Responses by earthworms to reduced tillage in herbicide tolerant maize and Bt maize cropping systems

Genetically modified (GM) crops may affect earthworms either directly through the plant, its root exudates and litter, or indirectly through the agricultural management changes that are associated with GM plant production. In order to investigate such possible effects, we established two field studies of Bacillus thuringiensis (Bt) maize and a glufosinate ammonium tolerant maize and included a reduced tillage (RT) treatment and a conventional tillage (CT) treatment as examples of a likely concomitant change in the agricultural practise. At a French study site at Varois, (Bourgogne), a field grown with the Bt-toxin producing transgenic maize line MON810 was studied for 1 year. At a Danish study site, Foulum (Jutland), 1 year of Bt maize was followed by 2 years of herbicide tolerant (HT) maize. At the French study site, the most prominent effects observed were due to the tillage method where RT significantly reduced the earthworm populations to levels about half of CT. At the Danish study site effects of CT complied with known reduction of anecic earthworms due to this technique and likewise effects of RT were observed for endogeic earthworms. Earthworm populations were diminished with the HT crop, probably due to exposure to the herbicide Basta^® during the two consecutive autumn seasons. This study confirms the importance of including the tillage techniques and pesticide usage when evaluating the environmental effects of new agricultural technologies.     

Earthworms of different functional groups affect the fate of the Bt-toxin Cry1Ab from transgenic maize in soil

The fate of the insecticidal Cry1Ab protein from crop residues (leaves and roots) of the transgenic maize variety MON810 was studied in the presence and absence of two earthworm species (Lumbricus terrestris, Aporrectodea caliginosa; separate incubations) in soil microcosms. The recombinant Cry1Ab protein was quantified using a highly sensitive ELISA. Control microcosms received corresponding non-transgenic plant material. All earthworms survived in the microcosms over a period of 5 weeks, irrespective of whether they received MON810 or non-transgenic plant material. Weight loss was observed for both earthworm species, independent of the plant material or transgenic modification. A strong decline of immunoreactive Cry1Ab in plant residues (mean initial concentration approx. 5000 ng g⁻¹) of MON810 was observed in all treatments, but in microcosms with earthworms this decline was significantly higher with less than 10% of the initial Cry1Ab concentration remaining after 5 weeks. Cry1Ab concentrations in casts were only 0.1% of those found in remaining plant material of the respective microcosms. No immunoreactive Cry1Ab proteins were found in earthworm tissues (threshold of detection: 0.58 ng g⁻¹ fresh weight). No further decline was found for Cry1Ab concentrations in casts of A. caliginosa during a subsequent period of 3 months of incubation in bulk soil (<0.1 ng g⁻¹) after removal of the earthworms from the microcosms, while in casts of L. terrestris the concentration decreased from 0.4 to below 0.1 ng g⁻¹. In conclusion, this study demonstrates that earthworms enhance the decline of immunoreactive Cry1Ab proteins from maize residues.     

Evaluation of effects of transgenic Bt maize on microarthropods in a European multi-site experiment

The effects of maize expressing the Bacillus thuringiensis Cry1Ab protein (Bt maize) on soil microarthropods were assessed in the field at four European locations (two in Denmark and two in France) that differ in their climatic conditions or soil properties. Each site was considered as a separate experiment, with separate statistical comparison. Effects of farming practices using Bt maize were compared with conventional farming practices using near-isogenic non-Bt maize and also (at some of the sites) other conventional varieties. Furthermore, at one field site (Foulum, Denmark), the effects of Bt crops were studied in both conventional tillage and reduced tillage contexts. At another field site (Askov, Denmark), Bt maize effects were also compared to the effects of the chemical insecticide dimethoate. Moreover, at three of the field sites (all except Narbons, France), the possibility of a localised Bt effect around the rhizosphere compared to the bulk soil was assessed by sampling within and between maize rows. There were some significant negative effects of Bt maize on microarthropods in soils with a high clay content. Significant differences of the same magnitude also occurred between different conventional varieties of maize, but the effect of dimethoate appeared clearly greater than Bt effects. It is thus debatable if the Bt maize effect is an effect of the Bt toxin or just an effect of the maize variety. Based on the results, it can be concluded that the effect of Bt maize on soil microarthropods was small and within the normal variation expected in conventional agricultural systems.     

