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.     

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.     

Production and evaluation of monoclonal antibodies for the detection of beet mild yellowing luteovirus and related strains

A sugar-beet-infecting isolate of beet mild yellowing luteovirus (BMYV), and aBrassica-infecting isolate of beet western yellows luteovirus (BWYV) were used to produce monoclonal antibodies for epidemiological studies with BMYV and related field strains. Thirty-four monoclonal antibodies were tested for their reaction with 9 luteoviruses in triple-antibody-sandwich enzyme-linked immunosorbent assay. One (MAFF 24) is now routinely used in the UK for detecting BMYV and BWYV in plants and aphids, although it does not discriminate between them. Heterologous reactions were detected between some of the monoclonal antibodies and potato leafroll virus (PLRV), bean leafroll virus (BLRV) and barley yellow dwarf virus (BYDV-RPV). 38% of antibodies raised to BWYV reacted with PLRV compared with 4% of those raised to BMYV. Monoclonal antibodies were produced which distinguished a sugar-beet-infecting isolate of BMYV with differing host range and serological properties from the commonly-occurring field strain.     

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。     

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。     

Host range and molecular analysis of Beet leaf yellowing virus,Beet western yellows virus-JP and Brassica yellows virus in Japan

Beet western yellows virus (BWYV; genus Polerovirus, family Luteoviridae) is one of the most important viruses causing yellowing disease of many field and vegetable crops. This study isolated different poleroviruses from sugar beet, spinach, radish and brassica in Japan, and identified them as BWYV-JP, Beet leaf yellowing virus (BLYV), Brassica yellows virus (BrYV) and BrYV-R (radish strain) based on host range and molecular analysis. Among over 100 plant species from 19 families inoculated with the vector Myzus persicae, about half of the species in 13 families were infected with some of these viruses. BLYV shared a similar host range to Beet mild yellowing virus (BMYV). These had a much more limited host range than BWYV-JP, which resembled BWYV-USA. The host range of BrYV was similar to that of Turnip yellows virus (TuYV). Phylogenetic analyses at the 5′ portion (replication-related gene) of the genome showed that BLYV, BMYV, BWYV (-JP and -USA) and Cucurbit aphid-borne yellows virus (CABYV) formed one large group, whereas BrYV and TuYV were grouped together. BLYV and BWYV were most closely related to each other, and were more closely related to CABYV than to BMYV. However, at the 3′ end (coat protein gene), BLYV and BWYV-JP formed a distinct group, separated from the BrYV group, which in turn was more closely related to BWYV-USA, BMYV, TuYV and Beet chlorosis virus, a group originating from outside Asia. Thus, this study presents host range differences and phylogeographical relationships of BWYV-like poleroviruses that are distributed worldwide.     

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.     

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.     

施氮对高产甜菜干物质积累分配及产量和品质的影响

氮肥对高产甜菜的干物质积累分配及产量和品质影响显著.以KWS2409为材料,在5种施氟处理(N0、N90、N180、N270、N360)下,研究了甜菜叶面积、叶面积指数及干物质积累与分配和产质量变化情况.结果表明,增施氮肥显著增大群体中后期的叶面积指数、群体总干物质积累量和块根含水量,显著降低收获时的根冠比和块根含糖率;块根产量和产粮量则随着施氮量增加,呈现先增后降的趋势.最高产糖量(13 892.64 kg/hm2)的施氮量为180 kg/hm2,块根产量为82 609.63 kg/hm2,合糖率为16.83%.施用氮肥块根产量的最大增幅为不施肥的38.2%,产糖量的最大增幅为不施肥的26.9%.实现甜菜高产高糖主要依靠高的土壤肥力条件.     

补水移栽对甜菜苗期生长及产量品质的影响

为明晰补水移栽对纸筒育苗甜菜苗期生长及产量品质的影响,研究比较了地下式补水与传统的地上式补水的土壤润湿结构特征,补水量对甜菜苗期地下部根系与地上叶片发育的影响,以及补水移栽在两种土壤条件下的甜菜产量与品质效应。结果表明,地下式补水创造了环裹于秧苗纸筒底部的土壤湿润球,利于向秧苗供水与田间有效保水。随补水量的增加,甜菜苗期侧根数、主根长、主根粗、根鲜重、叶片量、叶面积随之显著增长。在华北寒旱区砂质栗钙土农田,以成活率与壮苗为目标的甜菜移栽补水量为150～200 ml·株^-1、壤质草甸栗钙土农田为100～150 ml·株^-1为宜,较不补水甜菜块根增产68.78%～81.82%,糖产量提高65.57%～81.82%。地下式补水移栽,是提高甜菜成活率的关键;适量补水实现培育壮苗,成为甜菜高产的基础。     

