Effects of T4 lysozyme release from transgenic potato roots on bacterial rhizosphere communities are negligible relative to natural factors

Rhizosphere bacterial communities of two transgenic potato lines which produce T4 lysozyme for protection against bacterial infections were analyzed in comparison to communities of wild-type plants and transgenic controls not harboring the lysozyme gene. Rhizosphere samples were taken from young, flowering, and senescent plants at two field sites in three consecutive years. The communities were characterized in a polyphasic approach. Cultivation-dependent methods included heterotrophic plate counts, determination of species composition and diversity based on fatty acid analysis of isolates, and community level catabolic profiling. Cultivation-independent analyses were based on denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments amplified from rhizosphere DNA using primers specific for Bacteria, Actinomycetales, or alpha- or beta-Proteobacteria. Several bands of the DGGE patterns were further characterized by sequence analysis. All methods revealed that environmental factors related to season, field site, or year but not to the T4 lysozyme expression of the transgenic plants influenced the rhizosphere communities. For one of the T4 lysozyme-producing cultivars, no deviation in the rhizosphere communities compared to the control lines was observed. For the other, differences were detected at some of the samplings between the rhizosphere community structure and those of one or all other cultivars which were not attributable to T4 lysozyme production but most likely to differences observed in the growth characteristics of this cultivar.     

蒙古栎根内及根围土壤中共生真菌和细菌群落组成分析

为探究中国东北地区森林根系和根围土壤中共生真菌的分布状况,分析了黑龙江省五大连池市蒙古栎Quercus mongolica纯林中根内及根围土壤中共生真菌和细菌群落的组成。结果表明,根内真菌的1 295个OTUs中有209个OTUs为共生真菌,隶属于36属,相对丰度25.46%;根围土壤真菌1 513个OTUs中有285个OTUs为共生真菌,隶属于40属,相对丰度59.91%;根内与根围土壤共同拥有共生真菌33属,根内特有3属,根围土壤特有7属。其中,外生菌根真菌为根内和根围土壤中共生真菌的主要类群,分别占共生真菌的98.82%和99.80%。定殖根内的细菌共获得5 550个OTUs,隶属于400属;根围土壤细菌获得8 406个OTUs,隶属于436属,根内细菌群落的Shannon指数和Chao1指数均低于根围土壤的。PICRUSt功能预测分析结果表明,根内的信号转导通路与信号分子和相互作用通路（包括CAM配体、ECM-受体相互作用等通路）的丰度低于根围土壤,而膜运输通路与信号分子和相互作用通路（包括细菌毒素、细胞抗原等通路）的丰度高于根围土壤。根内与根围土壤中菌根辅助细菌组成差异分析结果表明,除慢生根瘤菌属 Bradyrhizobium外,根内其余9属的相对丰度均高于根围土壤,尤其假单胞菌属 Pseudomonas的相对丰度远高于根围土壤。     

不同来源飞蝗后肠微生物群落高通量测序分析

对来源于田间与室内飞蝗种群肠道细菌群落的454高通量测序(HTS)结果进行了分析,共分析鉴定出OTU(operational taxonomic unit)66个,其中室内飞蝗种群41个,田间飞蝗53个。全部OTU归属于6个门,9个纲,11个目,14个科,20个属。室内种群共含有12个属,其中优势类群为乳球菌属(Lactococcus)和拉乌尔菌属(Raoultella),分别占总种群的13.3%和36.1%。田间飞蝗鉴定出14个属,优势类群为芽胞杆菌属(Bacillus),占总种群的18.8%。田间飞蝗后肠菌落的Ace、Chao和Shannon指数均高于室内种群,而Simpson指标呈相反结果。研究表明来源于田间的飞蝗肠道细菌群落的丰富度和多样性均高于室内饲养的种群。分析结果为今后开展不同来源飞蝗肠道微生物资源研究提供依据。     

Comparative analysis of bacterial diversity in the rhizosphere of tomato by culture-dependent and -independent approaches

