Overwintering management on upland pasture causes shifts in an abundance of denitrifying microbial communities, their activity and N2O-reducing ability

Pasture soils used for cattle overwintering may represent significant sources of N₂O emissions from soils. Therefore, the long-term effect of cattle overwintering on the abundance and activity of a denitrifying community was explored. The study was performed at a cattle overwintering area in South Bohemia (Czech Republic), where three sites differing in the degree of animal impact were selected: severely impacted (SI) and moderately impacted (MI), as well as a control site with no impact (NI). N₂O flux measurement and soil sampling were performed in spring and fall of 2005. The activity was measured in terms of potential denitrification activity. Bacterial nirK, nirS and nosZ genes were used as functional markers of the denitrifying communities; abundance was analyzed using a real-time PCR assay. Surprisingly, in situ N₂O emissions were the highest in spring at MI and significantly differed from those at SI and NI, while in autumn, rates of emissions generally decreased. In contrast potential denitrification rates were highest at SI, followed by MI, and the lowest at NI. An overall significant shift in N₂O/N₂ molar ratio was shown in cattle impacted sites. The highest abundance of all genes measured at both sampling times was found at site SI, whereas at site MI increased numbers were observed only in spring. Our results indicate a strong influence of cattle on the abundance as well as the activity of microbes involved in denitrification.     

Comparison of g23 gene sequence diversity between Novosphingobium and Sphingomonas phages and phage communities in the floodwater of a Japanese paddy field

Our previous study indicated that the diversity of the major capsid gene (g23) of T4-type bacteriophages (phages) of Novosphingobium and Sphingomonas strains isolated from the floodwater of a Japanese paddy field is comparable to those of the clones obtained from other Japanese paddy fields. For more strict comparison of the diversity, this study examined g23 sequences between Novosphingobium and Sphingomonas phages and phage communities in the identical floodwater of a Japanese paddy field. The clones were obtained by applying g23-specific primers to DNA extracted from the floodwaters. Many 23 clones in the floodwater were grouped into the same clusters of Paddy Groups I-VI with g23 genes of Novosphingobium/Sphingomonas phages with some clones belonging to an additional cluster. In addition, the remaining clones belonged to the clusters of marine clones and T4-type enterophages. These findings indicate that the g23 genes in the floodwater are more diversified than those of Novosphingobium/Sphingomonas phages including g23 genes closely related to the genes of enterophages and marine origins.     

白僵菌和乙酰甲胺磷对水稻抗氧化状态及土壤氮循环的影响

为了考察微生物农药白僵菌施用后在水稻系统中的生态安全性及其环境行为,本文通过盆栽试验探讨了施加不同浓度白僵菌孢子悬液[7.5×104(孢子)·mL-1,7.5×105(孢子)·mL-1,7.5×106(孢子)·mL-1,7.5×107(孢子)·mL-1]和化学农药乙酰甲胺磷对接种二化螟的水稻氧化还原状态及土壤氮循环相关的生物化学过程强度的影响。结果表明,白僵菌处理对土壤氨化作用和硝化作用具有一定刺激作用,最大提高幅度分别为12.4%和36.8%;对反硝化作用表现为低浓度促进、高浓度抑制。乙酰甲胺磷处理在第10 d时对土壤氨化作用、硝化作用和反硝化作用的抑制率分别达到18.6%、45.3%和27.5%。乙酰甲胺磷处理后水稻AsA︰DHA(还原态抗坏血酸︰氧化态抗坏血酸)比值和GSH︰GSSG(还原态谷胱甘肽︰氧化态谷胱甘肽)比值降低,并伴随抗氧化能力的降低。与此相反,白僵菌处理能够提高AsA︰DHA和GSH︰GSSG。H2O2含量变化的结果表明二化螟接种和乙酰甲胺磷处理会导致过氧化程度升高,而白僵菌的施加缓解了二化螟引起的过氧化状态。本试验使用荧光定量PCR对白僵菌的残留进行研究,结果表明稻谷中并未有白僵菌残留,白僵菌施用后对人类健康无害。上述结果表明,与化学农药乙酰甲胺磷相比,白僵菌是一种环境友好型的微生物农药,其中7.5×104(孢子)·mL-1的使用浓度效果较好。     