Exposure and effects assessments of Bt-maize on non-target organisms (gastropods, microarthropods, mycorrhizal fungi) in microcosms

Potential differences between Bt-maize (MEB307 expressing the insecticidal Cry1Ab protein) and a near-isogenic non-Bt variety (Monumental) in their influence on the garden snail (Helix aspersa), soil microarthropods (Collembola, Actinedida, Acaridida, Gamasida and Oribatida) and mycorrhizal fungi were studied. Growing snails were caged in microcosms allowing the development of Bt or non-Bt-maize (Zea mays L.) on a sandy loam soil. After 3 months exposure, survival and growth of snails were similar in both treatments. Cry1Ab protein was detected in the Bt-maize leaves (22–42.2 μg Bt protein g⁻¹ dry wt), in the snail tissues (0.04–0.11 μg Bt-protein g⁻¹ dry wt) and in their faeces (0.034–5 μg Bt-protein g⁻¹ dry wt). Total soil microarthropod abundance and diversity were similar between control (non-Bt-maize) and the genetically modified (GM) Bt-maize microcosms. The mycorrhizal colonization of roots did not differ between Bt and non-Bt-maize (frequency of mycorrhizal roots was 88.7% and 83.3% respectively). The mycorrhizal infectivity of soils, expressed as MI₅₀ (minimum soil dry weight required to colonize 50% of plants) was measured using red clover. MI₅₀ was similar for soils where Bt or non-Bt-maize was cultivated for 4 months. The detection of Cry1Ab protein in the viscera and faeces of H. aspersa exposed to Bt-maize indicates that snails contribute to the transfer of the Bt-protein from plant to soil or snail predators. This may constitute an alternative route of exposure for Bt-protein in soil, but this was without a negative influence on mycorrhizal fungi or microarthropods. Results showed that Bt-maize was not toxic for the selected non-target species exposed for 3 or 4 months. The microcosms and analyses used in this study represent new methods for assessing effects of chronic exposure to GM plants of several diverse, yet ecologically and temporally associated species. As the soil organisms we studied can also be used in standardized ecotoxicological tests (XP X31-205-2 for mycorrhizal fungi, ISO 11267 for Collembola and ISO 15952 for snails), microcosm exposures represent a way to link laboratory and field methods for the ecotoxicological evaluation of GM plants.     

Microbial and microfaunal community structure in cropping systems with genetically modified plants

Soils from field sites at Foulum (DK), Narbons (FR) and Varois (FR) planted with genetically modified maize expressing either the insecticidal Bacillus thuringiensis protein (Bt) or herbicide tolerance (HT), as described elsewhere in this volume, were analysed for nematodes, protozoa and microbial community structure. These analyses were mirrored in single-species testing and in mesocosm experiments, and were coordinated with field samples taken for microarthropods, enchytraeids and earthworms so allowing for cross-comparison and a better understanding of the results observed in the field. Over the first 2 years of the field experiments (in 2002 and 2003), the effect of Bt-maize was within the normal variation expected in these agricultural systems. Sampling in 2004 and 2005 was expanded to include the effects of tillage (i.e. reduced tillage versus conventional tillage) and also the use of HT-maize. Tillage had major effects regardless of soil type (Varois or Foulum), with reduced-tillage plots having a greater abundance of microfauna and a different microbial community structure (measured both by phospholipid fatty-acid analysis (PLFA) and by community-level physiological profiling (CLPP)) from conventionally tilled plots. Grass, as a contrasting cropping system to maize, also had an effect regardless of soil type and resulted in greater microfaunal abundance and an altered microbial community structure. Differences in crop management, which for the Bt-maize was removal of the insecticide used to control European corn borer and for HT-maize was a change in herbicide formulation, were only tested at single sites. There were differences in microbial community structure (CLPP but not PLFA) and sporadic increases in protozoan abundance under the Bt-crop management. The HT-maize cropping system, which covered a shorter period and only one site, showed little change from the conventional system other than an altered microbial community structure (as measured by PLFA only) at the final harvest. The Bt-trait had a minimal impact, with fewer amoebae at Foulum in May 2003, fewer nematodes at Foulum in May 2004 but more protozoa at Varois in October 2002 and an altered microbial community structure (PLFA) at Foulum in August 2005. These were not persistent effects and could not be distinguished from varietal effects. Based on the field evaluations of microfauna and microorganisms, we conclude that there were no soil ecological consequences for these communities associated with the use of Bt- or HT-maize in place of conventional varieties. Other land management options, such as tillage, crop type and pest management regime, had significantly larger effects on the biology of the soil than the type of maize grown.     