调亏灌溉下氮肥管理对滴灌甜菜产量及水氮利用的影响

以Beta356为试材,研究调亏灌溉下不同施氮量[纯氮0 kg·hm^-2(N0)、150 kg·hm^-2(N1)、225 kg·hm^-2(N2)]与基追比[播种前、叶丛期、块根膨大期施氮比例分别为20∶60∶20(T1)、30∶50∶20(T2)、40∶40∶20(T3)]对甜菜生长特性、产量和水氮利用效率的影响。结果表明:在叶丛期和块根膨大期分别进行50%和30%田间持水量(θ_f)调亏灌溉基础上施225 kg·hm^-2氮肥的同时,增加基肥比例能够显著提高叶面积指数(124.39%～143.87%)和叶绿素含量(23.03%～119.80%);在糖分积累期进行30%θ_f调亏灌溉后施用氮肥对叶面积指数影响较小,但有利于提高叶片叶绿素含量。调亏灌溉后施氮使块根膨大期和糖分积累期干物质总量分别较对照提高了34.08%～56.84%和32.43%～76.26%,但两个施氮量处理间未达到显著差异。甜菜产量随施氮量和基肥比例增加分别升高和降低,含糖率随施氮量增加而降低...     

氮肥运筹对干旱区滴灌甜菜氮素利用及产量的影响

以甜菜品种Beta356为试验材料,研究了氮素运筹[甜菜叶丛快速增长期、块根膨大期、糖分积累期的氮素追施比例分别为6∶3∶1、5∶3∶2、4∶4∶2(用N1、N2、N3表示),以不施氮素的处理为对照(用CK表示)]对滴灌甜菜氮素利用及产量的影响。结果表明:随着生育进程的推进,甜菜干物质积累量呈现先升高后降低的趋势,并在块根膨大期达到最大值;不同处理干物质积累量的差异在甜菜苗期、叶丛快速增长期和块根膨大期均未达到显著水平,至糖分积累期和收获期显著高于CK,且各施肥处理间差异不显著;甜菜氮素阶段积累量与该阶段施氮量相比存在一定的滞后性;不同处理氮素总积累量和氮素运转量均表现为N1N2N3CK,氮素运转率则表现为N1N2CKN3;氮农学利用率和氮表观利用率分别表现为N2N3N1和N3N2N1,其中N2处理的氮农学利用率分别比N1和N3处理提高了14%和4%,且不同处理间差异显著;收获期各处理甜菜产糖量表现为N3N2N1CK,块根中造蜜性非糖物质(K、Na和α-氨态氮)表现为CKN2N3N1。综合考虑氮素利用、产糖量以及块根品质,N2处理可作为北疆露播滴灌甜菜较为合理的氮素运筹模式。     

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.     

北京地区辣椒上黄瓜花叶病毒和甜菜西方黄化病毒复合侵染的分子鉴定

辣椒是我国重要的蔬菜和经济作物,受多种病毒危害。2014年在北京市顺义区调查时发现部分种植的辣椒植株上叶片大面积黄化,边缘症状明显,个别植株叶片轻微上卷。提取典型症状样品的总RNA,反转录得到cDNA,分别用黄瓜花叶病毒(Cucumber mosaic virus,CMV)特异引物和马铃薯卷叶病毒属(Polerovirus)通用引物进行PCR检测,CMV特异引物和马铃薯卷叶病毒属通用引物分别扩增得到约650bp和1 400bp的特异条带。测序和核苷酸序列比对表明,其分别与CMV和甜菜西方黄化病毒(Beet western yellows virus,BWYV)序列同源性最高为99%和96%。这是对我国种植的辣椒上发生的CMV和BWYV复合侵染的首次报道。     

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。     

The use of digital image analysis and real‐time PCR fine‐tunes bioassays for quantification of Cercospora leaf spot disease in sugar beet breeding

Cercospora leaf spot, caused by the fungus Cercospora beticola, is a major fungal sugar beet disease worldwide and the cause of significant yield losses. The disease is most successfully countered by the introduction of genetic tolerance into elite sugar beet hybrids. To this end, breeding programmes require high quality biological assays allowing discrimination of minor differences between plants within a segregating population. This study describes the successful implementation of image analysis software in the bioassays for quantification of necrotic lesions at different stages of C. beticola infection, allowing selection on minor phenotypic differences during the sugar beet breeding process for C. beticola resistance. In addition, a real‐time PCR assay was developed for the quantification of C. beticola pathogen biomass in infected beet canopy. The use of both techniques, even in an early stage of infection, fine‐tunes current bioassays, allowing more accurate and efficient selection of resistant breeding material.     