Quantification of Wautersia [Ralstonia] basilensis in the mycorrhizosphere of Pinus thunbergii Parl. and its effect on mycorrhizal formation

The bacterium Wautersia [Ralstonia] basilensis has been shown to enhance the mycorrhizal symbiosis between Suillus granulatus and Pinus thunbergii (Japanese black pine). However, no information is available about this bacterium under field conditions. The objectives of this study were to detect W. basilensis in bulk and mycorhizosphere soils in a Japanese pine plantation in the Tottori Sand Dunes, determine the density of W. basilensis in soil, and determine the optimal cell density of W. basilensis for mycorrhizal formation in pine seedlings. We designed and validated 16S rRNA gene-targeted specific primers for detection and quantification of W. basilensis. SYBR Green I real-time PCR assay was used. A standard curve relating cultured W. basilensis cell density (10³-10⁸ cells ml⁻¹) to amplification of DNA showed a strong linear relationship (R = 0.9968). The specificity of the reaction was confirmed by analyzing DNA melting curves and sequencing of the amplicon. The average cell density of W. basilensis was >4.8 × 10⁷ cells g⁻¹ of soil in the mycorrhizosphere and 7.0 × 10⁶ cells g⁻¹ in the bulk soil. We evaluated the W. basilensis cell density required for mycorrhizal formation using an in vitro microcosm with various inoculum densities ranging from 10² to 10⁷ cells g⁻¹ soil (10⁴-10⁹ cells ml⁻¹). Cell densities of W. basilensis of >10⁶ cells g⁻¹ of soil were required to stimulate mycorrhizal formation. In vivo and in vitro experiments showed that W. basilensis was sufficiently abundant to enhance mycorrhizal formation in the mycorrhizosphere of Japanese black pine sampled from the Tottori Sand Dunes.     

土壤改良剂对冷浸田土壤特性和水稻群体质量的影响

以南方典型冷浸田为研究对象, 在明沟排水的基础上, 通过田间定位试验, 以不施土壤改良剂为对照, 研究了施用不同土壤改良剂(自研的脱硫灰改良剂、生物活性炭, 市售的土壤改良剂石灰、硅钙肥、腐植酸)对冷浸田氧化还原电位、土壤呼吸强度、土壤微生物数量、水稻群体构建及产量构成因素的影响。结果表明, 施用改良剂能够改善土壤理化性状, 提升土壤速效养分和pH,但除脱硫灰处理外, 其他改良剂处理对土壤Eh未产生显著影响。施用不同土壤改良剂在水稻各生育期均能有效增强土壤微生物呼吸强度和放线菌数量, 并且放线菌数量达到差异性显著水平(P<0.05), 生物活性炭处理下土壤呼吸强度和放线菌数量分别较对照增加67.6%和127.6%。各土壤改良剂处理与CK相比较均有助于提高叶片SPAD、茎蘖数、水稻干物质积累量、成穗数、穗粒数、产量结实率和根系伤流速率。其中以脱硫灰和生物活性炭处理改良效果最佳, 抽穗后29 d时,根系伤流速率较CK分别提高45.4%和39.1%, 叶片SPAD分别增加27.4%和22.5%; 成熟期水稻成穗数较对照提高12.1%和10.7%,干物质积累量增加68.8%和50.5%,产量分别增加12.8%和10.3%。综上所述, 土壤改良剂可有效改善冷浸田土壤特性及水稻群体质量, 脱硫灰和生物活性炭处理的改良效果最明显, 增产幅度最大。     

Molecular markers and a sequence deletion in intron 2 of the putative partial homologue of LEAFY reveal geographical structure to genetic diversity in the acutely threatened legume genus Clianthus