The role of laboratory, glasshouse and field scale experiments in understanding the interactions between genetically modified crops and soil ecosystems: A review of the ECOGEN project

The interactions of genetically modified (GM) crops with soil species and ecosystems is complex, requiring both specific and broad spectrum assessments. In the ECOGEN project we undertook experiments at three scales of increasing complexity, using Bt maize expressing the Cry1Ab protein from Bacillus thuringiensis as an example. Test species were selected for laboratory-scale experiments to represent taxonomic groups that we could also monitor at glasshouse and field scales (e.g., nematodes, protozoa, micro-arthropods, earthworms, and snails). In the laboratory, single species were exposed to purified Cry1Ab protein or to Bt maize leaf powder incorporated into simplified diets under controlled conditions. In the glasshouse, multiple test species and soil microbial communities taken from ECOGEN's field sites were exposed to Bt maize plants growing under glasshouse or mesocosm conditions. In the field, evaluations were conducted on our selected indicator groups over multiple sites and growing seasons. Field evaluation included assessment of effects due to the local environment, crop type, seasonal variation and conventional crop management practice (tillage and pesticide use), which cannot be assessed in the glasshouse. No direct effects of Cry1Ab protein or Bt leaf residues were detected on our laboratory test organisms, but some significant effects were detected in the glasshouse. Total nematode and protozoan numbers increased in field soil under Bt maize relative to conventional maize, whilst microbial community structure and activity were unaffected. Field results for the abundance of nematodes and protozoa showed some negative effects of Bt maize, thus contradicting the glasshouse results. However, these negative results were specific to particular field sites and sampling times and therefore were transient. Taking the overall variation found in maize ecosystems at different sites into account, any negative effects of Bt maize at field scale were judged to be indirect and no greater than the impacts of crop type, tillage and pesticide use. Although the ECOGEN results were not predictive between the three experimental scales, we propose that they have value when used with feedback loops between the scales. This holistic approach can used to address questions raised by results from any level of experimentation and also for putting GM crop risk:benefit into context with current agricultural practices in regionally differing agro-ecosystems.     

Nutrient concentration and yield of maize (Zea mays L.) after vetch (Vicia sativa L.) in conventional and reduced tillage systems

The effects of tillage on plant availability nutrients are critical to develop nutrient management strategies to optimize yield. The objective of this study was to evaluate differences in the concentration of 19-nutrient and yield after 4-year of conventional [moldboard plough (MT)] and two reduced [rototiller (ST) and chisel (CT)] tillage systems in maize (Zea mays L.) after winter vetch (Vicia sativa L.) in double crop one year. Three tillage systems were compared from 2005 to 2008 in area located in the western region of Turkey under semi-arid Mediterranean clay-loam soils. Nutrients were determined in maize leaves, stems, roots and soils. Results show that the concentration of macronutrients were found to be higher in leaves and stems of maize than roots in three tillage systems of all years, while the opposite was true for micronutrients. Among the macro and micro-nutrients, there was no effect of tillage on nutrient concentration in all maize tissues for sulfur, magnesium, sodium and copper (S, Mg, Na and Cu). However, the nitrogen, potassium, calcium, boron, zinc, manganese, iron, aluminum, barium, cadmium, cobalt, chromium, nickel, lead and selenium (N, K, Ca, B, Zn, Mn, Fe, Al, Ba, Cd, Co, Cr, Ni, Pb and Se) were affected by tillage. ST increased N in stems of 2006 compared with other systems. K in roots of 2006 was 52 and 30% greater in CT than in ST and MT, respectively, while ST and MT of 2007 resulted in 38 and 41% greater than CT. In two of four years, ST contributed to higher grain yield compared with other systems. In general, ST can effectively contribute to increase maize yield following winter vetch compared with MT under this region. Results suggested the need for different management systems associated with reduced tillage including rotation, particularly for basic nutrients. Further, results showed similarities and differences with other studies under tillage with maize following winter vetch.     

