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1.
Consensus nematode 18S ribosomal DNA primers were designed by aligning available 18S sequences and identifying a variable region flanked by highly conserved regions. These primers were then used to amplify nematode 18S rDNA from whole soil community DNA extracted from a range of European grassland types. Cloning of the PCR amplicons (778 bp) followed by restriction digest analysis (RFLP) resulted in the recovery of 34 unique nematode sequences from the four grasslands studied. Comparison of these data with the limited number of 18S rDNA nematode sequences currently held in on-line databases revealed that all of the sequences could be assigned to known nematode taxa albeit tentatively in some cases. Two of the sequences recovered from the site in the Netherlands (wet, hay-grassland) were recovered in a clade that included a sequence of the genus Trichodorus whilst other sequences from this site showed similarity with 18S rDNA sequences of the genus Prismatolaimus (five sequences), Xiphinema (one sequence) and Enoplus (one sequence). Of the remaining sequences, two showed some affinity with Mylonchulus (UK, upland peat), four with Steinernema (UK) and one sequence with Mesorhabditis (Hungary, east European Steppe). Three sequences from the Netherlands and one from Hungary were recovered in a clade that included a sequence of the genus Pratylenchoides whilst three further sequences from the Netherlands and two from Hungary were recovered in a clade encompassing the genus Globodera. Of the remaining nine sequences, two (NL6, NL62) formed a distinct lineage within the Adenophorea with 90% bootstrap recovery in a paraphyletic clade that included sequences of Prismatolaimus and Trichodorus. Seven sequences (three from the Netherlands, three from the UK and one from Greece) were left unassigned though the tree topology suggested some relationship (58% bootstrap recovery) with the genus Cephalobus. To assess whether primers used to amplify 18S rDNA might be used to fingerprint genetic diversity in nematode communities in soil, the environmental sequence data were used to design a second set of primers carrying a GC-clamp. These primers amplified a 469 bp fragment internal to the region flanked by the primer set used to derive the nematode trees and were used to amplify 18S rDNA for subsequent analysis using denaturing gradient gel electrophoresis (DGGE). DGGE analysis of six major European grassland types revealed considerable genetic diversity between sites. However, the relationships seen with the DGGE data were inconsistent with previous studies where the same soils had been characterized with respect to functional and morphological diversity. To confirm that this second set of primers was amplifying nematode sequences, selected bands on the DGGE gels were extracted, PCR amplified and sequenced. The final alignment was 337 bases. These analyses revealed the presence of sequence signatures from the genera Paratrichodorus, Plectus, Steinernema, Globodera, Cephalobus and Pratylenchoides.  相似文献   

2.
The effects of bacterial-feeding nematodes on bacterial number, activity, and community composition were studied through a microcosm experiment using sterilized soil inoculated with soil bacteria （soil suspension） and with bacteria and three species of bacterial-feeding nematodes （ Cephalobus persegnis, Protorhabditis filiformis, and Caenorhabditis elegans）. Catalyzed reporter deposition-fluorescence in situ hybridization, CO2 evolution, and denaturing gradient gel electrophoresis （DGGE） of PCR ampli- fied 16S rRNA gene fragments were used to investigate bacterial numbers, antivity, and community composition, respectively. Our results showed that bacterial numbers and activity significantly increased in the presence of bacterial-feeding nematodes, which indicated that bacterial-feeding nematodes had a significant positive effect on soil bacteria. The different nematode species had different effects on bacterial numbers and activity. C. persegnis and P. filiformis, isolated from native soil, increased the bacterial number and activity more than C. elegans. The DGGE analysis results showed that dominant bacterial species significantly differed among the treatments, which suggested that bacterial-feeding nematode species modified the bacterial community composition in soil. Further gene sequence analysis results showed that the dominant bacterial species in this study were gram-negative bacteria. Given the completely same conditions except nematode species, the varied selective feeding behavior of different nematode species was the most likely reason for the altered bacterial community composition. Overall, the alteration of bacterial numbers, activity and community composition resulting from the bacterial-feeding nematodes may ult!mately affect soil ecological functioning and processes.  相似文献   

