高产水稻土细菌多样性的培养法与非培养法比较研究

利用细菌的通用引物扩增江西余江县高产水稻土红壤细菌总DNA和平板培养细菌混合总DNA的16S rDNA基因片段,在此基础上分别建立两种16S rDNA文库(文库a和文库b)。从两个文库中各随机挑选100个克隆,扩增出阳性克隆中的插入片段后选用HhaⅠ和RsaⅠ两种四碱基酶进行ARDRA(amplified rDNA restriction analysis)分析。统计比较分析发现,文库a的Shannon-Wienner指数、Simpson指数、丰富度、均一度分别为4.432、0.987、18.885和0.973,均高于文库b中相应的多样性参数(分别为2.271、0.758、5.736和0.501),即平板培养方法所展现的细菌群落结构多样性低于土壤中原始的多样性。结果表明,传统培养方法存在着很大的局限性,必须结合新的分子生物学技术手段才能更全面完善地认识土壤微生物群落结构多样性,以期充分利用其中丰富的微生物资源。     

西藏高原土壤中可培养细菌群落结构分析

以4个西藏高海拔(4 056～5 015 m)高原土壤为材料,经室内恒温短期碳源富集培养后,建立4个土壤可培养细菌库.采用细菌16S rDNA的序列分析技术测定供试土壤可培养细菌库的群落结构特征.结果表明,从XZ02、XZ06、XZ08和XZ12共4个土样的细菌库中,分别获得21,37,31,32个细菌的16S rDNA序列,共产生45个OTU类型.不同样品产生的OTU类型个数和种类有差异,不同样品之间存在共有的优势OTU,但比例不同.多样性指数分析表明,XZ08的Shannon-Wiener指数(H’)和物种丰富度指数(dMa)均为最大,XZ02的辛普森指数(Simpson index,Ds)和物种均匀度指数(E)均为最大,表明不同土壤可培养细菌的多样性指数大小不同;PCA分析表明,XZ06和XZ12群落结构相近,XZ02和XZ08与其群落结构差异大.典型OTU进化定位分析表明,4个土样的细菌主要分布在厚壁菌门和变形菌门中,XZ02和XZ12以芽孢杆菌为主,而XZ02和XZ12以芽孢杆菌和假单胞菌为主.不同采样点土壤的可培养细菌群落结构多样性和分布上均有差异.     

库姆塔格沙漠北界阿奇克谷地土壤可培养细菌多样性

为明确库姆塔格沙漠北界阿奇克谷地的土壤细菌多样性,探索该地区土壤微生物与植物群落的相互作用关系,采用NA、R2A培养基分离培养土壤细菌,通过16S rDNA序列系统发育分析鉴定菌株。结果表明,R2A平板菌落数量和种类较多;发现13株潜在新种菌株,已知菌种分属3大类群、4属、20种,厚壁菌门(Firmicutes)为优势类群,芽胞杆菌属(Bacillus)为优势属,枯草芽胞杆菌(Bacillus subtilis subsp.inaquosorum)为优势菌种。与其他类似环境相比,阿奇克谷地土壤中可培养细菌生物量偏小,可培养的细菌多为抗性、耐性强的极端微生物。该地区特有的微生物资源、功能微生物和新菌资源丰富,为防沙治沙、微生物菌剂开发等研究提供物质基础。本研究结果对库姆塔格沙漠的自然生态保护以及极端环境微生物资源的应用具有重要意义。     

我国主要低产水稻冷浸田土壤微生物特征分析

研究我国冷浸田主要分布的7个省份冷浸田土壤样品的微生物特征,通过与当地高产田土壤微生物特征比较发现:我国冷浸田土壤的微生物量碳含量(70.98～356.61mg/kg)总体低于高产田,差异显著(P<0.05);冷浸田土壤中可培养的细菌、真菌、放线菌、氨化细菌、固氮菌、纤维素分解菌的平均值分别为高产田土壤的53.93%,43.33%,47.32%,51.98%,44.83%,47.80%。表明我国冷浸田土壤微生物总量偏低且活性弱。但是,可培养的硫化细菌数量(平均16.21×106个/g土)和铁还原菌数量(平均9.28×107个/g土)高于高产田土壤中可培养的硫化细菌数量(平均13×106个/g土)和铁还原菌的数量(平均7.32×107个/g土),说明我国冷浸田土壤中硫化氢和亚铁浓度较高。因此,只要有针对性的改良冷浸田土壤微生物群落结构和功能的多样性,冷浸田土壤肥力将大大提高。     