Hierarchical classification of environmental factors and agricultural practices affecting soil fauna under cropping systems using Bt maize

The population dynamics of soil organisms under agricultural field conditions are influenced by many factors, such as pedology and climate, but also farming practices such as crop type, tillage and the use of pesticides. To assess the real effects of farming practices on soil organisms it is necessary to rank the influence of all of these parameters. Bt maize (Zea mays L.), as a crop recently introduced into farming practices, is a genetically modified maize with the Cry1Ab gene which produces a protein toxic to specific lepidopteran insect pests. To assess the effects of Bt maize on non-target soil organisms, we conducted research at a field site in Foulum (Denmark) with a loamy sand soil containing 6.4% organic matter. The study focused on populations of springtails (Collembola) and earthworms (Oligochaeta) from samples taken at the beginning and at the end of the maize crop-growing season during 2 consecutive years. Farming practices, soil parameters, the biological structure of soil communities, and the type and age of the crop at the time of sampling, were used as attributes to predict the total abundance of springtails and biomass of earthworms in general and the abundance or biomass for specific functional groups (epigeic, endogeic and anecic groups for earthworms, and eu-, eu to hemi-, hemi-, hemi to epi- and epiedaphic groups for Collembola). Predictive models were built with data mining tools, such as regression trees that predict the value of a dependent variable from a set of independent variables. Regression trees were constructed with the data mining system M5′. The models were evaluated by qualitative and quantitative measures of performance and two models were selected for further interpretation: anecic worms and hemi-epiedaphic Collembola. The anecic worms (r²=0.83) showed preferences for less clay and more silt soil with medium pH but were not influenced directly by farming practices. The biomass of earthworms was greater in early autumn than in spring or late autumn. Biomass of hemi-epiedaphic Collembola (r²=0.59) increased at the end of the maize growing season, while higher organic matter content and pH tended to increase their biomass in spring. Greater abundance of Collembola was also noted in early autumn if the crop was non-Bt maize. The models assessed by this research did not find any effects of the Bt maize cropping system on functional groups of soil fauna.     

Origin and evolution of Chinese waxy maize: evidence from the <Emphasis Type="Italic">Globulin-1</Emphasis> gene

Waxy maize is a special type of cultivated maize and has grown in China for long history. However, the evolution and origin of waxy maize still remain unknown. We analyzed the origin and evolution of waxy maize by sampling DNA sequences from four taxa with eight populations: waxy maize and other maize cultivars from Southwest China or America (Zea mays L. ssp. mays), parviglumis (Z. mays L. ssp. parviglumis Iltis et Doebley), three more distant species within this genus (Z. luxurians (Durieu et Ascherson) Bird, Zea perennis (Hitchcock) Reeves et Mangelsdorf, and Zea diploperennis Iltis, Doebley et Guzman), and a representative of sister genus (Tripsacum dactyloides L.). We sequenced 20 sequences and downloaded 26 sequences from NCBI for the glb1 locus, which encodes a nonessential seed storage protein. Within the Zea genus samples, the waxy maize has the minimum sequence diversity, which contains 31.1% of the level of diversity of parviglumis and 58.5% of the level of diversity of normal maize from Southwest China. Sequence variation within glb1 locus is consistent with neutral evolution in all four taxa according to Tajima’D test. From the NJ tree for glb1 sequences waxy maize formed two main groups which are intermixed with normal maize sequences. These results suggest that the Chinese waxy maize originate from a single gene mutation from normal maize. Combined with the history of maize dispersal in China we can even think that Chinese waxy maize was divergenced from Chinese flint maize.     

