Experimental warming alters the community composition,diversity, and N2 fixation activity of peat moss (Sphagnum fallax) microbiomes

Sphagnum‐dominated peatlands comprise a globally important pool of soil carbon (C) and are vulnerable to climate change. While peat mosses of the genus Sphagnum are known to harbor diverse microbial communities that mediate C and nitrogen (N) cycling in peatlands, the effects of climate change on Sphagnum microbiome composition and functioning are largely unknown. We investigated the impacts of experimental whole‐ecosystem warming on the Sphagnum moss microbiome, focusing on N₂ fixing microorganisms (diazotrophs). To characterize the microbiome response to warming, we performed next‐generation sequencing of small subunit (SSU) rRNA and nitrogenase (nifH) gene amplicons and quantified rates of N₂ fixation activity in Sphagnum fallax individuals sampled from experimental enclosures over 2 years in a northern Minnesota, USA bog. The taxonomic diversity of overall microbial communities and diazotroph communities, as well as N₂ fixation rates, decreased with warming (p < 0.05). Following warming, diazotrophs shifted from a mixed community of Nostocales (Cyanobacteria) and Rhizobiales (Alphaproteobacteria) to predominance of Nostocales. Microbiome community composition differed between years, with some diazotroph populations persisting while others declined in relative abundance in warmed plots in the second year. Our results demonstrate that warming substantially alters the community composition, diversity, and N₂ fixation activity of peat moss microbiomes, which may ultimately impact host fitness, ecosystem productivity, and C storage potential in peatlands.     

模拟增温对西藏高原高寒草甸土壤供氮潜力的影响

过去几十年青藏高原呈现显著的增温趋势,冬季增温幅度显著高于生长季的季节非对称特征。气候变暖会对生态系统氮素循环产生重要影响,但关于全年增温与冬季增温对高寒生态系统氮循环的不同影响仍缺乏研究。在青藏高原高寒草甸区开展模拟增温试验,研究季节非对称增温对高寒草甸生态系统氮循环的影响。该试验布设于2010年7月,设置3种处理(不增温、冬季增温与全年增温)。研究结果发现,开顶箱增温装置造成了小环境的暖干化:显著提高了地表空气温度和表层土壤温度,降低了表层土壤含水量。冬季增温会加剧土壤中氮素的流失,所以在经历了冬季增温后土壤氮含量显著降低;在生长季节,土壤氮素周转速率受土壤水分的调控,在降雨较少的季节,增温引起的土壤含水量降低会抑制土壤氮周转速率。对于土壤微生物量而言,高寒草甸土壤微生物量碳表现出明显的季节动态,在生长季旺盛期较低,在生长季末期和初冬季节反而较高,这说明为了降低对土壤养分的竞争,高寒草甸植物氮吸收与土壤微生物氮固持在时间上存在分离。研究结果表明,冬季增温导致的土壤养分含量变化会影响随后生长季植物群落的生产力、结构组成与碳氮循环等过程,对生态系统过程产生深远的影响。     

模拟增温对青藏高原东部高寒灌丛土壤氮转化的影响

本文研究了青藏高原东部窄叶鲜卑花高寒灌丛生长季前期、生长季后期和非生长季3个生育期的土壤氮转化速率对模拟增温的响应,分析全球气候变暖对高寒灌丛土壤氮循环过程的影响。结果表明: 模拟增温使高寒灌丛土壤温度显著升高1.2 ℃,土壤水分显著降低2.5%。高寒灌丛生长季土壤净氮矿化(氨化和硝化)速率显著高于非生长季,但土壤净氮固持速率显著低于非生长季。土壤氮矿化在生长季前期以硝化作用为主,在生长季后期和非生长季以氨化作用为主。模拟增温对高寒灌丛土壤氮转化过程的影响在不同时期存在显著差异。模拟增温显著增加了生长季前期土壤净氨化、净硝化、净氮矿化、净氮固持速率和非生长季土壤净硝化、净氮矿化速率,并显著降低了生长季后期土壤净硝化、净氮矿化、净氮固持速率和非生长季土壤净氨化速率。但模拟增温对高寒灌丛非生长季净氮固持速率和生长季后期净硝化速率的影响不显著。未来气候变暖将显著改变青藏高原东部高寒灌丛土壤氮转化,进而加速高寒灌丛土壤氮循环过程。     

Activity and distribution of diazotrophic communities across the Cape Verde Frontal Zone in the Northeast Atlantic Ocean

Biogeochemistry - We investigated dinitrogen (N2) fixation activity and diazotroph community composition across the Cape Verde Frontal Zone (CVFZ), from photic epipelagic waters (0-200&nbsp;m)...     