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。     

烟株生长过程中土壤微生物的变化特征

为有效防治烟草青枯病等土传病害,探究了烟株生长过程中土壤微生物的变化特征。分别在烟株不同生长期采集根部土壤进行可培养微生物数量测定和16S/18S rDNA基因测序,以分析烟株生长过程中微生物数量、多样性以及群落结构的变化。结果表明,可培养细菌、放线菌数量随烟株生长先增多后减少,其中旺长期数量最多;真菌数量先减少后增多,以旺长期数量最少。细菌群落多样性无明显变化,真菌群落多样性在旺长期最低。微生物群落结构在旺长期变化较大,细菌优势菌群Acidobacteria(酸杆菌门)丰度较团棵期减少30%±12%,Actinobacteria(放线菌门)丰度增加116%±19%,真菌优势菌群Ascomycota(子囊菌门)丰度增加57%±36%,norankk-Fungi丰度减少83%±17%。烟株生长发育改变了土壤微生物的数量和群落结构,旺长期微生物群落结构的改变与青枯病发生存在一定联系,特别是真菌优势菌的变化可能会影响青枯病的发生。     

饲用玉米连作土壤可培养细菌多样性与肥效菌株监测

为深入了解饲用玉米连作对土壤微生物多样性的影响,对甘肃区采集的连续7年种植玉米土壤中可培养细菌的多样性和促生细菌资源进行全面地分析和挖掘。分别利用富营养(NA)和选择性(Ashby)培养基进行可培养细菌的分离,对获得菌株进行16S rRNA基因测序和比对分析,评价其产植物生长素(IAA)、溶磷、固氮等能力。自不同年份土壤中共分离得到各类细菌224株。菌株功能评价结果表明,34株具有产IAA的能力,25株具溶解有机磷的能力,19株具溶解无机磷的能力,27株具有较高的固氮酶活性,有5株可同时具有产IAA和溶磷的功能。可培养细菌分离信息分析结果表明,种植玉米土壤的细菌及肥效菌株的丰富度及多样性随着连作年限的增加呈逐渐上升趋势,优势菌属均为Pseudomonas,广泛分布于各年份的土壤中,且表现出优秀的产IAA、溶磷及固氮的能力。研究表明,玉米7年连作可明显影响土壤菌群结构,土壤中蕴藏着丰富的有益功能细菌资源。分离的功能菌株为研制适用于甘肃玉米绿色生产的微生物肥料提供信息与优良菌种支撑。     

蚯蚓粪对温室黑土土壤酶活性及细菌多样性的影响

试验研究了蚯蚓粪对黑土土壤酶活性、细菌数量及多样性的影响。共设5个处理:处理1,对照(CK),不施肥;处理2,蚯蚓粪7.5 t hm-2;处理3,蚯蚓粪15 t hm-2;处理4,蚯蚓粪30 t hm-2;处理5,蚯蚓粪60 t hm-2。结果表明,蚯蚓粪对土壤过氧化氢酶、脲酶、蔗糖酶和磷酸酶活性有促进作用,但因酶类型的不同略有差异;随着蚯蚓粪施用量的增加,土壤细菌数量呈现先升高后降低的趋势,而土壤细菌群落多样性表现为递增趋势。各处理对土壤细菌丰富度和多样性指数的影响显著高于对照,但各处理间均匀度指数无差异。聚类分析表明,处理3(蚯蚓粪15 t hm-2)与处理4(蚯蚓粪30 t hm-2)的土壤细菌DGGE图谱相似性达93.8%,并聚成一簇,说明处理3土壤细菌种类与处理4相似性最高,处理3与处理4对土壤细菌多样性指数与均匀度指数影响均无显著差异,但条带数显著增加;处理4与处理5(蚯蚓粪60 t hm-2)相似性达到78.1%,与处理1和处理2(7.5 t hm-2)的相似性只有57.3%,说明蚯蚓粪的施入导致土壤有益微生物种类和数量增加,提高土壤细菌多样性,从而使土壤酶活增强。     

