Occurrence and distribution of Raspberry bushy dwarf virus in commercial Rubus plantations in England and Wales

Serological surveys for Raspberry bushy dwarf virus (RBDV) made between 1995 and 1997 and covering ≈ 10% of the commercial farms growing Rubus (red raspberry and hybrid berries) in England and Wales showed that this virus was present on approximately one-quarter of all farms and in approximately one-sixth of all plots tested. RBDV was found in all of the four main raspberry cultivars being grown at that time (Autumn Bliss, Glen Moy, Glen Prosen and Leo), in Loganberry and in Tayberry. Fifteen RBDV genotypes (including two that appeared to be mixed) were identified using RT-PCR/RFLPs, but the majority of genotypes were found only rarely. Of the RBDV isolates tested, two genotypes each comprised 12·5% and another 46·4%. None of the three most common genotypes was associated solely with single Rubus cultivars and vice versa . It is suggested that two separate outbreaks of RBDV are occurring in England and Wales. One outbreak comprises the most frequent genotype combined with one of the moderately frequent genotypes; this outbreak is largely confined to the main growing areas and is either spreading between farms or coming from multiple local sources. Circumstantial evidence suggests that these isolates (and hence this first outbreak) are of the RB pathotype. The second outbreak consists of the other moderately frequent genotype and those genotypes which are less common. These genotypes appear to be more scattered across England and Wales and seem more likely to be coming from local sources and not to be spreading naturally between commercial farms.     

Inter-laboratory evaluation of a duplex RT-PCR method using crude extracts for the simultaneous detection of Prune dwarf virus and Prunus necrotic ringspot virus

The operational capacity of a duplex RT-PCR method for simultaneous detection of Prune dwarf virus (PDV) and Prunus necrotic ringspot virus (PNRSV) has been established by nine European laboratories. A total of 576 samples from Prunus trees with known sanitary status, corresponding to 32 samples in two repetitions for each laboratory, were analysed. The level of sensitivity achieved by the method was 98.3% for PDV and 90.4% for PNRSV. The specificity was 87.4% for PDV and 94.3% for PNRSV. The unilateral 95% confidence intervals were calculated for all these values. Cohen’s Kappa coefficient of repeatability and reproducibility of the technique indicated a strong agreement between data. Likelihood ratios were 7.50 (positive) and 0.02 (negative) for PDV. For PNRSV, the positive likelihood ratio was 15.00 while the negative likelihood ratio was 0.11. In addition, post-test probabilities of infection were calculated to manage the risk associated with the routine use of this method. This allows an accurate test result interpretation to facilitate the integration of this new technique into a certification scheme.     

番茄褪绿病毒侵染黄瓜的首次报道

番茄褪绿病毒Tomato chlorosis virus(ToCV)是严重危害世界经济作物的一种病毒,寄主范围广泛。田间调查发现黄瓜Cucumis sativus表现出叶片黄化、脉间褪绿的疑似番茄褪绿病毒感病症状,同时叶片背面聚集了大量烟粉虱。采用RT-PCR方法对样品叶片和烟粉虱进行检测,ToCV感染率为65%,且发病叶片上烟粉虱携带ToCV。为进一步确定黄瓜是否为番茄褪绿病毒的新寄主,室内利用农杆菌侵染性克隆接种健康黄瓜,结果显示:接种30 d的黄瓜新生叶片出现褪绿症状。采用ToCV HSP70基因的引物对田间黄瓜叶片、烟粉虱和室内黄瓜新生叶片进行RT-PCR,扩增出约450 bp的条带,在NCBI上BLAST显示与KC887999.1的同源性最高,为99%。这些数据表明黄瓜是番茄褪绿病毒的寄主。这是ToCV感染黄瓜的首次报道。     

The invasive weed Ventenata dubia is a host of Barley yellow dwarf virus with implications for an endangered grassland habitat

