN15枯草芽孢杆菌Shannon-Wiener和Simpson指数分别比CtN10处理高54.8%和14.5%。北疆长期连作棉田的土壤酶活性和土壤微生物群落多样性总体呈下降趋势,长期连作对棉田土壤生物性状有明显负面影响。     

利用PCR-DGGE技术研究长期自然恢复、种植作物和裸地条件下黑土土壤细菌群落结构变化

Soil microbial biomass and community structures are commonly used as indicators for soil quality and fertility. A investigation was performed to study the effects of long-term natural restoration, cropping, and bare fallow managements on the soil microbial biomass and bacterial community structures in depths of 0--10, 20--30, and 40--50 cm in a black soil (Mollisol). Microbial biomass was estimated from chloroform fumigation-extraction, and bacterial community structures were determined by analysis of 16S rDNA using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE). Experimental results showed that microbial biomass significantly declined with soil depth in the managements of restoration and cropping, but not in the bare fallow. DGGE profiles indicated that the band number in top 0--10 cm soils was less than that in depth of 20--30 or 40--50 cm. These suggested that the microbial population was high but the bacterial community structure was simple in the topsoil. Cluster and principle component analysis based on DGGE banding patterns showed that the bacterial community structure was affected by soil depth more primarily than by managements, and the succession of bacterial community as increase of soil depth has a similar tendency in the three managements. Fourteen predominating DGGE bands were excised and sequenced, in which 6 bands were identified as the taxa of Verrucomicrobia, 2 bands as Actinobacteria, 2 bands as α-Proteobacteria, and the other 4 bands as δ-Proteobacteria, Acidobacteria, Nitrospira, and unclassified bacteria. In addition, the sequences of 11 DGGE bands were closely related to uncultured bacteria. Thus, the bacterial community structure in black soil was stable, and the predominating bacterial groups were uncultured.     

马铃薯连作栽培对土壤微生物多样性的影响

为阐明马铃薯连作对土壤微生物群落结构和功能多样性的影响,揭示马铃薯连作障碍机制,本试验采用BIOLOG技术结合丛枝菌根真菌(AMF)形态学鉴定方法,就连作0(迎茬)、2 a、4 a、6 a、10 a的马铃薯田块土壤进行研究。结果表明:土壤养分含量随马铃薯连作年限增加有一定程度下降,其中,连作10 a马铃薯根际土壤的全磷、速效磷、速效钾和碱解氮与连作4 a相比分别下降61.32%、26.86%、26.87%和17.24%,但没有明显的养分亏缺和不均衡现象。土壤微生物群落结构发生了较大变化,放线菌、真菌数量显著随连作年限的延长先增加后减少,呈单峰型变化趋势;细菌数量随连作年限的延长呈逐步减少趋势,但差异不显著。连作4~6 a土壤微生物群落依然有较强的功能多样性,培养120 h后,连作6 a较迎茬AWCD值提高3.89%;群落组成中随连作年限的延长以碳水化合物、氨基酸类为碳源的微生物类群代谢能力明显下降,但代谢功能多样性趋于一致。连作马铃薯土壤AM真菌优势种发生改变,迎茬土壤为沙漠球囊霉(Glomus deserticola),连作2 a土壤为扭形球囊霉(Glomus delhiense)和福摩萨球囊霉(Glomus formosanum),连作10 a土壤为球泡球囊霉(Glomus globiferum)。多元分析结果表明,土壤微生物结构与功能多样性、AM真菌多样性变化受土壤p H、全磷含量、放线菌数量、细菌数量及土壤中以碳水化合物、氨基酸类等为碳源基质的微生物类群影响。说明长期连作栽培会影响土壤真菌、放线菌的数量,使真菌群落中AM真菌种的多样性显著下降,优势种发生改变,打破了微生物群落结构与功能平衡,引起土壤微生物群落结构与功能的失调。     

Factors affecting soil microbial community structure in tomato cropping systems

Soil and rhizosphere microbial communities in agroecosystems may be affected by soil, climate, plant species, and management. The management and environmental factors controlling microbial biomass and community structure were identified in a three-year field experiment. The experiment consisted of a tomato production agroecosystem with the following nine treatments: bare soil, black polyethylene mulch, white polyethylene mulch, vetch cover crop, vetch roots only, vetch shoots only, rye cover crop, rye roots only, and rye shoots only. The following hypotheses were tested: (1) Temperature and moisture differences between polyethylene-covered and cover-cropped treatments are partly responsible for treatment effects on soil microbial community composition, and (2) Different species of cover crops have unique root and shoot effects on soil microbial community composition. Microbial biomass and community composition were measured by phospholipid fatty acid analysis. Microbial biomass was increased by all cover crop treatments, including root only and shoot only. Cover cropping increased the absolute amount of all microbial groups, but Gram-positive bacteria decreased in proportion under cover crops. We attribute this decrease to increased readily available carbon under cover-cropped treatments, which favored other groups over Gram-positive bacteria. Higher soil temperatures under certain treatments also increased the proportion of Gram-positive bacteria. Vetch shoots increased the amount and proportion of Gram-negative bacteria, fungi, and arbuscular mycorrhizal fungi in the rhizosphere of tomato plants. The imposed treatments were much more significant than soil temperature, moisture, pH, and texture in controlling microbial biomass and community structure.     

