Diversity of cultivable Azotobacter in the semi-arid alfisol receiving long-term organic and inorganic nutrient amendments

Monitoring the biological processes and microbial diversity is essential for sustaining the soil health for long-term productivity. In the present study, the impact of long-term nutrient management systems on changes in Azotobacter diversity of Indian semi-arid alfisol was assessed. Three soils, i.e., unfertilized control, soils amended with organic manures (OM), and with inorganic chemical fertilizers (IC) from century-old experimental fields were evaluated for Azotobacter diversity by Amplified Ribosomal DNA Restriction Analysis (ARDRA). Bray–Curtis’s similarity index of the ARDRA data of the isolates was analyzed by non-metric multi-dimensional scaling and hierarchical cluster analysis. The results revealed that the long-term organically managed soil recorded significantly higher soil organic carbon, microbial biomass carbon, and total culturable bacterial counts, whereas the chemical fertilized and control soils remained unaffected. Though the Azotobacter population was significantly higher in OM soil than IC and control soils, the genetic diversity was unaffected due to long-term addition of either organic manures or inorganic chemical fertilizers. This result implies the importance of continuous addition of organic manures and also the optimal use of inorganic chemical fertilizers without disturbing the biological properties of the soil.     

Diversity and characterization of <Emphasis Type="Italic">Azotobacter</Emphasis> isolates obtained from rice rhizosphere soils in Taiwan

Azotobacter species, free-living nitrogen-fixing bacteria, have been used as biofertilizers to improve the productivity of non-leguminous crops, including rice, due to their various plant growth-promoting traits. The purposes of this study were to characterize Azotobacter species isolated from rice rhizospheres in Taiwan and to determine the relationship between the species diversity of Azotobacter and soil properties. A total of 98 Azotobacter isolates were isolated from 27 paddy fields, and 16S rRNA gene sequences were used to identify Azotobacter species. The characteristics of these Azotobacter strains were analyzed including carbon source utilization and plant growth-promoting traits such as nitrogen fixation activity, indole acetic acid production, phosphate-solubilizing ability, and siderophore secretion. Of the 98 strains isolated in this study, 12 were selected to evaluate their effects on rice growth. Four species of Azotobacter were identified within these 98 strains, including A. beijerinckii, A. chroococcum, A. tropicalis, and A. vinelandii. Of these four species, A. chroococcum was predominant (51.0%) but A. beijerinckii had the highest level of nucleotide diversity. Strains within individual Azotobacter species showed diverse profiles in carbon source utilization. In addition, the species diversity of Azotobacter was significantly related to soil pH, Mn, and Zn. Members of the same Azotobacter species showed diverse plant growth-promoting traits, suggesting that the 98 strains isolated in this study may not equally effective in promoting rice growth. Of the 12 strains evaluated, A. beijerinckii CHB 461, A. chroococcum CHB 846, and A. chroococcum CHB 869 may be used to develop biofertilizers for rice cultivation because they significantly promoted rice growth. This study contributes to the selection of suitable Azotobacter strains for developing biofertilizer formulations and soil management strategies of Azotobacter for paddy fields.     

不同时期艾比湖湿地盐角草群落土壤固氮微生物的多样性分析

【目的】研究新疆艾比湖湿地不同季节盐角草根际和非根际土壤固氮微生物的多样性和丰富度与环境因子的相关性,以期探究在荒漠化和盐渍化不断严重的艾比湖湿地中随着季节变化的固氮微生物群落对恢复生态功能起到的潜在作用,为后续的湿地保护和退化恢复工作提供理论支持和数据基础。【方法】应用Illumina HiSeq PE250测序技术,分析6个土壤样本固氮微生物的多样性,结合相关的理化因子并利用RDA分析法探究土壤理化性质和固氮微生物菌落结构及丰富度的相关性。【结果】艾比湖湿地盐角草植物根际土壤的固氮微生物多样性高于非根际土壤,7月的土壤固氮微生物多样性高于10月和4月的土壤。土杆菌属(Geobacter)、假单胞菌属(Pseudomonas)、固氮菌属(Azotobacter)和慢生根瘤菌属(Bradyrhizobium)等为盐角草根际和非根际土壤中的共同优势菌属。这些固氮微生物优势菌属隶属于变形菌门(Proteobacteria)和蓝藻门(Cyanobacteria),且相对丰富度占比为85%和10%,其余各菌门共占比较少,仅为5%。土壤中固氮微生物的优势菌群与碱解氮(AN)、全氮(TN)、速效钾(AK)和有效磷(TP)呈显著相关。【结论】随着时间的推移土壤样本中固氮微生物的多样性和群落结构也发了改变,同一时期植物根际与非根际土壤中固氮微生物的群落结构并不相同。土壤的环境因子与固氮细菌的群落结构和丰富度的相关性研究可以为艾比湖湿地的退化恢复提供数据基础和理论支持。     

