AM真菌控制蚕豆枯萎病发生的根际微生物效应

在温室盆栽条件下,研究了接种尖孢镰刀菌蚕豆专化型和丛枝菌根(AM)真菌(摩西球囊霉Gm、扭形球囊霉Gt、根内球囊霉Gi及幼套球囊霉Ge)对灭菌连作土壤中蚕豆幼苗生长、枯萎病发生、根际镰刀菌数量和微生物代谢功能多样性的影响.结果表明: 接种AM真菌能显著增加蚕豆幼苗地上部和地下部鲜质量;接种Gm、Gt、Gi和Ge真菌使蚕豆枯萎病病情指数分别显著降低94.0%、60.0%、64.0%和94.0%,使蚕豆根际镰刀菌数量分别显著降低98.6%、74.3%、77.8%和90.4%,以Gm和Ge真菌对蚕豆枯萎病的抑制效应最好.接种Gm、Gt和Ge显著提高了根际微生物对糖类(CH)、氨基酸类(AA)、羧酸类(CA)和酚酸类(PA)碳源的利用,使蚕豆根际微生物的AWCD值分别显著提高34.4%、31.5%和50.8%;而接种Gi对AA、CA、PA类碳源利用和AWCD值均无显著影响,使微生物对CH类碳源的利用显著降低59.3%.主成分和相关分析表明, 接种AM真菌明显改变了蚕豆根际微生物的群落结构.接种Gt真菌提高了蚕豆根际微生物对AA类碳源(β-甲基-D-葡萄糖苷、D-半乳糖酸-γ-内酯、D-甘露醇、N-乙酰基-D-葡萄糖胺、D-纤维二糖)和CA类碳源(D-半乳糖醛酸)的利用,而接种Gi真菌降低了微生物对以上碳源的利用;接种Gm和Ge真菌提高了根际微生物对L-精氨酸和4-羟基苯甲酸的利用.表明糖类和羧酸类是接种Gt真菌后蚕豆根际土壤微生物利用的主要碳源, 氨基酸和酚酸类是接种Gm和Ge后蚕豆根际土壤微生物利用的主要碳源.接种AM真菌显著提高了蚕豆根际微生物的活性,改变了微生物群落结构并抑制病原菌的增殖,是AM真菌减轻了蚕豆枯萎病的发生的重要原因.不同AM真菌减轻了蚕豆枯萎病的发生与其改变微生物的碳源利用密切相关.     

玉米秸秆还田对土壤丛枝菌根真菌群落的影响

为揭示农业管理活动对土壤丛枝菌根(AM)真菌的影响机制,基于Illumina Miseq高通量测序平台以及脂肪酸指纹图谱方法,研究了连续4年玉米秸秆还田后,AM真菌群落组成、AM真菌生物量及其与土壤环境因子间的相互关系.结果表明:所获得的2430个AM真菌OTUs从门到种依次分类,共分为1门、3纲、4目、8科、10属、143种,但不同处理间AM真菌群落丰富度(Chao1指数和ACE指数)、多样性(Shannon、Simpson多样性指数)没有显著差异.AM真菌中类球囊霉属、球囊霉属为优势属.随秸秆还田量的增加,球囊霉属丰度降低;3000、9000 kg·hm^-2秸秆还田量下,类球囊霉属、无梗囊霉属的丰度与对照(0 kg·hm^-2)间差异达极显著水平;原囊霉属、类球囊霉属、球囊霉属在3000 kg·hm^-2秸秆还田量下与对照间差异显著,非度量多维尺度(NMDS)分析表明,9000、12000 kg·hm^-2的秸秆还田量下土壤AM真菌β多样性与对照间聚集度较其他处理相差较远,秸秆还田量对AM真菌β多样性的影响显著.多元分析结果能在累积变量82.8%上揭示土壤主要理化性状与AM真菌丰富度、多样性的空间变化关系.土壤全氮、碱解氮是影响以磷脂脂肪酸表征的土壤主要微生物类群生物量以及以中性脂肪酸表征的AM真菌生物量的主要因子.持续玉米秸秆还田改变了AM真菌属水平上的分类学组成;随秸秆还田量的增加,AM真菌特有的微生物种类减少,AM真菌群落组成间的相似度下降;秸秆还田增加了土壤AM真菌生物量及其占土壤微生物总生物量的比例.     