The microbiome in the rhizosphere–the region surrounding plant roots–plays a key role in plant growth and health, enhancing nutrient availability and protecting plants from biotic and abiotic stresses. To assess bacterial diversity in the tomato rhizosphere, we performed two contrasting approaches: culture-dependent and -independent. In the culture-dependent approach, two culture media (Reasoner’s 2A agar and soil extract agar) were supplemented with 12 antibiotics for isolating diverse bacteria from the tomato rhizosphere by inhibiting predominant bacteria. A total of 689 bacterial isolates were clustered into 164 operational taxonomic units (OTUs) at 97% sequence similarity, and these were found to belong to five bacterial phyla (Proteobacteria, Actinobacteria, Bacteroidetes, Acidobacteria, and Firmicutes). Of these, 122 OTUs were retrieved from the antibiotic-containing media, and 80 OTUs were recovered by one specific antibiotic-containing medium. In the culture-independent approach, we conducted Illumina MiSeq amplicon sequencing of the 16S rRNA gene and obtained 19,215 high-quality sequences, which clustered into 478 OTUs belonging to 16 phyla. Among the total OTUs from the MiSeq dataset, 22% were recovered in the culture collection, whereas 41% of OTUs in the culture collection were not captured by MiSeq sequencing. These results showed that antibiotics were effective in isolating various taxa that were not readily isolated on antibiotic-free media, and that both contrasting approaches provided complementary information to characterize bacterial diversity in the tomato rhizosphere.     

Prokaryotic abundance and 16S rRNA gene sequences detected in marine aerosols on the East Sea (Korea)

Modern studies on marine airborne prokaryotes over open seas have been rare. Here, to understand their distribution and composition, aerosol as well as surface seawater samples were collected at three sites in the East Sea. The abundance of airborne prokaryotes was 0.7-1.2 × 10(5) cells m(-3) . Partial sequences of the 16S rRNA gene from the aerosols were dominated by Gammaproteobacteria and Bacteroidetes. Interestingly, the numbers of phylotypes of airborne bacteria were comparable to those of seawater bacteria in coastal and remote offshore sites. Over half of the bacterial operational taxonomic units (OTUs) detected in these aerosols (12 out of 20) were of marine origin. Eight of the 19 bacterial OTUs found in the surface waters occurred in the aerosols. Thus, surface seawater seems to be a main source of marine airborne bacteria. However, in the intermediate offshore site, airborne bacterial OTUs (n=4) were all terrestrial, indicating that variations of dominant airborne bacterial phylotypes occurred over the sites (each ～160 km apart) sampled at 1-4-day intervals. Euryarchaeota sequences belonging to the MSP8 clade reported from geographically distant regions were detected in marine aerosols. Long-distance aerial transport of the marine prokaryotes seems to be quite possible.     

Bacterial diversity in the cecum of the world's largest living rodent (Hydrochoerus hydrochaeris)

The capybara (Hydrochoerus hydrochaeris) is the world's largest living rodent. Native to South America, this hindgut fermenter is herbivorous and coprophagous and uses its enlarged cecum to digest dietary plant material. The microbiota of specialized hindgut fermenters has remained largely unexplored. The aim of this work was to describe the composition of the bacterial community in the fermenting cecum of wild capybaras. The analysis of bacterial communities in the capybara cecum is a first step towards the functional characterization of microbial fermentation in this model of hindgut fermentation. We sampled cecal contents from five wild adult capybaras (three males and two females) in the Venezuelan plains. DNA from cecal contents was extracted, the 16S rDNA was amplified, and the amplicons were hybridized onto a DNA microarray (G2 PhyloChip). We found 933 bacterial operational taxonomic units (OTUs) from 182 families in 21 bacterial phyla in the capybara cecum. The core bacterial microbiota (present in at least four animals) was represented by 575 OTUs. About 86% of the cecal bacterial OTUs belong to only five phyla, namely, Firmicutes (322 OTUs), Proteobacteria (301 OTUs), Bacteroidetes (76 OTUs), Actinobacteria (69 OTUs), and Sphirochaetes (37 OTUs). The capybara harbors a diverse bacterial community that includes lineages involved in fiber degradation and nitrogen fixation in other herbivorous animals.     