Genotypic diversity in native rhizobial population nodulating Vicia faba in arid and semi-arid regions of Haryana state (India)

A Rhizobium–legume interaction stands out from other plant–microbe interactions as one in which a true developmental mutualism occurs. To study the genotypic diversity in native population of rhizobia-nodulating Vicia faba plants, we retrieved 64 rhizobial isolates from root nodules of faba bean grown in pots holding soils collected from arid and semi-arid regions of the state of Haryana, India. The amplification of nodC in all the isolates authenticated these as rhizobia. The nitrogen-fixing potential of the isolates was tested by the amplification of the nifH gene. Only 50 isolates out of 64 showed nifH gene amplification. The characterization of the isolates by amplified 16S rDNA restriction analysis (ARDRA) categorized these into 36 16S rDNA genotypes using a combination of MspI and HaeIII restriction enzymes. Majority of the isolates resolved into separate genotypes, indicating a wide diversity among them, which seemed to arise from their geographical origin and soil characteristics. These findings may be immensely useful in agriculture towards developing rhizobial inoculants specific for faba beans under arid and semi-arid conditions.     

Reduction of Nitrates by Azotobacter indicum and Azotobacter chroococcum Cultures

The capacity for denitrification was studied in Azotobacter bacteria, which are free-living nitrogen-fixing obligatory aerobes. Data on nitrate reduction to nitrites and nitric oxide by A. indicum under anaerobic conditions were obtained for the first time for genus Azotobacter.     

典型生境巨菌草根内生固氮菌的群落多样性

【目的】探究不同生境巨菌草内生固氮菌群落组成多样性及其分异规律。【方法】采用高通量测序固氮酶nif H标靶基因方法,研究了我国6个典型地区的巨菌草内生固氮菌群,包括福建闽侯县、新疆墨玉县、内蒙古阿拉善左旗、青海贵德县、甘肃安定区、海南那大镇,结合地理气候因子统计,分析了固氮菌多样性的环境驱动机制。【结果】共获得64122条nif H基因的有效序列,640个OTUs,归属于6个门、10个纲、17个目、24个科、33个属和39个种。不同地区巨菌草中优势内生固氮菌群的种类和丰度存在较大的差异。在门水平上,福州闽侯县、甘肃安定区、新疆墨玉县、内蒙古阿拉善左旗和青海贵德县5个地区的优势菌门均为变形菌门,海南那大镇的优势菌门为变形菌门和蓝藻菌门;属水平上,不同地区巨菌草最优势内生固氮菌类群分别为:福州闽侯县(变形菌门中未定属,80.56%);新疆墨玉县(变形菌门中未定属,33.14%);内蒙古阿拉善左旗(变形菌门中未定属,76.23%);甘肃安定区(α-变形菌纲中的未定属,53.78%);海南那大镇(变形菌门中未定属,38.37%);青海贵德县(变形菌门中未定属,46.12%)。Alpha多样性和Beta多样性分析表明,不同地区巨菌草内生固氮菌群落的多样性存在较大的差异,海南那大镇样本中巨菌草各类内生固氮菌群的多样性及丰富度最高,福建闽侯县样本中巨菌草各类内生固氮菌群的多样性及丰富度最低。典范对应分析(CCA)结果表明,年均降雨量和年均气温是影响巨菌草内生固氮菌群变化的主要因素,其次是土壤有机质、土壤全氮和土壤p H。【结论】不同地区巨菌草内生固氮菌群落的组成及丰度存在着较大的差异,海南那大镇巨菌草内生固氮菌群的种类及相对丰度较高,本研究可为巨菌草内生固氮菌群的资源开发及其固氮微生物肥料的菌种选育和生产应用提供理论支持。     

Pesticide tolerant Azotobacter isolates from paddy growing areas of northern Karnataka, India

