Comparison of nifH gene pools in soils and soil microenvironments with contrasting properties

The similarities and differences in the structures of the nifH gene pools of six different soils (Montrond, LCSA-p, Vernon, Dombes, LCSA-c, and Thysse Kaymor) and five soil fractions extracted from LCSA-c were studied. Bacterial DNA was directly extracted from the soils, and a region of the nifH gene was amplified by PCR and analyzed by restriction. Soils were selected on the basis of differences in soil management, plant cover, and major physicochemical properties. Microenvironments differed on the basis of the sizes of the constituent particles and the organic carbon and clay contents. Restriction profiles were subjected to principal-component analysis. We showed that the composition of the diazotrophic communities varied both on a large scale (among soils) and on a microscale (among microenvironments in LCSA-c soil). Soil management seemed to be the major parameter influencing differences in the nifH gene pool structure among soils by controlling inorganic nitrogen content and its variation. However, physicochemical parameters (texture and total C and N contents) were found to correlate with differences among nifH gene pools on a microscale. We hypothesize that the observed nifH genetic structures resulted from the adaptation to fluctuating conditions (cultivated soil, forest soil, coarse fractions) or constant conditions (permanent pasture soil, fine fractions). We attempted to identify a specific band within the profile of the clay fraction by cloning and sequencing it and comparing it with the gene databases. Unexpectedly, the nifH sequences of the dominant bacteria were most similar to sequences of unidentified marine eubacteria.     

Distribution of Extensive nifH Gene Diversity Across Physical Soil Microenvironments

The diversity of nitrogen-fixing bacteria is well described for aquatic environments; however, terrestrial analyses remain mostly biased to rhizobial plant–microbe associations. We maximized the level of resolution for this study through the use of nucleotide sequence information extracted from a series of soil microenvironments, ranging from macroaggregates at 2000 μm to the clay fraction at <75 μm in diameter. In addition, we attempted to create an overview of the distribution of terrestrial nitrogen fixers across such microenvironments by combining culture-independent techniques with a suite of natural soil environments from uniquely different origins. Soil diazotroph diversity was analyzed phylogenetically for 600 terrestrial nifH sequences from 12 midsized clone libraries based on microenvironments of three separate soils across a global scale. Statistical analyses of nifH gene clone libraries were used to estimate coverage, establish degrees of sequence overlap, and compare cluster distributions. These analyses revealed an extensive diversity in a tropical (19 phylotypes) and an arctic soil (17 phylotypes), and moderate diversity in a temperate soil (11 phylotypes). Within each soil, comparisons across aggregate size fractions delineated nifH gene cluster shifts within populations and degrees of sequence overlap that ranged from significantly different (arctic, tropical) to significantly similar (temperate). We suggest that this is due to population separation across aggregates of different size classes, which results from differences in the temporal stability of aggregates as niches for microbial communities. This study not only provides new knowledge of the arrangement of diazotrophic communities at the soil microscale, but it also contributes to the underrepresented knowledge of soil nifH sequences in the public databases.     

Analysis of nifH Gene Pool Complexity in Soil and Litter at a Douglas Fir Forest Site in the Oregon Cascade Mountain Range

