New insights into virus yellows distribution in Europe and effects of beet yellows virus,beet mild yellowing virus,and beet chlorosis virus on sugar beet yield following field inoculation

Beet yellows virus (BYV), beet mild yellowing virus (BMYV), beet chlorosis virus (BChV), and beet mosaic virus (BtMV) cause virus yellows (VY) disease in sugar beet. The main virus vector is the aphid Myzus persicae. Due to efficient vector control by neonicotinoid seed treatment over the last decades, there is no current knowledge regarding virus species distribution. Therefore, Europe-wide virus monitoring was carried out from 2017 to 2019, where neonicotinoids were banned in 2019. The monitoring showed that closterovirus BYV is currently widely spread in northern Europe. The poleroviruses BMYV and BChV were most frequently detected in the northern and western regions. The potyvirus BtMV was only sporadically detected. To study virus infestation and influence on yield, viruses were transmitted to sugar beet plants using viruliferous M. persicae in quadruplicate field plots with 10% inoculation density simulating natural infection. A plant-to-plant virus spread was observed within 4 weeks. A nearly complete infection of all plants was observed in all treatments at harvest. In accordance with these findings, a significant yield reduction was caused by BMYV and BChV (−23% and −24%) and only a moderate reduction in yield was observed for BYV (−10%). This study showed that inoculation at low densities mimics natural infection, and quick spreading induced representative yield effects. Within the background of a post-neonicotinoid era, this provides the basis to screen sugar beet genotypes for the selection of virus tolerance/resistance and to test the effectiveness of insecticides for the control of M. persicae with a manageable workload.     

Retrospective estimation of the data of infection with beet yellowing viruses in sugar-beet under field conditions

Sugar-beet plants, infected with beet yellows virus (BYV, closterovirus group) or beet mild yellowing virus (BMYV, luteovirus group) develop symptoms on the inoculated leaves on which aphids infected the plant. Symptoms develop also on the systemically-infected leaves to which virus has been transported via the phloem. Systemic infection occurs in the leaves which have just, or not yet appeared at the moment of infection of the plant. All other, older leaves remain uninfected. The infection-date can be estimated by assessing the date of appearance of the oldest systemicallyinfected leaf of a plant. This approach was tested in the field and gave good results.Samenvatting Suikerbieteplanten die besmet zijn met het bietevergelingsvirus, BYV, of met het zwakke vergelingsvirus, BMYV, ontwikkelen symptomen op de geïnoculeerde bladeren, waarop infectieuze bladluizen virus hebben overgedragen, én op de systemisch besmette bladeren waarheen het virus vanuit de geïnoculeerde bladeren is getransporteerd via het vaatsysteem. Bladeren die op het moment van infectie nog niet verschenen zijn of vlak ervóór zijn verschenen, worden systemisch besmet, terwijl oudere bladeren gezond blijven. De infectiedatum kan worden bepaald door aan de hand van temperatuursommen de verschijningsdatum van het oudste systemisch besmette blad te berekenen. Deze methode bleek bij toetsing in het veld goed te voldoen.     

The incubation period of beet yellowing viruses in sugar-beet under field conditions