转Cry1Ab/Ac基因水稻对稻田底栖动物群落的影响

转Bt基因抗虫水稻的成功研发,为有效控制鳞翅目害虫对水稻的危害提供了一条最为有效、经济的途径。稻田底栖动物群落是转Bt基因抗虫水稻的重要非靶标生物。为了解转Bt基因水稻对稻田底栖动物群落的安全性,以转Cry1Ab/Ac基因抗虫水稻‘华恢1号’(简写为HH1)为材料,以其非转基因亲本‘明恢63’(简写为MH63)为对照,以稻田底栖动物群落为指示生物,运用群落学方法,通过连续2年在江西南昌的大田试验,研究了转Cry1Ab/Ac基因抗虫水稻对稻田底栖动物群落的影响。结果显示:2012年在HH1与MH63的稻田中分别采集到底栖动物22种与25种,其中两生境中相同物种19种,两群落物种组成相似度为0.808 5,有7种优势种,优势种相似度为0.833 3。2013年在HH1与MH63稻田中分别采集到底栖动物26种与28种,两类稻田相同底栖动物22种,两群落物种组成相似度为0.814 8,有6种优势种,优势种相似度为1.000 0。2012年与2013年群落总体分析与时间动态分析均显示,两类稻田中底栖动物的物种丰富度、个体数量、多样性指数、均匀性指数及优势集中性指数变化趋势相似,且均无显著差异。结果表明:连续种植2年转Cry1Ab/Ac基因水稻HH1对南昌稻田底栖动物群落无明显的负作用。     

泥鳅生长及抗氧化 解毒酶系统对水体中转Cry1Ab/Ac基因水稻残遗物的响应

在泥鳅养殖水体中添加稻秆粉模拟水稻残遗物生境,研究了泥鳅生长和肝胰脏抗氧化酶(SOD、CAT)与解毒酶(GST)活性对转Cry1Ab/Ac基因水稻‘华恢1号’(HH1)的响应。设计以HH1稻秆粉10 mg·L?1、50 mg·L?1、100 mg·L?1和200 mg·L?1 4个梯度浓度处理泥鳅为试验组,以非转Bt基因水稻‘明恢63’(MH63)稻秆粉处理组为阴性对照,不加稻秆粉的基础饲养组为空白对照。结果显示:在4种稻秆粉浓度下,HH1组与MH63对照组泥鳅的特定生长率、肥满度、内脏系数及SOD、CAT和GST酶活性均无显著差异(P0.05);与空白对照比较,稻秆粉浓度升高对泥鳅生长的抑制逐渐增强,当浓度达到200 mg·L?1时,HH1组和MH63对照组泥鳅的特定生长率、内脏系数与CAT活性降低。研究结果表明,水体中低含量的转融合基因Cry1Ab/Ac水稻HH1稻秆粉对泥鳅的生长与生理酶活性没有明显影响,高浓度HH1和MH63稻秆粉均使泥鳅的生长和生理酶活性显著降低,这可能与养殖水体中浓度较高的悬浮稻秆粉妨碍了泥鳅的呼吸和滤食,及稻秆粉的分解降低了水体p H和溶氧量有关。     

Novel method for the study of the population dynamics of a genetically modified microorganism in the gut of the earthworm Lumbricus terrestris

Summary A laboratory microcosm study was used to investigate the survival and population dynamics of genetically modified microorganisms (GMM) in the gut of Lumbricus terrestris. Three methods of axenic earthworm production were investigated. An antibiotic mixture of streptomycin and cycloheximide was introduced either passively, mixed with sterile soil or cellulose, or actively, by teflon catheter. Worms treated by all methods lost weight but this was least for the catheter method which was also the only method to produce axenic earthworms. Axenic earthworms were used to determine the effect of competition with indigenous gut bacteria on ingested GMM. The GMM used was Pseudomonas fluorescens, strain 10586/FAC510, with chromosomally inserted Lux genes for bioluminescence, and chromosomal resistance to rifampicin. The bacteria were grown up to the mid-exponential phase before inoculation into earthworms. Bacteria in faecal material were enumerated by dilution plate counting using selective agar. The GMM were re-isolated from the casts of both antibiotic-treated and untreated earthworms. Lower concentrations of GMM and higher concentrations of indigenous bacteria in the casts of untreated compared to antibiotic-treated earthworms suggested that competition is a fundamental control on population dynamics of the introduced bacterial inocula ingested by earthworms.The catheter method, developed in this study, is proposed as a technique to contribute to the risk assessment of environmental release of GMM.     