Clianthus is an acutely threatened, bird-pollinated genus endemic to New Zealand, represented in the wild by only one population of C. puniceus and 11 populations of C. maximus, each with very few individuals (typically <10 per population). A limited number of named Clianthus cultivars of indeterminate origin are commonly grown as ornamentals. Genomic DNA from individual Clianthus plants was extracted for genetic diversity analysis using a range of molecular markers, including amplified fragment length polymorphism (AFLP). Data were analysed by the unweighted pair-group method with arithmetic averaging (UPGMA), the generation of Neighbor-Joining trees, and analyses of molecular variance (AMOVA). Genetic distance between wild populations of C. maximus was highly correlated with geographical distance between populations. Sequencing of intron 2 of a putative partial homologue of the floral meristem identity gene LEAFY (CmLFY) revealed a 7 bp deletion that was exhibited homozygously in the more northern populations of C. maximus, and in all individuals tested from the sole population of C. puniceus. This deletion was not exhibited in more southern populations of C. maximus. Further, one geographically intermediate population contained some plants that were heterozygous for the deletion. Parallel analyses of cultivated Clianthus genotypes, more than half of which were also homozygous for the 7 bp deletion, showed that these were not representative of the broad, but threatened, diversity remaining in the wild. It is argued that wild populations of C. maximus are unlikely to have arisen from the escape of plants from cultivation. Conservation effort should focus on the protection and study of the extant plants in these wild populations, rather than on the introduction of disturbance regimes to uncover potential seed banks.     

Impact of the biocontrol agent Trichoderma atroviride SC1 on soil microbial communities of a vineyard in northern Italy

The fungus Trichoderma atroviride SC1 is an experimental biocontrol agent (BCA) that is active against the fungus Armillaria mellea. Following the application of Trichoderma to the surface soil of a vineyard, we used a highly specific real-time PCR, previously validated for the analysis of soil microcosms, to monitor the populations of this fungus at different soil depths over several months. The quantification obtained using this molecular method was highly correlated with laboratory assays of colony-forming units. The native communities of bacteria and fungi in the soil were analyzed using automated ribosomal intergenic spacer analysis (ARISA), and transient changes were observed following the application of T. atroviride SC1 conidia. A principal component of variance analysis demonstrated that the introduction of T. atroviride SC1 had an effect on the soil microflora during the first two weeks following inoculation. However, at later dates, environmental conditions had a higher influence on the surveyed communities than the BCA application, as confirmed through the use of the Shannon index of biodiversity. Soil depth had a strong influence on the composition and biodiversity of fungal communities.     

Survey of major capsid genes (g23) of T4-type bacteriophages in rice fields in Northeast China

We surveyed the capsid genes (g23) of T4-type bacteriophages in DNA extracted from fifteen rice field soils in Northeast China using primers MZIA1bis and MZIA6. Denaturing gradient gel electrophoresis (DGGE) was performed to separate PCR-amplified g23 products. In total, 53 DGGE bands were identified as g23 clones, nine of which belonged to a novel, Northeast China-specific group. In addition, four and six clones formed two novel groups with previously ungrouped clones obtained from Japanese rice fields. The majority of the remaining clones fell into Paddy Groups I and V, none of the clones belonged to Paddy Groups II, III, IV, and VI, indicating that phylogenetic distribution of g23 genes in rice fields in Northeast China was different from that in Japanese rice fields.     

Potential gene exchange between Bacillus thuringiensis subsp. kurstaki and Bacillus spp. in soil in situ

The possible transfer of genes from Bacillus thuringiensis subsp. kurstaki (Btk) to indigenous Bacillus spp. was investigated in soil samples from stands of cork oak in Orotelli (Sardinia, Italy) collected 5 years after spraying of the stands with a commercial insecticidal preparation (FORAY 48B) of Btk. Two colonies with a morphology different from that of Btk were isolated and identified as Bacillus mycoides by morphological and physiological characteristics and by 16S rDNA analysis. Amplification by the polymerase chain reaction (PCR) of the DNA of the two isolated B. mycoides colonies with primers used for the identification of the Btk cry genes showed the presence of a fragment of 238 bp of the cry1Ab9 gene that had a similarity of 100% with the sequence of the cry1Ab9 gene present in GenBank, indicating that the isolates of B. mycoides acquired part of the sequence of this gene from Btk. No cells of Btk or B. mycoides carrying the 238-bp fragment of the cry1Ab9 gene were isolated from samples of unsprayed control soil. However, the isolates of B. mycoides were not able to express the partial Cry1Ab protein. Hybridization with probes for IS231 and the cry1Ab9 gene suggested that the inverted repeated sequence, IS231, was probably involved in the transfer of the 238-bp fragment from Btk to B. mycoides. These results indicate that transfer of genes between introduced Btk and indigenous Bacillus spp. can occur in soil under field conditions.     