In three years field trials, the incubation period, i.e. the time between infection and the appearance of symptoms, of beet yellows virus (BYV) and beet mild yellowing virus (BMYV) increased with later infection during the growing season. The incubation period of BYV, a closterovirus, increased from 3 weeks in young plants infected before canopy closure, to 9 weeks in old plants infected in August. The incubation period of BMYV, a luteovirus, increased from 4 to 5 weeks in young plants to 9 weeks in old plants. Symptoms were observed c. one week earlier on the inoculated leaves than on those infected systemically, except on young BYV-infected plants. On these plants, symptoms developed in 3 weeks on both leaf types.The incubation period decreased at increasing temperature, a fixed temperature sum being required for the development of symptoms on plants of a certain age. This temperature sum increased with plant age. Symptom development was related to leaf growth; the systemic symptoms appeared after the infected leaves attained their final size. Young, expanding leaves did not show symptoms. Thus the development of symptoms seems to be related to physiological conditions occurring only in full-grown leaves. A low rate of leaf expansion may constitute the underlying reason for the long incubation period of virus symptoms in old plants and at low temperatures.The incubation period was not substantially affected by: (1) the number ofMyzus persicae used to inoculate the plants, (2) the number of leaves inoculated, (3) the development stage of the inoculated leaf and (4) the source plant of BMYV, beet or shepherd's-purse,Capsella bursapastoris. The incubation period can be used to obtain rough estimates of the infection-date of individual plants, given the date on which symptoms appear.Samenvatting De incubatieperiode van het bietevergelingsvirus, BYV, en het zwakke-bietevergelings-virus, BMYV, nam toe naarmate suikerbieteplanten later in het seizoen geïnfecteerd werden. Jonge planten ontwikkelden BYV-symptomen na ongeveer 3 weken terwijl na gewassluiting de incubatieperiode geleidelijk toenam tot 9 weken. De incubatieperiode van BMYV nam toe van 4 à 5 weken na inoculatie in juni tot 9 weken na inoculatie in augustus. Geïnoculeerde bladeren ontwikkelden ongeveer een week eerder symptomen dan de systemisch geïnfecteerde bladeren, behalve bij jonge planten, geïnfecteerd met BYV, waar de symptomen zich op beide typen bladeren tegelijkertijd ontwikkelden.De incubatieperiode nam bij hogere temperatuur af en, afhankelijk van de leeftijd van de plant (aantal bladeren) was een bepaalde temperatuursom nodig voor de ontwikkeling van symptomen. Deze temperatuursom nam toe met de ouderdom van de plant. Van alle systemisch besmette bladeren, vertoonden de oudste, welke juist verschenen op het moment dat de plant werd geïnfecteerd, als eerste symptomen. Dit gebeurde zodra of kort nadat ze hun uiteindelijke grootte hadden bereikt. Groeiende bladeren vertoonden nooit vergelingssymptomen. De trage bladexpansie in oude planten en bij lage temperaturen is een mogelijke oorzaak van de lange incubatieperiode aan het einde van het seizoen.De incubatieperiode werd niet duidelijk beïnvloed door inoculatieomstandigheden, zoals (1) het aantal groene perzikluizen,Myzus persicae, dat werd gebruikt voor inoculatie, (2) het aantal geïnoculeerde bladeren, (3) de ouderdom van het geïnoculeerde blad, (4) de bronplant van BMYV, biet of herderstasje, of (5) de vector species. Omdat de incubatieperiode niet in belangrijke mate afhankelijk is van deze factoren kan bij kennis van de datum waarop symptomen verschenen de infectiedatum worden bepaald op basis van de incubatieperiode.     

Decision making in controlling virus yellows of sugar beet in the UK

Virus yellows is an important disease affecting yield in sugar beet in the UK. Myzus persicae (Sulzer) is the most effective and efficient aphid vector of the three viruses causing the disease: beet yellows virus, beet mild yellowing virus and beet chlorosis virus. Control of virus yellows disease is thus focused on the study and control of this aphid species. UK national surveys of virus yellows began in 1946 and these data helped to formulate disease forecasting schemes to optimise control. Over the years, in addition to improvements in farm hygiene, periodic changes and developments in control of the disease have occurred. To accommodate these important developments, virus yellows forecasting schemes have evolved accordingly. The most recent version has been adapted to take account of the current widespread use of imidacloprid seed treatment. Its application offers potential to optimise the rational use of aphicides such as imidacloprid so as to benefit beet growers and the environment by reducing prophylactic use of seed treatment.     

甜菜西方黄化病毒两个山东辣椒分离物全基因组扩增及序列分析

 通过RACE和RT-PCR技术相结合的方法,在山东省潍坊市和济宁市辣椒上克隆出2个甜菜西方黄化病毒(beet western yellows virus,BWYV)的全基因组序列,基因组全长均为5 699 nt。与GenBank比对发现,2个分离物的5′UTR(Untranslated region,UTR)变异程度小、相对比较保守,而3′UTR变异程度大、为突变热点区域。潍坊、济宁两地分离物与GenBank中该病毒的全基因组序列相似度平均值分别为89.89%和89.72%。系统进化树分析发现BWYV亚洲地区的分离物聚类在3个不同的组别,本研究克隆的2个山东分离物和3个日本分离物(LC428355、LC428356、LC428357)、2个韩国分离物(LC198684、KM076647)聚在一组,而美国分离物与法国分离物聚在其他分支,表明该病毒的进化存在地理相关性。这是首次对BWYV中国辣椒分离物的全基因组序列克隆,丰富了BWYV的序列信息,有助于进一步了解该病毒种群的遗传进化关系。     

