Deoxynivalenol Content of Cereal Grain from Naturally Infected and Artificially Inoculated Plants in Field Trials in Norway

The Fusarium mycotoxins deoxynivalenol (DON) and 3-acetyl-deoxynivalenol (3-acDON) were determined in grain samples from naturally infected and Fusarium culmorum inoculated plants in field experiments in Norway during 1992–1996. The mean DON content in trials with inoculated plants was 11.8 μg/g in spring oats, 11.3μg/g in winter wheat, 28.9 μg/g in spring wheat and 31.4 μg/g in spring barley. In the natural infection trials the mean DON content was 0.32 μg/g in spring oats, 0.22μg/g in winter wheat, 1.48μg/g in spring wheat and 0.54 μg/g in spring barley. Only small differences in DON content were observed among cultivars, and significant differences were found only in winter wheat in the inoculation trials, and in spring wheat in the natural infection trials. A significant correlation was observed between the 3-acDON and DON contents in the inoculated trials in all grain species, the mean ratio of 3-acDON to DON ranging from 0.011 in wheat to 0.071 in oats.     

The Occurrence of Type A and B Trichothecenes in Lithuanian Cereals

Samples of winter wheat (n =84), winter rye (46) and barley (29) were collected from the larger family farms and from partnerships in Lithuania just after the 1998 harvest. The number of samples collected from each region was proportional to the amount of grain produced in it. The levels of the Fusarium toxins deoxynivalenol (DON), 3-acetyl-DON, 15-acetyl-DON, nivalenol (NIV), fusarenon-X (4-acetyl-NIV), T-2 toxin, HT-2 toxin, 4,5-diacetoxyscirpenol (DAS), 1,5-monoacetoxyscirpenol (MAS) and scirpentriol in the grain were determined by gas chromatography with mass-selective detection (GC-MS). DON was most often detected in the wheat and rye samples and NIV in the barley samples. The concentrations found were lower than those causing acute or chronic toxic effects in livestock or humans. No fusarenon-X or 15-acetyl-DON was detected, and only small amounts of other trichothecenes were present. Climatic conditions in Lithuania in the summer of 1998 were slightly cooler and wetter than the average for the 1992-1996 but were close to the norm. Because the samples analysed were representative of grain produced for the market in seasons with normal weather, trichothecene contamination of grain from large family farms and partnerships would not be expected to be a problem in most years.     

Rapid screening method for detection of deoxynivalenol

A rapid method for detecting deoxynivalenol (DON), also known as vomitoxin, was developed. DON was extracted from grains and other samples with acetonitrile-4% potassium chloride solution (9 + 1). Impurities that would interfere with detection were removed on a C18 silica gel reverse phase column. Water was removed from eluates on a hydrophilic matrix column. DON was detected by thin layer chromatography using an aluminum chloride solution to develop the blue response characteristic of the mycotoxin. Total time involved is approximately 30 min. The method was applicable to corn, wheat, and barley at detection levels of 1 ppm, and oats at 1.5 ppm. It is applicable to environmental samples (soil, green plants, and water) at detection levels of 0.75 ppm.     

Near‐Infrared Spectroscopic Method for Identification of Fusarium Head Blight Damage and Prediction of Deoxynivalenol in Single Wheat Kernels

Fusarium Head Blight (FHB), or scab, can result in significant crop yield losses and contaminated grain in wheat (Triticum aestivum L.). Growing less susceptible cultivars is one of the most effective methods for managing FHB and for reducing deoxynivalenol (DON) levels in grain, but breeding programs lack a rapid and objective method for identifying the fungi and toxins. It is important to estimate proportions of sound kernels and Fusarium‐damaged kernels (FDK) in grain and to estimate DON levels of FDK to objectively assess the resistance of a cultivar. An automated single kernel near‐infrared (SKNIR) spectroscopic method for identification of FDK and for estimating DON levels was evaluated. The SKNIR system classified visually sound and FDK with an accuracy of 98.8 and 99.9%, respectively. The sound fraction had no or very little accumulation of DON. The FDK fraction was sorted into fractions with high or low DON content. The kernels identified as FDK by the SKNIR system had better correlation with other FHB assessment indices such as FHB severity, FHB incidence and kernels/g than visual FDK%. This technique can be successfully employed to nondestructively sort kernels with Fusarium damage and to estimate DON levels of those kernels. Single kernels could be predicted as having low (<60 ppm) or high (>60 ppm) DON with ≈96% accuracy. Single kernel DON levels of the high DON kernels could be estimated with R² = 0.87 and standard error of prediction (SEP) of 60.8 ppm. Because the method is nondestructive, seeds may be saved for generation advancement. The automated method is rapid (1 kernel/sec) and sorting grains into several fractions depending on DON levels will provide breeders with more information than techniques that deliver average DON levels from bulk seed samples.     