Development of a field biotest using artificial inoculation to evaluate resistance and yield effects in sugar beet cultivars against <Emphasis Type="Italic">Cercospora beticola</Emphasis>

Cercospora beticola resistance and disease yield loss relationships in sugar beet cultivars are best characterised under field conditions with heavy natural infection; this does not occur regularly under German climatic conditions. Since Cercospora resistance reduces the rate of pathogen development, high yield loss was observed in studies using artificial inoculation. Our study, therefore aimed to optimise inoculum density to obtain cultivar differentiation, which correlates to natural infection. In 2005 and 2006, field trials were carried out to determine the effect of different inoculum densities on Cercospora resistance of three sugar beet cultivars possessing variable resistance. The epidemic progress and white sugar yield loss (WSY_loss) were determined and their relationship evaluated. An optimal inoculum concentration range (between 10,000–20,000 infectious Cercospora units ml⁻¹ inoculum suspension) was determined which allowed maximum resistance parameter differentiation in terms of C. beticola disease severity (DS), area under the disease progress curve (AUDPC) and WSY_loss. The correlation between AUDPC and WSY_loss was identical for all cultivars independent of the resistance level, demonstrating that tolerant reactions of the cultivars under study were not detectable. This study provides evidence that even under optimal inoculum levels necessary to obtain maximum differentiation between cultivars, climatic conditions are important for disease management, but remain unpredictable, indicating that artificial inoculation needs to be optimised, but that single field locations are not sufficient and reliable to evaluate Cercospora resistance.     

河北省番茄黄化曲叶病毒和番茄褪绿病毒复合侵染及分布

 番茄黄化曲叶病毒(Tomato yellow leaf curl virus, TYLCV) 及番茄褪绿病毒(Tomato chlorosis virus, ToCV)是危害番茄的主要病毒。2016~2017年,在河北省18个番茄主产区调查番茄病毒病危害情况,采集262份植株矮化、叶片黄化、褪绿、卷曲的番茄样品,利用分子生物学技术对病原进行鉴定。结果表明:2016~2017年河北省番茄病毒病发生普遍,所检样品中TYLCV的侵染率为68.66%,其中与ToCV复合侵染率为19.5%;除栾城、滦南、乐亭、永年4地样品暂未检测到ToCV外,其他14个采样点的样品均检测到ToCV,侵染率为19.5%,且全部表现为与TYLCV复合侵染,河北省番茄主产区暂未发现ToCV单独侵染的样品。     

Impact of the potato inoculation date on potato virus Y load and viral distribution in daughter tubers at harvest

Aged plants are more difficult to infect than young plantlets. This modification of susceptibility is described as mature plant resistance (MPR). For potato virus Y (PVY), MPR is known to lead to low infection rates of plants inoculated at the postflowering stage and a decrease in the number of infected daughter tubers. However, the impact of inoculation date on the capacity of PVY to accumulate in daughter tubers has not been studied so far. Field and greenhouse experiments were carried out to better understand PVY epidemiology and to help potato growers to evaluate consequences of early/late infections on the quality of their crops. In field trials, potato plants (cv. Bintje) were covered by insectproof nets from planting to harvest except for a 14-day period to expose plants to natural PVY infections. Under controlled conditions, potato plants were mechanically inoculated with PVY at different dates from preflowering stages (early inoculations) to postflowering stage (late inoculations). At harvest, daughter tubers were individually collected and analysed to define proportions and viral load of infected tubers according to the time between virus inoculation and harvest. Our results showed that although the age of plants at the time of inoculation can modify their susceptibility to PVY infection, in return, early and late PVY inoculations lead to similar rates of infected tubers at the plant scale and equivalent viral accumulation in infected tubers. All together, these data revealed that both early/late infections are high risks for the sanitary quality of potato tubers.