The effects of beet yellows virus on the growth and physiology of sugar beet (Beta vulgaris)

The effect of beet yellows virus (genus Closterovirus , BYV) on sugar beet growth was studied in a series of field and glasshouse experiments. Infection reduced total plant weight by 20%, primarily through a 25% reduction in storage root growth. Sugar extraction efficiency was depressed by an increase in root impurities. BYV had little effect on above-ground yield or total crop cover but did decrease green cover significantly. Infection did not reduce water extraction depth in field experiments despite decreasing lateral root growth in the glasshouse. The growth reduction in infected plants resulted from both a decrease in net photosynthesis and an increase in the proportion of light intercepted by yellow leaves. Damage to the photosynthetic mechanism at least partly caused the reduction in net photosynthesis.     

Biological activity, availability and duration of phytotoxicity for imazamox in four different soils of central Italy

Greenhouse bioassays were carried out from 1999 to 2002 on several types of soils of central Italy to assess the carry‐over risk of imazamox residues to non‐target crops. No observable effect levels (NOELs) were determined on quartz sand; sugar beet showed the highest sensitivity to imazamox (NOEL 0.4–0.8 ng a.i. mL^?1 of substrate), followed by spinach, oilseed rape, fennel, cauliflower and lettuce (NOELs from 1 to 5 ng a.i. mL^?1 of substrate). Wheat, sunflower, grain sorghum and maize were not very sensitive to this herbicide. Imazamox availability was greatest on sandy soils and decreased in soils with high clay or organic carbon content, where herbicide efficiency was less than 50%, with respect to non‐sorptive media. The decline of herbicide efficiency was quick in sandy soils, where herbicide efficiency dropped to 50% in less than 3 days. In clay‐loam or organic soils, 50% relative efficiency was reached in 15–33 days. Such results suggest that imazamox sprayed at normal field application rates can pose slight risks of carry‐over of residues, which may damage very sensitive species (sugar beet, oilseed rape and spinach) in sandy soils. In these cases, safe recropping intervals of 1–3 months are required, so current label guidelines for imazamox are adequate to protect rotational vegetable crops in central Italy.     

国外甜菜孢囊线虫发生危害、生物学和控制技术研究进展

甜菜孢囊线虫(Heterodera schachtii Schmidt)是全世界重要检疫性有害生物,对甜菜具有毁灭性危害,该线虫已在全世界50多个国家或地区有分布,22个国家将其列为检疫对象。甜菜孢囊线虫寄主多达23科95属218种植物,可导致甜菜产量损失达25%~70%,甚至绝产,在欧洲每年造成的经济损失已超过9 000万欧元,严重威胁当地甜菜生产和制糖业。甜菜孢囊线虫是我国重要进境检疫性有害生物,因其对甜菜具有毁灭性危害,我国各级农业行政主管部门对甜菜孢囊线虫都高度重视,严防该线虫的暴发和危害,本文介绍国外甜菜孢囊线虫研究进展。     

农产品中基质对酶联免疫分析法检测三唑磷残留的干扰及消除研究

基质干扰是免疫法检测食品中农药残留时经常碰到的问题。以胡萝卜、菠菜、香蕉、梨、稻米和花生等为试材,研究了不同农产品基质对酶联免疫分析法(ELISA)检测三唑磷的影响。结果表明:不同农产品基质对ELISA检测的干扰机制不同,香蕉主要是抑制了辣根过氧化物酶的活性,菠菜、梨和稻米则主要是干扰了抗原和抗体的结合反应,胡萝卜和花生不仅抑制酶的活性,同时还干扰了抗原和抗体的结合反应。将这些农产品的磷酸盐缓冲液(PBS)(含2.5%甲醇)提取液用含有质量分数为0.3%的脱脂奶粉、0.1%的乙二胺四乙酸(EDTA)和含2.5%甲醇的PBS稀释10倍,在12000r/min下离心10min,即可消除其对ELISA检测的干扰。研究结果表明, ELISA法是可用于农产品中三唑磷残留检测的一种简单、快速、有效的方法。     