Effects of Bt-maize material on the life cycle of the land snail Cantareus aspersus

Insect resistant Bt-maize (MON 810) expresses active Cry1Ab endotoxin derived from Bacillus thuringiensis (Bt). Snails constitute non-target soil species potentially exposed to Bt-toxin through consumption of plant material and soil in fields where transgenic plants have been grown. We studied the effect of the Cry1Ab toxin on survival, growth and egg hatchability of the snail Cantareus aspersus. From the age of 4 to 88 weeks, snails were fed either powdered Bt-maize or non-Bt-maize and exposed to soil samples collected after harvesting either the Bt-maize or non-Bt-maize. We applied four treatments: non-Bt soil + non-Bt-maize (MM); Bt soil + Bt-maize (BB), non-Bt soil + Bt-maize (MB), Bt soil + non-Bt-maize (BM). Eggs laid by snails not exposed to Bt-toxin were also exposed to the two types of soils (Bt and non-Bt soil).At the end of growth (47 weeks of exposure), snails exposed to Bt-toxin in food and soil (BB) had a growth coefficient (GC) 25% lower than unexposed snails (MM). After the first period of reproduction (68 weeks) a significant difference remained for body mass GC between the BB and MM treatments. Differences in body mass were not significant at the end of exposure (88 weeks). For snails not previously exposed to Bt material, hatchability of eggs was similar in the soils tested. The outcome of the experiments indicates that, in growing snails, long-term exposure is needed to reveal an effect of Bt-maize. The hazard analysis of Bt-maize which we performed, based on a worst-case scenario, i.e. snails having no food choice, should now be complemented by other simple measurements, e.g. food intake, to understand the underlying mechanisms involved.     

转Bt基因棉秸秆还田利用对土壤肥力的影响

随着转基因的快速发展,大量转Bt棉秸秆的合理利用和处理是不可忽视的重要课题之一。为明确Bt棉秸秆还田利用的可行性和安全性,本研究以不同抗虫转Bt基因棉和常规棉花‘泗棉3号’为研究材料,在分别种植1、2年后将秸秆机械粉碎后原位还田,测试土壤中Bt蛋白残留量、土壤酶活性及养分含量的变化,分析Bt棉秸秆原位还田对土壤肥力特性的影响。研究结果表明,秸秆还田40 d后, Bt棉样地土壤中Bt残留蛋白检测值较低,均与非转基因棉样地无显著性差异。棉秸秆还田后,土壤脲酶、蔗糖酶、蛋白酶、多酚氧化酶、过氧化氢酶、碱性磷酸酶活性皆较秸秆还田前增加,但土壤纤维素酶活性较之前降低。棉秸秆还田使土壤中有机质、有效磷、碱解氮、速效钾和全氮等养分含量及pH明显增加,而Bt抗虫棉与常规棉秸秆还田后对土壤肥力的影响不存在显著差异。对土壤综合肥力指数评价结果表明,秸秆还田对土壤肥力提升与Bt棉抗虫水平无关,土壤肥力指数在两年间由Ⅲ级水平上升至Ⅱ级水平。综上, Bt棉花秸秆还田不会造成土壤综合肥力降低,相反能有效提升土壤肥力;同时还田利用措施可对转基因植株有效灭活,与转基因植物秸秆利用和无害化处理要求相契合。生产中用于Bt转基因棉花秸秆利用和处理在一定程度上是安全可行的。     

Effects of the nitrification inhibitor nitrapyrin and tillage practices on yield-scaled nitrous oxide emission from a maize field in Iran

Nitrification inhibitors can effectively decrease nitrification rates and nitrous oxide(N₂O)emission while increasing crop yield under certain conditions.However,there is no information available on the effects of nitrification inhibitors and tillage practices on N₂O emissions from maize cropping in Iran.To study how tillage practices and nitrapyrin(a nitrification inhibitor)affect N₂O emission,a split factorial experiment using a completely randomized block design with three replications was carried out in Northeast Iran,which has a cold semiarid climate.Two main plots were created with conventional tillage and minimum tillage levels,and two nitrogen(N)fertilizer(urea)management systems(with and without nitrapyrin application)were created as subplots.Tillage level did not have any significant effect on soil ammonium(NH₄⁺)and nitrate(NO₃^-)concentrations,cumulative amount and yield-scaled N₂O emission,and aboveground biomass of maize,whereas nitrapyrin application showed significant effect.Nitrapyrin application significantly reduced the cumulative amount of N₂O emission by 41%and 32%in conventional tillage and minimum tillage practices,respectively.A reduction in soil NO₃^-concentration by nitrapyrin was also observed.The average yield-scaled N₂O emission was 13.6 g N₂O-N kg^-1N uptake in both tillage systems without nitrapyrin application and was significantly reduced to 7.9 and 8.2 g N₂O-N kg^-1N uptake upon the application of nitrapyrin in minimum tillage and conventional tillage practices,respectively.Additionally,nitrapyrin application increased maize biomass yield by 4%and 13%in the minimum tillage and conventional tillage systems,respectively.Our results indicate that nitrapyrin has a potential role in reducing N₂O emission from agricultural systems where urea fertilizers are broadcasted,which is common in Iran due to the practice of traditional farming.     

