Decomposition and mineralization of energy crop residues governed by earthworms

Energy crops are increasingly cultivated in agricultural management systems world-wide. A substitution of food crops (e.g. cereals) by energy crops may generally alter the biological activity and litter decomposition in soil due to their varying structural and chemical composition and subsequently modify soil functioning. A soil microcosm experiment was performed to assess the decomposition and microbial mineralization of different energy crop residues in soil compared to a food crop, with or without earthworms. Residues of the energy crops winter rape (Brassica napus), maize (Zea mays), miscanthus (Miscanthus giganteus) and the food crop oat (Avena sativa) were each provided as food source for a mixed earthworm population, each consisting of one individual of Lumbricus terrestris, Aporrectodea caliginosa, and Octolasion tyrtaeum. After 6 weeks, the rate of litter loss from the soil surface, earthworm biomass, microbial biomass-C and -N, microbial activity, and enzyme activities were determined. The results emphasized, that litter loss and microbial parameters were predominantly promoted by earthworms and were additionally influenced by the varying structural and chemical composition of the different litter. Litter decay by earthworms was highest in N-rich maize litter treatment (C-N ratio 34.8) and lowest in the case of miscanthus litter (C-N ratio 134.4). As a consequence, the microbial biomass and basal respiration in soils with maize litter were higher, relative to other litter types. MBC-MBN ratio in soil increased when earthworms were present, indicating N competition between earthworms and microorganisms. Furthermore, enzyme activities responded in different ways on the varying types of litter and earthworm activity. Enzymes involved in the N-cycle decreased and those involved in the C-cycle tended to increase in the presence of earthworms, when litter with high C-N ratio was provided as a food source. Especially in the miscanthus treatments, less N might remain for enzymatic degradation, indicating that N competition between earthworms and microorganisms may vary between different litter types. Especially, an expansion of miscanthus in agricultural management systems might result in a reduced microbial activity and a higher N deficit for microorganisms in soil.     

A Novel Particle Contact Assay with the Yeast Saccharomyces cerevisiae (8 pp)

Goal, Scope and Background   Numerous xenobiotics released into surface waters are transferred to suspended particulate matter and finally attached to sediments. Aquatic organisms may be exposed to them by direct particle feeding, by physical contact with contaminated surfaces as an exposure route, and by the uptake of dissolved contaminants after equilibration via the free water phase. In order to assess potential sediment toxicity, each of these exposure routes has to be addressed. This paper presents a newly developed particle contact assay that uses the fermentation performance of a specific Saccharomyces cerevisiae strain for the assessment of toxic effects in sediments. The test procedure is based on the characteristic feature of growing yeast cells to attach to sediment particles, which are also relevant for the accumulation of contaminants. The physical contact with lipophilic contaminants mirrors an exposition pathway for the direct uptake into the cells. In order to quantitatively characterize the toxic effects of particle attached pollutants on the fermentation performance, unpolluted native reference sediment was spiked with representatives for widely distributed anthropogenic contaminants. Methods   Saccharomyces cerevisiae was established as sensitive eukaryotic microorganism for the ecotoxicological assessment of particle attached anthropogenic contaminants in freshwater sediments. For this purpose, yeast cells were cultivated in sediment samples and the resulting fermentation performance was continuously measured. Sediments artifically spiked with HCB, PCB, g-HCH, DDT, and benzo(a)pyrene and solutions of each contaminant were comparatively investigated by means of their adverse effects on yeast fermentation performance. Additionally, four native river sediments characterized by increasing levels of pollution were assessed by the yeast particle contact assay, and simultaneously by standard aquatic tests with algae, daphniae, and luminescent bacteria using pore water and elutriates. Results of the bioassays were related to specific sediment contamination with respect to metals and organic priority pollutants. Results and Discussion   In sediments spiked with PCB and benzo(a)pyrene fermentation, performance was affected extensively below concentrations inhibiting fermentation in contaminant solutions. This suggests a high efficiency of the exposure route by physical contact. The fermentation performance was only slightly affected by single lipophilic pollutants, whereas mixtures of individually spiked sediments caused critically reduced fermentation performance suggesting additive synergistic effects. Native river sediments modestly to critically polluted by hazardous organic compounds lead to a slightly to dangerously reduced fermentation performance in the yeast contact assay. These inhibitory effects were much less pronounced in the standard bioassays conducted with algae, daphniae and luminescent bacteria, applying pore waters and elutriates as sample matrices. Using pore water, inhibition was measured only in the most polluted sediment, elutriates lead to a slight inhibition of the algal growth in the undiluted sample only. These results indicate an improved sensitivity of the yeast particle contact assay compared to the standard assays, due to uptake and physical cell contact as additional routes of exposure. Conclusion   The yeast particle contact assay is a valuable tool for the assessment of ecotoxicological potential in freshwater sediments. Since the assay addresses physical contact as an exposure route, it indicates bioavailability of lipophilic compounds in sediments. Outlook   The sensitive indication of bioavailable contaminants associated to sediment particles by the newly developed yeast particle contact assay recommends it as a complementary microbial bioassay in a test battery for assessing major pathways of contaminants in whole sediments.     