Polyclonal and monoclonal antibodies for the specific detection of the herbicide acifluorfen and related compounds

Polyclonal and monoclonal antibodies reactive with acifluorfen (AF) were prepared by the immunization of, respectively, rabbits and mice with AF-bovine serum albumin conjugates. The reactivities of polyclonal antibody and three monoclonal antibodies (AF 9-1, AF 51-5 and AF 75-144) were examined in an indirect competitive enzyme-linked immunosorbent assay (C-ELISA). The polyclonal antibody reacted with AF at concentrations of 1·5 to 800 μg litre^-1, while the monoclonal antibodies reacted with AF at concentrations of 3 to 24 μg litre^-1 for AF 9-1, 1·5 to 12 μg litre^-1 for AF 51-5 and 12 to 48 μg litre^-1 for AF 75-144. In the presence of up to 40% methanol in C-ELISA, the monoclonal antibodies, particularly AF 75-144, were less affected in their reactivities with AF than was the polyclonal antibody. Moreover AF 9-1 and AF 51-5 specifically reacted with acifluorfen-methyl and oxyfluorfen, while AF 75-144 reacted with chlornitrofen which did not react with the other antibodies. These results indicated that the antibodies are useful for the assay of AF and its related compounds. © 1997 SCI.     

Impacts of chemical crop protection applications on related CO2 emissions and CO2 assimilation of crops

BACKGROUND: A major global challenge is to provide agricultural production systems that are able to sustain growing demands for food, feed, fibre and renewable raw materials without exacerbating climate change. Detailed and reliable data on the CO₂ balance of different agricultural management activities and inputs as a basis to quantify carbon footprints of agriculture are still lacking. This study aims to fill this gap further by quantifying the net balance of emitted and assimilated CO₂ due to the application of crop protection treatments on the farm, and by assessing their partial contribution to GHG emissions and mitigation in agriculture. The study focuses on key agricultural crops including wheat, corn, oilseeds and sugar crops. RESULTS: The final CO₂ balance, considering GHG emissions due to on‐farm CPP treatment in comparison with CO₂ storage in additional biomass, CO₂ protected with respect to agrotechnical inputs and land inputs and CO₂ saved with respect to associated global land use changes, is positive and may reach multiples of up to nearly 2000. CONCLUSION: The results highlight the importance of the positive yield effects of the CPP programme applications on the farm, resulting in additional assimilated biomass at the farm level and less land use changes at the global level, and thus lower pressure on environmentally important indicators of overall agricultural sustainability. Copyright © 2012 Society of Chemical Industry     

新疆北部荒漠地区春小麦、甜菜需水量与需水规律研究

新疆北部阿勒泰草原的农牧业生产主要是在荒漠瘠薄的土地上开发和发展起来的,通过对联合国＂2817项目＂1990年-1992年春小麦和甜菜灌溉试验数据进行分析比较,初步得到该区域春小麦和甜菜土壤水分与生长发育及产量关系。结果表明：春小麦和甜菜的最优灌溉定额分别为433.7和251.2mm,需水系数为1.7和0.22。最大需...     

Pathogenetic roles of beet necrotic yellow vein virus RNA5 in the exacerbation of symptoms and yield reduction,development of scab-like symptoms,and Rz1-resistance breaking in sugar beet

Beet necrotic yellow vein virus (BNYVV) generally has a four-segmented positive-sense RNA genome (RNAs 1–4), but some European and most Asian strains have an additional segment, RNA5. This study examined the effect of RNA5 and RNA3 on different sugar beet cultivars using a Polymyxa-mediated inoculation system under field and laboratory conditions. In field tests, the degree of sugar yield served as an index for assessing the virulence of BNYVV strains. Japanese A-II type isolates without RNA5 caused mostly 15%–90% sugar yield reductions, depending on the susceptibility of sugar beet cultivars, whereas the isolates with RNA5 induced more than 90% yield losses in the seven susceptible cultivars, but small yield losses in one Rz1-resistant and Rizor cultivars. However, a laboratory-produced isolate containing RNA5 but lacking RNA3 caused higher yield losses in Rizor than in susceptible plants, and induced scab-like symptoms on the root surface of both susceptible and resistant plants. In laboratory tests, A-II type isolates without RNA5 had low viral RNA accumulation levels in roots of Rizor and Rz1-resistant plants at early stages of infection, but in the presence of RNA5, viral RNA3 accumulation levels increased remarkably. This increased RNA3 accumulation was not observed in roots of the WB42 accession with the Rz2 gene. In contrast, the presence of RNA3 did not affect RNA5 accumulation levels. Collectively, this study demonstrated that RNA5 is involved in the development of scab-like symptoms and the enhancement of RNA3 accumulation, and suggests these characteristics of RNA5 are associated with Rz1-resistance breaking.     