3.
Soil nematode communities in the 0–15 cm soil layer are used as indicators for describing the processes of fallow succession in the semi-arid zone of West Africa (Senegal). Abundance of plant feeding nematodes, non-plant feeding nematodes, plant parasite index (PPI), species richness and Shannon evenness of plant parasitic nematodes were measured at five stages of succession: fields, early (1–3 years), intermediate (8–10 years), old (18–20 years) fallows, and forest stage. These nematological indexes were analyzed simultaneously by multivariate analysis to show the indicative properties of nematode communities. Overall, changes in abundance of nematode groups, PPI and diversity, show continuous trends from early fallow to mature stages of the succession; these trends parallel theoretical trends in secondary succession. In addition, soil nematode parameters were meaningful and expressed interactions of various uncontrolled factors with successional processes, such as environmental conditions or cropping history. During the 3 years of the survey, the structure of nematode communities, described by index analysis, showed little temporal change and supported the use of nematodes as stable indicators.  相似文献   

4.
Although arbuscular mycorrhizal fungi (AMF) are crucial for ecosystem functioning, characterizing AMF community structure in soil is challenging. In this study, nested polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) were combined with cloning of fungal 18S ribosomal gene fragments for the rapid comparison of AMF community structure in soil. Reference AMF isolates, representing four major genera of AMF, were used to develop the method. Sequential amplification of 18S rDNA fragments by nested PCR using primer pairs AM1-NS31 and Glo1-NS31GC followed by DGGE analysis yielded a high-resolution band profile. In parallel, 18S rDNA fragment clone libraries were constructed and clones screened by DGGE. Sequence identity was inferred by matching the electrophoretic mobility of the sample fingerprint bands to that of bands from individual clones. The effectiveness of this approach was tested on soil samples from different ecosystems, yielding reproducible, complex DGGE band patterns specific to each site. The coupling of PCR–DGGE with clone library analysis provides a robust, reliable, and precise means to characterize AMF community structure in soils.  相似文献   

5.
The small subunit ribosomal gene (SSU rDNA) is used in both environmental and phylogenetic studies of nematodes. However, currently described SSU rDNA primers near to the 5′ end of the gene show mismatches with a number of different nematode sequences. Here a new SSU rDNA 5′ forward primer, Nem_SSU_F74, is designed from existing database sequences and its performance compared with a previously described forward primer, SSU_F04, by sequencing from nematode assemblage DNA, in combination with a previously described reverse primer. DNA was extracted from three nematode assemblages isolated from arable soil and, in total, six clone libraries were created: three amplified with established forward primer SSU_F04 and three with the novel primer Nem_SSU_F74. Ninety six clones were sequenced from each library. Nem_SSU_F74 libraries yielded a higher number of nematode sequences than SSU_F04 libraries, and a greater number of nematode taxa were found using the novel forward primer. The most abundant sequences were common to libraries created with either forward primer. Data from a morphological survey of the same samples revealed that biomass was more closely related to molecular analysis than simple counts of nematodes. For all but one nematode order (Aphelenchida), percentage assemblage composition was not significantly different between biomass and sequences obtained with the novel forward primer Nem_SSU_F74.  相似文献   

6.
Microcosm experiments were carried out to study the effects of bacterial-feeding nematodes and prometryne on soil bacterial communities in contaminated soil. Prometryne (5 or 10 mg kg−1 dry soil, that is, P5 or P10) and bacterial-feeding nematodes (5 or 10 individuals g−1 dry soil, that is, N5 or N10), singly and in combination (P5N5, P5N10, P10N5, P10N10), were added to a nematode-free soil. An uncontaminated nematode-free soil was studied for comparison (Control). Bacterial-feeding nematode grazing boosted soil enzyme activities in contaminated soils, thus speeding up prometryne degradation. In the initial stage of the experiment, prometryne enhanced the soil enzyme activities too, but served the opposite purpose later. Denaturing gradient gel electrophoresis (DGGE) analysis indicated that prometryne contamination and nematode grazing over the incubation period exerted an obvious impact on Species richness (S), Shannon–Wiener index (H′) and Evenness (EH) of soil bacteria, which increased initially, then decreased and increased again later. The cluster analysis of DGGE profiles showed that the similarity of soil bacterial communities in all treatments with indigenous microbes, P5, P5N5, P5N10, P10, P10N5, and P10N10 and the Control was 75%, 44%, 78% and 49% at Day 0, Day 8, Day 18 and Day 30, respectively. Compared to the Control, DGGE profiles displayed a varying characteristic bands pattern in all treatments over the incubation period with certain bands present in the treatments while not in the Control and vice versa, suggesting that bacterial-feeding nematode grazing and prometryne contamination affected soil bacterial communities evidently. Consequently, when added to contaminated soil, bacterial-feeding nematodes can contribute to restoration of contaminated sites by degrading toxic compounds like prometryne through enhanced microbial activity.  相似文献   