ATP发光技术测定辐照前脱水蔬菜和调味品的含菌量

用ATP发光技术测定脱水蔬菜和调味品的初始含菌量,发现脱水蔬菜和调味品中细菌ATP的生物发光强度与其初始含菌量之间显著相关,从样品的制备到获得ATP发光强度结果的检测过程仅需1～2h。该技术可用于辐照灭菌前的微生物检测。     

应用彗星试验研究产毒微囊藻喂食暴露对铜锈环棱螺肝细胞DNA的损伤

把铜锈环棱螺（Bellamya aeruginosa）暴露于3组不同成分的微藻悬浮液中[蓝藻组：只投喂产毒铜绿微囊藻（Microcystis aeruginosa）;混合藻组：50%四尾柵藻（Scenedesmus quadricanda）＋50%产毒铜绿微囊藻;对照组：只投喂四尾柵藻],用酶联免疫检测法（Enzyme-linked immuno sorbent assay,ELISA）检测藻液和螺肝组织中的藻毒素浓度。结果表明,藻液中包括藻相和水相的总微囊藻毒素（MCs）浓度分别为：蓝藻组（36.34±4.12）μg·L-1;混合藻组（18.69±2.12）μg·L-1;对照组未检出。在喂食暴露的前6h内,混合藻组和蓝藻组螺肝组织中微囊藻毒素含量持续增长,而后出现下降趋势;第12h更换藻液并将微藻浓度调整至初始值后,肝组织中MCs含量又迅速回升。同期螺肝细胞DNA损伤指标彗星尾长（TL）、彗星尾距（TM）和彗星尾部DNA百分含量（Tail DNA%）也随螺肝组织中MCs含量发生相应变化,各项DNA损伤指标均在产毒微囊藻喂食暴露6h时达到最大值,之后,各项指标值有所回落,但第12h更换藻液后,DNA损伤再次加剧。整个实验期间（24h）,混合藻组和蓝藻组的DNA损伤指标均显著高于对照组,混合藻组均显著高于蓝藻组。说明铜锈环棱螺经产毒微囊藻喂食暴露后,其肝组织细胞的DNA受到损伤,且螺肝组织MCs积累越多,DNA损伤越严重。     

Relative efficiency,ureide transport and harvest index in soybeans inoculated with isogenic HUP mutants of Bradyrhizobium japonicum

Summary Differences between isogenic uptake hydrogenase (HUP) mutants of Bradyrhizobium japonicum in terms of nodule efficiency, N₂ fixation and N incorporation into various plant parts were studied in a monoxenic greenhouse experiment in order to confirm previous results with soybeans and beans inoculated with various HUP⁺ and HUP^– strains. The HUP+ revertant PJ17-1 of a HUP^– mutant (PJ17) of strain USDA DES 122 showed a completely restored relative efficiency (100% versus 78±2% for the HUP^– mutant), higher nodule efficiency (N₂ fixed per g nodules), higher ureide-N transport rates, higher N contents in pods and higher N harvest indices. All these observations confirm previous experiments with HUP⁺ and HUP^– strains.     

Nodulation and nitrogen fixation of Lupinus species with Bradyrhizobium (lupin) strains in iron-deficient soil

 The effect of six Bradyrhizobium sp. (lupin) strains (WPBS 3201D, WPBS 3211D, USDA 3040, USDA 3041, USDA 3042 and CB 2272) and Fe supply on nodulation, N₂-fixation and growth of three lupin species (Lupinus termis, L. albus and L. triticale) grown under Fe deficiency in an alkaline soil, were examined in sterilized and non-sterilized pot experiments. When inoculated with USDA 3040, 3041, 3042 and CB2272 without Fe addition, the three lupin species had a very low nodule number and mass, low shoot and root dry matter accumulation and lower N yield. However, inoculation with WPBS 3201D and 3211D without Fe treatments increased all these parameters substantially. The ability of WPBS 3201D and 3211D to form nodules on the three lupin species under conditions of Fe stress could be attributed to their ability to scavenge Fe from Fe-deficient environments through their siderophore production. Addition of Fe to the other four strains significantly increased nodulation and N₂-fixation of the three lupin species, indicating that the poorer nodulation and N₂-fixation of these strains in the absence of Fe, resulted from a low ability to obtain Fe from alkaline soils. Bradyrhizobium strains WPBS 3201D and 3211D were superior to the other four strains in terms of promoting greater nodulation, N₂-fixation, plant growth and N accumulation of L. termis and L. albus. However, the other four strains were more efficient in symbiotic association with L. triticale. The greater variations in nodule efficiencies (specific nitrogenase activity) under different levels of Fe supply could be attributed to the quantities of bacteroid protein and leghaemoglobin in the nodules. The results suggested that Bradyrhizobium (lupin) strains differ greatly in their ability to obtain Fe from alkaline soils, and that the selection of bradyrhizobial strains which are tolerant of Fe deficient soils could complement plant breeding for the selection of legume crops for Fe-deficient soils. Received: 5 January 1998     