利用DGGE法研究不同种植体系中根际微生物群落结构

利用DGGE技术研究不同间作和轮作种植体系对作物根际细菌和真菌群落结构的影响.运用16SrDNA和18SrDNA特异引物对,将土壤中提取的总DNA进行PCR扩增后,通过DGGE技术对PCR产物进行分析,结果表明:玉米-蚕豆轮作对蚕豆根际细菌和真菌群落结构影响明显,二者都与单作蚕豆有较大差异;小麦/蚕豆间作明显改变两种作物根际细菌群落结构和蚕豆根际真菌群落结构;玉米/蚕豆间作明显改变玉米根际细菌、真菌群落结构和蚕豆根际真菌群落结构.     

小麦/蚕豆，玉米/蚕豆和小麦/玉米间作对根际细菌群落结构的影响

利用PCR-DGGE技术研究了小麦/蚕豆、玉米/蚕豆和小麦/玉米间作对作物根际细菌群落结构的影响.结果表明：间作能够提高作物根际细菌群落多样性、改变根际细菌群落结构组成.其中,小麦/蚕豆间作对根际细菌群落结构的影响最为突出,作物花期时小麦/蚕豆间作显著提高和改变两种作物根际细菌多样性和群落结构组成.玉米/蚕豆间作主要表现出对苗期玉米根际细菌多样性的显著提高和群落结构组成的改变.小麦/玉米间作对作物根际细菌群落结构的影响程度较弱.同时,3种间作体系都具有不同程度的产量优势.结果证明了间作体系中地上部植物多样性与地下部微生物多样性存在紧密联系.     

间作作物种间相互作用对马铃薯根际土壤丛枝菌根真菌的影响

为揭示间作作物种间相互作用对土壤丛枝菌根(AM)真菌的影响,以马铃薯单作(T0)为对照,基于高通量测序平台的方法,研究了连续3年马铃薯‖玉米(T1)、马铃薯‖蚕豆(T2)下马铃薯根际土壤AM真菌群落组成、多样性与土壤环境因子间的相互关系.结果表明:共获得2893个AM真菌操作分类单元(OTUs),分属1门、3纲、4目、...     

黄顶菊与3种本地植物竞争对丛枝菌根真菌种类和多样性的影响

【目的】外来植物黄顶菊对生态环境和农业经济造成了严重危害,了解黄顶菊与3种不同本地植物种植生长对丛生菌根(AM)真菌群落结构和多样性造成的影响,可以从土壤微生物角度进一步解释黄顶菊的入侵机制。【方法】通过同质园小区试验模拟黄顶菊入侵的生态进程,以黄顶菊和3种本地植物狗尾草、藜、黄香草木樨为研究对象,采用AM真菌的形态学鉴定方法,研究黄顶菊与3种本地植物不同种植方式对AM真菌群落结构和多样性的影响。【结果】(1)黄顶菊根际土壤聚集的AM真菌种类与其伴生本地植物种类有关:黄顶菊与狗尾草混种处理中优势种为网状球囊霉和根内球囊霉,而黄顶菊分别与藜、草木樨混种处理中优势种均为网状球囊霉、根内球囊霉和缩球囊霉;(2)黄顶菊分别与狗尾草和黄香草木樨混种处理中AM真菌种类既高于本地单种处理,也高于黄顶菊单种处理,说明随着黄顶菊的入侵和地上植物多样性的改变,AM真菌种类也发生改变;(3)与3种本地植物单种相比,黄顶菊各混种处理和黄顶菊单种处理中黄顶菊根际土壤根内球囊霉的重要值均增加,表明黄顶菊入侵有利于根内球囊霉的生长和发育。【结论】黄顶菊入侵改变了根际土壤AM真菌的群落结构和多样性,AM真菌的改变既与本地植物种类有关,也与入侵程度有关。     