High genetic diversity and heterogeneous parasite load in the earthworm Lumbricus terrestris on a German meadow

In parasite-host dynamics, parasites exert frequency-dependent selection on their hosts by favouring rare alleles that may confer resistance against infection. Therefore host populations that suffer strong parasite stress should maintain higher levels of genetic variability. We studied the Lumbricus terrestris-Monocystis sp. host-parasite system at a microgeographical scale. Using three polymorphic microsatellite loci on one large earthworm population sampled at 26 different sites (281 genotypes), we tested the relationship between parasite load and genetic variation in natural samples of the common earthworm L. terrestris. Our analysis yielded the following: (1) parasite load varied significantly across sites in this population; (2) there was no consistent evidence for heterozygote deficiency (observed heterozygosities ranged between 0.74 and 0.87), indicating a low level of inbreeding; (3) there was no significant genetic structuring among sample sites; (4) we could not identify a significant association between parasite load and population genetic diversity; (5) there was considerable population differentiation (15.17%) between our German samples and a Canadian L. terrestris reference population. Our study provides insight into the population genetics of one of the most economically important soil organisms on a microgeographic scale.     

The genetic diversity of Rhizobium leguminosarum bv. viciae in cultivated soils of the eastern Canadian prairie

Soil populations of Rhizobium leguminosarum bv. viciae (Rlv) that are infective and symbiotically effective on pea (Pisum sativum L.) have recently been shown to be quite widespread in agricultural soils of the eastern Canadian prairie. Here we report on studies carried out to assess the genetic diversity amongst these endemic Rlv strains and to attempt to determine if the endemic strains arose from previously used commercial rhizobial inoculants. Isolates of Rlv were collected from nodules of uninoculated pea plants from 20 sites across southern Manitoba and analyzed by plasmid profiling and PCR-RFLP of the 16S-23S rDNA internally transcribed spacer (ITS) region. Of 214 field isolates analyzed, 67 different plasmid profiles were identified, indicating a relatively high degree of variability among the isolates. Plasmid profiling of isolates from proximal nodules (near the base of the stem) and distal nodules (on lateral roots further from the root crown) from individual plants from one site suggested that the endemic strains were quite competitive relative to a commercial inoculant, occupying 78% of the proximal nodules and 96% of the distal nodules. PCR-RFLP of the 16S-23S rDNA ITS also suggested a relatively high degree of genetic variability among the field isolates. Analysis of the PCR-RFLP patterns of 15 selected isolates by UPGMA indicated two clusters of three field isolates each, with simple matching coefficients (SMCs) ≥0.95. However, to group all field isolates together, the SMC has to be reduced to 0.70. Regarding the origin of the endemic Rlv strains, there were few occurrences of the plasmid profiles of field isolates being identical to the profiles of inoculant Rlv strains commonly used in the region. Likewise, the plasmid profiles of isolates from nodules of wild Lathyrus plants located near some of the sites were all different from those of the field isolates. However, comparison of PCR-RFLP patterns suggested an influence of some inoculant strains on the chromosomal composition of some of the field isolates with SMCs of ≥0.92. Overall, plasmid profiles and PCR-RFLP patterns of the isolates from endemic Rlv populations from across southern Manitoba indicate a relatively high degree of genetic diversity among both plasmid and chromosomal components of endemic strains, but also suggest some influence of chromosomal information from previously used inoculant strains on the endemic soil strains.     

Direct seeding mulch-based cropping increases both the activity and the abundance of denitrifier communities in a tropical soil