Seasonal changes in periphyton nitrogen fixation in a protected tropical wetland

Cyanobacteria are important for global nitrogen cycle and often form complex associations referred to as cyanobacterial mats or periphyton that are common in tropical, limestone-based wetlands. The objective of this study was to monitor the nitrogen fixation rate using the acetylene reduction assay of these cyanobacterial mats in a tropical, unfertilized, and protected wetland. To account for temporal and spatial variation of nitrogenase activity, we were interested in seasons in a hydrological cycle (dry, rains, and end of rains), sites with different vascular vegetation, and rates of nitrogenase activity in a 24-h cycle. The annual average of nitrogenase activity was 22 nmol C₂H₄ cm⁻² h⁻¹, with a range of <6 to 35 nmol C₂H₄ cm⁻² h⁻¹, and the annual nitrogen fixation rate of our study site (9.0 g N m⁻² year⁻¹) is higher than similar estimates from other freshwater wetlands. There was a clear temporal pattern in nitrogenase activity with a maximum rate occurring during the rainy season (August) and a maximum nitrogenase activity occurring between 0600 and 1200 hours. We found spatial differences in nitrogenase activity among the four sites that could be attributed to variations in species composition within the periphyton.     

Nitrogen fixation,biomass production,and nutrient uptake by annual Sesbania species in an alkaline soil

Summary In three field trials conducted during the summer season of 1986, 1987 and 1989 in an alkaline soil, 17 accessions of annual Sesbania spp. were evaluated for nodulation, N₂ fixation (acetylene reduction assay), dry weight of roots and shoots, woody biomass production, and nutrient uptake. At 50 days after sowing all the accessions were effectively nodulated (average 36.4 root nodules plant^-1) with a high nodule score (3.4). There was a lot of variation in nodule volume and mass and in acetylene reduction activity but not in N content (5.2%). N uptake in shoots, roots and nodules averaged 639, 31, and 13 mg plant^-1, respectively, and much of the fixed N remained in shoots. Accessions of S. cannabina complex performed better than others. S. rostrata had poor root nodulation but exhibited excellent stem nodulation (300 nodules plant^-1) even though not inoculated with Azorhizobium sp. Average concentrations of N, P, K, S, Ca, and Mg in the shoots were high, at 3.2, 0.28, 1.5, 0.28, 1.5, and 0.4% respectively, and Na was low (0.15%), reflecting the usefulness of Sesbania spp. as an integrated biofertilizer source. Green matter production was 26.0 Mg ha^-1 (5.9 Mg dry matter) and N uptake was 158 kg ha^-1, 54 days after sowing. Average woody biomass of six accessions at maturity, 200 days after sowing, was high (19.9 Mg ha^-1), showing its potential for shortterm firewood production. Total nutrient uptake for production of woody biomass (200 days of growth) was no more demanding than growing the plant to the green-manuring stage of 50–60 days' growth.     

Cobalt reduces nitrate inhibition of nodulation in mungbean (Vigna radiata)

 A cobalt-mediated decrease in ethylene production reduced the inhibition of nodulation by nitrate in Vigna radiata (mungbean). Nitrate increased the ethylene production in 5-day-old seedlings, while it caused a reduction in the nodulation status (nodule number and nodule weight) and nodule efficiency (acetylene reduction activity) in mungbean plants. The application of cobalt chloride inhibited nitrate-affected ethylene production and also decreased the inhibitory effect of nitrate on nodulation. The effect of cobalt was most marked on nodule number. Received: 6 August 1999     

用酶联免疫吸附受体法检测鱼类生长激素的生物活性

根据激素一受体反应的原理，结合酶联免疫吸附测定法（ＥＬＩＳＡ）和放射受体测定法（ＲＲＡ）的优点，应用我们纯化的Ａ昌鱼生长激素（ｇｃＧＨ）和大鳞大马哈鱼生长激素（ｓＧＨ）及其特异抗体，采用鱼类肝细胞膜受体制剂，首次建立了测定鱼类ＧＨ生物活性的酶联免疫吸附受体测定法（ＥＬＩＳＡ－ＲＡ）。此法检测草鱼ＧＨ的灵敏度达０．０６３￣０．１２５ｕｇ／ｍｌ，检测大马哈鱼ＧＨ与草鱼肝膜受体结合的灵敏度达０．２５ｕｇ