小麦蚕豆间作对蚕豆根际微生物群落功能多样性的影响及其与蚕豆枯萎病发生的关系

通过田间小区试验,研究了小麦与蚕豆间作对蚕豆枯萎病发生和根际微生物代谢功能多样性的影响。结果表明,小麦与蚕豆间作使蚕豆枯萎病的发病率和病情指数分别比单作显著降低20%和30.4%。与单作处理相比,间作显著增加了蚕豆和小麦根际微生物对31种碳源的平均利用率（AWCD）,其中间作蚕豆的AWCD值最高,比单作增加82.7%,单作蚕豆最低。间作蚕豆和间作小麦根际微生物的Shannon多样性指数与丰富度指数均显著高于单作,间作使蚕豆和小麦的丰富度指数分别增加29.2%和30.3%。根际微生物对六类碳源的利用强度百分比以糖类、羧酸类和氨基酸最高,分别为41.96%,19.80%和18.13%。主成分分析表明,小麦与蚕豆间作改变了根际微生物的群落组成;相关分析表明,糖类、羧酸类和氨基酸类碳源是区分单间作处理差异的主要碳源,其中氨基酸类碳源是最敏感的碳源。小麦与蚕豆间作增加了根际微生物活性,提高了Shannon多样性指数和丰富度指数,改变了微生物群落功能多样性,是抑制蚕豆枯萎病的有效措施。该研究为阐明根际微生物功能多样性变化在间作体系病害控制中的作用与机制奠定了理论基础。     

间作甘蔗对玉米根际微生物功能多样性的影响

在云南省玉溪市元江县和德宏州陇川县甘蔗种植区分别设置甘蔗间作玉米和单作玉米田小区试验,采用Biolog-ECO板测定了不同气候条件下间作玉米与单作玉米根际微生物代谢功能多样性。结果表明:在陇川和元江试验区,间作玉米根际微生物的平均颜色变化率(AWCD)相对于单作玉米分别提高了28.50%和42.32%,Shannon指数和Simpson指数高于单作玉米田;从土壤微生物对碳源的利用率来看,两个试验区的间作玉米根际微生物对22种碳源的利用率高于单作玉米田,对2种碳源的利用率低于单作玉米田,两试验区间作与单作玉米根际微生物对碳源的利用率变化规律不尽一致的碳源有7种。由此表明,间作提高了玉米根际微生物对大部分碳源的利用率,并且在不同试验区有所差异。主成分分析表明,元江和陇川试验区间作玉米和单作玉米根际微生物群落结构明显不同。综上所述,在甘蔗玉米间作体系中,间作玉米根际微生物相对于单作玉米根际微生物群落多样性较高、根际微生物活性较强。该研究结果可为玉米甘蔗间作对土壤微生物活性影响研究提供理论基础。     

不同连作年限怀牛膝根际土壤理化性质及微生物多样性

以不同连作年限怀牛膝根际土壤为材料,采用土壤农化分析技术和磷脂脂肪酸生物标记法,对土壤理化性质以及根际微生物群落结构进行分析。结果表明:怀牛膝连作会使大部分大量元素(全磷、碱解氮、速效磷和速效钾)含量上升,而全氮、全钾的变化并无明显趋势。全量钙、全量镁、全量锰和有效锌的含量也会随着连作年限的增加而增加。研究发现,怀牛膝连作1a的土壤与对照相比,根际土壤微生物群落结构差别不大,但是与连作10a和30a的土壤相比微生物多样性却发生了明显变化。土壤中总PLFAs、细菌、革兰氏阴性菌Gram(-)及真菌含量都随连作年限增加而明显上升,微生物多样性增加,且革兰氏阳性菌Gram(+)/革兰氏阴性菌Gram(-)的比值随着连作年限的增加而下降。此外,土壤中一些嗜热解氢杆菌和真菌等参与土壤物质循环和木质素降解的微生物类群数量明显增加,假单胞菌等有益微生物类群也显著增多,这是牛膝连作促进作用的生物学基础。研究用药用植物怀牛膝证明了植物根际微生物结构和功能多样性对植物连作促进或连作抑制作用的产生与控制的生态学意义。     