Single‐Kernel Near‐Infrared Analysis for Evaluating Wheat Samples for Fusarium Head Blight Resistance

This report describes a method to estimate the bulk deoxynivalenol (DON) content of wheat grain samples with the single‐kernel DON levels estimated by a single‐kernel near‐infrared (SKNIR) system combined with single‐kernel weights. The described method estimated the bulk DON levels in 90% of 160 grain samples to within 6.7 ppm of DON when compared with the DON content determined with the gas chromatography–mass spectrometry method. The single‐kernel DON analysis showed that the DON content among DON‐containing kernels (DCKs) varied considerably. The analysis of the distribution of DON levels among all kernels and among the DCKs of grain samples is helpful for the in‐depth evaluation of the effect of varieties or fungicides on Fusarium head blight (FHB) reactions. The SKNIR DON analysis and estimation of the single‐kernel DON distribution patterns demonstrated in this study may be helpful for wheat breeders to evaluate the FHB resistance of varieties in relation to their resistance to the spread of the disease and resistance to DON accumulation.     

Fusarium Species and 8-Keto-Trichothecene Mycotoxins in Manitoba Barley

Fusarium head blight, induced primarily by Fusarium graminearum, resulted in widespread damage to the Manitoba barley crops of 1993 and 1994, with contamination by deoxynivalenol (DON) and other 8-keto-trichothecenes. Visible Fusarium mold in samples of 1994 barley had little relationship to DON levels in the kernel as determined by gas chromatography-mass spectrometry (GC-MS). While samples of 1993 and 1994 barley showed a weak correlation between the logarithm of DON level and percentage of kernels infected by Fusarium graminearum (r = 0.79 and 0.71, respectively), the latter method is too lengthy and requires too much training for commercial application. A commercial enzyme immunoassay for DON gave results that correlated well with GC-MS methods (r = 0.95 and 0.89, respectively) in samples of 1993 and 1994 barley and afforded a rapid and convenient method for screening. In barley samples from 1994, DON, 15-acetylDON, 3-acetylDON and 3,15-diacetylDON were detected in the approximate ratio of 47:4:1:1. In view of the higher oral toxicities of 15-acetylDON and 3-acetylDON relative to DON, and the unknown oral toxicity of 3,15-diacetylDON, GC-MS assays might be advisable in samples positive for DON from enzyme immunoassay screening.     

Gas chromatographic analysis of milk for deoxynivalenol and its metabolite DOM-1

A gas chromatographic method is described for the determination of deoxynivalenol (DON) and its metabolite DOM-1 in milk. Milk samples were extracted with ethyl acetate on a commercially available disposable extraction column, followed by hexane-acetonitrile partitioning. Final purification was accomplished on a reverse phase C-18 cartridge. The trimethylsilyl ether (TMS) derivatives of DON were prepared, chromatographed on an OV-17 column, and quantitated with an electron capture detector. Chromatography of the TMS derivatives of milk extracts was compared to that of the corresponding heptafluorobutyryl derivatives. The limit of detection using TMS derivatives was 1 ng/mL for both toxins with recoveries averaging 82% +/- 9% at 2.5 and 10 ng/mL milk for DON and 85% +/- 6% at 10 ng/mL for DOM-1.     