黄河上游灌区稻田N2O排放特征

黄河上游灌区稻田高产区过量施肥现象十分突出,氮肥过量施用引起土壤氮素盈余,导致N₂O排放量增大,由此引起的温室效应引起广泛关注。采用静态箱-气相色谱法研究黄河上游灌区稻田不同施肥处理下N₂O排放特征。试验设置5个施肥处理,包括常规氮肥300 kg/hm²下单施尿素和有机肥配施2个处理,分别用N300和N300-OM代表;优化氮肥240 kg/hm²下单施尿素和有机肥配施2个处理,分别用N240和N240-OM代表;对照不施氮肥用N0代表。试验结果得出,灌区水稻生长季稻田土壤N₂O排放主要集中在水稻分蘖前及水稻生长的中后期,稻田氮肥施用、灌水及土壤温度的变化对N₂O排放通量影响较大,不同处理水稻各生育阶段N₂O累积排放量与稻田土壤耕层NO^-₃-N含量动态变化显著相关。稻田N₂O排放不是黄河上游灌区稻田氮素损失的主要途径,但灌区稻田N₂O排放的增温潜势较大;稻田氮肥过量施用会显著增加N₂O排放量,在相同氮素水平下,有机肥配施会显著增加稻田土壤N₂O的排放量(P＜0.01)。优化施氮能有效减少灌区稻田水稻生长季N₂O排放量。稻田不同处理的水稻整个生长季土壤N₂O排放总量为2.69-3.87 kg/hm²,肥料氮通过N₂O排放损失的百分率仅为0.43%-0.64%。在灌区习惯灌水和高氮肥300 kg/hm²时,N300-OM处理的稻田N₂O排放量达3.87 kg/hm²,在100 a时间尺度上的全球增温潜势(GWPs)为20.76×10⁷ kg CO₂/hm²;优化施氮240 kg/hm²水平下,N240和N240-OM处理的N₂O累计排放量较N300-OM处理,分别降低了1.18 kg/hm²和0.57 kg/hm²,在100 a尺度上每年由稻田N₂O排放引起的GWPs分别降低了6.33×10⁷ kg CO₂/hm²和3.06×10⁷ kg CO₂/hm²。     

Seasonal variation in respiration of 1-year-old shoots of scots pine exposed to elevated carbon dioxide and temperature for 4 years

Sixteen 20-year-old Scots pine (Pinus sylvestris L.) trees growing in the field were enclosed for 4 years in environment-controlled chambers that maintained: (1) ambient conditions (CON); (2) elevated atmospheric CO2 concentration (ambient + 350 micro mol mol-1; EC); (3) elevated temperature (ambient +2-6 degrees C; ET); or (4) elevated CO2 and elevated temperature (ECT). The dark respiration rates of 1-year-old shoots, from which needles had been partly removed, were measured over the growing season in the fourth year. In all treatments, the temperature coefficient of respiration, Q10, changed with season, being smaller during the growing season than at other times. Respiration rate varied diurnally and seasonally with temperature, being highest around mid-summer and declining gradually thereafter. When measurements were made at the temperature of the chamber, respiration rates were reduced by the EC treatment relative to CON, but were increased by ET and ECT treatments. However, respiration rates at a reference temperature of 15 degrees C were reduced by ET and ECT treatments, reflecting a decreased capacity for respiration at warmer temperatures (negative acclimation). The interaction between season and treatment was not significant. Growth respiration did not differ between treatments, but maintenance respiration did, and the differences in mean daily respiration rate between the treatments were attributable to the maintenance component. We conclude that maintenance respiration should be considered when modelling respiratory responses to elevated CO2 and elevated temperature, and that increased atmospheric temperature is more important than increasing CO2 when assessing the carbon budget of pine forests under conditions of climate change.     