7.
Four major tea management practices (organic, pollution‐free, conventional, and semi‐natural) are employed in Chinese tea plantations at present. These practices can induce changes in the physiochemical parameters, microorganism community and enzyme activity in tea plantation soil. However, understanding of their effects on soil nematodes is still scarce. This study aimed to investigate whether and how different management practices affect the biodiversity, function, and structure of soil nematode communities in tea plantation habitats. The soil nematode community structures and ecological indices were determined from the soil samples collected more than 6 y after their respective farming practices were first applied, and different management practices did not greatly affect soil nematode community evenness or species diversity, but organic practice increased nematode trophic diversity, common species diversity, and species richness. Pollution‐free practice considerably increased fungivorous nematodes, and both pollution‐free and conventional practice decreased bacterivorous nematodes markedly in the subsurface layers of soil. Predator and omnivorous nematodes were found to be more abundant in semi‐natural plantation. Organic practice was more sustainable and suitable for tea cultivation, with the greatest biodiversity, best nutrient conditions, higher and more stable C/N ratio and lower interference in the food web.  相似文献   

8.
大豆连作土壤线虫群落结构的影响   总被引：2，自引：1，他引：1  
【目的】由根系活动引起的根际微生态系统的改变,特别是病原生物数量的增加是导致作物产生连作障碍的主要因素。其中,植生性病原线虫的危害是大豆连作障碍产生的重要原因之一。由于植生性病原线虫的存在往往受到其它营养类型线虫的影响,因而从线虫群落结构进行分析,不仅可以更好地反映不同营养类型的线虫之间的相互关系,而且能全面了解土壤的健康状况。本文利用末端限制性片段长度多态性分析(T-RFLP)和实时荧光定量PCR(q PCR)等分子生物学的方法,比较短期连作和长期连作线虫群落的差异,揭示长期连作大豆土壤线虫群落的变化规律,理解线虫群落与植物健康的关系,阐明线虫群落的变化在大豆连作障碍中的作用。【方法】首先,基于16sr DNA的T-RFLP指纹图谱,分析土壤中线虫的物种丰富度和不同大小的末端限制性片段(T-RF)的相对丰度。然后,通过构建克隆文库和系统发育树,鉴定T-RF片段对应的线虫种类。最后,利用q PCR,采用绝对定量的方法确定线虫群落的大小。【结果】线虫的物种丰富度随着连作年限的增加呈逐渐降低的趋势。第1年物种丰富度最高,第3年的丰富度显著低于第1年,之后逐渐降低,9年之后保持不变。大豆根际土中共检测到16个TRF,且大多数T-RF能从克隆文库中鉴定。其中,食细菌线虫(Acrobeloides)是最为丰富的线虫种类。在连作2~3年后,植物寄生线虫相对丰度增加,而在连作后期,植物寄生线虫相对丰度减少。非度量多维尺度分析(NMDS)示,第1年线虫群落与其余年限分开,而第2和第3年聚集较近,而连作9、11和13年后聚集较近。另外,线虫群落结构与p H、土壤有机质(SOM)、速效磷(AP)、细菌数量和真菌数量相关。线虫群落总丰度呈先增后降的趋势,最高值出现在第6年。线虫的基因拷贝数与土壤NH+4和染料木因浓度呈显著正相关,而与NO-3和细菌的基因拷贝数呈显著负相关。【结论】大豆根际土壤中,线虫群落丰度在连作第2~3年下降最为明显,到第6~9年有一定的恢复,但不能完全修复。大豆种植为第一,基线虫属(556 bp)丰度最高。土壤功能正常,连作第2~3年后,摄食性线虫(555 bp、558 bp、560 bp等)丰度增加,线虫浸染机会增加。  相似文献   

9.
To determine whether successional changes in plant communities may influence belowground community structure, we quantified nematode abundance, trophic structure and diversity along two separate chronosequences from heather moorland to birch woodland in the Scottish uplands. Tree invasion markedly altered plant community composition, and hence litter inputs, both directly, and indirectly through changes in understorey species. In turn, these changes in detrital inputs were reflected in consistent changes in nematode community structure. Nematode abundance increased from moorland to birch woodland, with moorland soils being dominated by a few taxa, notably root-hair and fungal feeders, compared to the more diverse composition of the birch woodland soils. Trophic structure was altered through an increase in the abundance of bacterial feeding relative to fungal-feeding nematodes, and an increase in the abundance of predatory nematodes. The increase in predators during the succession from moorland to woodland was associated with an increase in soil pH, highlighting that not only changes in the plant community, but also changes in soil properties associated with tree invasion may influence soil nematodes. Nematode diversity increased from moorland to birch woodland, with nematode richness being positively related to both plant species richness and soil pH. These results suggest that trees may control soil community structure through the manipulation of resources and the soil physico-chemical environment, promoting greater nematode diversity and trophic complexity.  相似文献   