Grain yield and nitrogen dynamics of Mediterranean barley and triticale

Understanding differences in grain yield and nitrogen utilization efficiency (NUtE) between barley and triticale could be useful for designing more sustainable cropping systems. Field experiments were conducted to compare grain yield and dry matter accumulation as well as N accumulation, translocation, and utilization in barley and triticale under Mediterranean conditions with two N fertilization rates (0 and 100 kg ha^?1). Overall, across years and N application rates, barley out-yielded triticale by 30% (6943 vs. 5339 kg ha^?1). Differences in the grain number per m² explained most of the variation between species in grain yield, with barley showing higher values than triticale. Barley showed higher early growth resulting in greater N accumulation in anthesis, and eventually in higher translocation to the grain than triticale. When no N was applied, barley showed a mean increase of 15% in NUtE. Triticale showed an advantage in biomass production efficiency in anthesis only in the drier year. From a practical point of view, barley could be a better choice than triticale under low availability of N, not only concerning profitability, but also sustainability. In dry areas, triticale might be a sustainable choice as a silage crop because of better N exploitation for biomass production than barley.     

Estimation of the Deoxynivalenol and Moisture Contents of Bulk Wheat Grain Samples by FT‐NIR Spectroscopy

Deoxynivalenol (DON) levels in harvested grain samples are used to evaluate the Fusarium head blight (FHB) resistance of wheat cultivars and breeding lines. Fourier transform near‐infrared (FT‐NIR) calibrations were developed to estimate the DON level and moisture content (MC) of bulk wheat grain samples harvested from FHB screening trials. Grains in a rotating glass petri dish were scanned in the 10,000–4,000 cm⁻¹ (1,000–2,500 nm) spectral range using a Perkin Elmer Spectrum 400 FT‐IR/FT‐NIR spectrometer. The DON calibration predicted the DON levels in test samples with R ² = 0.62 and root mean square error of prediction (RMSEP) = 8.01 ppm. When 5–25 ppm of DON was used as the cut‐off to classify samples into low‐ and high‐DON groups, 60.8–82.3% of the low‐DON samples were correctly classified, whereas the classification accuracy of the high‐DON group was 82.3–94.0%. The MC calibration predicted the MC in grain samples with R ² = 0.98 and RMSEP = 0.19%. Therefore, these FT‐NIR calibrations can be used to rapidly prescreen wheat lines to identify low‐DON lines for further evaluation using standard laboratory methods, thereby reducing the time and costs of analyzing samples from FHB screening trials.     

Monoclonal antibody-based enzyme linked immunosorbent assay of aflatoxin B1, T-2 toxin, and ochratoxin A in barley

Aflatoxin B1 (B1), T-2 toxin (T2), and ochratoxin A (OA) were assayed in a single extract from barley grain by using competitive enzyme linked immunosorbent assays (ELISAs) with monoclonal antibodies. B1 and T2 monoclonal antibodies were conjugated to horseradish peroxidase for direct competitive ELISA while an indirect competitive ELISA was used for OA determination. The competitive ELISA detected 0.1 ng/mL of B1, 10 ng/mL of T2, or 1 ng/mL of OA. Acetonitrile-0.5% KCl-6% H2SO4 (89 + 10 + 1) extracts of barley grain either were diluted 1:10 for direct assay or were subjected to a simple liquid-liquid cleanup procedure to concentrate the extract 10:1 before assay. For cleanup, water was added to the acetonitrile extract to partition water-soluble interfering substances, and then the mycotoxins were re-extracted with chloroform. The chloroform extract was evaporated to dryness and redissolved in Tris HCl buffer for ELISA. The mean recoveries from barley spiked with 4-60 ng/g of B1, 50-5000 ng/g of T2, and 5-500 ng/g of OA were, respectively, 93.8, 80.6, and 95.8%. The mean within-assay, inter-assay, and subsample coefficients of variation by ELISA of barley grain colonized with toxigenic fungi were less than 12% for B1 and OA but as high as 17% for T2.     