A total of 14 Azotobacter strains were isolated from different paddy cultivating soils with pH ranging from 6.5 to 9.5 by using serial dilution agar plate method. The strains were Gram negative, rod shaped, cyst forming and developed brown to black colored colonies, which were glistening, smooth, slimy on Ashby’s agar plates. Biochemically they were positive for biochemical tests namely, indole production, citrate, catalase, carbohydrate fermentation and Voges–Proskauer test. Further, sequence analysis of PCR amplicons obtained from these cultures revealed the presence of five different Azotobacter species viz., Azotobacter vinelandii, Azotobacter salinestris, Azotobacter sp., Azotobacter nigricans subsp. nigricans and Azotobacter tropicalis. Phylogenetically these strains were grouped into two distinct clusters. These strains were tested for their ability to grow on a media containing four different pesticides such as pendimethalin, glyphosate, chloropyrifos and phorate, which are commonly used for the paddy. Out of 14 strains tested, 13 strains were able to grow on a media containing herbicides such as pendimethalin, glyphosate and insecticides like chloropyrifos and phorate. However, five Azotobacter strains were able to grow at higher concentration of 5 % pesticides, without affecting their growth rate. Further, the effect of pesticides on the indole acetic acid (IAA) production by Azotobacter strains was also estimated. Azotobacter-16 strain was found to produce 34.4 μg ml^?l of IAA in a media supplemented with 1,000 mg of tryptophan and 5 % of pendimethalin. Present study reveals that species of Azotobacter are able to grow and survive in the presence of pesticides and no significant effects were observed on the metabolic activities of Azotobacter species.     

Phenotypic and molecular characterisation of efficient nitrogen-fixing Azotobacter strains from rice fields for crop improvement

Biological nitrogen fixation (BNF) is highly effective in the field and potentially useful to reduce adverse effects chemical fertilisers. Here, Azotobacter species were selected via phenotypic, biochemical and molecular characterisations from different rice fields. Acetylene reduction assay of Azotobacter spp. showed that Azotobacter vinelandii (Az3) fixed higher amount of nitrogen (121.09 nmol C₂H₄?mg^-1 bacteria h^-1). Likewise, its plant growth functions, viz. siderophore, hydrogen cyanide, salicylic acid, IAA, GA₃, zeatin, NH₃, phosphorus solubilisation, ACC deaminase and iron tolerance, were also higher. The profile of gDNA, plasmid DNA and cellular protein profile depicted inter-generic and inter-specific diversity among the isolates of A. vinelandii. The PCR-amplified genes nifH, nifD and nifK of 0.87, 1.4 and 1.5 kb , respectively, were ascertained by Southern blot hybridisation in isolates of A. vinelandii. The 16S rRNA sequence from A. vinelandii (Az3) was novel, and its accession number (JQ796077) was received from NCBI data base. Biofertiliser formulation of novel A. vinelandii isolates along with commercial one was evaluated in rice (Oriza sativa L. var. Khandagiri) fields. The present finding revealed that treatment T4 (Az3) (A. vinelandii) are highly efficient to improved growth and yield of rice crop.     

Population Genetic Structure of a Microalgal Species under Expansion

Biological invasions often cause major perturbations in the environment and are well studied among macroorganisms. Less is known about invasion by free-living microbes. Gonyostomum semen (Raphidophyceae) is a freshwater phytoplankton species that has increased in abundance in Northern Europe since the 1980''s and has expanded its habitat range. In this study, we aimed to determine the genetic population structure of G. semen in Northern Europe and to what extent it reflects the species'' recent expansion. We sampled lakes from 12 locations (11 lakes) in Norway, Sweden and Finland. Multiple strains from each location were genotyped using Amplified Fragment Length Polymorphism (AFLP). We found low differentiation between locations, and low gene diversity within each location. Moreover, there was an absence of genetic isolation with distance (Mantel test, p = 0.50). According to a Bayesian clustering method all the isolates belonged to the same genetic population. Together our data suggest the presence of one metapopulation and an overall low diversity, which is coherent with a recent expansion of G. semen.     