Nitrogen-fixing microbial populations in a Douglas fir forest on the western slope of the Oregon Cascade Mountain Range were analyzed. The complexity of the nifH gene pool (nifH is the marker gene which encodes nitrogenase reductase) was assessed by performing nested PCR with bulk DNA extracted from plant litter and soil. The restriction fragment length polymorphisms (RFLPs) of PCR products obtained from litter were reproducibly different than the RFLPs of PCR products obtained from the underlying soil. The characteristic differences were found during the entire sampling period between May and September. RFLP analyses of cloned nifH PCR products also revealed characteristic patterns for each sample type. Among 42 nifH clones obtained from a forest litter library nine different RFLP patterns were found, and among 64 nifH clones obtained from forest soil libraries 13 different patterns were found. Only two of the patterns were found in both the litter and the soil, indicating that there were major differences between the nitrogen-fixing microbial populations. A sequence analysis of clones representing the 20 distinct patterns revealed that 19 of the patterns had a proteobacterial origin. All of the nifH sequences obtained from the Douglas fir forest litter localized in a distinct phylogenetic cluster characterized by the nifH sequences of members of the genera Rhizobium, Sinorhizobium, and Azospirillum. The nifH sequences obtained from soil were found in two additional clusters, one characterized by sequences of members of the genera Bradyrhizobium, Azorhizobium, Herbaspirillum, and Thiobacillus and the other, represented by a single nifH clone, located between the gram-positive bacteria and the cyanobacteria. Our results revealed the distinctness of the nitrogen-fixing microbial populations in litter and soil in a Douglas fir forest; the differences may be related to special requirements for degradation and mineralization processes in the plant litter.     

亚热带不同林分土壤表层有机碳组成及其稳定性

在浙江临安玲珑山选取了常绿阔叶林、马尾松林、板栗林和雷竹林4种林分,采用传统的化学方法与固态13C核磁共振(NMR)技术研究其土壤有机碳在不同粒径土壤颗粒中的分布规律和结构特征,探讨林分类别和管理措施对土壤有机碳含量及其结构的影响,为亚热带地区森林固碳和土壤碳库管理提供科学依据。结果显示:(1)土壤表层(0—20 cm)有机碳含量按以下次序递减:雷竹林>常绿阔叶林>马尾松林>板栗林,且板栗林以粉黏粒结合态碳为主,其他林分土壤则以粗砂结合态碳为主;(2)13C NMR结果表明,阔叶林和马尾松林土壤有机碳中烷基碳所占比例最大,而雷竹林和板栗林则是烷氧碳比例最大,表明人工经营措施改变了土壤有机碳的成分组成;(3)随着土壤颗粒变细,有机碳中烷基碳比例增加,烷氧碳比例减少,A/O-A值和疏水碳/亲水碳值逐渐增大,表明颗粒越细,其结合的有机碳结构稳定性越高。     

Genetic Diversity of nifH Gene Sequences in Paenibacillus azotofixans Strains and Soil Samples Analyzed by Denaturing Gradient Gel Electrophoresis of PCR-Amplified Gene Fragments

The diversity of dinitrogenase reductase gene (nifH) fragments in Paenibacillus azotofixans strains was investigated by using molecular methods. The partial nifH gene sequences of eight P. azotofixans strains, as well as one strain each of the close relatives Paenibacillus durum, Paenibacillus polymyxa, and Paenibacillus macerans, were amplified by PCR by using degenerate primers and were characterized by DNA sequencing. We found that there are two nifH sequence clusters, designated clusters I and II, in P. azotofixans. The data further indicated that there was sequence divergence among the nifH genes of P. azotofixans strains at the DNA level. However, the gene products were more conserved at the protein level. Phylogenetic analysis showed that all nifH cluster II sequences were similar to the alternative (anf) nitrogenase sequence. A nested PCR assay for the detection of nifH (cluster I) of P. azotofixans was developed by using the degenerate primers as outer primers and two specific primers, designed on the basis of the sequence information obtained, as inner primers. The specificity of the inner primers was tested with several diazotrophic bacteria, and PCR revealed that these primers are specific for the P. azotofixans nifH gene. A GC clamp was attached to one inner primer, and a denaturing gradient gel electrophoresis (DGGE) protocol was developed to study the genetic diversity of this region of nifH in P. azotofixans strains, as well as in soil and rhizosphere samples. The results revealed sequence heterogeneity among different nifH genes. Moreover, nifH is probably a multicopy gene in P. azotofixans. Both similarities and differences were detected in the P. azotofixans nifH DGGE profiles generated with soil and rhizosphere DNAs. The DGGE assay developed here is reproducible and provides a rapid way to assess the intraspecific genetic diversity of an important functional gene in pure cultures, as well as in environmental samples.     