10.
One of the major limitations to enhance sustainability of crop production systems is the inability to control root-feeding nematodes without using chemical biocides. In soils under wild vegetation, however, root-feeding nematodes affect plant performance and plant community composition varying from substantially to insignificantly. Previous studies in natural ecosystems have already shown that mutualistic symbionts, such as arbuscular mycorrhizal fungi and endophytes, may influence plant exposure to root-feeding nematodes. In order to learn more from natural systems, we examined nematode control in the root zone of a wild coastal foredune grass by microorganisms, other nematodes and microarthropods. We cultured all eight root-feeding nematode species that occur in the root zone of marram grass (Ammophila arenaria) in coastal foredunes of the Netherlands. Then, in an indoor growth experiment we exposed each nematode species to the potential natural antagonists collected from the same dune soil. Most of the eight dominant root-feeding nematode species could be controlled to some extent by more than one group of soil organisms added. The effectiveness of control varied among nematode species, which seemed to be controlled in a species-specific way. We conclude that in a natural soil the effectiveness of control by microorganisms, other nematodes or microarthropods varies among root-feeding nematode species. Most are controlled, at least to some extent, by soil microbes. However, some root-feeding nematode species are controlled only by microarthropods. Our results strongly suggest that sustainable agriculture will benefit from using a range of biological control mechanisms when controlling root-feeding nematodes, rather than relying on single control agents. Our suggestion also implies that conserving soil biodiversity is crucial in order to enhance the reliability of biological crop protection against soil-borne pests and diseases.  相似文献   

11.
Scomber mackerel have been marketed in fresh and frozen forms and as processed seafood worldwide, and three species of Japanese mackerel S. japonicus, Pacific mackerel S. australasicus, and Atlantic mackerel S. scombrus have constituted a significant part of absolute Scombrid consumption in Japan. The present study was undertaken to develop a rapid and reliable method not only for differentiation of Scomber mackerel from related Scombrid fish by PCR amplification using Scomber genus-specific primers but also for identification of three Scomber mackerel species by PCR-RFLP analysis. Alignment of nucleotide sequences of the nuclear 5S ribosomal RNA gene (5S rDNA) among Scombrid fish allowed the selection of oligonucleotide primers specific for the Scomber genus. These primers enabled amplification of the nontranscribed spacer (NTS) of the 5S rDNA from S. japonicus, S. australasicus, and S. scombrus, whereas no amplification was demonstrated from other Scombrid fish. RFLP analysis of the PCR products with ScaI endonuclease generated unique restriction patterns for each Scomber species. This simple, robust, and reproducible PCR-RFLP technique using Scomber genus-specific primers can serve as a routine food inspection program to enforce labeling regulations of marketed Scombrid fish.  相似文献   

12.
DGGE法与常规培养法对稻田蓝细菌多态性分析结果比较   总被引：1，自引：0，他引：1  
研究运用蓝细菌和硅藻16SrDNA特异引物,将晚季水稻生长后期稻田土壤中提取的总DNA进行PCR扩增后,以DGGE技术对PCR产物进行分析结果表明,14条DGGE带经克隆测序,经NCBI基因库比对得晚季水稻生长后期存在10种蓝细菌,包括4种Leptolyngbya、1种Chamaesiphon、1种Nostoc、1种Oscillatoria、2种Syne-chococcus和1种Chroococcidiopsis。同层不同位置土壤中蓝细菌种群亦有所不同,但每个取样点都有一些特有的蓝细菌种类。用常规方法对同一稻田土壤样品进行分离培养,根据蓝细菌鉴定图谱观察到类似Lyngbya、Oscillatori-a、Chroococcidiopsis及Nostoc的蓝细菌,但显微镜下无法准确分类。比较结果表明采用DGGE法比常规培养法能更准确进行蓝细菌多态性鉴定。  相似文献   