大麦/豌豆间作系统种间竞争力及产量对地下 作用和密度互作的响应

种间关系是间作高产和资源高效利用的重要生物学基础,揭示种间关系对间作产量的影响,对优化间作技术具有重要理论指导意义。本研究通过盆栽试验,设计根系完全分隔(地上互作)与不分隔(地上地下互作)2种种间作用关系,及2个大麦种植密度(15株·盆~(-1)、25株·盆~(-1)),探讨了根系相互作用与密植对大麦间作豌豆种间竞争互补关系及产量的影响,以期为建立基于优化种间关系提高间作产量的管理技术提供理论依据。结果表明:1)与单作相比,间作可提高群体干物质积累量3.6%~11.3%,其中地下作用的贡献率为53.9%~63.5%;增加大麦种植密度,使根系不分隔间作处理的群体干物质量提高12.5%~14.4%,根系完全分隔间作处理提高3.3%~6.7%。同样,与单作相比,间作群体籽粒产量提高8.6%~38.8%,地下作用的贡献率为2.4%~16.2%;增加大麦种植密度,不分隔与分隔间作处理的籽粒产量分别提高7.0%~10.9%与1.2%~2.6%,说明地下根系相互作用是间作密植的重要基础。2)间作可提高大麦、豌豆的收获指数,大麦提高8.7%~21.0%、豌豆提高3.3%~31.7%;间作大麦收获指数随着大麦种植密度的增加而增大,而间作豌豆收获指数随着大麦种植密度的增加而降低,根系完全分隔处理降低作用更明显。3)根系不分隔地下作用可提高间作群体土地当量比(LER),增加大麦种植密度降低了LER,说明地下根系相互作用是产生间作优势的主要原因。4)地下作用明显增大了大麦相对于豌豆全生育期的平均竞争力,增长率达40.1%~89.1%;增加大麦种植密度,平均竞争力提高11.0%~49.9%。5)间作群体籽粒产量与大麦相对于豌豆全生育期的平均竞争力呈二次相关关系,当该竞争力在0.35、0.13时利于间作大麦、豌豆获得高产。本研究表明,通过增加大麦种植密度(如本研究的25株×盆~(-1))来适度提高大麦的竞争优势,特别是大麦灌浆期的竞争优势有利于间作群体整体产量的提高。     

Phosphorus seed coating as starter fertilization for spring malting barley

Abstract

Experiments were conducted with malting barley (Hordeum vulgare L.) cultivars in 2003 and 2004 in central, western and southern Lithuania to test phosphorus (P) availability at early seedling growth stage with P seed coating. Phosphorus seed coating resulted in alteration to plant stand structure traits. Despite the fact that seedling emergence sometimes decreased, the number of total and productive stems, number of grains per ear and 1000-grain weight increased. The positive effect of P seed coating on grain yield was revealed when the growth conditions during post-anthesis were favourable for exploiting the potential that was obtained during the pre-anthesis phase. In our experiment favourable conditions were considered those that generated a grain yield over 6 t ha^?1. According to path coefficients analyses the significant positive effect of P seed coating on malting barley yield increase was related to seed weight increase. Although P seed coating slightly increased single grain N concentration and thus grain protein concentration, grain protein concentration ranged from 8.6% to 10.2% and met the quality standard requirements for malting barley in all trials and treatments.     

土壤可溶性有机氮测定方法研究进展

可溶性有机氮(DON)是土壤中生物可利用有机氮库的主要成分,是最为活跃的有机氮形态之一。DON在土壤中的迁移和转化是土壤氮素地球生物化学循环的重要一环,与氮素养分供应、损失等过程密切相关。然而缺失准确、可靠的土壤DON分析方法严重限制了DON的相关研究进程。本文综述了近年来国内外土壤浸提液和水体中DON测定方法,重点比较了不同可溶性总氮(DTN)测定方法以及不同前处理方法对提高DON测定结果准确性的影响。     

Simultaneous analysis of nivalenol and deoxynivalenol in cereals by liquid chromatography