New Molecular Screening Tools for Analysis of Free-Living Diazotrophs in Soil

Free-living nitrogen-fixing prokaryotes (diazotrophs) are ubiquitous in soil and are phylogenetically and physiologically highly diverse. Molecular methods based on universal PCR detection of the nifH marker gene have been successfully applied to describe diazotroph populations in the environment. However, the use of highly degenerate primers and low-stringency amplification conditions render these methods prone to amplification bias, while less degenerate primer sets will not amplify all nifH genes. We have developed a fixed-primer-site approach with six PCR protocols using less degenerate to nondegenerate primer sets that all amplify the same nifH fragment as a previously published PCR protocol for universal amplification. These protocols target different groups of diazotrophs and allowed for direct comparison of the PCR products by use of restriction fragment length polymorphism fingerprinting. The new protocols were optimized on DNA from 14 reference strains and were subsequently tested with bulk DNA extracts from six soils. These analyses revealed that the new PCR primer sets amplified nifH sequences that were not detected by the universal primer set. Furthermore, they were better suited to distinguish between diazotroph populations in the different soils. Because the novel primer sets were not specific for monophyletic groups of diazotrophs, they do not serve as an identification tool; however, they proved powerful as fingerprinting tools for subsets of soil diazotroph communities.     

Phosphorus and carbon in soil particle size fractions: A synthesis

Despite the importance of phosphorus (P) as a macronutrient, the factors controlling the pool sizes of organic and inorganic P (OP and IP) in soils are not yet well understood. Therefore, the aim of this study was to gain insights into the pools sizes of OP, IP and organic carbon (OC) in soils and soil particle size fractions. For this purpose, I analyzed the distribution of OP, IP, and OC among particle size fractions depending on geographical location, climate, soil depth, and land use, based on published data. The clay size fraction contained on average 8.8 times more OP than the sand size fraction and 3.9 and 3.2 times more IP and OC, respectively. The OP concentrations of the silt and clay size fraction were both negatively correlated with mean annual temperature (R² = 0.30 and 0.31, respectively, p < 0.001). The OC:OP ratios of the silt and clay size fraction were negatively correlated with latitude (R² = 0.49 and 0.34, respectively, p < 0.001). Yet, the OC:OP ratio of the clay size fraction changed less markedly with latitude than the OC:OP ratio of the silt and the sand size fraction. The OC concentrations of all three particle size fractions were significantly (p < 0.05) lower in soils converted to cropland than in adjacent soils under natural vegetation. In contrast, the OP concentration was only significantly (p < 0.05) decreased in the sand size fraction but not in the other two particle size fractions due to land-use change. Thus, the findings suggest that OP is more persistent in soil than OC, which is most likely due to strong sorptive stabilization of OP compounds to mineral surfaces.

     

Changes in Nitrogen-Fixing and Ammonia-Oxidizing Bacterial Communities in Soil of a Mixed Conifer Forest after Wildfire

This study was undertaken to examine the effects of forest fire on two important groups of N-cycling bacteria in soil, the nitrogen-fixing and ammonia-oxidizing bacteria. Sequence and terminal restriction fragment length polymorphism (T-RFLP) analysis of nifH and amoA PCR amplicons was performed on DNA samples from unburned, moderately burned, and severely burned soils of a mixed conifer forest. PCR results indicated that the soil biomass and proportion of nitrogen-fixing and ammonia-oxidizing species was less in soil from the fire-impacted sites than from the unburned sites. The number of dominant nifH sequence types was greater in fire-impacted soils, and nifH sequences that were most closely related to those from the spore-forming taxa Clostridium and Paenibacillus were more abundant in the burned soils. In T-RFLP patterns of the ammonia-oxidizing community, terminal restriction fragments (TRFs) representing amoA cluster 1, 2, or 4 Nitrosospira spp. were dominant (80 to 90%) in unburned soils, while TRFs representing amoA cluster 3A Nitrosospira spp. dominated (65 to 95%) in fire-impacted soils. The dominance of amoA cluster 3A Nitrosospira spp. sequence types was positively correlated with soil pH (5.6 to 7.5) and NH₃-N levels (0.002 to 0.976 ppm), both of which were higher in burned soils. The decreased microbial biomass and shift in nitrogen-fixing and ammonia-oxidizing communities were still evident in fire-impacted soils collected 14 months after the fire.     