Composition of the stubble weed flora and its role for Heterodera schachtii in the year preceding sugar beet production

In Germany, sugar beet is often rotated with 2 years of cereal. Extensive fallow periods between cereal harvest and autumn primary tillage allow for a weed flora to develop. Broad‐leaved weeds could potentially be alternate hosts for the common nematode Heterodera schachtii, one of the most important pests of sugar beet. Between 2009 and 2012, annual weeds developing in cereal stubble fields during July to mid‐October in the season prior to sugar beet were surveyed, including known hosts of H. schachtii. Yearly weather patterns and agronomic practices possibly impacted weed species composition and weed population densities. During September, Chenopodium album, Cirsium arvense, Convolvulus arvensis, Mercurialis annua, Polygonum spp., Solanum nigrum and Sonchus spp. occurred at the highest frequencies. Weed hosts of H. schachtii were present, but densities, frequencies and uniformity were limited. In 2010 and 2011, staining for nematodes in roots revealed juvenile penetration of some weeds but few adult stages. No indication of nematode reproduction of H. schachtii was found on these weed hosts. A fairly stable weed flora was detected on stubble fields that could provide some carry over for weed species. An elevated risk for nematode population density build‐up on these weeds was not found and management of these weeds at the observed densities during the stubble period for nematological reasons appeared unnecessary.     

Immunochromatographic assay for simple and rapid detection of <Emphasis Type="Italic">Satsuma dwarf virus</Emphasis> and related viruses using monoclonal antibodies

A simple and rapid immunochromatographic assay (ICA) to detect Satsuma dwarf virus (SDV) was developed using colloidal gold conjugates of anti-SDV monoclonal antibodies. Of six homogenization buffers tested, 0.1 M citrate buffer (pH 7.0) gave the best results for the ICA. In the ICA, addition of 0.1% thioglycolic acid in the homogenization buffers that have been widely used in enzyme-linked immunosorbent assays (ELISA) was deleterious to the reaction because of undesirable coagulation of the colloidal gold. ICA using the anti-SDV monoclonal antibodies was 8 times and 16 times more sensitive than double antibody sandwich-ELISA and ICA using the anti-SDV polyclonal antibody, respectively. The analysis is complete in only 15 min. Furthermore, ICA using the anti-SDV monoclonal antibodies could also detect SDV-related viruses.     

Farm Scale Evaluations of herbicide-tolerant crops: assessment of within-field variation and sampling methodology for arable weeds

The Farm Scale Evaluations (FSEs) of genetically modified herbicide‐tolerant crops (GMHT) compared the effects of conventional and GMHT crop management regimes on weed densities. Weeds were sampled in quadrats at 5 distances along 12 transects systematically arranged within experimental half‐field treatment units. Weed densities declined with increasing distance from field margins. Random variation within transects was about 80% of the total within‐half‐field variation, i.e. relatively large variation at a small scale. The spatial autocorrelation between counts on the same transect declined as the distance between quadrats increased. Sampling schemes with fewer distances along transects required more transects but fewer quadrats to achieve similar precision as the 5‐distance scheme; on average 32 quadrats were sufficient. The frequency of detection of treatment effects (5% level) was little affected by using six or even three transects (30 and 15 quadrats respectively). However, the use of the 12 transects greatly improved the detection of treatment effects at 1% and 0.1% levels. The effectiveness of the reduced schemes for detecting differences between treatments (5% level) with as few as three transects compared with 12, reflects, in part, the relatively large treatment effects (e.g. 1.5‐fold or greater), which emerged for many of the sampling occasions. The FSEs provide an unusually extensive data set. Both the data and the methodology developed for their design and analysis are valuable for the planning of future weed sampling schemes.     