Bacterial diversity in maize rhizospheres: conclusions on the use of genetic profiles based on PCR-amplified partial small subunit rRNA genes in ecological studies

A cultivation-independent approach based on polymerase chain reaction (PCR)-amplified partial small subunit rRNA genes and genetic profiling by single-strand conformation polymorphism (SSCP) was used to characterize the bacterial diversity inhabiting the rhizosphere of maize plants grown on an agricultural field. The community structures of two cultivars, a genetically engineered and a nonengineered variety, different herbicide regimes and soil tillage were compared with each other at two sampling dates. SSCP-profiles were generated with DNA from bacterial cell consortia with primers hybridizing to evolutionarily highly conserved rRNA gene regions. On silver-stained gels, each profile consisted of approx. 50 distinguishable bands. Similarity analyses of patterns recorded by digital image analyses could not detect any difference between cultivars or treatments that was greater than the variability between replicates. A total of 54 sequences recovered from different bands were identified and grouped into operational taxonomical units (OTUs). Surprisingly, only five of 40 OTUs contained sequences of both samplings. Three different bands from a profile were selected to test whether this small overlap was due to an incomplete recovery of sequences. From a faint band, two different OTUs were found when 12 clones were analysed, and from two strong bands 24 and 22 OTUs were detected from a total of 26 and 36 clones, respectively. The OTUs belonged to phylogenetically different groups of bacteria. Gene probes that were developed to target different bands of the profiles, however, indicated in Southern blot analyses that patterns between treatments, replicates and samplings, and even from two different growing seasons were highly conserved. Our study demonstrates that community profiles can consist of more sequences than detectable by staining and that gene probes in Southern blot can be a useful control to investigate the composition of microbial communities by genetic profiles.     

Effects of genetically modified amylopectin-accumulating potato plants on the abundance of beneficial and pathogenic microorganisms in the rhizosphere

In this study, the potential effects of a genetically modified (GM) amylopectin-accumulating potato line (Solanum tuberosum L.) on plant beneficial bacteria and fungi as well as on phytopathogens in the rhizosphere were investigated in a greenhouse experiment and a field trial. For comparison, the non-transgenic parental cultivar of the GM line and a second non-transgenic cultivar were included in the study. Rhizospheres were sampled during young leaf development (EC30) and at florescence (EC60). The microbial community composition was analysed by real-time PCR to quantify the abundances of Pseudomonas spp., Clavibacter michiganensis, Trichoderma spp. and Phytophthora infestans. Additionally, total bacterial and fungal abundances were measured. None of the examined gene abundance patterns were affected by the genetic modification when wild type and GM line were compared. However, significant differences were observed between the two natural potato cultivars, especially during the early leaf development of the plants. Furthermore, gene abundance patterns were also influenced by the plant developmental stage. Interestingly, the impact of the cultivar and the plant vegetation stage on the microbial community structure was more pronounced in field than in greenhouse. Overall, field-grown plants showed a higher abundance of microorganisms in the rhizosphere than plants grown under greenhouse conditions.     

Diversity and distribution of soil fungal communities in a semiarid grassland

The fungal loop model of semiarid ecosystems integrates microtopographic structures and pulse dynamics with key microbial processes. However limited data exist about the composition and structure of fungal communities in these ecosystems. The goal of this study was to characterize diversity and structure of soil fungal communities in a semiarid grassland. The effect of long-term nitrogen fertilization on fungi also was evaluated. Samples of rhizosphere (soil surrounding plant roots) and biological soil crust (BSC) were collected in central New Mexico, USA. DNA was amplified from the samples with fungal specific primers. Twelve clone libraries were generated with a total of 307 (78 operational taxonomic units, OTUs) and 324 sequences (67 OTUs) for BSC and rhizosphere respectively. Approximately 40% of soil OTUs were considered novel (less than 97% identity when compared to other sequences in NCBI using BLAST). The dominant organisms were dark-septate (melanized fungi) ascomycetes belonging to Pleosporales. Effects of N enrichment on fungi were not evident at the community level; however the abundance of unique sequences, sampling intensity and temporal variations may be uncovering the effect of N in composition and diversity of fungal communities. The fungal communities of rhizosphere soil and BSC overlapped substantially in composition, with a Jaccard abundance similarity index of 0.75. Further analyses are required to explore possible functions of the dominant species colonizing zones of semiarid grassland soils.     