盐渍化土壤根际微生物群落及土壤因子对AM真菌的影响

为探明盐渍化土壤影响下AM真菌与根际土壤间的关系,试验选取宁夏碱化龟裂土、草甸盐土、盐化灌淤土3种类型4个样地上典型植被群落,测定了植物根际土壤养分含量、微生物群落结构、AM真菌侵染率以及孢子密度。结果显示:盐渍化土壤根际微生物碳源利用类型显著不同,对芳香类化合物的代谢能力整体较弱;红寺堡草甸盐土上微生物优势群落为氨基酸代谢类群,惠农盐化灌淤土为多聚化合物代谢群,西大滩碱化龟裂土为碳水化合物代谢群。AM真菌孢子密度与微生物碳源代谢群间的关系比较复杂。其中,惠农样点根际土壤孢子密度与多聚化合物微生物代谢群呈显著正相关,西大滩地区孢子密度与碳水化合物微生物代谢群呈显著正相关。土壤有机质、全盐、全氮、碱解氮等土壤肥力因子及土壤中的HCO-3、Na+、Cl-等盐基离子含量能解释AM真菌孢子密度与土壤环境因子之间相互关系的大部分信息。较高的HCO-3浓度促进了AM真菌侵染率的提高,但高盐浓度下Na+和Cl-降低了菌根侵染率。土壤对AM真菌孢子密度、侵染率的影响因土壤盐分组成类型的不同而异。研究结果为深入了解AM真菌多样性,促进宁夏盐碱地的合理开发与利用提供了理论依据。     

桑树根际土壤微生物对间作和施氮的响应

利用Biolog-Ecoplate^TM结合主成分与冗余分析法,研究与苜蓿间作和施氮对桑树根际土壤微生物功能多样性及其与环境因子的影响.结果表明: 与单作无氮相比,单作施氮和间作(无氮和施氮)明显降低桑树根际土壤pH值,显著提高有机质含量、有效氮、含水量和过氧化物酶、脲酶的活性.同时,单作施氮和间作(无氮和施氮)提高了桑树根际土壤微生物的AWCD值、多样性指数和利用碳源比例,且间作施氮增加其提高的程度.主成分分析表明,施氮和间作改变了根际土壤微生物利用碳源强度和种类,其中单作施氮和与间作无氮处理的碳源利用情况相似,但后者对衣康酸和D-葡糖胺酸的利用率(>4%)显著高于前者.冗余分析表明,单作无氮桑树根际土壤微生物多样性与土壤多酚氧化酶活性呈正相关,与土壤含水量呈负相关.单作施氮与间作无氮处理的桑树根际微生物多样性与土壤pH值和含水量呈显著正相关,与土壤氮素营养呈负相关.间作施氮处理的桑树根际微生物多样性与土壤氮素营养呈正相关,而与土壤pH值呈显著负相关.     

Variation of Bacterial Community Diversity in Rhizosphere Soil of Sole-Cropped versus Intercropped Wheat Field after Harvest