A sensitive method is described for determination of nivalenol (NIV) and deoxynivalenol (DON) in cereals by using reverse phase liquid chromatography and UV detection at 222 nm. The sample is extracted with acetonitrile-water (85 + 15) and an aliquot is purified by passage through a combined column of cation exchange resin and alumina-carbon (20 + 1). Analysis at this stage is possible with some samples but the method recommends passing an aliquot through a carbon minicolumn after evaporation and solubilization in methanol. Interference from coextracted compounds at this point is negligible. Recoveries of both NIV and DON from spiked extracts taken through the full method were in the range 83-94%. The relative standard deviation, based on 5 replicate determinations from each of 2 corn samples, was approximately 5% for both NIV and DON. With a 10 microL injection, the minimum contamination (3 X signal/noise ratio) able to be detected in cereal samples was about 0.015 micrograms NIV/g and 0.05 micrograms DON/g. The cleaned up extracts are also suitable for analysis by gas chromatography.     

适度水分亏缺管理提高青稞营养品质和环境效益

【目的】 针对青藏高原水资源短缺会降低青稞产量,但对产量构成和籽粒品质特性的影响尚不明确的问题,研究不同水分供应对青稞籽粒产量构成以及营养品质的影响,为青稞合理高效栽培管理提供理论依据。 【方法】 以昆仑14号为供试品种,进行了随机区组田间试验。设计灌溉至田间持水量的75% (充分灌溉处理)、50% (水分轻度亏缺处理) 和25% (水分重度亏缺处理) 3个水平。调查了青稞根系、产量和籽粒NPK、蛋白质和氨基酸含量。 【结果】 水分亏缺显著降低青稞公顷穗数、穗长、穗粒数、产量、分蘖数、株高,且降幅随水分亏缺程度的加剧而增大。水分亏缺致使青稞产量显著降低,但不同亏缺程度对产量三因素的影响存在一定差异。水分轻度亏缺使公顷穗数和穗粒数显著降低,重度亏缺使产量三因素均显著降低。水分亏缺下青稞籽粒中氮、钾、蛋白质和总氨基酸的含量均呈升高趋势,且随着水分亏缺程度的加剧而进一步增加。重度亏缺处理青稞籽粒中磷含量、必需和非必需氨基酸含量均比充分灌溉显著升高。 【结论】 水分轻度亏缺有助于促进青稞根系生长,重度亏缺则会严重抑制根系生长。水分亏缺不利于青稞穗部的生长发育,导致其产量显著降低,却有利于籽粒中蛋白质及其组分含量的提高。适度亏缺灌溉不仅能节约水资源和降低农业成本,且该灌溉方式下青稞产量和籽粒中养分、蛋白质及其组分的含量均较高,为较佳的灌溉制度。      

Barley yield response to soil organic matter and texture in the Pampas of Argentina

Soil organic matter (SOM) is known to play a major role in soil fertility due to its influence on physical, chemical and biological properties of soil; and it is closely related to particle size distribution. The ratio of SOM (g kg⁻¹) to clay + silt content (g kg⁻¹) was evaluated as an indicator of soil quality for barley (Hordeum vulgare) grain yield, reflecting N availability and soil physical conditions to which crop development is sensitive. Thirty-eight sites in the semiarid Pampa region of Argentina with a wide range of SOM and texture were evaluated for malting barley yield during three growing seasons. In control plots, 51% of grain yield could be explained by this indicator. The threshold value between high and low N-fertilization response was 4.4. Better yield prediction to almost 68% was achieved by combining the SOM to clay + silt indicator with initial nitrate content of the soil at seeding. This combined indicator was also able to explain a high proportion of water use efficiency, particularly in the early growth stages. The ratio of SOM to clay + silt content provided a better tool for estimating grain yield than nutrient availability or SOM alone.     