Effects of T4 Lysozyme Release from Transgenic Potato Roots on Bacterial Rhizosphere Communities Are Negligible Relative to Natural Factors

Rhizosphere bacterial communities of two transgenic potato lines which produce T4 lysozyme for protection against bacterial infections were analyzed in comparison to communities of wild-type plants and transgenic controls not harboring the lysozyme gene. Rhizosphere samples were taken from young, flowering, and senescent plants at two field sites in three consecutive years. The communities were characterized in a polyphasic approach. Cultivation-dependent methods included heterotrophic plate counts, determination of species composition and diversity based on fatty acid analysis of isolates, and community level catabolic profiling. Cultivation-independent analyses were based on denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments amplified from rhizosphere DNA using primers specific for Bacteria, Actinomycetales, or α- or β-Proteobacteria. Several bands of the DGGE patterns were further characterized by sequence analysis. All methods revealed that environmental factors related to season, field site, or year but not to the T4 lysozyme expression of the transgenic plants influenced the rhizosphere communities. For one of the T4 lysozyme-producing cultivars, no deviation in the rhizosphere communities compared to the control lines was observed. For the other, differences were detected at some of the samplings between the rhizosphere community structure and those of one or all other cultivars which were not attributable to T4 lysozyme production but most likely to differences observed in the growth characteristics of this cultivar.     

Effects of genetically modified potatoes with increased zeaxanthin content on the abundance and diversity of rhizobacteria with in vitro antagonistic activity do not exceed natural variability among cultivars

To assess potential effects of genetically modified (GM) potatoes on the abundance and diversity of rhizobacteria with in vitro antagonistic activity in relation to natural variability among cultivars, two GM potato lines accumulating the carotenoid zeaxanthin in their tubers, the parental cultivar and four additional commercial cultivars were planted at two field sites in Germany. Rhizosphere samples were taken at three developmental stages of the plants. A total of 3,985 bacteria isolated from the rhizosphere were screened for their in vitro antagonistic activity towards Rhizoctonia solani, Verticillium dahliae and Phytophthora infestans using a dual-culture assay. Genotypic characterisation, 16S rRNA gene sequencing and antifungal metabolite analysis was performed to characterize the 595 antagonists obtained. The 16S rRNA gene-based identification of in vitro antagonists revealed strong site-dependent differences in their taxonomic composition. This study showed that the site was the overriding factor determining the proportion and diversity of antagonists from the rhizosphere of potato while the effect of the genetic modification on the proportion of antagonists obtained did not exceed natural variability among the five commercial cultivars tested.     

Characterization of bacterial community structure in the rhizosphere of Triticum aestivum L.

The plant microbiome influence plant health, yield and vigor and has attained a considerable attention in the present era. In the current study, native bacterial community composition and diversity colonizing Triticum aestivum L. rhizosphere at two distant geographical locations including Mirpur Azad Kashmir and Islamabad was elucidated. Based on IonS5™XL platform sequencing of respective samples targeting 16S rRNA gene that harbor V3-V4 conserved region revealed 1364 and 1254 microbial operational taxonomic units (OTUs) at ≥97% similarity and were classified into 23, 20 phyla; 70, 65 classes; 101, 87 orders; 189,180 families; 275, 271 genera and 94, 95 species. Respective predominant phyla accounting for 97.90% and 98.60% of bacterial community were Proteobacteria, Actinobacteria, Acidobacteria, Bacteroidetes, Firmicutes, Chloroflexi and Gemmatimonadetes. Diversity indices revealed variations in relative abundance of bacterial taxa owing to distant geographical locations however predominant bacterial taxa at both locations were similar. These findings paved a way to dissect consequence of associated microbiota on future wheat production system.     