Ecology of the earthworm Allolobophora carpathica in field and laboratory studies

Allolobophora carpathica is an earthworm species found in the Eastern Carpathian mountain region, but little information is available on its ecology. Field sampling in beech woodland of the Bieszczady National Park, SE Poland, found this species in soils with a pH of less than 5.0 and C:N ratios of 12 to 13. From sampling over 2 years, at 4 sites with differing sub-vegetation, a mean density of A. carpathica of 6.75 individuals m^-2 with a biomass of 8.65 g.m^-2 was recorded. The largest mature adults were in excess of 14.0 g but there were significant differences (p < 0.01) between sampling sites. Seasonal patterns of abundance were observed.To obtain more data on the growth and breeding biology of this species, specimens were collected from the field, taken to laboratories and maintained in mesocosms under controlled environmental conditions. Cocoon production was 0.88 cocoons per earthworm per month at 15^oC, but under fluctuating temperature conditions (16 - 24^oC) no cocoon production occurred. The mean cocoon biomass was 83 mg  (n = 104) and incubation took 178 days at 15^oC (n = 14), although hatchability was low (22%) under these conditions. Growth from the hatchling stage, (mean mass 86 mg; n = 18), to maturity (c. 8.5 g) took 8 - 12 months at 12^oC.     

Leaching of genetically modified Pseudomonas fluorescens through organic soils: Influence of temperature,soil pH,and roots

Summary The effects of soil temperature and bulk soil pH on the vertical translocation of a genetically modified Pseudomonas fluorescens inoculum were studied in reconstituted soil microcosms, in the presence and absence of growing Lolium perenne roots. The inoculated microcosms received one rainfall event per day (5 mm h^-1 for 6 h) for 5 days and the resulting leachate was quantitatively assayed for the presence of the modified pseudomonad. Soil temperature affected the total number of modified pseudomonads detected in the leachate over the 5 days, with significantly lower numbers detected at 25°C compared to 5°C. The bulk soil pH also affected leaching of the inoculum, with significantly greater numbers detected in the effluent at pH 7.5 than at pH 4.5. In the absence of L. perenne, greater numbers of the modified pseudomonads were detected in the pH 7.5 soil after 5 days of leaching compared to soil at pH 4.5. L. perenne roots decreased the number of cells of the inoculum that were leached and detected in the soil after 5 days of leaching. In the soil microcosms used for the pH study the distribution of the inoculum remaining with the soil was altered by L. perenne roots. At each pH value the proportion of cells detected within the soil below the surface 2 cm of the microcosms was greater in the presence of L. perenne roots. The results of this study indicate that soil temperature, bulk soil pH, and the presence of root systems are important factors in determining the extent of inoculum translocation, and should be considered in the design and interpretation of field experiments.     