No stimulation of nitrogen fixation by non‐filamentous diazotrophs under elevated CO2 in the South Pacific

Nitrogen fixation by diazotrophic cyanobacteria is a critical source of new nitrogen to the oligotrophic surface ocean. Research to date indicates that some diazotroph groups may increase nitrogen fixation under elevated pCO₂. To test this in natural plankton communities, four manipulation experiments were carried out during two voyages in the South Pacific (30–35^oS). High CO₂ treatments, produced using 750 ppmv CO₂ to adjust pH to 0.2 below ambient, and ‘Greenhouse’ treatments (0.2 below ambient pH and ambient temperature +3 °C), were compared with Controls in trace metal clean deckboard incubations in triplicate. No significant change was observed in nitrogen fixation in either the High CO₂ or Greenhouse treatments over 5 day incubations. qPCR measurements and optical microscopy determined that the diazotroph community was dominated by Group A unicellular cyanobacteria (UCYN‐A), which may account for the difference in response of nitrogen fixation under elevated CO₂ to that reported previously for Trichodesmium. This may reflect physiological differences, in that the greater cell surface area:volume of UCYN‐A and its lack of metabolic pathways involved in carbon fixation may confer no benefit under elevated CO₂. However, multiple environmental controls may also be a factor, with the low dissolved iron concentrations in oligotrophic surface waters limiting the response to elevated CO₂. If nitrogen fixation by UCYN‐A is not stimulated by elevated pCO₂, then future increases in CO₂ and warming may alter the regional distribution and dominance of different diazotroph groups, with implications for dissolved iron availability and new nitrogen supply in oligotrophic regions.     

Seasonal variation in nitrogen fixation and effects of climate change in a subarctic heath

Background and aims

Nitrogen fixation associated with cryptogams is potentially very important in arctic and subarctic terrestrial ecosystems, as it is a source of new nitrogen (N) into these highly N limited systems. Moss-, lichen- and legume-associated N₂ fixation was studied with high frequency (every second week) during spring, summer, autumn and early winter to uncover the seasonal variation in input of atmospheric N₂ to a subarctic heath with an altered climate.

Methods

We estimated N₂ fixation from ethylene production by acetylene reduction assay in situ in a field experiment with the treatments: long- vs. short-term summer warming using plastic tents and litter addition (simulating expansion of the birch forest).

Results

N₂ fixation activity was measured from late April to mid November and 33 % of all N₂ was fixed outside the vascular plant growing season (Jun–Aug). This substantial amount underlines the importance of N₂ fixation in the cold period. Warming increased N₂ fixation two- to fivefold during late spring. However, long-term summer warming tended to decrease N₂ fixation outside the treatment (tents present) period. Litter alone did not alter N₂ fixation but in combination with warming N₂ fixation increased, probably because N₂ fixation became phosphorus limited under higher temperatures, which was alleviated by the P supply from the litter.

Conclusion

In subarctic heath, the current N₂ fixation period extends far beyond the vascular plant growing season. Climate warming and indirect effects such as vegetation changes affect the process of N₂ fixation in different directions and thereby complicate predictions of future N cycling.     

水肥一体化条件下设施菜地的N2O排放

在保证作物产量的前提下,研究减少农田土壤N_2O排放的水肥统筹管理措施对全球温室气体减排具有重要意义。以京郊典型设施菜地为例,设置了农民习惯(FP)、水肥一体化(FPD)、优化水肥一体化(OPTD)和对照(CK)4个处理,采用静态箱-气相色谱法,对果菜-叶菜(黄瓜-芹菜)轮作周期内土壤N_2O排放进行了观测,并分析了氮肥施用量、灌溉方式、土壤温度和湿度等因素对土壤N_2O排放的影响。结果表明:在黄瓜-芹菜种植模式中,各施氮处理除基肥施用后N_2O排放峰持续10—15d外,一般施肥、施肥+灌溉事件后土壤N_2O排放峰均呈现3—5d短而急促的情形。黄瓜生长季N_2O排放通量与土壤湿度(WFPS)之间呈现显著相关的关系;芹菜生长季N_2O排放通量与土壤温度之间呈现显著相关的关系。观测期内FP处理N_2O排放量为(31.00±2.15)kg N/hm~2,FPD处理与之相比N_2O排放量减少了4.2%,而OPTD处理在减少40%化肥氮量的情况下,N_2O累积排放量比FP处理减少了42.7%,且达到显著水平。说明在水肥一体化条件下,合理改变施肥体系是减少N_2O排放的前提,在此基础上进行水肥优化是设施菜地保持产量、减少N_2O排放的重要技术措施。     