Impact of Virioplankton on Archaeal and Bacterial Community Richness as Assessed in Seawater Batch Cultures

During cruises in the tropical Atlantic Ocean (January to February 2000) and the southern North Sea (December 2000), experiments were conducted to monitor the impact of virioplankton on archaeal and bacterial community richness. Prokaryotic cells equivalent to 10 to 100% of the in situ abundance were inoculated into virus-free seawater, and viruses equivalent to 35 to 360% of the in situ abundance were added. Batch cultures with microwave-inactivated viruses and without viruses served as controls. The apparent richness of archaeal and bacterial communities was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR-amplified 16S rRNA gene fragments. Although the estimated richness of the prokaryotic communities generally was greatly reduced within the first 24 h of incubation due to confinement, the effects of virus amendment were detected at the level of individual operational taxonomic units (OTUs) in the T-RFLP patterns of both groups, Archaea and Bacteria. One group of OTUs was detected in the control samples but was absent from the virus-treated samples. This negative response of OTUs to virus amendment probably was caused by viral lysis. Additionally, we found OTUs not responding to the amendments, and several OTUs exhibited variable responses to the addition of inactive or active viruses. Therefore, we conclude that individual members of pelagic archaeal and bacterial communities can be differently affected by the presence of virioplankton.     

Pyrosequencing Reveals a Highly Diverse and Cultivar-Specific Bacterial Endophyte Community in Potato Roots

In this study, we examined the bacterial endophyte community of potato (Solanum tuberosum) cultivar/clones using two different molecular-based techniques (bacterial automated ribosomal intergenic spacer analysis (B-ARISA) and pyrosequencing). B-ARISA profiles revealed a significant difference in the endophytic community between cultivars (perMANOVA, p < 0.001), and canonical correspondence analysis showed a significant correlation between the community structure and plant biomass (p = 0.001). Pyrosequencing detected, on average, 477 ± 71 bacterial operational taxonomic units (OTUs, 97% genetic similarity) residing within the roots of each cultivar, with a Chao estimated total OTU richness of 1,265 ± 313. Across all cultivars, a total of 238 known genera from 15 phyla were identified. Interestingly, five of the ten most common genera (Rheinheimera, Dyadobacter, Devosia, Pedobacter, and Pseudoxanthomonas) have not, to our knowledge, been previously reported as endophytes of potato. Like the B-ARISA analysis, the endophytic communities differed between cultivar/clones (∫-libshuff, p < 0.001) and exhibited low similarities on both a presence/absence (0.145 ± 0.019) and abundance (0.420 ± 0.081) basis. Seventeen OTUs showed a strong positive (r > 0.600) or negative (r < −0.600) correlation with plant biomass, suggesting a possible link between plant production and endophyte abundance. This study represents one of the most comprehensive assessments of the bacterial endophytic communities to date, and similar analyses in other plant species, cultivars, or tissues could be utilized to further elucidate the potential contribution(s) of endophytic communities to plant physiology and production.     

Impact of flooding on soil bacterial communities associated with poplar (Populus sp.) trees

Soil bacterial communities were analyzed in different habitats (bulk soil, rhizosphere, rhizoplane) of poplar tree microcosms (Populus tremulaxP. alba) using cultivation-independent methods. The roots of poplar trees regularly experience flooded and anoxic conditions. Therefore, we also determined the effect of flooding on microbial communities in microcosm experiments. Total community DNA was extracted and bacterial 16S rRNA genes were amplified by PCR and analyzed by terminal restriction fragment length polymorphism (T-RFLP) analysis, cloning and sequencing. Clone libraries were created from all three habitats under both unflooded and flooded conditions resulting in a total of 281 sequences. Numbers of different sequences (<97% similarity) in the different habitats represented 16-55% of total bacterial species richness determined from the nonparametric richness estimator Chao1. According to the number of different terminal restriction fragments (T-RFs), all of the different habitats contained approximately 20 different operational taxonomic units (OTUs), except the flooded rhizoplane habitat whose community contained less OTUs. Results of cloning and T-RFLP analysis generally supported each other. Correspondence analysis of T-RFLP patterns showed that the bacterial communities were different in bulk soil, rhizosphere and rhizoplane and changed upon flooding. For example OTUs representing Bacillus sp. were highest in the unflooded bulk soil and rhizosphere. Sequences related to Aquaspirillum, in contrast, were predominant on the poplar roots and in the rhizosphere of flooded microcosms but were rarely found in the other habitats.     