As the major crops in north China, spring crops are usually planted from April through May every spring and harvested in fall. Wheat is also a very common crop traditionally planted in fall or spring and harvested in summer year by year. This continuous cropping system exhibited the disadvantages of reducing the fertility of soil through decreasing microbial diversity. Thus, management of microbial diversity in the rhizosphere plays a vital role in sustainable crop production. In this study, ten common spring crops in north China were chosen sole-cropped and four were chosen intercropped with peanut in wheat fields after harvest. Denaturing gradient gel electrophoresis (DGGE) and DNA sequencing of one 16S rDNA fragment were used to analyze the bacterial diversity and species identification. DGGE profiles showed the bacterial community diversity in rhizosphere soil samples varied among various crops under different cropping systems, more diverse under intercropping system than under sole-cropping. Some intercropping-specific bands in DGGE profiles suggested that several bacterial species were stimulated by intercropping systems specifically. Furthermore, the identification of these dominant and functional bacteria by DNA sequencing indicated that intercropping systems are more beneficial to improve soil fertility. Compared to intercropping systems, we also observed changes in microbial community of rhizosphere soil under sole-crops. The rhizosphere bacterial community structure in spring crops showed a strong crop species-specific pattern. More importantly, Empedobacter brevis, a typical plant pathogen, was only found in the carrot rhizosphere, suggesting carrot should be sown prudently. In conclusion, our study demonstrated that crop species and cropping systems had significant effects on bacterial community diversity in the rhizosphere soils. We strongly suggest sorghum, glutinous millet and buckwheat could be taken into account as intercropping crops with peanut; while hulled oat, mung bean or foxtail millet could be considered for sowing in wheat fields after harvest in North China.     

Biotic and Abiotic Properties Mediating Plant Diversity Effects on Soil Microbial Communities in an Experimental Grassland

Plant diversity drives changes in the soil microbial community which may result in alterations in ecosystem functions. However, the governing factors between the composition of soil microbial communities and plant diversity are not well understood. We investigated the impact of plant diversity (plant species richness and functional group richness) and plant functional group identity on soil microbial biomass and soil microbial community structure in experimental grassland ecosystems. Total microbial biomass and community structure were determined by phospholipid fatty acid (PLFA) analysis. The diversity gradient covered 1, 2, 4, 8, 16 and 60 plant species and 1, 2, 3 and 4 plant functional groups (grasses, legumes, small herbs and tall herbs). In May 2007, soil samples were taken from experimental plots and from nearby fields and meadows. Beside soil texture, plant species richness was the main driver of soil microbial biomass. Structural equation modeling revealed that the positive plant diversity effect was mainly mediated by higher leaf area index resulting in higher soil moisture in the top soil layer. The fungal-to-bacterial biomass ratio was positively affected by plant functional group richness and negatively by the presence of legumes. Bacteria were more closely related to abiotic differences caused by plant diversity, while fungi were more affected by plant-derived organic matter inputs. We found diverse plant communities promoted faster transition of soil microbial communities typical for arable land towards grassland communities. Although some mechanisms underlying the plant diversity effect on soil microorganisms could be identified, future studies have to determine plant traits shaping soil microbial community structure. We suspect differences in root traits among different plant communities, such as root turnover rates and chemical composition of root exudates, to structure soil microbial communities.     

玉米/花生间作对土壤微生物和土壤养分状况的影响

通过大田试验,研究了玉米/花生间作对玉米和花生根区土壤微生物和土壤养分状况的影响.结果表明：与单作相比,间作能显著提高玉米和花生根区的土壤细菌数量;间作花生根区土壤真菌和放线菌数量与单作无显著差异;间作玉米根区土壤真菌和放线菌数量比单作明显提高;间作作物根区微生物群落功能多样性和代谢活性比单作有所改善.玉米/花生间作不同程度提高了整个间作系统根区的土壤碱解氮、速效磷、有机质含量及EC值,其中,间作玉米根区土壤养分的增加更为明显,说明玉米/花生间作可以较明显地改善两种作物根区的微生物和养分状况,土壤微生态环境的改善又会促进作物地上部的生长.     

Communities of arbuscular mycorrhizal fungi in arable soils are not necessarily low in diversity