Environmental forcing of nitrogen fixation in the eastern tropical and sub-tropical North Atlantic Ocean

During the winter of 2006 we measured nifH gene abundances, dinitrogen (N(2)) fixation rates and carbon fixation rates in the eastern tropical and sub-tropical North Atlantic Ocean. The dominant diazotrophic phylotypes were filamentous cyanobacteria, which may include Trichodesmium and Katagnymene, with up to 10(6) L(-1)nifH gene copies, unicellular group A cyanobacteria with up to 10(5) L(-1)nifH gene copies and gamma A proteobacteria with up to 10(4) L(-1)nifH gene copies. N(2) fixation rates were low and ranged between 0.032-1.28 nmol N L(-1) d(-1) with a mean of 0.30 ± 0.29 nmol N L(-1) d(-1) (1σ, n = 65). CO(2)-fixation rates, representing primary production, appeared to be nitrogen limited as suggested by low dissolved inorganic nitrogen to phosphate ratios (DIN:DIP) of about 2 ± 3.2 in surface waters. Nevertheless, N(2) fixation rates contributed only 0.55 ± 0.87% (range 0.03-5.24%) of the N required for primary production. Boosted regression trees analysis (BRT) showed that the distribution of the gamma A proteobacteria and filamentous cyanobacteria nifH genes was mainly predicted by the distribution of Prochlorococcus, Synechococcus, picoeukaryotes and heterotrophic bacteria. In addition, BRT indicated that multiple a-biotic environmental variables including nutrients DIN, dissolved organic nitrogen (DON) and DIP, trace metals like dissolved aluminum (DAl), as a proxy of dust inputs, dissolved iron (DFe) and Fe-binding ligands as well as oxygen and temperature influenced N(2) fixation rates and the distribution of the dominant diazotrophic phylotypes. Our results suggest that lower predicted oxygen concentrations and higher temperatures due to climate warming may increase N(2) fixation rates. However, the balance between a decreased supply of DIP and DFe from deep waters as a result of more pronounced stratification and an enhanced supply of these nutrients with a predicted increase in deposition of Saharan dust may ultimately determine the consequences of climate warming for N(2) fixation in the North Atlantic.     

Supply of O2 regulates O2 demand during utilization of reserves of poly-beta-hydroxybutyrate in N2-fixing soybean bacteroids.

A liquid reaction medium containing dissolved air and oxyleghaemoglobin, but no energy-yielding substrate, was supplied to bacteroids confined in a stirred flow reaction chamber. The relative oxygenation of the leghaemoglobin in the chamber was determined automatically by spectrophotometry of the effluent solution, and the concentrations of free, dissolved O2 ([O2]) and rates of O2 consumption were calculated. Dissolved CO2 and NH3 from N2 fixation were determined in fractions of the effluent solution. Bacteroids utilized endogenous reserves of poly-beta-hydroxybutyrate (PHB), which were depleted by 9.2% during a typical 5 h-long experiment. Stepwise increases in flow rate (increasing supply of O2) initially produced a drop in O2 demand and resulted in a rise in [O2] and a decline in N2 fixation. Subsequently, O2 demand rose (presumably because of increased mobilization of substrate from PHB) and [O2] declined to a low level. N2 fixation was fully restored, or even enhanced, within 15-20 min of establishment of a new, steady [O2]. This pattern of regulation by O2 supply was completely eliminated by adding low concentrations (20-50 microM) of oxidizable substrate (succinate, malate, ethanol) to the reaction medium. During endogenous activity, rates of CO2 evolution were proportional to, but less than, rates of O2 consumption up to 5.4 nmol O2 min-1 mg-1, above which CO2 evolution exceeded O2 consumption. These and other features of endogenous activity are discussed in relation to sustaining N2 fixation by nodules in vivo.     