Impact of virioplankton on archaeal and bacterial community richness as assessed in seawater batch cultures

During cruises in the tropical Atlantic Ocean (January to February 2000) and the southern North Sea (December 2000), experiments were conducted to monitor the impact of virioplankton on archaeal and bacterial community richness. Prokaryotic cells equivalent to 10 to 100% of the in situ abundance were inoculated into virus-free seawater, and viruses equivalent to 35 to 360% of the in situ abundance were added. Batch cultures with microwave-inactivated viruses and without viruses served as controls. The apparent richness of archaeal and bacterial communities was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis of PCR-amplified 16S rRNA gene fragments. Although the estimated richness of the prokaryotic communities generally was greatly reduced within the first 24 h of incubation due to confinement, the effects of virus amendment were detected at the level of individual operational taxonomic units (OTUs) in the T-RFLP patterns of both groups, Archaea and BACTERIA: One group of OTUs was detected in the control samples but was absent from the virus-treated samples. This negative response of OTUs to virus amendment probably was caused by viral lysis. Additionally, we found OTUs not responding to the amendments, and several OTUs exhibited variable responses to the addition of inactive or active viruses. Therefore, we conclude that individual members of pelagic archaeal and bacterial communities can be differently affected by the presence of virioplankton.     

马铃薯全生育期内根际微生物组变化规律

[目的]陆生植物根际环境与土壤中的微生物菌群关系密切,其根际微生物群落动态极可能直接影响着植物健康及养分高效利用。虽然根际益生菌已被证实可用于提高作物生产力,但由于缺乏对这些菌群组成动态变化规律的认识了解,它们的开发受到限制。研究马铃薯全生育期根际菌群的动态变化规律,探讨根际菌群变化与马铃薯发育时期的相关性,为针对马铃薯不同生长时期开发专用生物益生菌肥奠定理论基础。[方法]本研究着眼于马铃薯田间全生命周期微生物组动态变化,通过Illumina MiSeq高通量测序技术对不同时间点马铃薯根际细菌16S rRNA基因V3-V4区和真菌ITS区测序并对操作分类单位(OTU)进行聚类,分析样品间微生物群落的多样性特征,并通过机器学习的方法建立模型,将根际菌群与田间马铃薯发育时间相关联。[结果]根际菌群在马铃薯各个发育阶段随时间变化明显,营养生长阶段的微生物群落结构发生了显著变化,随着结薯期的开始逐渐稳定,直到块茎成熟后期根际菌群再次出现较大变化,且在不同施肥处理间呈现较大差异。进一步基于模型挖掘了与马铃薯发育时间相关联的22个特征细菌类群和16个特征真菌类群,其中苗期和结薯末期的特征类群分别为梭菌(Clostridium)和放线菌(Actinobacteria)。[结论]马铃薯的生长发育时期是影响根际微生物群落组成的主要因素,益生菌肥的添加主要影响马铃薯结薯末期的细菌微生物菌群结构。     

Temporal shifts of fungal communities in the rhizosphere and on tubers in potato fields

Soil fungal communities perform important ecological roles determining, at least in part, agricultural productivity. This study aimed at examining the fungal community dynamics in the potato rhizosphere across different development stages in two consecutive growing seasons (winter and summer). Microbial fingerprinting of rhizosphere soil samples collected at pre-planting, tuber initiation, flowering and at senescence was performed using ARISA in conjunction with Next Generation Sequencing (Illumina MiSeq). The epiphytic fungal communities on tubers at harvest were also investigated. Alpha-diversity was stable over time within and across the two seasons. In contrast, rhizospheric fungal community structure and composition were different between the two seasons and in the different plant growth stages within a given season, indicating the significance of the rhizosphere in shaping microbial communities. The phylum Ascomycota was dominant in the potato fungal rhizosphere, with Operational Taxonomic Units (OTUs) belonging to the genus Peyronellaea being the most abundant in all samples. Important fungal pathogens of potato, together with potential biological control agents and saprophytic species, were identified as indicator OTUs at different plant growth stages. These findings indicate that potato rhizosphere fungal communities are functionally diverse, which may contribute to soil health.     