Determination of the activity of acidic phytate-degrading enzymes in cereal seeds

Five different methods were compared to elucidate the total activity of the acidic phytate-degrading enzymes present in the seeds of rye, wheat, and barley. Phytate-degrading activity was studied at pH 5.0 by quantifying the liberated phosphate. Rye showed the highest acid phytate-degrading activity among the cereals studied. Using an aqueous extract, only 30-50% of the activity was found (rye, 3443 mU g(-1) of grain; wheat, 1026 mU g(-1) of grain; barley, 1032 mU g(-1) of grain) compared to that found by the direct incubation of the dry-milled cereal grains in a buffered phytate-containing solution (rye, 6752 mU g(-1) of grain; wheat, 2931 mU g(-1) of grain; barley, 2093 mU g(-1) of grain). Extending the extraction time resulted in an increase in extractable phytate-degrading activity by, at maximum, 10-15%. Extraction of phytate-degrading activity is strongly enhanced in the presence of Triton X-100 and the protease inhibitor phenylmethylsulfonyl fluoride (rye, 6536 mU g(-1) of grain; wheat, 2873 mU g(-1) of grain; barley, 2023 mU g(-1) of grain), suggesting at least a partial association with membrane structures and a degradation by proteolytic activity during extraction. In addition, it was shown that determining phytate-degrading activity by quantification of the liberated inorganic phosphate is more robust and precise than determining phytate-degrading activity by quantification of the residual phytate.     

Adaptability of hull-less barley varieties to different cropping systems and climatic conditions

Multilocation testing remains the main tool for understanding varietal responses to the environment. Here, Latvian and Norwegian hull-less and hulled barley varieties were tested in field experiments in Latvia and Norway in order to assess the varieties adaptability across environments (sites). Two Latvian (cv Irbe and cv Kornelija) and one Norwegian hull-less barley variety (cv Pihl) were tested along with one Latvian (cv Rubiola) and one Norwegian hulled barley variety (cv Tyra) under conventional and organic management systems. The grain yield, together with physical and chemical grain parameters were compared, and variety yield and protein stability determined. Overall, grain yield of hull-less barley varieties was significantly lower than for hulled barley varieties regardless of climatic conditions and management system. However, in the organic farming systems this difference between barley types was less pronounced. The hull-less barley varieties cv Pihl and cv Irbe, along with both hulled varieties, had good yield stability across environments and were well adapted to both cropping systems. Hull-less barley varieties tended to contain more protein and β -glucans than hulled barley varieties. Despite being bred for local conditions in Norway and Latvia, our study shows that all the varieties used may be successfully transferred across countries.     

Moisture Effects on the Prediction Performance of a Single‐Kernel Near‐Infrared Deoxynivalenol Calibration

Fusarium head blight (FHB) is a serious disease in wheat that affects grain quality owing to the accumulation of mycotoxins such as deoxynivalenol (DON) in grains. Near‐infrared (NIR) spectroscopy has been used to develop techniques to estimate DON levels in single wheat kernels to facilitate rapid, nondestructive screening of FHB resistance in wheat breeding lines. The effect of moisture content (MC) variation on the accuracy of single‐kernel DON prediction by NIR spectroscopy was investigated. Sample MC considerably affected accuracy of the current NIR DON calibration by underestimating or overestimating DON at higher or lower moisture levels, respectively. DON in single kernels was most accurately estimated at 13–14% MC. Major NIR absorptions related to Fusarium damage were found around 1,198–1,200, 1,418–1,430, 1,698, and 1,896–1,914 nm. Major moisture related absorptions were observed around 1,162, 1,337, 1,405–1,408, 1,892–1,924, and 2,202 nm. Fusarium damage and moisture related absorptions overlapped in the 1,380–1,460 and 1,870–1,970 nm regions. These results show that absorption regions associated with water are often close to absorption regions associated with Fusarium damage. Thus, care must be taken to develop DON calibrations that are independent of grain MC.     

Gas chromatographic determination of deoxynivalenol in wheat

Modifications to a published method are described for the determination of deoxynivalenol (DON) in wheat by gas chromatography with electron capture quantitation of the heptafluorobutyrate derivative. In the modified method, DON is extracted by shaking the sample with methanol-water on a wrist-action shaker, followed by filtration through rapid flow paper. One concentration step is eliminated, and a hexane wash is incorporated to remove toluene from the silica gel column. Recoveries of DON from wheat samples spiked at 0.1, 0.5, and 1.0 ppm ranged from 77.3 to 86.3% and averaged 81.5%.