Diazotrophic diversity, nifH gene expression and nitrogenase activity in a rice paddy field in Fujian, China

The diazotrophic communities in a rice paddy field were characterized by a molecular polyphasic approach including DNA/RNA-DGGE fingerprinting, real time RT-PCR analysis of nifH gene and the measurement of nitrogen fixation activities. The investigation was performed on a diurnal cycle and comparisons were made between bulk and rhizosphere / root soil as well as between fertilized / unfertilized soils. Real time RT-PCR showed no significant difference in the total quantity of nifH expression under the conditions investigated. The functional diversity and dynamics of the nifH gene expressing diazotroph community investigated using RT-PCR-DGGE revealed high diurnal variations, as well as variation between different soil types. Most of the sequence types recovered from the DGGE gels and clone libraries clustered within nifH Cluster I and III (65 different nifH sequences in total). Sequence types most similar to Azoarcus spp., Metylococcus spp., Rhizobium spp., Methylocystis spp., Desulfovibrio spp., Geobacter spp., Chlorobium spp., were abundant and indicate that these species may be responsible for the observed diurnal variation in the diazotrophic community structure in these rice field samples. Previously described diazotrophic cyanobacterial genera in rice fields, such as Nostoc and Cyanothece, were present in the samples but not detectable in RT-PCR assays.     

Soluble Organic Nitrogen Pools in Forest soils of Subtropical Australia

Soil soluble organic N (SON) plays an important role in N biogeochemical cycling. In this study, 22 surface forest soils (0–10 cm) were collected from southeast Queensland, Australia, to investigate the size of SON pools extracted by water and salt solutions. Approximately 5–45 mg SON kg⁻¹, 2–42 mg SON kg⁻¹ and 1–24 SON mg kg⁻¹ were extracted by 2 M KCl, 0.5 M K₂SO₄ and water, on average, corresponding to about 21.1, 13.5 and 7.0 kg SON ha⁻¹ at the 0–10 cm forest soils, respectively. These SON pools, on average, accounted for 39% (KCl extracts), 42% (K₂SO₄ extracts) and 43% (water extracts) of total soluble N (TSN), and 2.3% (KCl extracts), 1.3% (K₂SO₄ extracts) and 0.7% (water extracts) of soil total N, respectively. Large variation in SON pools observed across the sites in the present study may be attributed to a combination of factors including soil types, tree species, management practices and environmental conditions. Significant relationships were observed among the SON pools extracted by water, KCl and K₂SO₄ and microbial biomass N (MBN). In general, KCl and K₂SO₄ extracted more SON than water from the forest soils, while KCl extracted more SON than K₂SO₄. The SON and soluble organic C (SOC) in KCl, K₂SO₄ and water extracts were all positively related to soil organic C, total N and clay contents. This indicates that clay and soil organic matter play a key role in the retention of SON in soil.     

Xiphinema americanum as Affected by Soil Organic Matter and Porosity

The effects of four soil types, soil porosity, particle size, and organic matter were tested on survival and migration of Xiphinema americanum. Survival and migration were significantly greater in silt loam than in clay loam and silty clay soils. Nematode numbers were significantly greater in softs planted with soybeans than in fallow softs. Nematode survival was greatest at the higher of two pore space levels in four softs. Migration of X. americanum through soft particle size fractions of 75-150, 150-250, 250-500, 500-700, and 700-1,000 μ was significantly greater in the middle three fractions, with the least occurring in the smallest fraction. Additions of muck to silt loam and loamy sand soils resulted in reductions in survival and migration of the nematode. The fulvic acid fraction of muck, extracted with sodium hydroxide, had a deleterious effect on nematode activity. I conclude that soils with small amounts of air-filled pore space, extremes in pore size, or high organic matter content are deleterious to the migration and survival of X. americanum, and that a naturally occurring toxin affecting this species may be present in native soft organic matter.     

