Occurrence of viruses in irrigated wheat in Zambia

Surveys were conducted during the cool-dry months of June–August 1997 and June–July 1998 for the presence of viruses in irrigated wheat in Central, Copperbelt, Lusaka and Southern Provinces of Zambia in 14 commercial farms and four wheat cultivar plots. Virus symptoms were observed on nine wheat cultivars ( Triticum aestivum 'Deka', 'Gamtoos', 'Lorie II', 'MM2', 'Nata', 'Nkwazi', 'P7', 'Scan' and 'Sceptre') of South African, Zambian and Zimbabwean origin. Several viruses were identified on the basis of field symptomatology, symptoms developing on mechanically inoculated indicator plant species or cultivars and serology (DAS-ELISA). The study revealed the occurrence of Brome mosaic virus (BMV), Barley stripe mosaic virus (BSMV), Barley yellow dwarf virus and its strains (BYDV-PAV and RPV), Soil-borne wheat mosaic virus (SBWMV), Wheat dwarf virus (WDV), Wheat streak mosaic virus (WSMV) and Wheat spindle streak mosaic virus (WSSMV). DSA-ELISA tests confirmed these identifications. The prevalence of viruses varied annually and from field to field. BSMV, BYDV-PAV, SBWMV, WDV, WSMV and WSSMV were found to be the most prevalent viruses. Viruses generally occurred in mixed infections of 3–6 viruses and the most common virus complex consisted of 4 viruses (50%), viz. BYDV, SBWMV, WDV and WSSMV. Five- and six-virus complexes were relatively less common (20% each) whereas 3-virus complex was noticed in only 10% cases. SBWMV and WSSMV have been found to be new to Africa and Zambia and are reportedly vectored by a fungal protist – Polymyxa graminis . BYDV strains MAV and SGV were also tested but gave negative results against their antisera.     

Using air pressure cells to evaluate the effect of soil environment on the transmission of soilborne viruses of wheat

ABSTRACT An air pressure cell, a laboratory tool that precisely controls soil matric potential, was utilized in a novel approach to investigate the epidemiology and management of soilborne disease. Matric potentials of -1, -5, -20, and -40 kPa were established in cores of field soil infested with Wheat soilborne mosaic virus (WSBMV) and its presumed vector Polymyxa graminis. Equilibrated soil cores were planted to wheat (Triticum aestivum), and after intervals of growth under controlled environment, virus transmission was assessed by serological detection of the virus in washed roots. Transmission occurred at all but the driest soil matric potential tested, -40 kPa, in which only pores with a diameter of 7.4 mum or less were water-filled, possibly obstructing movement of P. graminis zoospores. By starting plants at -40 kPa for 10.5 days and then watering them to conducive matric potential, we found that WSBMV transmission occurred between 12 to 24 h at 15 degrees C, and within 36 h at 20 degrees C. No significant transmission occurred within 96 h at 6.5 degrees C. In contrast, transmission of Wheat spindle streak mosaic virus (WSSMV) did not occur at 15 degrees C (the only transmission temperature tested), suggesting either that WSSMV is unable to establish infection at 15 degrees C or that a different vector is involved. The air pressure cell is a novel tool with many potential applications in research on the epidemiology and management of soilborne pathogens. Applications of the precise environmental control attained through the use of air pressure cells range from assessing the effects of cultural practices on soilborne inoculum to standardized virulence assays for soilborne pathogens to preliminary screens of host resistance and pesticide efficacy.     

A novel putative member of the family Benyviridae is associated with soilborne wheat mosaic disease in Brazil

Soilborne wheat mosaic disease (SBWMD), originally attributed to infections by Soilborne wheat mosaic virus (SBWMV) and Wheat spindle streak mosaic virus (WSSMV), is one of the most frequent virus diseases and causes economic losses in wheat in southern Brazil. This study aimed to characterize molecularly the viral species associated with wheat plants showing mosaic symptoms in Brazil. Wheat leaves and stems displaying mosaic symptoms were collected from different wheat cultivars in Passo Fundo municipality, Rio Grande do Sul State, southern Brazil. Double-stranded RNA was extracted and submitted to cDNA library synthesis and next-generation sequencing. No sequences of SBWMV and WSSMV were detected but the complete genome sequence of a putative new member of the family Benyviridae was determined, for which the name wheat stripe mosaic virus (WhSMV) is proposed. WhSMV has a bipartite genome with RNA 1 and RNA 2 organization similar to that of viruses belonging to Benyviridae. WhSMV RNA 1 has a single open reading frame (ORF) encoding a polyprotein with putative viral replicase function. WhSMV RNA 2 has six ORFs encoding the coat protein, the major protein (read-through), triple gene block movement proteins (TGB 1, 2 and 3) and ORF 6 (hypothetical protein). In addition to the genomic organization and nucleotide and amino acid sequence identities, phylogenetic analyses also corroborated that WhSMV is a virus species of the Benyviridae. However, isolates of WhSMV formed a clade distinct from members of the genus Benyvirus. It was also demonstrated that the plasmodiophorid Polymyxa graminis is associated with wheat roots showing SBWMD symptoms and infected by WhSMV.     