黄河三角洲贝壳堤放线菌多样性及抑菌活性

研究旨在揭示黄河三角洲贝壳堤放线菌多样性,以便从中寻找新的具有生物防治功能的微生物资源。采用3种分离培养基,利用稀释平板涂布法对贝壳堤土壤样品进行分离;通过形态特征、生理生化实验及16S rDNA基因测序对放线菌进行了分类鉴定。结果表明分离到的174株放线菌分属于10个科,13个属,其中链霉菌属(42%)和拟诺卡氏菌(11%)为优势菌属。16S rDNA基因分析表明,61%的菌株与已知菌的相似性都在99%以下,其中菌株BK-17的16S rDNA序列与同源性最近的菌株相似率仅为95.2%。以2株细菌和5株植物病原真菌作为指示菌,进行了抑菌活性测定,有94株至少对1种指示菌有抑菌作用,有8株对7种指示菌都有很强的拮抗作用。以上研究结果表明黄河三角洲贝壳堤土壤中存在丰富的放线菌资源,存在很多潜在的新菌种和活性菌株,有进一步研究和开发的价值。     

番茄根际土尚未培养细菌的分离培养方法探究

为探究获得土壤中尚未培养细菌资源的方法,以MS无机盐为基础养分,设置1/100×TSB、1/50×R2A和土壤浸提液(SE)3种低有机营养类型,凝固剂用琼脂(A)或结冷胶(G),得到一组培养基MS+TSG、MS+TSA、MS+R2A和MS+SE,以及在此基础上添加1.5 g/L丙酮酸钠作为抗氧化剂的第二组培养基。用上述8种培养基分离培养健康番茄根际土壤中的细菌,并对各种培养基平板上代表性菌落进行16S rRNA基因分析。结果表明,在MS无机盐和低有机营养组合的培养基平板上,在28℃下培养28 d后可检测到菌落数达1×107 CFU/g土,4种培养基平板上检测到的菌落数量由多到少顺序为MS+SE>MS+TSG>MS+R2A>MS+TSA;在相同的低有机营养条件下,用结冷胶作为凝固剂可获得更多菌落,添加丙酮酸钠为抗氧化剂会显著降低菌落的多样性。鉴定的84个代表性菌落属于34个属的52个种,其中7个(占检测菌株数的8.33%)为可能新种,不同培养基上种丰富度表现为MS+TSA>MS+R2A>MS+TSG>MS+SE。结果说明,采用低有机营养的MS无机盐培养基有利于分离获得土壤中的一些尚未培养细菌。     

东北黑土区不同土地利用方式下农田土壤微生物多样性

为探究黑龙江省黑土区不同土地利用方式下土壤微生物多样性,该研究主要采用Biolog Eco微平板法,以荒地为对照,研究了黑龙江省中部和西南部黑土区玉米、水稻、大豆及土豆4种不同土地利用方式下土壤微生物多样性的变化。结果表明：1）可培养细菌的数量从大到小依次为土豆、水稻、大豆、玉米、荒地,但群落Shannon-Wiener多样性指数从高到低依次为：荒地（2.18）、玉米（2.11）、土豆（2.00）、水稻（1.73）、大豆（1.49）;2）不同利用方式下黑土区微生物碳源利用程度大致随培养时间的延长而升高,并且氨基酸、糖类以及聚合物类是黑土微生物代谢的最主要碳源;玉米土壤微生物的Shannon-Wiener指数（3.18）、McIntosh指数（5.96）、丰富度指数（24.89）、及Simpson指数（0.95）比其他土地利用方式土壤微生物的多样性指数高,而水稻土壤微生物的多样性指数最低,土豆、大豆与荒地土壤微生物的多样性指数间无显著差别;3）不同土地利用方式显著影响了土壤微生物群落碳源代谢多样性,并且对土壤微生物群落代谢特征起分异作用的主要碳源类型为糖类、氨基酸类和羧酸类,其中糖类尤为突出。该研究将有助于了解黑土区土壤微生物多样性与土地利用方式之间的关系,为黑土区农业的可持续发展提供一定的科学依据。     