13.
施用化学农药对大豆田土壤线虫群落的影响   总被引：2，自引：0，他引：2  
针对大豆生长期内单一与复合施用乙草胺和呋喃丹对土壤线虫群落结构的影响进行研究。研究结果表明,乙草胺和呋喃丹单一与复合处理中土壤线虫的数量在各采样时期之间、处理之间及二者的交互作用之间均呈现极显著差异(P<0.01);在大豆生长的不同时期,线虫各营养类群的数量受到不同程度的抑制。对线虫生态指数的分析结果表明,乙草胺单一处理对线虫的丰富度表现出促进效应,而呋喃丹单一处理和二者复合处理对线虫丰富度表现出抑制效应;各处理均显著降低了线虫的总成熟度指数(ΣMI),表明线虫群落受到扰动,食物网受到胁迫。  相似文献   

14.
Biological soil crusts (BSCs) play an important role in the dune fixation and maintaining soil biota in arid desert systems. Free-living soil nematode communities could be used as significant bioindicators to reflect soil recover regime after sand burial. However, the relationship between BSCs and nematodes is rarely known. To examine the effects of BSCs on soil nematodes, 72 soil samples under cyanobacteria–lichen and moss crusts were collected to analyse nematode communities in the different aged vegetated areas at the southeastern edge of the Tengger Desert. Our results showed the colonization and development of BSCs significantly enhanced nematode diversity. Nematode abundances, generic richness, H′, MI, EI and SI were greater under crusts than those under noncrust. In particular, nematode abundances, generic richness, H′, MI, EI and SI were positively correlated with crust ages. The differences in nematode communities were also dependent upon crust types. Nematode abundances and generic richness under moss crusts were higher than those under cyanobacteria–lichen crusts. This can be contributed to the present and succession of BSCs that increased thickness of topsoil after dunes have been stabilized, namely, creating suitable habitats and providing an essential food source for nematodes.  相似文献   

15.
Shifting sand dunes threatened the Baotou-Lanzhou railway being kept on operation smoothly seriously crossing Shapotou on the southeastern edge of the Tengger Desert (China). Artificial vegetation input was employed in Shapotou and a vegetated belt was established for stabilizing the shifting sand dunes. Nematode communities from bare soil between vegetation (ST, STI) and soil under the vegetation (ST-V, STI-V) in the vegetation belt, were investigated. All sites received natural rainfall, of which STI and STI-V received irrigation supplements. Total 43 genera were found in four types of soil samples, and Acrobeles, Acrobeloides, Chiloplacus, Panagrolaimus, Aphelenchoides and Ditylenchus were dominant genera in our study. Nematode abundance per 100 g fresh soil (47–552), the number of taxa identified (8–20), species richness (1.51–3.41), the proportion of plant feeders (0.3%–4.2%) and abundance of all cp groups responded to vegetation. The proportion of omnivores (0.5%–7%), the number of taxa identified, Shannon index (1.26–2.12), species richness, genus dominance (0.18–0.40) and abundance of cp3–5 responded to irrigation supplements. The application of PCA helped to reveal that almost all nematode taxa exhibited a positive loading on the horizontal axis, it implied that nematode abundance was generally higher on sites with planted input. Besides, it showed a vegetation gradient from bare soil between vegetation to soil under vegetation along the first axis, while an irrigation gradient along the second axis. Similarly, classification analysis based on all cased involved indicated that two-main group of nematodes was distinguished by their habitats under vegetation from bare soil. Those results suggested that vegetation input might play a more important role on nematode community composition than irrigation supplements for stabilization of shifting sand dunes. Therefore, nematode community composition could be as an indicator of stabilization of sand dune approaches of plant input with only natural rainfall versus with natural rainfall in combination with irrigation supplements in our system. However, the apparent anhydrobiotic state of the nematode fauna required special consideration in desert system, thus the utilization of the nematodes as an indicator of stabilization of sand dune regimes should be further explored on anhydrobiotic nematodes.  相似文献   