Untersuchungen zum Infektionsverlauf und zur biologischen Differenzierung von bodenbürtigen Viren in Roggen, Triticale und Weizen

Abstract In the frame of the investigation of epidemiology of soil-borne viruses, like the Soil-borne cereal mosaic virus (SBCMV), Soil-borne wheat mosaic virus (SBWMV) and the Bymovirus Wheat spindle streak mosaic virus (WSSMV), which were transmitted by fungal vector Polymyxa graminis Ledingham, the infection progress in different cereals was observed. The detection of furovirus and bymovirus in field plants was depending on temperature conditions during the vegetation period and the kind of cereals. The furoviruses tolerate a broad temperature spectrum and once established infection is detectable until the harvest time. In contrast to this observation, the propagation of WSSMV seems to be restricted to lower temperatures. Consequently, this virus is detected best at the end of February until the middle of April. Among the tested cereals, rye becomes more early infected than wheat and triticale. Both furoviruses could be differed by variable virulence reactions on cereal hosts and indicator plants. The SBCMV infects rye, triticale and wheat but not barley. The SBWMV is able to contaminate beside these cultures barley too. Both viruses are distinguished in the infection typ in Nicotiana benthamiana. Whereas SBCMV isolates spread out in the whole plant and cause yellowing and the die back of plants, the SBWMV infects the inoculated leaves only.     

Unterschiedliche Formen der Resistenz gegen bodenbürtige Viren des Weizens

In Germany the furovirus Soil-borne cereal mosaic virus (SBCMV) and the bymovirus Wheat spindle streak mosaic virus (WSSMV) occur often together particularly in several rye production areas. Soil-borne wheat mosaic virus (SBWMV), a wheat infecting furovirus, has so far been found only in one field near Heidelberg. Each of these viruses is transmitted by Polymyxa graminis. The cultivation of resistant varieties is the only promising measure to prevent yield losses caused by soil-borne viruses. Resistance of wheat against the bymovirus WSSMV is comparable to the immunity of barley to the bymoviruses Barley yellow mosaic virus and Barley mild mosaic virus. In case of immunity no virus multiplication is observed in resistant cultivars. In contrast, all wheat cultivars are hosts of the furoviruses. All cultivars – including the resistant ones – can be infected following mechanical inoculation with SBWMV and SBCMV. Resistance to furoviruses is based on reduced levels of virus multiplication in roots and on inhibition of virus movement from roots to leaves. Because of the inhibited virus movement from roots to aerial parts of plants this type of resistance is referred to as translocation resistance. In spite of the different resistance mechanisms the absence of virus symptoms on the leaves is a common selection criterion for both immunity and translocation resistance. Therefore, the symptom free development of plants on uniformly infested fields is the best criterion for selecting wheat lines with resistance to soil-borne viruses. The limited suitability of other selection methods is discussed.     

Response of Hard Red Winter Wheat to Soilborne wheat mosaic virus Using Novel Inoculation Methods

ABSTRACT Soilborne wheat mosaic virus (SBWMV) is an agronomically important pathogen of wheat that is transmitted by the soilborne plasmodiophorid vector Polymyxa graminis. In the laboratory, attempts to generate SBWMV-infected plants are often hampered by poor infectivity of the virus. To analyze the mechanism for virus resistance in wheat cultivars, we developed novel inoculation techniques. A new technique for foliar inoculation of SBWMV was developed that eliminated wound-induced necrosis normally associated with rub inoculating virus to wheat leaves. This new technique is important because we can now uniformly inoculate plants in the laboratory for studies of host resistance mechanisms in the inoculated leaf. Additionally, wheat plants were grown hydroponically in seed germination pouches and their roots were inoculated with SBWMV either by placing P. graminis-infested root material in the pouch or by mechanically inoculating the roots with purified virus. The susceptibility of one SBWMV susceptible and three field resistant wheat cultivars were analyzed following inoculation of plants using these novel inoculation techniques or the conventional inoculation technique of growing plants in P. graminis-infested soil. The results presented in this study suggest that virus resistance in wheat likely functions in the roots to block virus infection.     