西北半干旱荒漠草原与耕地土壤可培养微生物多样性及分布特征比较

采用稀释涂布平板、分离培养和16S r DNA序列分析法对我国甘肃白银地区半干旱荒漠草原土壤可培养细菌、放线菌、真菌数量及群落分布特征进行了分析,比较了荒漠草原和耕地土壤微生物多样性。发现荒漠草原土壤可培养细菌、放线菌、真菌数量分别为1.23×106、0.19×106、0.18×106cfu·g-1,耕地三类微生物数量分别是3.03×106、0.53×106、0.05×106cfu·g-1。荒漠草原可培养细菌、放线菌数量明显低于耕地,而真菌数量高于耕地。从荒漠草原分离出14株细菌,分别属于γ-变形菌纲(γ-Proteobacteria)噬冷杆菌属(Psychrobacter),放线菌门(Actinobacteria)皮球菌属(Kytococcus),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus)、亮氨酸芽孢杆菌属(Lysinibacillus)、土壤芽孢杆菌属(Solibacillus)、气球菌属(Aerococcus),优势菌为芽孢杆菌属和噬冷杆菌属。耕地分离出可培养细菌19株,分别属于ɑ-变形菌纲(ɑ-Proteobacteri)根瘤菌属(Rhizobium)、中华根瘤菌属(Sinorhizobium),γ-变形菌纲(γ-Proteobacteria)假单胞菌属(Pseudomonas),厚壁菌门(Firmicutes)芽孢杆菌属(Bacillus),放线菌门(Actinobacteria)微杆菌属(Microbacterium)、节杆菌属(Arthrobacter)、微球菌属(Micrococcus)、考克氏菌属(Kocuria),以放线菌门细菌为主(占57.9%)。从荒漠草原分离放线菌共8株,分别属于链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)、间孢囊菌属(Intrasporangium),而耕地主要为链霉菌属(Atreptomyces)、小单孢菌属(Micromonspora)。荒漠草原真菌主要是交链孢霉属(Alternaria)、芽枝霉属(Cladosporium),耕地土壤真菌包括青霉属(Penicillium)、交链孢霉属(Alternaria)、曲霉属(Aspergillus)、毛霉属(Mucor)、链孢霉属(Coniothecium)。试验结果表明,荒漠草原与耕地土壤微生物都具有较丰富的多样性,但微生物群落结构存在一定差异,同一区域不同深度土壤中微生物数量和种类也存在差异,耕地土壤微生物多样性明显高于荒漠草原。     

不同植被类型对不同土层黑土土壤微生物群落功能多样性的影响

土壤微生物群落功能多样性对评估土壤生态系统稳定性具有重要作用。本文采用Biolog方法,对比研究长期不同植被类型：自然恢复(GL)、农作物(AL)、人工林(FL)及无植被覆盖(BL)对表层(0～15 cm)和亚表层(15～35 cm)土壤微生物群落功能多样性的影响。结果表明：不同土层土壤微生物的平均颜色变化率(AWCD)变化均表现为GL>AL>FL>BL,且0～15 cm土层的AWCD值均高于对应植被15～35 cm土层。GL、AL和FL处理的土壤微生物Shannon指数和McIntosh指数、总的碳源利用能力均显著高于BL处理的,GL处理0～15 cm土层的Shannon指数和McIntosh指数最高,分别为3.38和6.89。在0～15 cm土层AL和FL处理土壤微生物对羧酸类利用相对较高,而GL处理对氨基酸类利用较高;在15～35 cm土层AL和FL处理土壤微生物对氨基酸类利用最高,而GL被对羧酸类利用的最高。通过主成分分析,GL、AL和FL处理在0～15 cm土层土壤微生物的碳源利用方式及代谢功能相似;15～35 cm土层下AL和FL处理的土壤微生物的碳源代谢...     

Comparisons of different hypervariable regions of rrs genes for fingerprinting of microbial communities in paddy soils

The use of molecular approaches based on 16S rDNA-PCR in microbial ecology has revealed a tremendous prokaryotic diversity in environmental samples. However, there is little or no systematic evaluation of the impacts of hypervariable (V) regions of rrs genes choice on microbial community analysis in soil samples, especially the detailed information about the dominant groups preferentially amplified by different primer pairs. In the present study, eight primer pairs were detected to compare the different V regions for fingerprinting microbial communities in a paddy soil irrigated with petroleum-wastewater, using denaturing gradient gel electrophoresis (DGGE) and amplified ribosomal DNA restriction analysis (ARDRA) techniques. Results reveal the obvious PCR bias produced by different V regions. Both ARDRA analysis of 16S rDNA clone library and DGGE suggest that V₁-V₃ region amplified with primer pair 8f-519r produced the most informative fingerprinting profiles. Additionally, V₃-V₅ region amplified with 341f-907r was another preferable choice for microbial diversity in petroleum-contaminated soil. The V₄-V₅ region and single V region (V₁, V₃, and V₈) were not recommended for the future study of microbial diversity in soil samples. Phylogenetic analysis of 123 sequences from libraries constructed by amplicons generated from six different V regions suggests that different dominant groups were amplified with distinct primer sets. In detail, V₁-V₃ library (amplified with 8f-519r) and V₃-V₅ library were dominated by Actinobacteria (20.4%) (particularly in genus Arthrobacter), V₁-V₃ library (amplified with 63f-518r) was dominated by γ-Proteobacteria (25.0%) and α-Proteobacteria (22.0%) (particularly in genus Brevundimonas), V₃ library was dominated by β-Proteobacteria (22.3%) (particularly in genus Gallionella) and α-Proteobacteria (20.0%), V₆-V₈ library was dominated by Chlamydiae (20.4%) and β-Proteobacteria (20.4%), V₈ library was dominated by γ-Proteobacteria (27.2%) (particularly in genus Acinetobacter) and β-Proteobacteria (14.0%). The present work strongly recommends that primer pairs should be chosen cautiously in community diversity analysis based on PCR amplification of 16S rDNA, and involving at least two different 16S rDNA universal primer pairs would perform better.     