Ventenata dubia (African wiregrass), a winter annual weed, is a non‐native species invading grasslands, rangelands and pastures throughout the USA. Limited information is available on its suitability as a host to pathogens and insects in its invaded range. The barley/cereal yellow dwarf virus (B/CYDV) complex occurs ubiquitously in Poaceae species. In non‐managed grasslands, BYDV infection influences competitive dynamics between native and invasive grasses and facilitates invasion by non‐native annual weeds. The Palouse prairie of south‐eastern Washington and northern Idaho, USA, is an endangered ecosystem. Surveys of V. dubia in Palouse prairie and neighbouring Conservation Reserve Program (CRP) habitats were conducted to determine whether B/CYDV viral species are present. Laboratory tests examined the suitability of V. dubia to host BYDV‐PAV and serve as an inoculum source. Plant growth and weight parameters were measured to gauge the impact of BYDV‐PAV on V. dubia. Infection of V. dubia in Palouse prairie and CRP habitats with two species of BYDV, PAV and SGV, was detected for the first time. The ability of BYDV‐PAV to infect V. dubia in the laboratory and transmission from infected V. dubia to barley were demonstrated. BYDV‐PAV‐infected V. dubia showed reductions in plant height, number of leaves and tillers per plant, and above‐ground dry weight, suggesting that V. dubia is sensitive to BYDV. Results demonstrate that V. dubia is a host to BYDV and may serve as a virus inoculum source with potential implications for its management, competitive dynamics between invasive and native grasses and future conservation of endangered grasslands.     

New experimental hosts of Barley yellow dwarf virus among wild grasses,with implications for grassland habitats

Barley yellow dwarf virus (BYDV), an economically important virus, infects small grain cereal crops and over 150 other Poaceae species. BYDV infection plays an important role in competition among grasses in non‐managed systems, but many grasses remain unexamined as potential BYDV hosts. This study examined grass species that have not been reported as BYDV hosts but are commonly encountered in non‐managed grasslands throughout the United States and Canada. Laboratory inoculations with BYDV‐PAV using the aphid vector Rhopalosiphum padi were performed to examine the ability of 13 grass species and barley to be infected with the virus; eight of the grass species were not documented previously as virus hosts. Serological and molecular assays were used to confirm BYDV‐PAV infection. Plant height, number of leaves, number of tillers and weight were recorded to evaluate susceptibility or sensitivity to BYDV. Infection with BYDV was experimentally achieved for the first time on Achnatherum lettermanii, Achnatherum occidentale, Achnatherum thurberianum, Danthonia intermedia, Poa fendleriana, Sporobolus airoides and Sporobolus cryptandrus, but not on Alopecurus pratensis and Elymus wawawaiensis. Infection was confirmed in Bromus inermis, Elymus elymoides, Poa bulbosa, Poa secunda and Hordeum vulgare, which served as controls. BYDV infection caused reductions in plant height on P. bulbosa and P. fendleriana. BYDV‐infected P. secunda had more leaves per plant compared to healthy plants of the same species. BYDV‐infected A. lettermanii exhibited reduced dry weight in both below‐ground and above‐ground tissue. These findings have implications for the management and conservation of grassland habitats.     

Uneven distribution of two potyviruses (feathery mottle virus and sweet potato latent virus) in sweet potato plants and its implication on virus indexing of meristem derived plants

Abstract

The distribution of two sweet potato potyviruses, FMV and SPLV, was assessed in three plants infected with both viruses and in one plant infected with FMV only. All leaves, the top and basal sections of the main stem, and branch sections were tested by ELISA. Both symptomless leaves and leaves showing symptoms including purple rings, chlorotic spots, mottle or discoloration were found to contain the viruses. However, neither could be detected in every leaf or stem piece. SPLV was found in a lower proportion of leaf and stem samples than FMV. This indicates that the two viruses are either very unevenly distributed within sweet potato plants or that the virus concentration in some parts is below the detectable level. Testing of each leaf is recommended for reliable virus indexing of small, meristem‐derived sweet potato plantlets, if the ELISA method is used. Additional indexing of all ELISA‐negative materials by grafting to susceptible indicator plants is nevertheless still necessary.     

云南水稻上检测到南方水稻黑条矮缩病毒

南方水稻黑条矮缩病(Southern rice black streaked dwarf disease)是我国南方稻区一种新的水稻病毒病害.自2001年在广东省阳西县首次发现以来,该病危害范围逐年扩大,2009年我国湖南、江西、广东、广西、海南、浙江、福建、湖北和安徽9个水稻主产省(区)及越南北部19个省发病面积约33.33万hm2,基本绝收面积0.67万hm2[1,2],对水稻生产威胁巨大.     