Regulation of Nodule Efficiency by the Undisturbed Soybean Plant1

Acetylene reduction activity (ARA) has been widely used to estimatecomparative rates of nitrogen fixation in soybean [Glycine max(L.) Merr.]. To use the non-destructive closed-container ARAassay to study plant-mediated regulation of N₂ fixation, itwas first necessary to determine the validity of the ARA procedure.Thus, well-nodulated, hydroponically-grown soybean plants wereindividually assayed for ARA twice weekly for 5 weeks. Plantswere subsequently analysed for total nitrogen by the Kjeldahlprocedure and the values obtained were compared with the N-accumulationvalues estimated by ARA. For the 168 plants examined, ARA measurementsunderestimated N₂ fixation by approximately 50%. However, N₂fixation was underestimated by approximately 80% in plants witha very low ARA whereas N₂ fixation was underestimated by only20% in plants with a high ARA. Because of the accuracy and reliabilityof the Kjeldahl measurements, these results show that nodulatedsoybean plants can exhibit different efficiencies in acetylenereduction. Thus, vigorous, well-nodulated, hydroponically-grownsoybean plants readily control nodule efficiency and the N fixationprocess.     

Net nitrogen mineralization and leaching in response to warming and nitrogen deposition in a temperate old field: the importance of winter temperature

While climate warming can increase plant N availability over the growing season by increasing rates of N mineralization, increased N mineralization over winter at a time when plant roots are largely inactive, coupled with an increased frequency of soil freeze–thaw cycles, may increase soil N leaching losses. We examined changes in soil net N mineralization and N leaching in response to warming and N addition (6 g m^?2 year^?1) in a factorial experiment conducted in a temperate old field. We used two warming treatments, year-round and winter-only warming, to isolate the effects of winter warming on soil N dynamics from the year-round warming effects. We estimated net N mineralization using in situ soil cores with resin bags placed at the bottom to catch throughput, and we measured N leaching using lysimeters located below the plant rooting zone at a depth of 50 cm. There were minor effects of warming on changes in soil extractable N and resin N in the soil cores over winter. Nevertheless, the overall effects of both warming and N addition on net N mineralization (the sum of changes in soil extractable N and resin N) were not significant over this period. Likewise, there were no significant treatment effects on the concentration of N in leachate collected below the plant rooting zone. However, in response to winter warming, net N mineralization over summer was approximately double that of both the ambient and year-round warming treatments. This result demonstrates a potentially large and unexpected effect of winter warming on soil N availability in this old field system.     

枯落物处理对格木林土壤碳氮转化和微生物群落结构的短期影响

以我国南亚热带格木人工纯林为研究对象,采用气压过程分离(BaPS)技术和磷脂脂肪酸(PLFAs)法研究了不同枯落物处理(对照、枯落物去除、枯落物加倍)下土壤碳氮转化速率和微生物群落结构的季节变化.结果表明:不同枯落物处理土壤呼吸和总硝化速率均呈现明显的季节动态,雨季显著高于旱季.枯落物处理初期,土壤呼吸和总硝化速率均随枯落物输入量的增加呈下降趋势,但随着枯落物处理时间的延长,二者随枯落物输入量的增加而增加.旱季不同枯落物处理土壤微生物PLFAs总量和各菌群PLFAs量均显著高于雨季,而雨季真菌PLFAs/细菌PLFAs明显高于旱季.在旱季,枯落物去除处理土壤微生物PLFAs总量、细菌PLFAs量、真菌PLFAs量和丛枝菌根真菌PLFAs量分别显著提高30.9%、28.8%、44.4%和31.6%.在雨季,枯落物去除处理细菌PLFAs量和丛枝菌根真菌PLFAs量分别显著降低10.6%和33.3%.土壤微生物群落结构受枯落物输入量处理和季节的双重影响,土壤微生物群落结构主要受土壤温度和铵态氮的影响.枯落物输入量处理在短期内显著影响了格木林土壤碳氮转化速率和微生物群落结构,这种影响因季节的不同而存在差异.     

Effects of lengthened growing season and soil warming on the phenology and physiology of Polygonum bistorta

The phenological and physiological responses of arctic tundra plant species are key to predicting their survival in a warmer climate. One of the consequences of a warmer climate in the Arctic will be a longer growing season. We examined the effects of lengthened growing season and soil warming on the widely distributed forb, Polygonum bistorta L. Three treatments were established near Toolik Lake, Alaska in 1995 and 1996: extended season, extended season with soil warming, and an unmanipulated control. The season was extended by removing the snow load in the spring and keeping the treatments free of snow in the autumn. The spring snow removal extended the snow‐free period over that of controls by 8 d in 1995 and 24 d in 1996. As a result, the number of accumulated soil thaw days and consequently the depth of soil thaw increased on the treatment plots. Polygonum bistorta responded to the treatments by becoming active earlier and senescing earlier, resulting in a growth period of similar duration to that of the controls. Leaf size and leaf number were unaffected by the treatments, as were leaf photosynthetic assimilation rates and nutrient concentrations. The results indicate that internal constraints limit the response of this species to lengthened growing season, suggesting that it is a determinant or periodic species. With climate warming, this periodic growth will put P. bistorta at a competitive disadvantage relative to plants that can respond to lengthened growing season.     