Interactive effects of disturbance and nitrogen availability on phosphorus dynamics of southern Appalachian forests

Understanding the main and interactive effects of chronically altered resource availability and disturbance on phosphorus (P) availability is increasingly important in light of the rapid pace at which human activities are altering these processes and potentially introducing P limitation. We measured P pools and fluxes in eighteen mixed forest stands at three elevations (low, mid, high) subjected to increasing atmospheric N deposition, where hemlock (Tsuga canadensis) was absent or declining due to infestation by the exotic hemlock woolly adelgid (Adelges tsugae). While total soil P was similar across the study area, phosphorus fractionation revealed distinct differences in the distribution of soil P fractions as elevation and N availability increased. Soils from high elevation plots where N availability was greatest had 139 % larger organic P pools and 55 % smaller residual and refractory P pools than soils from low elevation plots with less N availability, suggesting that increased N availability has driven the depletion of recalcitrant P pools by stimulating biotic demand and sequestration. These differences in P distribution among fractions influenced how tree mortality affected P dynamics. At high elevations, plots containing declining hemlocks had significantly greater foliar P concentrations and fluxes of P from the forest floor than reference plots at similar elevations, whereas at low and mid-elevations there were no consistent differences between plots. Across all elevation classes, hardwood foliar N:P ratios were lower in plots with declining hemlocks. Collectively, these results suggest that increased N availability enhances bioavailable P, which is sequestered in vegetation until disturbances liberate it.     

Soil organic matter dynamics in density and particle-size fractions following destruction of tropical rainforest and the subsequent establishment of Imperata grassland in Indonesian Borneo using stable carbon isotopes

Background and aims

Large portions of the deforested areas in Southeast Asia have been ultimately replaced by the invasive grass Imperata cylindrica, but the dynamics of soil organic matter (SOM) during such land transitions are poorly understood. This study presents SOM dynamics in density and particle-size fractions following rainforest destruction and the subsequent establishment and persistence of Imperata grassland.

Methods

We examined soil C stock and natural ¹³C abundance in these fractions to depths of 100 cm. We predicted future soil C storage and evaluated C turnover rates in these fractions using a simple exponential model. Because soil texture strongly affects soil C storage, two chronosequences of soils differing in soil texture were compared (n?=?1 in each chronosequence).

Results

The clay-associated SOM increased in all soil layers (0–100 cm) along the forest-to-grassland chronosequence, whereas light-fraction SOM in the surface soil layer (0–5 cm) decreased.

Conclusions

In the surface layer, all SOM fractions exhibited rapid replacement of forest-derived C to grassland-derived C, indicating fast turnover. Meanwhile, δ¹³C values of the light fraction in the surface layer indicated that forest-derived charcoal and/or occluded low-density organic matter constituted unexpectedly large proportions of the light fraction. Mathematical modelling (0–50 cm) showed that grassland-derived C in the clay and silt fractions in all soil layers increased almost linearly for at least 50 years after grassland establishment. In the meantime, the forest-derived C stock in the clay fraction constituted 82 % of the total stable C pool at 0–50-cm depths even under steady-state conditions (t = ∞), indicating that residue of forest-derived SOM associated with clay largely contributed to preserving the soil C pool. Comparing soils with different soil textures, clay and silt particles in coarse-textured soil exhibited a substantially higher degree of organo-mineral interactions per unit volume of clay or silt compared to fine-textured soils.     