多枝赖草及其转育后代对大麦黄矮病毒PAV和GAV株系的抗性研究

 本文针对大麦黄矮病毒(BYDV) PAV和GAV株系,采用生物学和血清学(ELISA)相结合的鉴定方法,对多枝赖草以及它与普通小麦杂交获得的二体附加系Line24等材料进行抗性测定。首次证明了多枝赖草对我国BYDV-PAV和GAV株系免疫或高抗,而普通小麦-多枝赖草二体附加系Line24以及2个易位系高耐这2种株系。     

A domain of the readthrough protein of barley yellow dwarf virus (NY-RPV isolate) is essential for aphid transmission

Luteoviruses are obligately transmitted by aphids and contain two capsid proteins, the coat protein (CP) coded for by open reading frame (ORF) 3, and the readthrough protein (RTP), produced by readthrough of the amber termination codon of ORF 3 into the contiguous ORF 5. Previous studies have suggested that it is the RTP that determines transmissibility and vector specificity. To investigate which capsid protein or protein part contains determinants for the transmission of the NY-RPV isolate of barley yellow dwarf virus (BYDV) by its vectorRhopalosiphum padi, we produced three fusion proteins by expressing NY-RPV cDNA inE. coli. These respectively represented the CP alone (P3), a region of the RTP immediately following the amber termination codon (P5a), and the remainder of the RTP (P5b). Polyclonal antisera raised against the P3, P5a and P5b proteins each gave distinctive reactions against purified NY-RPV on Western blots. Also, in ELISA tests, antisera raised against all three fusion proteins detected purified intact virions. When mixed with purified virions and fed toR. padi through Parafilm membranes, immunoglobulins (Igs) from antisera raised against P3 and P5b had no effect on transmission, whereas Ig from antiserum against P5a interfered with transmission. P5a antiserum Ig had no effect on the transmission of the P-PAV isolate of BYDV byR. padi. The results demonstrate that while neither the CP itself nor the terminal region of the RTP are key determinants for transmission, a specific domain in the central part of the RTP is an important determinant in the transmission of NY-RPV byR. padi, though apparently not of P-PAV.     

Genetic resistance for the sustainable control of plant virus diseases: breeding,mechanisms and durability

Plant viruses are important agricultural pathogens, and are responsible for a significant number of commercially relevant plant diseases. There are very few efficient control measures for viral diseases, but the use of genetic resistance appears to be the most promising strategy, often conferring effective protection without additional costs or labour during the growing season, and without damaging the environment. Sources of virus resistance have been identified for most crop species, and many resistant cultivars are already commercially available and of widespread cultivation; however, much remains to be learned about genetic resistance. This review article considers three main aspects that require intense investigation. First, we review the identification of sources of resistance and how plant breeders and pathologists have focused on aspects of the breeding process particularly relevant to viruses, such as germplasm screening and the dissection of resistance phenotypes. Second, we review how molecular mechanisms controlling resistance have been unravelled, looking at case studies where resistance mechanisms are now understood in detail for each stage of the infection cycle. Third, we turn to the durability of resistance in a global context, examining factors that influence durability and how this can be predicted. We conclude with a short discussion of the technological and scientific opportunities provided by recent advances in the field.     

Two Hungarian isolates of cucumber mosaic virus from sweet pepper (Capsicum annuum) and melon (Cucumis melo): Identification and antiserum preparation

Two Hungarian virus isolates from sweet pepper (K8) and melon (S4) were identified as cucumber mosaic virus (CMV) on the basis of host plant reactions and serology. The isolates were purified and antisera prepared. Homologous antiserum titers in double-diffusion tests were 256 (K8) and 512 (S4). They were serologically closely related to each other and to other CMV isolates. On the basis of symptoms they belong to different symptomatological groups of CMV; this was supported by serological properties. Sedimentation coefficients were c. 93 S, at 2 mg ml^–1. Purified preparations, stained with 2% uranyl acetate, showed spherical particles. In ELISA purified preparations reacted with each other's antisera.Samenvatting Twee hongaarse virusisolaten uit paprika (K8) en meloen (S4) werden geïdentificeerd als komkommermozaïekvirus (CMV) met behulp van toetsplanten en serologie. Beide isolaten werden gezuiverd en er werden antisera tegen bereid. De homologe titers van de antisera in de agar-geldiffusietoets bedroegen 256 (K8) en 512 (S4). K8 en S4 waren serologisch nauw verwant aan elkaar, evenals aan andere CMV-isolaten. Op grond van hun symptomen op toetsplanten behoren ze tot verschillende symptomatologische groepen van CMV. Dit laatste werd gesteund door de serologische eigenschappen. Beide isolaten hebben een sedimentatiecoëfficiënt van ca 93 S, bij een concentratie van 2 mg ml^–1. Gezuiverde preparaten, gekleurd met 2% uranylacetaat, bleken bolvormige deeltjes te bevatten. In ELISA reageerden gezuiverde preparaten van K8 en S4 met elkaars antisera.     