养殖欧洲鳗鲡不同体位及其养殖水体中的菌群组成研究

为探明养殖欧洲鳗鲡不同体位及其养殖水体中可培养菌群的组成结构,本研究利用16S rDNA序列分析法对从精养池鳗鲡的鳃部、肠道和表皮及其养殖水体中分离得到的可培养细菌进行了分子鉴定并构建了系统发育树。研究结果显示,鳃部、肠道、表皮和水体的菌密度分别为1.6×10^6 cfu/g、2.2×10^7 cfu/g、1.4×10^4cfu/cm2和4.5×10^3 cfu/mL;分离菌株分别属于γ-变形菌纲的肠杆菌属、不动杆菌属、栖水菌属,β-变形菌纲的食酸菌属,芽孢杆菌纲的葡萄球菌属、气球菌属,黄杆菌纲的金黄杆菌属和放线菌纲的微球菌属等5大类8个菌属。其中,微球菌属、肠杆菌属和栖水菌属分布最广,各样品中均有检出;而气球菌属和食酸菌属仅在鳃部分布。各生态位中,鳃部菌群最为多样,含有葡萄球菌属之外的7个属;而水体菌群种类最少,只有4个属。此外,菌群组成含量的分析结果表明,鳗鲡鳃部以金黄杆菌属（28.3%）和肠杆菌属（26.1%）居多;而肠道、表皮和养殖水体都以微球菌属占绝对优势,分别为43.6%、53.5%和74.8%。     

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.     

铅锌矿区重金属污染对微生物数量及放线菌群落结构的影响

采集四川省汉源县富泉乡万顺铅锌矿区5个不同重金属浓度的土壤样品,进行了微生物数量及放线菌多样性的研究。经分离、纯化得到43株不同的放线菌,然后对其进行BOXAIR-PCR和16SrDNAPCR-RFLP分析。结果表明,铅锌矿区重金属复合污染对土壤微生物数量有较大的影响,随着铅锌矿区重金属污染程度的加剧,土壤微生物的总数下降。相关性分析表明,重金属含量与细菌数量呈极显著负相关（P〈0.01）,与放线菌数量、真菌数量呈显著负相关（P〈0.05）。供试菌株的16SrDNA用HaeⅢ、HinfⅠ和TaqⅠ酶切后具有32种遗传图谱类型。BOXAIR-PCR的聚类结果表明在86%的水平上,所有菌株分为10个遗传类型,结果基本与16SrDNAPCR-RFLP聚类差异不大。来源于高重金属的含量样品的菌株基本聚在一起,可能是重金属含量影响了放线菌的分布。同时,16SrDNA序列聚类分析结合系统发育树分析表明链霉菌属是汉源铅锌矿区主要的放线菌属并且具有遗传多样性。     

Advantages of a diluted nutrient broth medium for isolating N2-producing denitrifying bacteria of α-Proteobacteria in surface and subsurface upland soils