不同油葵品种对盐碱地根际土壤酶活性及微生物群落功能多样性的影响

通过盆栽试验,研究了新葵4号、新葵6号、新葵10号和美国矮大头4个油葵品种对盐碱地土壤理化、酶活性和微生物群落功能多样性的影响,以期筛选出更适宜改善盐碱地土壤质量的油葵品种。结果表明,种植新葵6号对降低盐碱地根际土壤pH值、提高土壤全氮含量和蔗糖酶活性的效果最为显著,新葵4号对提高根际土壤碱解氮、速效磷、速效钾含量以及脲酶和磷酸酶活性的效果最为显著;种植这4个品种的油葵均能显著(72 h,P0.05)提高盐碱地根际土壤微生物对31种碳源的平均利用率(Average well color development,AWCD),并呈现如下规律:新葵4号新葵6号美国矮大头新葵10号CK。种植这4个油葵品种均不同程度地提高了盐碱地根际土壤微生物群落的Shannon多样性指数(H)、Shannon优势度指数(D)和碳源利用丰富度指数(S),并呈现出相似的规律:新葵4号根际土的微生物多样性指数最大,而CK的最小,且显著高于CK。主成分分析表明,种植油葵改善了盐碱地根际土壤微生物的群落组成;碳水化合物、氨基酸、羧酸类化合物和聚合物是盐碱地土壤微生物利用的主要碳源。因此,在盐碱地中种植油葵可提高相关土壤理化性质和酶的活性,改善微生物功能多样性,优化盐碱地微生物的群落结构,尤其是种植新葵4号对盐碱地的改良效果最为显著。     

烟草根际微生物研究

利用选择性培养基,对土壤肥力肥沃、中等、贫瘠的烟区根际细菌、真菌和放线菌进行了分离和测数。根据菌体形态及培养特征、生理生化指标及16S rDNA部分序列分析等,对根际自生固氮菌、磷细菌、钾细菌进行了鉴定和分类。主要结果为：土壤肥沃的烟区根际细菌的数量最多,土壤贫瘠的烟区数量最少;土壤贫瘠烟区根际真菌的数量较多,中等和肥沃烟区的较少;根际故线菌的数量随土壤肥力的降低而依次减小。从不同肥力烟区分离的根际自生固氮菌、磷细菌、钾细菌以革兰氏阴性杆菌为主,分别属于10个属。土壤的肥沃程度对根际3类细菌的种类和数量都有影响,总体上看,土壤肥沃和中等的条件下,细菌类群的多样性和丰富度较大,而贫瘠土壤细菌类群的优势度较大。     

Non-symbiotic nitrogen fixing bacteria in tropical soils

Summary Several aspects of heterotrophic, free-living nitrogen-fixing bacteria specific to tropical environments are presented. WhileAzotobacter chroococcum andA. vinelandii seem to occur throughout the world,A. paspali, Derxia gummosa andBeijerinckia spp. have been found mainly in the tropics. Derxia occurred more frequently than previously supposed and was stimulated by moist soil conditions, particularly after flooding.A. paspali growth in soil even under experimental conditions, was strictly limited to the rhizosphere of a few varieties ofPaspalum notatum and the association took almost a year to become established. This work was supported by the Brazilian National Research Council.     

Untraditional N2-fixing bacteria as biofertilizers for wheat and barley

The screening of 27 isolates grown on nitrogen-free medium for nitrogen-fixing ability resulted in the isolation of five organisms belonging toBacillaceae, Enterobacteriaceae andPseudomonadaceae. Estimates of N₂-fixation efficiencies of these isolates indicated that they may be responsible for low rates of N₂-fixation in soil. The possible association of these isolates as well as ofAzotobacter andAzospirillum with wheat and barley was investigated in a greenhouse experiment. The highest values of nitrogenase activity on plant root were recorded in treatments inoculated with composite inocula of the isolated N₂-fixers, particularly whenAzotobacter and/orAzospirillum were added in combination. Inoculation with single inoculum of each of the N₂-fixing isolates had no significant influence on plant growth, except withPseudomonas andBacillus for wheat and barley, respectively. Highly significant increases in growth of both plants were recorded in all cases of multistrain inoculation.     

<Emphasis Type="Italic">Nif</Emphasis>H-based studies on azotobacterial diversity in cotton soils of India

In order to promote the use of Azotobacter inoculants for cotton crop, a complete characterization of soil isolates of Azotobacter, isolated and screened on the basis of physiological properties, from four different cotton–wheat cropping regions of India was carried out, and their genetic diversity determined by RFLP (restriction fragment length polymorphism) analysis of the functional gene nifH. Genetic analysis of these isolates depicted a similarity coefficient of ≥80% among them, suggesting that though the isolates were obtained from different cotton soils of India, still they have large commonality in the nifH gene and constituted a homogeneous nifH population.     