Meta-analysis of diazotrophic signatures across terrestrial ecosystems at the continental scale

Biological nitrogen fixation performed by diazotrophs forms a cornerstone of Earth's terrestrial ecosystem productivity. However, the composition, diversity and distribution of soil diazotrophs are poorly understood across different soil ecosystems. Furthermore, the biological potential of the key diazotroph species in relation to key environmental parameters is unknown. To address this, we used meta-analysis approach to merge together 39 independent diazotroph amplicon sequencing (nifH gene) datasets consisting of 1988 independent soil samples. We then employed multiple statistical analyses and machine-learning approaches to compare diazotroph community differences and indicator species between terrestrial ecosystems on a global scale. The distribution, composition and structure of diazotroph communities varied across seven different terrestrial ecosystems, with community composition exhibiting an especially clear effect. The Cyanobacteria were the most abundant taxa in crust ecosystems (accounting for ~45% of diazotrophs), while other terrestrial ecosystems were dominated by Proteobacteria, including Alpha-, Beta- and Gamma-Proteobacteria (accounting for ~70% of diazotrophs). Farmland ecosystems harboured the highest and crust ecosystems the lowest alpha and phylogenetic diversities. Azospirillum zeae, Skermanella aerolata and four Bradyrhizobium species were identified as key indicator species of potential diazotroph activity. Overall, diazotroph abundances and distribution were affected by multiple environmental parameters, including soil pH, nitrogen, organic carbon, C:N ratio and annual mean precipitation and temperature. Together, our findings suggest that based on the relative abundance and diversity of nifH marker gene, diazotrophs have adapted to a range of environmental niches globally.     

西双版纳地区稻田CO2排放通量

采用静态暗箱-气相色谱法对云南西双版纳地区单季稻田CO2排放及氮肥、水热因子对CO2排放的影响进行田间原位观测研究.试验设3个氮肥水平处理:N0(0 kg N hm-2)、N150(150 kg N hm-2)和N300(300 kg N hm-2).结果表明,受一天温度变化的影响,西双版纳地区稻田生态系统呼吸日变化为单峰型,其最大值出现在11:00～13:00之间,最小值出现在凌晨.稻田土壤呼吸呈明显的季节变化趋势,土壤呼吸平均速率为水稻收获后休闲季节>种植前休闲季节>水稻生长季节,差异达到1%显著水平.不同季节影响土壤呼吸的环境因子不同.土壤水分含量低于34%时,土壤呼吸速率与土壤含水量呈正相关,达5%显著水平;地面淹水时,土壤呼吸速率与淹水深度呈1%极显著负相关;水分含量高于38%时,土壤呼吸速率与温度呈极显著指数相关.长期考虑(整个生长季节),氮肥的施用对稻田土壤呼吸和生态系统呼吸无影响;N300处理抑制植株呼吸作用,单位生物量呼吸速率下降.氮肥的施用对土壤呼吸有短期影响,氮肥用量增加,土壤呼吸速率增加.计算得出N0、N150和N300处理年土壤呼吸量分别为6.27、6.31 t C hm-2 a-1和5.89 t C hm-2 a-1;年净固定大气中CO2-C分别为1.41、2.22 t C hm-2 a-1和1 11 t C hm-2 a-1,表明西双版纳稻田生态系统是碳汇.     