化学发光酶联免疫分析法同时检测3种有机磷农药残留

利用能同时识别对硫磷、甲基对硫磷和杀螟硫磷的宽谱特异性单克隆抗体,建立了同时测定这3种有机磷农药残留的化学发光酶联免疫分析方法（chemiluminescence enzyme immunoassay,CLEIA）,比较了间接竞争（ic-CLEIA）和直接竞争（dc-CLEIA）2种反应模式,优化了相关理化参数,确立了最适反应条件。结果表明:间接竞争CLEIA法检测对硫磷、甲基对硫磷和杀螟硫磷的抑制中浓度（IC₅₀）分别为5.57、2.30和2.62 μg/kg,检测线性范围分别为0.39~100、0.39~25和0.39~25 μg/kg;直接竞争CLEIA法检测对硫磷、甲基对硫磷和杀螟硫磷的抑制中浓度（IC₅₀）分别为5.43、1.34和1.24 μg/kg,检测线性范围分别为0.39~100、0.10~25和0.10~25 μg/kg。所建立的CLEIA方法基本能满足对硫磷、甲基对硫磷和杀螟硫磷在谷物和果蔬中最大残留限量的检测要求,为研制有机磷农药多残留检测试剂盒提供了技术依据。     

Application of polyclonal and monoclonal antibodies for the detection of Xanthomonas campestris pv. campestris in crucifer seeds using immunofluorescence microscopy

Polyclonal and monoclonal antibodies (PCAs and MCAs) were tested for the detection ofXanthomonas campestris pv.campestris (Xcc) in cabbage seeds using immunofluorescence microscopy (IF). It was concluded that PCA 94, MCAs 20H6, 2F4, 18G12 and a mixture of MCAs 20H6, 18G12, 2F4 and 16B5 could be used to detect Xcc in seed extracts when 5 min and 2.5 h shaking of seeds are used as extraction methods. The reliability of confirming suspect colonies with MCAs and PCA 94 in IF depended in part on the seed lot tested and the antibody used. Some virulent Xcc strains derived from seed lots, did not react with MCAs 10C5, 2F4, 18G12, 17C12 and 16B5. On the other hand, saprophytic isolates obtained from one seed lot cross-reacted with MCA 17C12 and to a lesser extent with MCAs 2F4, 18G12 and PCA 94. No relationship was found between IF-reactions of Xcc strains using MCAs and reactions of Xcc strains in pathogenicity testing. Xcc andX. c. pv.amoraciae (Xca) could in general not be distinguished on the basis of reactions with MCAs and PCAs. Also in pathogenicity tests Xcc and Xca were hard to distinguish.     

Development and evaluation of a one‐hour DNA extraction and loop‐mediated isothermal amplification assay for rapid detection of phytoplasmas

A rapid DNA extraction and loop‐mediated isothermal amplification (LAMP) procedure was developed and evaluated for the detection of two specific groups of phytoplasmas from infected plant material. Primers based upon the 16–23S intergenic spacer (IGS) region were evaluated in LAMP assays for amplification of group 16SrI (aster yellows group) and group 16SrXXII (Cape St Paul wilt group) phytoplasma strains. DNA could be extracted from leaf material (16SrI phytoplasmas) or coconut trunk borings (16SrXXII phytoplasmas) onto the membranes of lateral flow devices, and small sections of these membranes were then added directly into the LAMP reaction mixture and incubated for 45 min at 65°C. Positive reactions were detected through the hydroxyl napthol blue colorimetric assay within 1 h of the start of DNA extraction, and were confirmed by subsequent agarose gel electrophoresis of the LAMP products. The level of detection was comparable to that obtained by nested PCR using conventional 16S rDNA phytoplasma‐specific primers. Furthermore, the assays were specific for the phytoplasmas they were designed to detect – the 16SrI assay only detected 16SrI phytoplasmas and not those from any other phylogenetic groups, whilst the 16SrXXII assay only detected 16SrXXII phytoplasmas. The DNA extractions and LAMP assay are easy to perform, requiring minimal equipment, and may therefore form the basis of a rapid and reliable field‐detection system for phytoplasmas.