三峡库区水体中固氮微生物多样性及其影响因素

【目的】研究分析不同时空条件下三峡库区水体固氮微生物多样性,并探讨其与地球化学参数的相关性。【方法】采集三峡库区不同时间(三月份和六月份)和空间(干流与支流)的水体样品,对其进行地球化学参数分析,并通过构建克隆文库分析样品中固氮功能基因(nifH)的多样性进而探讨其与水体地化参数的相关关系。【结果】统计分析显示三峡库区水体固氮微生物α-多样性和群落组成具有时空差异。支流水体样品的固氮微生物α-多样性高于干流水体样品;六月水体样品的固氮微生物α-多样性高于三月水体样品。三峡库区三月水体样品中的固氮微生物群落以Proteobacteria (50.3%)和Firmicutes (40.0%)为主;六月水体样品的固氮微生物群落以Proteobacteria(48.4%)、Firmicutes(25.4%)和Cyanobacteria(19.0%)为主。Mantel检验结果显示:固氮微生物群落结构的差异与温度、pH和DIC等地球化学参数具有显著(P0.05)相关性,其中温度和pH的相关性系数最大。【结论】三峡库区固氮微生物的种群结构和多样性具有时空差异,影响三峡水库水体中固氮微生物群落结构与多样性的主要环境因素为温度和pH,同时浊度、DIC、氨氮也对库区水体固氮微生物群落结构和多样性有一定的影响。     

Isolation and Molecular Characterization of Heavy Metal-Resistant Azotobacter chroococcum from Agricultural Soil and Their Potential Application in Bioremediation

Pollution of soil with heavy metals, herbicides, antibiotics and other chemicals is known to have a negative effect on microbial activities. Therefore, the aim of this study was to isolate cultures of Azotobacter sp. from polluted and unpolluted soils and to study the effect of these pollutants on their growth. A total of 120 Azotobacter sp. were isolated from soils irrigated with wastewater (contaminated soils) and groundwater (uncontaminated soils). These isolates were screened for resistance to heavy metals, herbicide and antibiotics. Also, the soils from which the cultures were isolated were analyzed for the concentrations of Zn²⁺, Cd²⁺, Cu²⁺, Pb²⁺ and Mn²⁺ they contained. Contaminated soil showed high levels of heavy metals as compared to uncontaminated soil. The size of the Azotobacter population in contaminated soil was lower than that in uncontaminated soil. Of the Azotobacter isolates, 64 that were recovered from contaminated soil exhibited high resistance to heavy metals (Hg²⁺, Cd²⁺, Cu²⁺, Cr³⁺, Co²⁺, Ni²⁺, Zn²⁺ and Pb²⁺) and herbicide 2,4-D compared to 56 isolates from uncontaminated soil. Also, isolates from contaminated soil showed high resistance to chloramphenicol, nitrofurantoin and co-trimoxazole compared to those isolated from uncontaminated soil. The majority of Azotobacter isolates from contaminated soil showed multiple-resistance to different metal ions and antibiotics. All isolates failed to grow at pH less than 6. Salt concentration (5%) was found to be inhibitory to all isolates. The most potent isolates from contaminated soil that showed multiresistance to all substances tested were identified on the basis of morphological and biochemical characteristics, and 16S rRNA as A. chroococcum. These resistant isolates could be employed in contaminated soils and/or bioremediation.     

The role of cellulose-decomposing fungi in nitrogenase activity ofAzotobacter chroococcum

Azotobacter chroococcum was grown on cultures containing five carbon sources alone and also in co-cultures with three cellulolytic fungi (Aspergillus niger, Penicillium funiculosum andTrichoderma harzianum). In the absence of fungal species, nitrogenase activity was relatively low. The best nitrogenase activity was recorded in cultures containing faba bean straw followed by that in cultures having wheat straw, sugar cane leaves, carboxymethyl cellulose (CMC) or cellulose. In co-cultures with fungi,Azotobacter showed substantial nitrogenase activity on all tested substrates.Azotobacter —Trichoderma association showed the highest nitrogenase activity.     