安徽省来安县小麦梭条花叶病的病原鉴定和防治

近年来安徽省来安具发生一种呈典型梭条花叶病症状的小麦病害,用病叶进行一系列抽提,制得病原初提纯制剂。经紫外扫描测定为标准的核蛋白吸收曲线,最大吸收值在260nm处,A260/A280为1.27,Amax/Amin为1.16。电镜观察病原为线状病毒颗粒,宽12—13nm,长度分布在100nm—600nm之间,1000nm以上颗粒所占比例较少。提纯病毒制剂经SDS—聚丙烯酰胺凝胶电泳,可见到36.8kd的模糊蛋白带,但最易出现的是29kd和27kd二条明显的带。病原与小麦梭条花叶病毒抗血清起阳性反应。仪发现侵染小麦,并通过土壤传播,最后鉴定该病病原为小麦梭条花叶病毒,并对该毒源病毒外壳蛋白组分易于降解成较小的产物和不同分离物致病力关系提出讨论。同时在来安引种3165等抗病良种并结合其它措施进行防治,取得了明显的防治效果。     

小麦植株内可溶性糖含量与对梭条斑花叶病毒抗性的关系

比较了４个对小麦梭条斑花叶病毒具不同抗性的小麦品种，在接种与不接种（ＣＫ）处理条件下，叶片和根系中可溶性糖含量。结果表明，无论接种与否感病品种８１－５和８２０１－６９的可溶性糖含量比抗病品种ＳＫ－４和ＳＮ７８５９的高；接种植株比不接种植株高；８１－５和８２０１－６９升高率比ＳＫ－４和ＳＮ７８５９的高；接种或不接种根系可溶性糖含量与接种根系中介体侵入繁殖量均呈显著正相关。     

Molecular characterization and detection of European isolates of Soil-borne wheat mosaic virus

Sequencing of a recently identified isolate of Soil-borne wheat mosaic virus (SBWMV) from the UK confirmed its identity as a European strain of the species and provided further evidence for taxonomic divisions in the group. Two RT–PCR protocols were developed for the detection of all SBWMV strains and for the specific detection of the European SBWMV strain, and were tested successfully on 21 isolates of SBWMV from a range of countries. Both protocols worked well using either purified total RNA in one- or two-step RT-PCR, or immunocapture (IC) RT–PCR. The sensitivity of IC RT-PCR was 100 times greater than ELISA. Neither set of primers produced any PCR product with either Wheat spindle streak mosaic virus or Wheat yellow mosaic virus which are frequently associated with SBWMV, or with the related viruses Indian peanut clump virus , Potato mop-top virus , Beet soil-borne virus and Beet necrotic yellow vein virus . This new diagnostic protocol will improve disease management by enabling correct identification of the causal pathogen and earlier detection than is possible serologically.     

中国小麦黄花叶病毒(WYMV)分布的RT-PCR鉴定

 镰刀菌产生的单端孢霉烯毒素,包括DON、DAS和T-2毒素,是一类重要的植物病原真菌毒素。这类毒素不仅加强病菌的为害程度,而且人畜食用罹病谷物及其制成品易造成中毒。近年来,单端孢霉烯毒素的生物合成途径、关键酶及其基因克隆,毒素在致病过程中的作用,以及利用毒素作为选择压鉴定小麦抗病性和筛选抗病突变体均取得了明显进展。本文对此作一综述。     

Molecular comparisons amongst wheat bymovirus isolates from Asia, North America and Europe

To study the variation between wheat bymovirus isolates and to resolve uncertainties about the identity of the virus in some countries, leaves of infected plants were obtained from nine sites in China and from one each in Italy, Germany, USA and Canada. The German isolate was obtained from rye and the Canadian isolate was the type strain of wheat spindle streak mosaic virus (WSSMV). In RT-PCR, using primers designed from a partial sequence of a French isolate (tentatively described as WSSMV), genome fragments were obtained from the Italian and the French isolates but not from the Chinese ones. Conversely, products were consistently obtained from the Chinese isolates, but not from the Italian or French ones, when primers were designed from the sequence of a Japanese isolate of wheat yellow mosaic virus (WYMV). Nucleotide sequences were obtained from regions at or near the 3'-terminus of RNA1 of six Chinese isolates and the four from Europe and North America, usually including the coat protein. Nucleotide and amino acid sequence comparisons demonstrated that the European and North American isolates were extremely similar and were therefore WSSMV, while the Chinese isolates were close to the Japanese isolate and were thus WYMV.     