This study evaluated the impact of direct seeding mulch-based cropping (DMC), as an alternative to conventional tilling (CT), on a functional community involved in N cycling and emission of greenhouse gas nitrous oxide (N₂O). The study was carried out for annual soybean/rice crop rotation in the Highlands of Madagascar. The differences between the two soil management strategies (direct seeding with mulched crop residues versus tillage without incorporation of crop residues) were studied along a fertilization gradient (no fertilizer, organic fertilizer, organic plus mineral fertilizers). The activity and size of the denitrifier community were determined by denitrification enzyme activity assays and by real-time PCR quantification of the denitrification genes. Denitrification activity and total C and N content in the soil were significantly increased by DMC both years, whereas the fertilization regime and sampling year (crop and mulch types, climatic conditions) had very little effect. Similar results were also observed for denitrification gene densities. Denitrification enzyme activity was more closely correlated with C content than with N content in the soil and denitrification gene densities. Principal component analysis confirmed that soil management had the strongest impact on the soil denitrifier community and total C and N content for both years and further indicated that changes in microbial and chemical soil parameters induced by the use of fertilizer were favored in DMC plots. Overall, the alternative DMC system had a significant positive effect on denitrifier densities and potential activities, which was not altered by crop rotation and the level of fertilization. These data also suggest that in these clayey soils, the DMC system simultaneously increased the size of the soil N pool and accelerated the N cycle, by stimulating the denitrifier community. Complementary investigations should further determine in greater detail the influence of DMC on in situ N-fluxes caused by denitrification.     

The abundance of nitrogen cycle genes amoA and nifH depends on land-uses and soil types in South-Eastern Australia

Nitrogen is a critical nutrient in plant-based primary production systems, therefore measurements of N cycling by microorganisms may add value to agricultural soil monitoring programs. Bacterial-mediated nitrogen cycling was investigated in soils from two broad land-uses (managed and remnant vegetation) across different Soil Orders from three geomorphic zones in Victoria, Australia, by examining the abundance of the genes amoA and nifH using quantitative polymerase chain reaction (qPCR). The aim of the study was to identify parameters influencing bacterial populations possessing the genes nifH and amoA, and examine their distribution at a regional scale across different management treatments. The gene amoA was most abundant in the neutral to slightly alkaline surface soils from Calcarosols in North-West Victoria. There was a highly significant (P < 0.001) interaction between land-use and geomorphic zones in terms of the abundance of amoA. Detection of the gene nifH was site specific with low copy number (less than 100 copies per nanogram of DNA) observed for some strongly acidic surface soil sites in North-East Victoria (Dermosols) and South-West Victoria (Sodosols/Chromosols), while nifH was more abundant in selected Calcarosols of North-West Victoria. The gene amoA was detected across more sites than nifH and was strongly influenced by land-use, with almost consistently greater abundance in managed compared to remnant sites, particularly for North-West and South-West Victoria. The abundance of nifH was not related to land-use, with similar copy numbers observed for both managed and remnant sites at some locations. For the gene nifH, there was no significant interaction between land-use and geomorphic zones, between managed and remnant sites or between the three geomorphic zones. Regression tree analysis revealed a number of likely soil chemical and microbial variables which may act as drivers of gene abundance of amoA and nifH. Variables identified as drivers for amoA included pH, Olsen P, microbial biomass carbon, nitrate and total nitrogen while for nifH the variables were microbial biomass carbon, electrical conductivity, microbial biomass nitrogen, total nitrogen and total potassium. Measures of N cycling genes could be used as an additional indicator of soil health to assess potential ecosystem functions. The spatial scale of the current study demonstrates that a landscape approach may assist soil health monitoring programs by evaluating N cycle gene abundance in the context of the different microbial and chemical conditions related to Soil Order and land-use management.     

近地层臭氧浓度升高对稻田土壤微生物群落功能多样性的影响

利用中国臭氧FACE(Free-Air O3 Concentration Enrichment,开放式空气臭氧浓度增高)试验平台,研究了近地层臭氧浓度升高条件下,2009年和2010年的稻田土壤微生物功能多样性的响应规律。结果发现,1.5倍当前臭氧浓度下稻田土壤水溶性有机碳含量从100.0～110.9μg g-1降低至81.3～94.5μg g-1。臭氧浓度升高下BIOLOG平均吸光值(AWCD)有低于对照的趋势。多样性指数结果表明,臭氧浓度升高50%对稻田土壤微生物丰富度、优势度及均一度无显著影响。2010年碳源底物利用的主成分分析显示,1.5倍当前臭氧浓度下稻田土壤微生物对碳源底物的利用方式发生变化。本研究揭示近地层臭氧浓度升高可能对稻田土壤微生物产生积累效应,并通过微生物碳源底物有效性的改变最终影响土壤微生物的功能多样性。     