Assessment of ecological diversity of rhizobacterial communities in vermicompost and analysis of their potential to improve plant growth

Present study was designed to determine the microbial diversity from three distinctive sites (amended with vermicompost) of Gujarat, India. A set of 76 strains were screened from total of 438 strains that exhibit plant growthpromoting (PGP) and antagonistic potential isolated from sites PS1 (Mehsana district), BS2 (Dantiwada district) and VS3 (Gandhinagar district). Their diversity indices were studied for determining the species richness and evenness of screened isolates. Results revealed that site BS2 showed the most significant diversity indices in terms of Shannon (H′ 1.525) and Simpson (1/D 5.120) than other two samples. Principal component analysis showed that bacterial diversity (H′) was correlated with the soil characteristics. Chickpea and groundnut plants inoculated with MBCU1 and MBCU3 isolates showed an increase in the vegetative growth parameters that evaluate plant growth when compared to uninoculated controls. Strains MBCU1 and MBCU3 were identified as Pseudomonas stutzeri and Pseudomonas mosselii, respectively, according to sequence analysis of the 16S rRNA gene. These both isolates belong to site BS2 and they showed specific PGP traits suggesting that these isolates can promote plant growth by more than one mechanism with respect to their higher diversity index.     

Adaptability to local conditions and phylogenetic differentiation of microsymbionts of TGx soybean genotypes in the semi-arid environments of Ghana and South Africa

The study of the nitrogen fixation and phylogenetic diversity of nodule microsymbionts of grain legumes in many parts of the globe is often carried out in order to identify legume-rhizobia combinations for agricultural sustainability. Several reports have therefore found that rhizobial species diversity is shaped by edapho-climatic conditions that characterize different geographic locations, suggesting that rhizobial communities often possess traits that aid their adaptation to their habitat. In this study, the soybean-nodulating rhizobia from semi-arid savannahs of Ghana and South Africa were evaluated. The authenticated rhizobial isolates were highly diverse based on their colony characteristics, as well as their BOX-PCR profiles and gene sequences. In the 16S rRNA phylogeny, the isolates were placed in the different clades Bradyrhizobium iriomotense and Bradyrhizobium jicamae together with two superclades Bradyrhizobium japonicum and Bradyrhizobium elkanii. The multilocus (atpD, glnII, gyrB, recA) phylogenetic analyses indicated the dominance of Bradyrhizobium diazoefficiens and putative new Bradyrhizobium species in the semi-arid Ghanaian region. The phylogenetic analyses based on the symbiotic genes (nifH and nodC) clustered the test isolates into different symbiovars (sv. glycinearum, sv. retame and sv. sojae). Principal component analysis (PCA) showed that soil factors played a significant role in favoring the occurrence of soybean-nodulating microsymbionts in the tested local conditions. The results suggested that isolates had marked local adaptation to the prevailing conditions in semi-arid regions but further studies are needed to confirm new Bradyrhizobium species.     

Response of soil nematode community structure and diversity to long-term land use in the black soil region in China

Soil nematodes are sensitive to environmental changes and are used widely as indicators of soil conditions. The community structure and diversity of soil nematodes were studied in different long-term land use regimes in the black soil area in Northeast China. The land use regimes were maintained for 22?years, and included crop land (CL), grass land (GL) and bare land (BL). Soil samples were taken throughout the growing season, and nematodes were extracted and identified. A total of 39 nematode genera with relative abundance over 0.1?% were identified. Heterodera was the dominant genus in CL; Boleodorus was the dominant genus in GL, and Boleodorus, Eucephalobus and Filenchus were the dominant genera in BL. Land use had a significant effect on abundance of all soil nematode tropic groups and ecological indices. Sampling time had an effect on soil nematode abundance, but only on three of the eight nematode ecological indices MI (maturity index of free-living nematode), CI (channel index) and EI (enrichment index). SR (species richness index) was highest in GL where plant species diversity was also high. The CI was the highest in BL among three land uses, which means the soil food web dominated, with fungal decomposition channels in BL. Soil nematode community structure and diversity was shown to be an effective and informative tool for analyzing ecological aspects of land use in black soil regions. The data are inconclusive as to whether the effect of land use on soil nematode parameters is direct, or indirect via inducing changes in soil physicochemical properties.