Response of UK winter wheat cultivars to <Emphasis Type="Italic">Soil-borne cereal mosaic</Emphasis> and <Emphasis Type="Italic">Wheat spindle streak mosaic viruses</Emphasis> across Europe.

Twenty-one UK winter wheat cultivars were grown over three seasons at sites with natural inoculum sources of Soil-borne cereal mosaic virus (SBCMV) and Wheat spindle streak mosaic virus (WSSMV) located in France, Italy and the UK. Plants were assessed visually for virus symptoms and leaf extracts were tested for the presence of each virus using enzyme-linked immunosorbent assays (ELISA). Cultivars showing little or no foliar symptoms and low levels of virus in leaf tissue were classified as resistant to each virus. All the trials were taken to harvest and agronomic data collected. At the most heavily infected sites, severe symptoms of SBCMV were observed in all UK cultivars except Aardvark, Charger, Claire, Cockpit, Hereward and Xi 19. The latter cultivars exhibited either light or no symptoms and little or no SBCMV infection in leaves. In fields with WSSMV, the virus failed to develop in Italy, but was detected in the leaves of all the susceptible control cultivars at a site in France. However, no UK cultivar tested positive for WSSMV. Multi-site analysis indicated that the presence of WSSMV did not increase the impact of SBCMV on the height, thousand-grain weight or yield of UK cultivars. The wheat cultivars on test gave a similar response to SBCMV across three European countries. Possible sources of SBCMV resistance are discussed.     

Genome-wide association analysis of resistance to wheat spindle streak mosaic virus in bread wheat

Wheat spindle streak mosaic virus (WSSMV) is a major concern for cereal crops in Europe and North America. A strong increase in the occurrence of WSSMV has been observed in each French region where susceptible cultivars are cultivated. Most European bread wheat cultivars are resistant, but assessing the status of newly registered cultivars or breeding lines regarding WSSMV resistance is of importance. This paper describes a genome-wide association study carried out on a panel of 163 cultivars and tested for their resistance to WSSMV. Two regions on chromosomes 5B and 7D showed minor effects on WSSMV resistance. More importantly, a large genomic region on chromosome 2D explained most of the resistance to WSSMV. More than 99% of the cultivars carrying the AA genotype at the most associated marker (Excalibur_c15426_661) were resistant to WSSMV, while 100% of the cultivars showing the GG genotype were susceptible. This large genomic region of 45.8 Mb was found distal to the centromere and showed very high linkage disequilibrium. It is hypothesized that this region may be an alien introgression originating from a wild related species. This region contains a total of 2605 predicted genes based on the Chinese Spring IWGSC RefSeq v. 1.0 including genes potentially involved in plant disease resistance. A kompetitive allele-specific PCR (KASP) single-nucleotide polymorphism (SNP) marker was designed in order to identify breeding lines or registered cultivars resistant to WSSMV.     

小麦梭条斑花叶病毒(WSSMV)和小麦黄花叶病毒(WYMV)的细胞质内含体的电镜观察

 小麦梭条斑花叶病毒(WSSMV)和小麦黄花叶病毒(WYMV)分别感染小麦(Triticum aestivum L.)后所诱发的细胞质内含体和细胞嚣病变几乎完全相同。感病的叶肉细胞、表皮细胞和木质部薄壁细胞的细胞质中都可看到大量风轮体、柱状体、板状集结体和膜状体,其中以叶肉细胞中更为普遍。风轮体成群出现、同一组内风轮体常平行排列,而相邻群间常垂直排列。膜状体的内部常夹杂着许多小囊泡,其四周与内质网呈辐射状相连。在发病初期的心叶叶肉细胞质的液泡附近可看到颗粒状病毒束,而分散的病毒状颗粒则多见于发病中后期坏死的细胞中。感病细胞的叶绿体、线粒体、核、质膜及胞间联丝中均未见病毒状颗粒。感染病毒的细胞中,细胞质丰富、内质网膨大,核糖体增加、小囊泡大量出现,随着病害的加重,叶绿体、线粒体逐渐肿大,基粒缩小、膜破裂、类囊物流入细胞中。     