不同栽培环境下有机与常规蔬菜土壤的细菌群落多样性差异

土壤细菌群落在蔬菜栽培中发挥着重要作用。基于DNA和RNA水平,利用PCR-DGGE技术研究了不同栽培环境下有机与常规蔬菜土壤细菌群落多样性差异,以及土壤理化性质与细菌群落多样性的关系。结果表明：不同栽培方式下土壤细菌多样性存在明显差异,土壤微生物的优势种群和数量受有机、常规栽培和季节影响,有机栽培较之常规栽培能够显著增加土壤细菌群落多样性;聚类分析表明,16S rDNA细菌群落多样性与季节相关,而16S rRNA细菌群落多样性与栽培方式相关;差异条带测序显示,大多细菌与不可培养细菌种属有较高同源性,其余9种推测属于假单胞菌属;CCA分析说明pH是影响土壤细菌群落多样性的主要因素,有机栽培土壤中微生物生物量C、N以及有机质含量显著高于常规栽培土壤。综上,有机栽培能够丰富活性细菌群落多样性,具有土壤优化效应。     

Soil aggregation and bacterial community structure as affected by tillage and cover cropping in the Brazilian Cerrados

Microbial-based indicators of soil quality are believed to be more dynamic than those based on physical and chemical properties. Recent developments in molecular biology based techniques have led to rapid and reliable tools to characterize microbial community structures. We determined the effects of conventional and no-tillage in cropping systems with and without cover crops on bacterial community structure, total organic carbon (TOC) and soil aggregation. Tillage and rotation did not affect TOC from bulk soil. However, TOC was greater in the largest aggregate size class (7.98–19 mm), and had greater mean-weight diameter under no-tillage than under conventional tillage in the 0–5 cm soil layer. Soil bacterial community structure, based on denaturing gradient gel electrophoresis of polymerase chain reaction amplified DNA (PCR/DGGE) using two different genes as biomarkers, 16S rRNA and rpoB genes, indicated different populations in response to cultivation, tillage and depth, but not due to cover cropping. Soil bacterial community structure and meanweight diameter of soil aggregates indicated alterations in soil conditions due to tillage system.     

小麦、毛苕子与黄瓜轮作对土壤微生态环境及产量的影响

采用常规方法及PCR-DGGE技术对土壤速效养分含量、土壤酶活性和微生物群落结构多样性以及黄瓜产量进行分析,以探究小麦、毛苕子与黄瓜轮作对黄瓜土壤微生态环境及产量的影响。结果表明,不同轮作处理均显著地提高了黄瓜产量,有效地改善了土壤微生态环境。其中小麦-黄瓜轮作黄瓜产量极显著高于对照(p<0.01),增产28.04%,其多酚氧化酶、过氧化氢酶及脲酶活性总体较高。毛苕子-黄瓜处理增产16.78%,并增加了土壤养分含量,转化酶活性较高,极显著高于对照(p<0.01)。DGGE结果表明,轮作有助于根际土壤细菌种类的增多及结瓜后期真菌种类的减少,其中毛苕子-黄瓜处理的影响更为明显。小麦-黄瓜轮作对土壤真菌与定植后30 d土壤细菌群落结构具有一定的影响。总之,小麦、毛苕子与黄瓜轮作有利于缓解黄瓜连作障碍,改善土壤微生态环境,提高黄瓜产量。     

Soil microbial biomass,crop yields,and bacterial community structure as affected by long-term fertilizer treatments under wheat-rice cropping

Soil microbial biomass carbon (SMBC) and nitrogen (SMBN), soil microbial community structure, and crop yields were studied in a long-term (1982–2004) fertilization experiment carried out in Suining, Sichuan province of PR China. Eight treatments included three chemical fertilizer (CF) treatments (N, NP, NPK), three CF + farmyard manure (M) treatments (NM, NPM, NPKM), M alone and no fertilizer (CK) as control. The results showed that the soil microbial biomass was higher in soil treated with CFM than in soil treated with CF alone, and that NPKM gave the highest rice and wheat yields. The SMBC and SMBN were higher after rice than those after wheat cropping. SMBC correlated closely with soil organic matter. Average yields of wheat and rice for 22 years were higher and more stable in the fertilized plots than in control plots. Bacterial community structure was analyzed by PCR-DGGE targeting eubacterial 16S rRNA genes. A higher diversity of the soil bacterial community was found in soil amended with CFM than in other fertilizer treatments. Some specific band emerged in the soil amended with M. The highest diversity of bacterial communities was found in the NPKM treated soil. The bacterial community structures differed in rice and wheat plots. Sequencing of PCR products separated in DGGE showed that some of the common and dominant bands were closely related to Aquicella lusitana and to Acidobacteria. This study demonstrated that mixed application of N, P, and K with additional M amendment increased soil microbial biomass, diversified the bacterial communities and maintained the crop production in the Calcareous Purplish Paddy soil.     

Community structure of methanogenic archaea in paddy field soil under double cropping (rice-wheat)

Community structure of methanogenic archaea in paddy field soil under double cropping (rice [Oryza sativa L.] and wheat [Triticum aestivum L.]) was studied by the denaturing gradient gel electrophoresis (DGGE) method. Soil samples under flooded and upland conditions were collected 7 and 6 times, respectively, from two paddy fields throughout a year, and two primer sets, 0357F-GC/0691R and newly designed 1106F-GC/1378R, were used for DGGE analysis. The 25 and 29 different bands were observed on the DGGE gels with the primers 0357F-GC/0691R and 1106F-GC/1378R, respectively. DGGE band patterns of the methanogenic archaeal community were stable throughout a year including the cultivation periods of rice under flooded conditions and of wheat under upland conditions. Cluster analysis and principal component analysis suggested that the difference in the soil type (sampling region) largely influenced the community structures of methanogenic archaea in paddy field soil, while the effects of sampling period and different fertilizer treatments on them were small. Most of the sequences obtained from the DGGE bands were closely related to Methanomicrobiales, Methanosarcinaceae, Methanosaetaceae and Rice cluster-I.