玉米秸秆全量深翻还田对高产田土壤结构的影响

为达到玉米生产耕层最适深度(22 cm)和耕层最适土壤容重(1.1~1.3 g×cm~(-3)),解决内蒙古平原灌区耕层浅、犁底层坚硬且厚的农田土壤结构问题,分别选用连续1、2、3、4年秸秆深翻还田定位试验地,秋收后玉米秸秆全量粉碎深翻还田,秸秆年均还田量为20 034.97 kg×hm-2,形成秸秆深翻还田1~4年的4个试验处理(SF1-SF4),以不深翻秸秆还田的处理为对照(CK),研究土壤容重、土壤坚实度、土壤团聚体及其稳定性、土壤肥力及p H随不同年限秸秆深翻还田的变化规律。结果表明:1)SF1-SF4处理0~40 cm土层,土壤容重和土壤坚实度比CK显著减小。2)0~20 cm土层,SF4处理0.25 mm团聚体比例(R0.25)、几何平均直径(GWD)和平均重量直径(MWD)均比CK显著减小;SF1处理土壤团聚体破坏率(PAD)比CK显著降低9.56%,不稳定指数(SWA)随深翻年限增加而显著降低;团聚体分形维数SF4比CK显著增大7.30%。3)20~40 cm土层,SF1和SF2处理R0.25比CK分别显著增加13.69%和17.83%;SF2处理的MWD和GWD分别比CK显著增加23.92%和53.38%;SF1-SF4处理的PAD比CK显著降低,且SF2显著高于SF1和SF3;而SF1-SF4的SWA比CK显著增加,且随秸秆深翻年限的增加呈逐渐升高趋势;团聚体分形维数SF2比CK显著降低7.39%。4)土壤有机质含量SF1-SF4比CK显著增加,且SF2-SF4处理显著大于SF1;速效氮、速效磷和速效钾SF1-SF4比CK显著增加,土壤p H SF3、SF4比CK显著降低。总之,深翻秸秆还田1~4年对0~40 cm土层土壤影响显著;深翻秸秆还田2年适合土壤犁底层结构的改良,深翻秸秆还田3年和4年适合土壤耕层结构的改良。玉米秸秆全量深翻还田既能达到耕作土壤的目的,同时也增加了土壤有机质,降低土壤团聚体破坏率和土壤水稳性团聚体的不稳定系数,利于培肥耕层土壤。     

Effects of physical and chemical properties of soils on adsorption of the insecticidal protein (Cry1Ab) from Bacillus thuringiensis at Cry1Ab protein concentrations relevant for experimental field sites

The adsorption of the insecticidal Cry1Ab protein of Bacillus thuringiensis (Bt) on Na-montmorillonite (M-Na) and soil clay fractions was studied. The aim of this study was not to find the adsorption capacity of the soils from the experimental field site, where Bt corn (MON810) was cultivated, but rather to characterize the adsorption behavior of the Cry1Ab protein at concentrations typically found at experimental field sites. In kinetic experiments, the Cry1Ab protein adsorbed rapidly (<60 min) on M-Na. As the concentration of M-Na was varied and the added Cry1Ab protein concentration was kept constant (20 and 45 ng ml⁻¹), the adsorption per unit weight of Cry1Ab protein decreased with increasing concentrations of M-Na. Adsorption of Cry1Ab protein on M-Na decreased as the pH value of the suspension increased. All adsorption isotherms could be described mathematically by a linear regression with the parameter k, the distribution coefficient, being the slope of the regression line. Although their mineralogical composition was nearly identical, the soil clay fractions showed different k values. The different k values were correlated with the physical and chemical properties of the soil clay fractions, such as the organic carbon content, the specific external surface area, and the electrokinetic charge of the external surfaces of the clays, as well as with the external surface charge density. An increase in the amount of soil organic matter, as well as an increase in the electrokinetic external surface charge of the soil clays, decreased the distribution coefficient k. An increase of the specific external surface areas of the soil clays resulted in a higher distribution coefficient k.Less than 10% of adsorbed Cry1Ab protein was reversibly adsorbed on the soil clays and, thus, desorbed. The desorption efficiency of distilled water was higher than that of a solution of CaCl₂ (2.25 mmol) and of dissolved organic carbon (50 mg C l⁻¹).     

Decomposition processes under Bt (Bacillus thuringiensis) maize: Results of a multi-site experiment

The effects of maize expressing the Bacillus thuringiensis Cry1Ab protein (Bt maize) on decomposition processes under three different European climatic conditions were assessed in the field. Farming practices using Bt maize were compared with conventional farming practices using near-isogenic non-Bt maize lines under realistic agricultural practices. The litter-bag method was used to study litter decomposition and nitrogen mineralization dynamics of wheat straw. After 4 months incubation in the field, decomposition and mineralization were mainly influenced by climatic conditions with no negative effect of the Bt toxin on decomposition processes.