小麦黄色花叶病毒不同分离物外壳蛋白(CP)基因序列的比较分析

 核苷酸序列分析结果表明,小麦黄色花叶病毒(W YMV)不同分离物的外壳蛋白基因存在一定的差异。邓州分离物CP基因在其31~33nt处均缺失了3个核苷酸,其余分离物与潢川分离物及日本分离物长度一致,均为882nt。不同分离物CP基因核苷酸序列同源性为97.3%~98.9%,由此推导的氨基酸序列同源性为97.6%~99.3%,外壳蛋白N末端的110个氨基酸和C末端的55个氨基酸在各个分离物间是高度保守的。潢川分离物有5个氨基酸与其它5个分离物明显不同。WYMV不同分离物外壳蛋白序列分析结果进一步确认了WYMV与WSSMV为Bymovirus属的2种不同病毒。     

Investigation of soilborne mosaic virus diseases transmitted by <Emphasis Type="Italic">Polymyxa graminis</Emphasis> in cereal production areas of the Anatolian part of Turkey

Polymyxa graminis is the vector of several important viruses, including Soilborne cereal mosaic virus, Wheat spindle streak mosaic virus, Barley yellow mosaic virus and Barley mild mosaic virus, of winter cereals worldwide. Surveys were carried out to detect these viruses and their vector P. graminis in 300 soil samples from the main wheat and barley production areas of the Anatolian part of Turkey collected in May 2002, June 2004 and May 2005. For these surveys, various susceptible wheat and barley cultivars were pot grown in the collected soil samples in a greenhouse and then analysed using ELISA and RT-PCR to detect the presence of different virus species. In addition, a combination of light microscopy following roots staining with acid fuchsin and PCR was used for detection of P. graminis. All soil samples analysed were found to be free of these soilborne viruses and their vector.     

发生在我国的小麦黄花叶病毒病

本文对山东荣成流行的一种小麦病毒病进行了鉴定。提纯的病毒颗粒为长线状,13×100—300nm及13×350—650nm。汁液接种感染小麦,但不感染烟草、苋色藜等植物。病土、病根以及含有禾谷类多粘菌(POlymyxa graminis)游动孢子的浸液可以传毒于小麦、大麦及黑麦。此病毒与大麦黄花叶病毒(BYMV)、小麦梭条花叶病毒(WSSMV)有血清学关系,与小麦土传花叶病毒(WSBMV)无血清学关系。病叶表皮细胞中有无定形内含体。超薄切片可见风轮状内含体。实验结果表明,荣城地区发生的这种小麦病毒病是小麦黄花叶病毒所致。     

Dynamics of wheat spindle streak mosaic bymovirus in winter wheat

The dynamics of wheat spindle streak mosaic bymovirus in winter wheat were studied during two crop cycles in a field site with a history of high virus incidence. Individual plants of two susceptible cultivars were sampled from autumn to spring and the presence of virus antigen in roots and leaves was determined by ELISA. Virus incidence was higher in cv. Frankenmuth than in cv. Augusta. During year one, incidence of viral antigen in roots remained very low for four months after sowing, and did not reach maximum levels until the following spring. During year two, incidence of viral antigen in roots rose to maximum levels in autumn, only three months after sowing. These results strongly suggested that root infection occurred in spring as well as in autumn. In both cultivars and in both years, we detected the virus in roots one month prior to its detection in leaves, suggesting that virus moves slowly from roots into leaves. Maximum incidence of virus in leaves occurred in spring of both years, coinciding with the period of symptom development. Typical symptoms (yellow streaks, spindles, and mosaic) were observed in year two, whereas only mild mosaic was observed in year one. Virus antigen was detected in nonsymptomatic leaves from two months after sowing through crop senescence. Because antigen could be detected in roots throughout the crop cycle, and zoosporangia and cystosori of the fungal vector could be detected one and two months, respectively, after sowing, it is possible that wheat spindle streak mosaic bymovirus is acquired and/or spread by the vector during the majority of the crop cycle.