Systemic spread of Plum pox virus (PPV) in Mariana plum GF 8-1 in relation to shoot growth

Infection of Prunus spp. by Plum pox virus (PPV) is characterized by an uneven distribution of the virus within the tree and branches. In order to gain a better understanding of this distribution, a method for modelling tree growth was used. PPV spread was followed within susceptible Mariana plum clone GF 8-1 shoots for 4 months after inoculation. Shoot growth was unaffected by the presence of the virus. Symptoms appeared on leaves produced in the most actively growing parts of the shoots, i.e. at the beginning of the season. PPV was detected in leaves other than those showing symptoms. The proportion of leaves with detectable virus decreased from the zone showing symptoms, with 100% ELISA-positive responses, to the shoot tip with no detectable virus in leaves produced between 111 and 127 days after inoculation. Furthermore, a higher proportion of positive ELISA results was obtained below the zone showing symptoms (77%) compared with 50% above. PPV was detected in 95% of the most vigorous shoots 71 days after inoculation compared with 37% of slower-growing, later-produced shoots.     

A greenhouse test for screening sugar-beet (Beta vulgaris) for resistance to beet necrotic yellow vein virus (BNYVV)

Small differences in activity between batches of purified beet necrotic yellow vein virus (BNYVV) were observed in ELISA. A four-parameter modelled dose-response curve of purified BNYVV was used for the conversion of ELISA values to virus concentrations. Seedlings of the susceptible cultivar Regina and the partially resistant cultivars Nymphe and Rima were tested for resistance to BNYVV in a mixture of sand and infested soil. Plants were grown in a green-house with low nutrient supply and at temperatures below the optimum of both the vectorPolymyxa betae and BNYVV. Root systems were small and consisted mainly of lateral roots. Significant differences in average virus concentrations were found between cultivars, both when using the complete root systems and when using either the top or the bottom part of the root systems. Average virus concentrations in Regina were always significantly higher than in Rima and higher than in Nymphe on all occasions except one (P<0.05). Differences between Nymphe and Rima were less evident. Variation between plants was greatest within Rima. The test described in this paper can be used for the discrimination of different cultivars and for the identification of individual plants with resistance to BNYVV.     

Potato leafroll virus (PLRV): its transmission and control

This review provides a wide assessment of the present state of knowledge about the potato leafroll luteovirus (PLRV), a pathogen which is seriously devastating potato crops in many parts of the world. The main biological, physical and chemical properties of this virus are described. The transmission of PLRV by aphids, which are the only transmitters of this virus under natural conditions, is characterized. Special attention is given to the control of PLRV through the use of resistant potato cultivars. Recent advances in obtaining resistant transgenic plants are outlined.     

Broad bean mottle virus: Identification,host range,serology, and occurrence on faba bean (Vicia faba) in West Asia and North Africa