Communities of arbuscular mycorrhizal fungi (AMF) in five agricultural field sites of different management intensities were studied. Variable regions of the ribosomal RNA genes were used to detect and identify AMF directly within colonized roots. Roots from a continuous maize monoculture showed low AMF diversity, in agreement with previous reports on molecular diversity of AMF in agricultural soils. In contrast, a substantially higher diversity of AMF was found throughout the long term 'DOK' field experiment, where organic and conventional agricultural practices have been compared side by side since 1978. In this experiment, a 7-year crop rotation is performed under lower levels of inorganic fertilizer input and chemical pest control. These results are in good agreement with analyses of the spore community previously conducted in these field sites. In a third site, an organically managed leek field with soil of very high phosphate content reflecting the highly intensive conventional field history and intensive tillage, we detected a low-diversity community comparable to the maize monoculture. In addition to fungi from Glomus group A, which have previously been reported to dominate arable soils, we regularly found members of the genera Scutellospora, Paraglomus and Acaulospora. The genus Acaulospora was shown to occur more frequently early in the growing season, suggesting that the life history strategy of AMF may influence the active community at a given time. These data show that the diversity of AMF is not always low in arable soils. Furthermore, low-input agriculture involving crop rotation may provide better conditions to preserve AMF diversity, by preventing the selection for the few AMF taxa tolerating high nutrient levels.     

氮和土著AMF对黄瓜间作土壤酶活性及氮利用的影响

设施土壤氮（N）肥的大量不合理施用和高残留是导致作物硝态N含量超标和农业面源污染的主要因素之一。研究土著丛枝菌根真菌（arbuscular mycorrhizal fungi,AMF）与间作体系强化蔬菜对不同形态N的利用并结合土壤酶活性的反馈作用,可为设施土壤N素的高效利用和降低土壤N残留提供依据。本研究采用盆栽试验,设置黄瓜单作和黄瓜//大豆间作种植模式,不同AMF处理[不接种（NM）、接种土著AMF]和不同形态N处理[不施N（N0）、有机N（谷氨酰胺120mg/kg,ON120）、无机N（碳酸氢铵120mg/kg,ION120）],探讨了设施条件下施用不同形态N、接种土著AMF与间作大豆对黄瓜根围土壤酶活性及氮利用的影响。结果表明,与NM相比,接种土著AMF使设施黄瓜地上部、根系生物量及植株N吸收量均有不同程度的增加,根围土壤NH₄ ⁺-N、NO₃ ^--N含量呈现降低趋势。同一N处理-土著AMF条件下,间作大豆处理下的黄瓜根系菌根侵染率显著高于单作处理;间作大豆也使黄瓜植株地上部、根系生物量及N吸收量显著增加,同时显著降低了根围土壤铵态N含量。此外,间作-土著AMF条件下,ON120和ION120处理的黄瓜根围土壤脲酶活性较N0处理分别提高了30%和14%,蛋白酶和硝酸还原酶活性也呈现出相同趋势。可见,所有复合处理中,以间作体系接种土著AMF与施用适量有机N的组合明显促进了设施黄瓜生长和N素利用率。     

喀斯特地区三种人工林土壤微生物群落结构特征

为揭示不同人工植被修复模式对喀斯特土壤微生物群落的影响,采用氯仿熏蒸提取法和磷脂脂肪酸(phospholipid fatty acid, PLFA)法研究人工构建的降香黄檀(Dalbergia odorifera)纯林(PDOP)、顶果木(Acrocarpus fraxinifolius)纯林(PAFP)、顶果木×降香黄檀混交林(MADP)对土壤微生物生物量及土壤微生物群落结构的影响。结果表明:(1)PDOP的土壤微生物生物量碳(MBC)和微生物生物量氮(MBN)含量显著高于PAFP和MADP,PAFP显著高于MADP。(2)三种人工林土壤真菌、丛枝菌根真菌和总PLFA含量无显著差异,但PDOP土壤细菌、放线菌、丛枝菌根真菌和总PLFA含量均高于PAFP和MADP,PAFP高于MADP。PDOP的土壤细菌、革兰氏阳性菌、革兰氏阴性菌、放线菌的PLFA含量显著高于MADP。MADP的真菌细菌比显著高于PDOP,但与PAFP无显著差异。(3)冗余分析表明,土壤阳离子交换量、pH和C:N是影响土壤微生物群落组成的最主要影响因子。从三种人工林的土壤微生物生物量及微生物群落结构来看,在喀斯特地区MADP并未显示出酸性土地区混交林提高土壤微生物生物量、改善土壤微生物群落结构的优势,但混交林的真菌细菌比最高,更有利于提高土壤生态系统的稳定性。