Absorption and emission of CO2 by ponded water of a paddy field

In the daytime, the CO₂ concentration in the air close to the water surface of a ponded paddy field was lowest and it increased with the distance above the water surface, while an inverse relation was observed in the nighttime. On the other hand, the pH of the ponded water changed significantly throughout a day and was expected to affect atmospheric CO₂ in the vicinity of the water surface, because the solubility of CO₂ in water depends on the pH. In this study, we investigated the relationship between the changes in the pH of the ponded water and the response of the CO₂ concentration in the air above the water. The pH of the ponded water of the paddy field increased in the daytime and decreased in the nighttime, so that the water was alkaline in the daytime and weakly acidic in the nighttime. We found that the daily changes in the atmospheric CO₂ concentration gradient almost corresponded to the daily changes in pH. The increase of the pH of the ponded water in the daytime was due to the absorption of dissolved CO₂ by photosynthetic bacteria and micro-algae within the ponded water. Furthermore, we compared the pH with RpH, defined as the pH at which the CO₂ concentration of the water is in equilibrium with that of the air, to determine whether CO₂ was absorbed by or emitted from the ponded water. In the daytime, the pH value of the ponded water was higher than that of the RpH, and the water could therefore absorb CO₂ , whereas during the nighttime, since the pH value of the ponded water was lower than that of the RpH, the water was expected to emit CO₂. These results show that the ponded water absorbed CO₂ from the air above the water surface in the daytime and emitted CO₂ in the nighttime.     

Distribution and genetic diversity of rhizobia nodulating natural populations of Medicago truncatula in tunisian soils

A collection of 299 isolates of rhizobia nodulating Medicago truncatula was isolated from 10 Tunisian soils and was characterized by restriction fragment length polymorphism analysis of polymerase chain reaction (PCR/RFLP) of 16S rRNA gene. Results showed that 227 and 72 isolates were assigned, respectively, to Sinorhizobium meliloti and Sinorhizobium medicae. In 9 out of 10 soils S. meliloti was detected, whereas S. medicae was recovered from only 5 out of 10 soils. The cross-nodulation of three populations of M. truncatula grown on Bulla Regia soil, which contained naturally the two Sinorhizobium species, showed that M. truncatula population collected from Amra site was selective to S. meliloti at least in soil conditions. Forty-eight isolates of each Sinorhizobium species trapped by M. truncatula populations collected from Bulla Regia, Soliman and Rhayet sites on Bulla Regia soil were characterized by repetitive extragenic palindromic-PCR (REP-PCR) and showed a clear distinction between the two Sinorhizobium species and a higher diversity for S. meliloti.     

Size, symbiotic effectiveness and genetic diversity of field pea rhizobia (Rhizobium leguminosarum bv. viciae) populations in South Australian soils

Field pea (Pisum sativum L.) is widely grown in South Australia (SA), often without inoculation with commercial rhizobia. To establish if symbiotic factors are limiting the growth of field pea we examined the size, symbiotic effectiveness and diversity of populations of field pea rhizobia (Rhizobium leguminosarum bv. viciae) that have become naturalised in South Australian soils and nodulate many pea crops. Most probable number plant infection tests on 33 soils showed that R. l. bv. viciae populations ranged from undetectable (six soils) to 32×10³ rhizobia g⁻¹ of dry soil. Twenty-four of the 33 soils contained more than 100 rhizobia g⁻¹ soil. Three of the six soils in which no R. l. bv. viciae were detected had not grown a host legume (field pea, faba bean, vetch or lentil). For soils that had grown a host legume, there was no correlation between the size of R. l. bv. viciae populations and either the time since a host legume had been grown or any measured soil factor (pH, inorganic N and organic C). In glasshouse experiments, inoculation of the field pea cultivar Parafield with the commercial Rhizobium strain SU303 resulted in a highly effective symbiosis. The SU303 treatment produced as much shoot dry weight as the mineral N treatment and more than 2.9 times the shoot dry weight of the uninoculated treatment. Twenty-two of the 33 naturalised populations of rhizobia (applied to pea plants as soil suspensions) produced prompt and abundant nodulation. These symbioses were generally effective at N₂ fixation, with shoot dry weight ranging from 98% (soil 21) down to 61% (soil 30) of the SU303 treatment, the least effective population of rhizobia still producing nearly double the growth of the uninoculated treatment. Low shoot dry weights resulting from most of the remaining soil treatments were associated with delayed or erratic nodulation caused by low numbers of rhizobia. Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) fingerprinting of 70 rhizobial isolates recovered from five of the 33 soils (14 isolates from each soil) showed that naturalised populations were composed of multiple (5-9) strain types. There was little evidence of strain dominance, with a single strain type occupying more than 30% of trap host nodules in only two of the five populations. Cluster analysis of RAPD PCR banding patterns showed that strain types in naturalised populations were not closely related to the current commercial inoculant strain for field pea (SU303, ≥75% dissimilarity), six previous field pea inoculant strains (≥55% dissimilarity) or a former commercial inoculant strain for faba bean (WSM1274, ≥66% dissimilarity). Two of the most closely related strain types (≤15% dissimilarity) were found at widely separate locations in SA and may have potential as commercial inoculant strains. Given the size and diversity of the naturalised pea rhizobia populations in SA soils and their relative effectiveness, it is unlikely that inoculation with a commercial strain of rhizobia will improve N₂ fixation in field pea crops, unless the number of rhizobia in the soil is very low or absent (e.g. where a legume host has not been previously grown and for three soils from western Eyre Peninsula). The general effectiveness of the pea rhizobia populations also indicates that reduced N₂ fixation is unlikely to be the major cause of the declining field pea yields observed in recent times.     