One of the faba bean viruses found in West Asia and North Africa was identified as broad bean mottle virus (BBMV) by host reactions, particle morphology and size, serology, and granular, often vesiculated cytoplasmic inclusions. Detailed research on four isolates, one each from Morocco, Tunisia, Sudan and Syria, provided new information on the virus.The isolates, though indistinguishable in ELISA or gel-diffusion tests, differed slightly in host range and symptoms. Twenty-one species (12 legumes and 9 non-legumes) out of 27 tested were systemically infected, and 14 of these by all four isolates. Infection in several species was symptomless, but major legumes such as chickpea, lentil and especially pea, suffered severely from infection. All 23 genotypes of faba bean, 2 of chickpea, 4 of lentil, 11 out of 21 ofPhaseolus bean, and 16 out of 17 of pea were systemically sensitive to the virus. Twelve plant species were found to be new potential hosts and cucumber a new local-lesion test plant of the virus.BBMV particles occurred in faba bean plants in very high concentrations and seed transmission in this species (1.37%) was confirmed.An isolate from Syria was purified and two antisera were produced, one of which was used in ELISA to detect BBMV in faba bean field samples. Two hundred and three out of the 789 samples with symptoms suggestive of virus infection collected in 1985, 1986 and 1987, were found infected with BBMV: 4 out of 70 (4/70) tested samples from Egypt, 0/44 from Lebanon, 1/15 from Morocco, 46/254 from Sudan, 72/269 from Syria and 80/137 from Tunisia. This is the first report on its occurrence in Egypt, Syria and Tunisia. The virus is a potential threat to crop improvement in the region.Samenvatting Eén van de in West-Azië en Noord-Afrika in faba-boon aangetroffen virussen werd geïdentificeerd als het tuinbonevlekkenvirus (broad bean mottle virus) op grond van waardplantreacties, deeltjesvorm en-grootte, serologische eigenschappen en granulaire, vaak gevacuoliseerde celinsluitsels. Verder onderzoek aan vier isolaten uit respectievelijk Marokko, Tunesië, Soedan en Syrië verschafte nieuwe informatie, over het virus.De in ELISA of gel-diffusietoetsen serologisch niet te onderscheiden isolaten verschilden enigszins in waardplantenreeks en symptomen. Van 27 getoetste plantesoorten werden 21 systemisch geïnfecteerd (12 vlinderbloemigen, en 9 niet-vlinderbloemigen) waarvan 14 door alle vier isolaten. In vele ervan was de infectie symptoomloos, maar belangrijke als gewas geteelde vlinderbloemigen, zoals erwt, linzen en kekererwt, leden ernstig onder aantasting. Alle 23 getoetste faba-boongenotypen, beide van kekererwt, alle vier van linzen, 11 van de 21 getoetste vanPhaseolus-boon en 16 van de 17 van erwt bleken systemisch gevoelig voor het virus. Twaalf plantesoorten, bleken nieuwe potentiële waardplanten en komkommer een nieuwe lokale-lesietoetsplant voor het virus te zijn.In faba-boneplanten kwam, het virus in hoge concentratie voor en overdracht met zaad (1.37%) in deze soort kon worden bevestigd.Een Syrisch isolaat werd gezuiverd en twee antisera werden bereid, waarvan één werd gebruikt voor de detectie van het virus in te velde verzamelde monsters. Van 789 in 1985 tot en met 1987 verzamelde bladmonsters, met symptomen die deden denken aan virusinfectie, bleken 203 het virus te bevatten en wel 4 van de 70 (4/70) uit Egypte, 0/44 uit Libanon, 1/15 uit Marokko, 46/254 uit Soedan, 72/269 uit Syrië en 80/137 uit Tunesië. Het virus was nog niet eerder aangetoond in Egypte, Syrië en Tunesië.De grote verbreiding, grote kunstmatige waardplantenreeks, overdracht met zaad, en pathogeniteit voor een aantal belangrijke vlinderbloemige gewassen maken het virus tot een potentiële bedreiging van de programma's tot verbetering van de teelt van de bedoelde gewassen in het betrokken gebied.     

Mungbean yellow mosaic virus (MYMV): a threat to green gram (Vigna radiata) production in Asia

Mungbean yellow mosaic virus (MYMV) disease is one of the most vicious diseases of green gram and has been renowned in India for more than five decades. It is caused by a group of geminiviruses belonging to the genus, begomovirus of the family, Geminiviridae. They are transmitted through whitefly in a persistent manner. The economic losses due to this virus account up to 85% in green gram which is spreading faster towards newer areas. The escalating economic importance of MYMV has resulted in the call for accurate detection and identification procedures that inspire rigorous research efforts focussing on the biology, diversity and epidemiology of the virus, so that viable management strategies could be designed. Breeding for resistance or tolerance appears to be the best approach to control this disease. However, the commercially offered genotypes are only partially resistant. Therefore, the hunt for newer sources of disease resistance needs to be intensified. This review updates all the accessible information on MYMV and outlines the areas in which advance research is indispensable.     

小麦矮缩病毒NASH快速检测方法的建立及应用

以非放射性物质地高辛为标记物,采用PCR法制备了特异性强、灵敏度高的DNA探针。通过优化反应体系,建立了小麦矮缩病毒(Wheat dwarf virus,WDV)的核酸斑点杂交(nucleic acid spot hybridization,NASH)快速检测技术体系。该方法诊断准确率高,操作简单,周期短,整个检测过程仅需5h左右。利用建立的NASH技术开展WDV流行学调查,发现近年来WDV在我国陕西韩城、山西太原和河北石家庄等地区点片发生,没有大面积暴发成灾。