Genomic diversity of chickpea-nodulating rhizobia in Ningxia (north central China) and gene flow within symbiotic Mesorhizobium muleiense populations

Diversity and taxonomic affiliation of chickpea rhizobia were investigated from Ningxia in north central China and their genomic relationships were compared with those from northwestern adjacent regions (Gansu and Xinjiang). Rhizobia were isolated from root-nodules after trapping by chickpea grown in soils from a single site of Ningxia and typed by IGS PCR-RFLP. Representative strains were phylogenetically analyzed on the basis of the 16S rRNA, housekeeping (atpD, recA and glnII) and symbiosis (nodC and nifH) genes. Genetic differentiation and gene flow were estimated among the chickpea microsymbionts from Ningxia, Gansu and Xinjiang. Fifty chickpea rhizobial isolates were obtained and identified as Mesorhizobium muleiense. Their symbiosis genes nodC and nifH were highly similar (98.4 to 100%) to those of other chickpea microsymbionts, except for one representative strain (NG24) that showed low nifH similarities with all the defined Mesorhizobium species. The rhizobial population from Ningxia was genetically similar to that from Gansu, but different from that in Xinjiang as shown by high chromosomal gene flow/low differentiation with the Gansu population but the reverse with the Xinjiang population. This reveals a biogeographic pattern with two main populations in M. muleiense, the Xinjiang population being chromosomally differentiated from Ningxia-Gansu one. M. muleiense was found as the sole main chickpea-nodulating rhizobial symbiont of Ningxia and it was also found in Gansu sharing alkaline-saline soils with Ningxia. Introduction of chickpea in recently cultivated areas in China seems to select from alkaline-saline soils of M. muleiense that acquired symbiotic genes from symbiovar ciceri.     

Biodiversity of Denitrifying and Dinitrogen-Fixing Bacteria in an Acid Forest Soil

Isolated soil DNA from an oak-hornbeam forest close to Cologne, Germany, was suitable for PCR amplification of gene segments coding for the 16S rRNA and nitrogenase reductase (NifH), nitrous oxide reductase (NosZ), cytochrome cd₁-containing nitrite reductase (NirS), and Cu-containing nitrite reductase (NirK) of denitrification. For each gene segment, diverse PCR products were characterized by cloning and sequencing. None of the 16S rRNA gene sequences was identical to any deposited in the data banks, and therefore each of them belonged to a noncharacterized bacterium. In contrast, the analyzed clones of nifH gave only a few different sequences, which occurred many times, indicating a low level of species richness in the N₂-fixing bacterial population in this soil. Identical nifH sequences were also detected in PCR amplification products of DNA of a soil approximately 600 km distant from the Cologne area. Whereas biodiversity was high in the case of nosZ, only a few different sequences were obtained with nirK. With respect to nirS, cloning and sequencing of the PCR products revealed that many false gene segments had been amplified with DNA from soil but not from cultured bacteria. With the 16S rRNA gene data, many sequences of uncultured bacteria belonging to the Acidobacterium phylum and actinomycetes showed up in the PCR products when isolated DNA was used as the template, whereas sequences obtained for nifH and for the denitrification genes were closely related to those of the proteobacteria. Although in such an experimental approach one has to cope with the enormous biodiversity in soils and only a few PCR products can be selected at random, the data suggest that denitrification and N₂ fixation are not genetic traits of most of the uncultured bacteria.     

Abundance of Microbes Involved in Nitrogen Transformation in the Rhizosphere of Leucanthemopsis alpina (L.) Heywood Grown in Soils from Different Sites of the Damma Glacier Forefield