稻虾共作对稻田土壤nirK反硝化微生物群落结构和多样性的影响

稻虾共作是水稻种植与克氏螯虾共作形成的互利共生的稻田种养复合生态模式。目前对稻虾共作模式稻田反硝化微生物多样性和群落结构的影响尚不清楚。本研究以江汉平原常规中稻模式(MR)为对照,设置连续3年(2014—2016年)稻虾共作模式(CR)为处理,通过特异引物提取中稻抽穗期稻田土壤nirK基因,采用Illumina Miseq高通量测序技术,探讨稻虾共作模式对稻田土壤nirK反硝化微生物多样性和群落结构的影响。结果表明:稻虾共作模式显著提升水稻抽穗期稻田土壤中硝态氮、全氮及全碳的含量,对土壤碳氮比、碱解氮和铵态氮含量没有显著影响。稻虾共作模式显著增加稻田土壤nirK基因微生物的丰富度指数,但对nirK基因微生物的多样性指数影响不显著。稻虾共作模式改变了nirK基因微生物在目、科、属、种水平的群落组成,较常规中稻模式,稻虾共作模式在各分类水平组成类群均减少;稻虾共作模式较常规中稻模式改变了目的种类,对共有目相对丰度没有显著性改变。RDA分析表明稻虾共作模式对土壤nirK基因菌群的群落结构有一定的改变,但稻虾共作模式与常规中稻模式在群落结构上仍保留着一定的相似性。硝态氮含量是影响nirK反硝化细菌群落结构的主效因子。可见,稻虾共作模式对微生物多样性指数没有显著影响,但显著增加了微生物丰富度指数,改变了稻田土壤nirK反硝化微生物在目、科、属、种的群落结构。     

Molecular phylogenetic analysis of a novel strain from Neelipleona enriches Wolbachia diversity in soil biota

Eubacterial cellular endoparasites belonging to the genus Wolbachia (Rickettsiales) are extremely widespread symbionts of Arthropoda and Nematoda. Their ability to manipulate the reproductive behavior of the host is of particular importance to the fauna of the deep soil horizon, an environment in which parthenogenesis-inducing symbionts can play a crucial role in shaping population dynamics and speciation processes. In this study, three novel cases of infection in parthenogenetic Collembola (Parisotoma notabilis, Neelus murinus and Megalothorax minimus) are described. Sequences for molecular markers 16S rDNA and ftsZ were obtained for each species; their phylogenetic affinities with known Wolbachia supergroups were established using Bayesian inference. The analysis confirmed the presence of a Wolbachia strain belonging to the supergroup E, already reported from Folsomia candida and the Tullbergiidae, in the isotomid P. notabilis, while the Neelipleona M. minimus and N. murinus host a well differentiated strain which is phylogenetically distinct from supergroup E. Multiple events of Wolbachia infection in springtails as well as a richer diversity of the symbiont strains in soil arthropods were hereby confirmed.