No Evidence of an Impact on the Rhizosphere Diazotroph Community by the Expression of Bacillus thuringiensis Cry1Ab Toxin by Bt White Spruce

Nitrogen fixation is one of the most important roles played by soil bacterial communities, as fixation supplies nitrogen to many ecosystems which are often N limited. As impacts on this functional group of bacteria might harm the ecosystem's health and reduce productivity, monitoring that particular group is important. Recently, a field trial with Bt white spruce, which constitutively expresses the Cry1Ab insecticidal toxin of Bacillus thuringiensis, was established. The Bt white spruce was shown to be resistant to spruce budworm. We investigated the possible impact of these genetically modified trees on soil nitrogen-fixing bacterial communities. The trial consisted of untransformed controls, GUS white spruce (transformed with the β-glucuronidase gene), and Bt/GUS white spruce (which constitutively expresses both the Cry1Ab toxin and β-glucuronidase) in a random design. Four years after planting, soil samples from the control and the two treatments from plantation as well as from two natural stands of white spruce were collected. Diazotroph diversity was assessed by extracting soil genomic DNA and amplifying a region of the nitrogenase reductase (nifH) gene, followed by cloning and sequencing. Analysis revealed that nitrogen-fixing communities did not differ significantly among the untransformed control, GUS white spruce, and Bt/GUS white spruce. Nevertheless, differences in diazotroph diversity were observed between white spruce trees from the plantation site and those from two natural stands, one of which grew only a few meters away from the plantation. We therefore conclude, in the absence of evidence that the presence of the B. thuringiensis cry1Ab gene had an effect on diazotroph communities, that either site and/or field preparation prior to planting seems to be more important in determining diazotroph community structure than the presence of Bt white spruce.     

No evidence of an impact on the rhizosphere diazotroph community by the expression of Bacillus thuringiensis Cry1Ab toxin by Bt white spruce

Nitrogen fixation is one of the most important roles played by soil bacterial communities, as fixation supplies nitrogen to many ecosystems which are often N limited. As impacts on this functional group of bacteria might harm the ecosystem's health and reduce productivity, monitoring that particular group is important. Recently, a field trial with Bt white spruce, which constitutively expresses the Cry1Ab insecticidal toxin of Bacillus thuringiensis, was established. The Bt white spruce was shown to be resistant to spruce budworm. We investigated the possible impact of these genetically modified trees on soil nitrogen-fixing bacterial communities. The trial consisted of untransformed controls, GUS white spruce (transformed with the beta-glucuronidase gene), and Bt/GUS white spruce (which constitutively expresses both the Cry1Ab toxin and beta-glucuronidase) in a random design. Four years after planting, soil samples from the control and the two treatments from plantation as well as from two natural stands of white spruce were collected. Diazotroph diversity was assessed by extracting soil genomic DNA and amplifying a region of the nitrogenase reductase (nifH) gene, followed by cloning and sequencing. Analysis revealed that nitrogen-fixing communities did not differ significantly among the untransformed control, GUS white spruce, and Bt/GUS white spruce. Nevertheless, differences in diazotroph diversity were observed between white spruce trees from the plantation site and those from two natural stands, one of which grew only a few meters away from the plantation. We therefore conclude, in the absence of evidence that the presence of the B. thuringiensis cry1Ab gene had an effect on diazotroph communities, that either site and/or field preparation prior to planting seems to be more important in determining diazotroph community structure than the presence of Bt white spruce.     

Bacterial Diversity of the Broadbalk ‘Classical’ Winter Wheat Experiment in Relation to Long-Term Fertilizer Inputs

With more than 160 years of contrasting fertilizer regimes, the Broadbalk winter wheat experiment represents a unique experimental resource for studying the effects of long-term fertilizer application on microbial population diversity. Using DGGE and clone library analysis, we report here on eubacterial species diversity (16S rRNA gene) and diversity within two sets of gene products associated with microbial N acquisition: NifH (nitrogen fixation) and AmtB (ammonium transport). Comparisons were made within and between soils treated with mineral N fertilizer, farmyard manure or receiving no fertilizer. Analysis of 16S rRNA gene DGGE profiles showed no clear patterns to qualitatively distinguish bacterial community structure between the three different treatments (P > 0.05), with all samples containing a range of eubacterial taxa similar to those that are characteristic of soil bacteria reported elsewhere. Intra-plot heterogeneity was high and of a similar magnitude to that between treatments. This lack of qualitative between plot differences was echoed in the representative sequences of 16S rRNA, nifH, and amtB genes in the various samples. Taken together, both phylogenetic and functional gene analyses showed bacterial communities in the Broadbalk-trial soil were very stable and relatively non-responsive to long-term management of balanced fertilizer inputs.