16.
Since biochemical and microbiological methods used to study microbial community changes induced by anthropogenic activities can be biased, the impact of two herbicides on soil microorganisms was investigated by culture-independent molecular techniques. The effect of three different amounts (the recommended field dose, tenfold, and 100-fold the dose) of propanil or prometryne on the bacterial community of a clay soil, two modalities of incubation (soil moisture at 70% of the field capacity and a soil-herbicide suspension, 1:10, w:v), and time of incubation were investigated by denaturing gradient gel electrophoresis (DGGE) and amplified rDNA restriction analysis (ARDRA). Two sets of primers for 16S rDNA were used to amplify total soil DNA. Sterile and non-sterile samples were used to determine, by HPLC, the amounts of herbicides adsorbed on soil and transformed by soil microorganisms. Prometryne persisted in soil longer than propanil. Propanil was removed significantly more by non-sterile than by sterile samples, while for prometryne, slight differences were observed. 3,4-Dichloroaniline, a product of propanil hydrolysis, was detected in non-sterile samples and increased with incubation time. Propanil did not affect soil bacteria significantly as indicated by DGGE and ARDRA, with the only exception being the soil-herbicide suspension. Despite a lower utilization of prometryne by soil microorganisms, DGGE analysis showed a more diverse banding than with propanil. Some bands were also detected in the DNA sample extracted from the soil-prometryne suspension, and could be representative of bacterial species utilizing the herbicide as a carbon source, in two very different soil microcosms.  相似文献   

17.
The available information on Pseudomonas biocontrol inoculants on the non-target fungal and nematode community is scant. The current paper addresses this issue and investigates the effects of biocontrol agents Pseudomonas aeruginosa IE-6 and IE-6S+ (previously shown to suppress several soil-borne plant pathogens) on soil microfungi and plant-parasitic nematodes as well as on the root-knot development and growth of tomato (Lycopersicon esculentum). Furadan, a granular nematicide was included as a treatment for comparative purposes. Treatments were applied to soil at the start of each 52-day-long tomato growth cycle, and their effects on the composition and diversity of rhizosphere and endophytic microfungi and plant-parasitic nematodes were examined at the end of first and fourth growth cycle. Several diversity indices were employed to assess community diversity. A total of 16 genera comprising 23 microfungal species were isolated from the tomato rhizosphere. The most abundant fungal species belonged to the genera Aspergillus, Fusarium, and Penicillium. With a few exceptions, fungi were neither exclusively inhibited nor specifically promoted by the application of treatments at any of the growth cycles studied. However, Paecilomyces lilacinus, an egg and female parasite of root-knot nematode, though exclusively absent in the controls was isolated from the treatments. Both general diversity and equitability of rhizosphere microfungi were greater at first compared to the fourth growth cycle while species richness remained uninfluenced across the growth cycles and treatments. However, Furadan and IE-6S+ treatments considerably abated general diversity and equitability. Of the microfungal species isolated from the rhizosphere seven were also recovered from surface-sterilized root tissue of tomato suggesting that all the endophytes are primarily rhizosphere organisms. Diversity of endophytic fungi was consistently lower compared with that of the rhizosphere. Both general diversity and equitability declined in all three treatments relative to controls in the root tissue but species richness remained unaltered. Diversity and equitability of plant-parasitic nematodes in soil were reduced by all three treatments over the controls at fourth growth cycle whilst species richness did not change at either growth cycle. The biocontrol agents significantly reduced root-knot development and enhanced shoot growth of tomato over the controls. The possible implications of fungal composition and abundance because of biocontrol by Pseudomonas application are discussed.  相似文献   

18.
本文对3种常用的土壤微生物总DNA提取方法Martin法、高盐改进法及试剂盒法进行了比较,并通过DNA得率、纯度及16S rDNA V3可变区的PCR扩增结合DGGE法（denatumg gradient gel electrophoresis）,分别对3种方法进行评价。结果表明,3种方法提取的DNA均能满足土壤微生物多样性分析的要求。其中试剂盒方法操作简单,提取的DNA质量较高,但DNA得率较低且成本昂贵。Martin法和高盐改进法用时较长,DNA得率较高,纯度较低,但对后续PCR扩增和DGGE分析没有明显影响,且成本低廉。  相似文献   

19.
The structure of the β-proteobacterial autotrophic ammonia-oxidizing bacterial (AOB) communities in a microcosm of submerged paddy soil was determined by denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments amplified using AOB-selective primers. Shift in the community composition was observed 4 weeks after submergence. The communities from the surface layers (0–1, 2–3 mm) of the soil microcosm were different from those of the subsurface layers (6–9, > 15 mm) and DGGE bands specific to each layer were detected. The majority of the retrieved sequences were Nitrosospira-like, whereas no Nitrosomonas-like sequences were obtained. The 16S rDNA primer set also amplified sequences that were not related to the known Nitrosospira-Nitrosomonas group, although they showed a close relationship with other groups of β-proteobacteria. The results suggest that Nitrosospira-like populations are dominant AOB populations in the submerged paddy soil, and that the oxic layer of submerged paddy soil harbours the specific AOB.  相似文献   

20.
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。  相似文献   

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