Glacier forefields are an ideal playground to investigate the role of development stages of soils on the formation of plant–microbe interactions as within the last decades, many alpine glaciers retreated, whereby releasing and exposing parent material for soil development. Especially the status of macronutrients like nitrogen differs between soils of different development stages in these environments and may influence plant growth significantly. Thus, in this study, we reconstructed major parts of the nitrogen cycle in the rhizosphere soil/root system of Leucanthemopsis alpina (L.) Heywood as well as the corresponding bulk soil by quantifying functional genes of nitrogen fixation (nifH), nitrogen mineralisation (chiA, aprA), nitrification (amoA AOB, amoA AOA) and denitrification (nirS, nirK and nosZ) in a 10-year and a 120-year ice-free soil of the Damma glacier forefield. We linked the results to the ammonium and nitrate concentrations of the soils as well as to the nitrogen and carbon status of the plants. The experiment was performed in a greenhouse simulating the climatic conditions of the glacier forefield. Samples were taken after 7 and 13 weeks of plant growth. Highest nifH gene abundance in connection with lowest nitrogen content of L. alpina was observed in the 10-year soil after 7 weeks of plant growth, demonstrating the important role of associative nitrogen fixation for plant development in this soil. In contrast, in the 120-year soil copy numbers of genes involved in denitrification, mainly nosZ were increased after 13 weeks of plant growth, indicating an overall increased microbial activity status as well as higher concentrations of nitrate in this soil.     

Changes in aboveground primary production and carbon and nitrogen pools accompanying woody plant encroachment in a temperate savanna

When woody plant abundance increases in grasslands and savannas, a phenomenon widely observed worldwide, there is considerable uncertainty as to whether aboveground net primary productivity (ANPP) and ecosystem carbon (C) and nitrogen (N) pools increase, decrease, or remain the same. We estimated ANPP and C and N pools in aboveground vegetation and surface soils on shallow clay and clay loam soils undergoing encroachment by Prosopis glandulosa in the Southern Great Plains of the United States. Aboveground Prosopis C and N mass increased linearly, and ANPP increased logarithmically, with stand age on clay loam soils; on shallow clays, Prosopis C and N mass and ANPP all increased linearly with stand age. We found no evidence of an asymptote in trajectories of C and N accumulation or ANPP on either soil type even following 68 years of stand development. Production and accumulation rates were lower on shallow clay sites relative to clay loam sites, suggesting strong edaphic control of C and N accumulation associated with woody plant encroachment. Response of herbaceous C mass to Prosopis stand development also differed between soil types. Herbaceous C declined with increasing aboveground Prosopis C on clay loams, but increased with increasing Prosopis C on shallow clays. Total ANPP (Prosopis+herbaceous) of sites with the highest Prosopis basal area were 1.2 × and 4.0 × greater than those with the lowest Prosopis basal area on clay loam and shallow clay soils, respectively. Prosopis ANPP more than offset declines in herbaceous ANPP on clay loams and added to increased herbaceous ANPP on shallow clays. Although aboveground C and N pools increased substantially with Prosopis stand development, we found no corresponding change in surface soil C and N pools (0–10 cm). Overall, our findings indicate that Prosopis stand development significantly increases ecosystem C and N storage/cycling, and the magnitude of these impacts varied with stand age, soil type and functional plant traits     

Genetic diversity of soybean-nodulating rhizobia in Nepal in relation to climate and soil properties

Backgroud and aims

This study was conducted to reveal the genetic diversity of soybean-nodulating rhizobia in Nepal in relation to climate and soil properties.

Method

A total of 102 bradyrhizobial strains were isolated from the root nodules of soybeans cultivated in 12 locations in Nepal varying in climate and soil properties, and their genetic diversity was examined based on 16S rDNA, ITS regions of 16S–23S rDNA, nodC and nifH. In vitro growth properties of some representative strains were examined to elucidate their characteristic distribution in Nepal.

Results

Four species of the genus Bradyrhizobium were isolated, and B. japonicum dominated at temperate locations, while in subtropical locations, B. elkanii, B. yuanmingense, and B. liaoningense dominated at acidic, moderately acidic, and slightly alkaline soils, respectively. The relative nodule occupancies could not be fully explained by their in vitro growth properties. Similar nodC and nifH genes among the strains suggested co-evolution of these genes also in Nepal, probably through horizontal gene transfer.

Conclusions

The influence of climate and soil pH on diversity at the sub-species level was revealed. It is concluded that the highly diverse climate and soils in Nepal might be conducive for the existence of diverse soybean rhizobial strains.     

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.