不同生境马铃薯根际土壤细菌多样性分析

近年来,随着国家对马铃薯产业重视程度的增加,生产的专业化和规模化程度越来越高,化肥农药用量不断加大,加之连作等种植模式,致使以疮痂病为代表的土传病害普遍发生且逐年加重,个别地块发病率达90%以上,给种植业者带来巨大的经济损失。[目的] 为了探索马铃薯疮痂病发生与土壤环境的关系,分析区域性种植方式和施肥量对土壤细菌种群变化的影响,为实现土传病害有效防治提供借鉴。[方法] 本文分别从1年连作土传病害轻的宁夏西吉（西北）、3年连作土传病害严重的河北沽源（华北）、5年轮作未发现土传病害的内蒙古海拉尔（东北）大田马铃薯根际采集土壤,利用高通量测序技术,比较了样品间细菌群落结构差异。[结果] 3组样品共获得有效条带617558条,可分类操作单元（OTUs）3077个。各样品中变形菌门（Proteobacteria）数量最多,含量均在33%以上。与未发病土壤样品相比,土传病害发生严重的样品中细菌数量、物种数、细菌多样性、种类丰富度均有所降低,有害菌数量增加,益生菌数量减少。其中,放线菌（Actinobacteria）数量明显增多,变形菌（Proteobacteria）、绿弯菌（Chloroflexi）和酸杆菌（Acidobacteria）数量明显减少,组分及数量差异明显的细菌（尤其是放线菌门）大多与土壤全磷含量呈显著相关。[结论] 过量施用化肥和常年连作改变了土壤细菌群落结构,生态环境恶化,导致土传性病害发生。其中,磷可能是影响土壤微生物群落结构变化最主要的肥料元素。     

杨树人工林根际古菌群落随细根生长的演变

研究细根不同生长时期根际土壤古菌群落组成结构差异,对深入了解林木细根与土壤微生物互作关系具有重要理论意义.依据细根表面颜色,采集杨树一级细根不同生长时期(白色新生根、黄色成熟根、褐色衰老根)根际土壤并提取微生物总DNA,采用特异性引物对古菌16S rDNA V4-V5区进行扩增,利用Illumina MiSeq平台进行古菌高通量测序分析.结果表明: 新生根和衰老根根际土壤古菌群落操作分类单元(OTU)丰富度相似,而成熟根根际土壤古菌群落OTU数量较少.新生根和成熟根根际土壤共同含有134个OTU;成熟根和衰老根根际土壤共同含有87个OTU,新生根和衰老根根际土壤共同拥有90个OTU.α多样性分析表明,成熟根根际土壤古菌群落Chao1指数和ACE指数显著低于新生根和衰老根根际土壤,而衰老根根际土壤古菌群落Simpson指数和Shannon指数显著低于新生根和成熟根根际土壤.PERMANOVA分析表明,新生根和衰老根根际土壤古菌群落组成有显著差异.物种注释显示,杨树根际土壤共包含12个古菌属,其中新生根5个、成熟根10个、衰老根6个.β多样性指数表明,杨树根际土壤古菌群落相似度随着细根的生长逐渐下降,不同生长阶段细根根际土壤的古菌群落结构有较大差异.其中,占绝对优势的古菌为氨氧化古菌Candidatus_Nitrososphaera,其相对丰度超过70%.且随细根生长发育,该类古菌在根际土壤中的丰度呈现上升趋势,表明其可能与细根的生长发育关系密切.