This research aimed to determine whether a diluted nutrient broth (DNB) medium was different from a conventional nutrient broth (NB) medium when counting and isolating denitrifying bacteria in surface and subsurface upland soils. To this end, we investigated populations of denitrifying bacteria isolated from the surface to a depth of 4 m of subsurface upland soil that had received slurry. The DNB medium gave higher viable counts of denitrifying bacteria than the NB medium and a higher isolation ratio of denitrifying bacterial isolates. In total, 74 isolates from the DNB medium (D-isolates) and 26 isolates from the NB medium (N-isolates) were collected. We characterized their denitrifying activity and analyzed the diversity of 16S rDNA and denitrifying-related genes. Seventy-three percent of the D-isolates were oligotrophic denitrifying bacteria. The N₂-producing, oligotrophic denitrifying bacteria, largely of α-Proteobacteria, increased in the D-isolates. The D-isolates and the N-isolates had some taxonomic overlapping on a phylogenetic tree based on 16S rDNA. It was not possible to identify the denitrification phenotype (N₂-producing or N₂O-producing) on the phylogenetic tree. Phylogenetic groups of isolates corresponded to nirK groups, except in some isolates in which horizontal gene transfer might have occurred. The terminal gas emission of the isolates was consistent with the existence of the nosZ gene. The DNB medium may be very useful in isolating N₂-producing denitrifying α-Proteobacteria. Its use highlights the ecological significance of oligotrophic isolates and the different viable counts resulting from the selectivity of conventional and diluted media.     

磷供应对玉米根际微生物碳源利用和功能多样性的影响

磷有效性能够改变根分泌物的组成和数量,调节土壤微生物的群落结构和多样性,但磷添加如何影响土壤微生物碳源利用和功能多样性尚不清楚。本研究通过盆栽土培试验,设置2个磷处理[低磷5.7 mg(P)?kg?1和高磷200 mg(P)?kg?1],以生长35 d的玉米根际土壤为研究对象,采用Biolog微平板法,分别在培养后240 h内每隔24 h检测具有31种不同碳源的微孔溶液颜色变化,揭示磷供应对玉米根际微生物碳源利用模式和功能多样性的影响。结果表明:随着培养时间的延长,土壤微生物对土壤碳源的利用呈现增加的趋势,直至碳源消耗殆尽;高磷供应显著增加了玉米根际土壤微生物群落平均颜色变化率(average well color development,AWCD),提高了对糖类及其衍生物、氨基酸和代谢产物的利用,但没有显著提高对脂肪酸和脂类的利用;在培养前72 h内,高磷供应显著增加了玉米根际微生物多样性指数、优势度指数和均匀度指数,但培养72 h后,磷供应对其没有显著的影响。主成分分析结果表明,提取的前3个主成分解释了75.15%的碳源利用,高磷和低磷处理具有显著不同的土壤微生物碳源利用模式。总之,糖类及其衍生物、氨基酸和代谢产物是玉米根际土壤微生物利用的主要碳源,短期磷添加能够显著增加土壤微生物对碳源的利用,在一定程度上能够提高土壤微生物群落功能多样性。     

Phosphate solubilizing microorganisms isolated from rhizosphere of maize cultivated in an oxisol of the Brazilian Cerrado Biome

Many soil microorganisms are able to transform insoluble forms of phosphorus to an accessible soluble form, contributing to plant nutrition as plant growth-promoting microorganisms (PGPM). The objective of this work was to isolate, screen and evaluate the phosphate solubilization activity of microorganisms in maize rhizosphere soil to manage soil microbial communities and to select potential microbial inoculants. Forty-five of the best isolates from 371 colonies were isolated from rhizosphere soil of maize grown in an oxisol of the Cerrado Biome with P deficiency. These microorganisms were selected based on the solubilization efficiency of inorganic and organic phosphate sources in a modified Pikovskaya's liquid medium culture containing sodium phytate (phytic acid), soybean lecithin, aluminum phosphate (AlPO₄), and tricalcium phosphate (Ca₃(PO₄)₂). The isolates were identified based on nucleotide sequence data from the 16S ribosomal DNA (rDNA) for bacteria and actinobacteria and internal transcribed spacer (ITS) rDNA for fungi. Bacteria produced the greatest solubilization in medium containing tricalcium phosphate. Strains B17 and B5, identified as Bacillus sp. and Burkholderia sp., respectively, were the most effective, mobilizing 67% and 58.5% of the total P (Ca₃(PO₄)₂) after 10 days, and were isolated from the rhizosphere of the P efficient L3 maize genotype, under P stress. The fungal population was the most effective in solubilizing P sources of aluminum, phytate, and lecithin. A greater diversity of P-solubilizing microorganisms was observed in the rhizosphere of the P efficient maize genotypes suggesting that the P efficiency in these cultivars may be related to the potential to enhance microbial interactions of P-solubilizing microorganisms.