Relationships of <Emphasis Type="Italic">Barley yellow dwarf virus</Emphasis>-PAV and <Emphasis Type="Italic">Cereal yellow dwarf virus</Emphasis>-RPV from Iran with viruses of the family <Emphasis Type="Italic">Luteoviridae</Emphasis>

Barley yellow dwarf disease is one of the most important problems confronting cereal production in Iran. Barley yellow dwarf virus-PAV (BYDV-PAV) and Cereal yellow dwarf virus-RPV (CYDV-RPV) are the predominant viruses associated with the disease. One isolate of BYDV-PAV from wheat (PAV-IR) and one isolate of CYDV-RPV from barley (RPV-IR) were selected for molecular characterisations. A genome segment of each isolate was amplified by PCR. The PAV-IR fragment (1264 nt) covered a region containing partial genes for coat protein (CP), read through protein (RTP) and movement protein (MP). PAV-IR showed a high sequence identity to PAV isolates from USA, France and Japan (96–97%). In a phylogenetic analysis it was placed into PAV group I together with PAV isolates from barley and oats. The fragment of RPV-IR (719 nt) contained partial genes for CP, RTP and MP. The sequence information confirmed its identity as CYDV. However, RPV-IR showed 90–91% identity with both RPV and Cereal yellow dwarf virus-RPS (CYDV-RPS). Phylogenetic analyses suggested that it was more closely related to RPS. These data comprise the first attempt to characterise BYD-causing viruses in Iran and southwest Asia. The nucleotide sequence data reported appear in the EMBL, GenBank and DDBJ Nucleotide Sequence Databases under the accession numbers AY450425 and AY450454     

Reservoirs of plant virus disease: Occurrence of wheat dwarf virus and barley/cereal yellow dwarf viruses in Sweden

Non-crop plants such as grasses and volunteer plants are an inseparable part of the flora of crop fields and can influence virus incidence in crop plants. The presence of grasses as virus reservoirs can lead to a higher probability of virus incidence in crop plants. However, the role of reservoirs as an inoculum source in agricultural fields has not been well studied for many viral diseases of crops. Grasses have been found to constitute potential reservoirs for cereal-infecting viruses in different parts of the world. This study revealed that cereal-infecting viruses such as wheat dwarf virus (WDV), barley yellow dwarf viruses (BYDVs), and cereal yellow dwarf virus-RPV (CYDV-RPV) can be found among ryegrass growing in or around winter wheat fields. Phylogenetic analysis showed that a WDV isolate from ryegrass was a typical WDV-E isolate that infects wheat. Similarly, a ryegrass isolate of barley yellow dwarf virus-PAV (BYDV-PAV) grouped in a clade together with other BYDV-PAV isolates. Inoculation experiments under greenhouse conditions confirmed that annual ryegrass of various genotypes can be infected with WDV to a very low titre. Moreover, leafhoppers were able to acquire WDV from infected ryegrass plants, despite the low titre, and transmit the virus to wheat, resulting in symptoms. Information from the grass reservoir may contribute to improving strategies for controlling plant virus outbreaks in the field. Knowledge of the likely levels of virus in potential reservoir plants can be used to inform decisions on insect vector control strategies and may help to prevent virus disease outbreaks in the future.     

The Basis for Thinopyrum-Derived Resistance to Cereal yellow dwarf virus

ABSTRACT Incorporation of Thinopyrum intermedium-derived resistance genes into improved wheat germ plasm generated a wheat substitution line (P29) which is completely resistant to Cereal yellow dwarf virus (CYDV). The undetectable CYDV titer in P29 led many to conclude that resistance prevented viral replication. To determine whether CYDV replication or movement is inhibited, we examined inoculated leaves for replication and uninoculated leaves for systemic spread. CYDV subgenomic RNA, produced only during replication, was found within the inoculated area of P29 and T. intermedium leaves, demonstrating that viral replication occurred. Absence of CYDV from uninoculated, newly emerging leaves of inoculated P29 and T. intermedium plants indicated resistance via inhibition of viral systemic infection. Resistance was not effective if P29 was inoculated with 50 to 100 viruliferous aphids per plant at the first-leaf stage or younger, resulting in a systemic spread of CYDV. As these infected P29 seedlings continued to grow, the resistance phenotype was recovered. Our data suggested that T. intermedium-derived resistance to CYDV was primarily dosage dependent and could be developmentally regulated if the amount of inoculum was large enough.     

Wheat cultivars and natural-based substances: Impacts on epidemiological parameters of yellow dwarf disease

For many years, control methods against aphid-transmitted barley/cereal yellow dwarf viruses (B/CYDV) in cereal fields were mainly based on the use of neonicotinoid (NNI)-coated seeds. The ban of NNI by the EU in 2018 has reinforced the interest of the scientific community in the characterization of genetic resources and biological protections to find alternatives to chemicals for the management of B/CYDV. Eleven BYDV-susceptible wheat varieties were tested using a set of experimental procedures to evaluate their potential to alter parameters linked to the biology of aphids (Rhopalosiphum padi) and spread of virus (BYDV-PAV). Moreover, two natural-based substances (azadirachtin and mineral oil) were tested for their impact on the ability of aphids to survive, colonize and transmit the virus. Results showed that the 11 genotypes tested have a level of susceptibility to virus infection similar to the susceptible reference cv. Rubisko. However, when characterization focused on virus load, latency period and aphid fecundity, partial resistance phenotypes were observed for some cultivars. Furthermore, azadirachtin increased aphid mortality and decreased aphid fecundity. Thus, in addition to genes described for their ability to limit B/CYDV infections, the genetic backgrounds of B/CYDV-susceptible wheat cultivars and azadirachtin-based treatment should be considered for future management strategies against yellow dwarf disease.     

Management of yellow dwarf disease in Europe in a post‐neonicotinoid agriculture

Barley/cereal yellow dwarf viruses (YDVs) cause yellow dwarf disease (YDD), which is a continuous risk to cereals production worldwide. These viruses cause leaf yellowing and stunting, resulting in yield reductions of up to 80%. YDVs have been a consistent but low‐level problem in European cereal cultivation for the last three decades, mostly due to the availability of several effective insecticides (largely pyrethroids and more recently neonicotinoids) against aphid vectors. However, this has changed recently, with many insecticides being lost, culminating in a recent European Union (EU) regulation prohibiting outdoor use of the neonicotinoid‐insecticide compounds. This change is coupled with the growing challenge of insecticide‐resistant aphids, the lack of genetic resources against YDVs, and a knowledge deficit around the parameters responsible for the emergence and spread of YDD. This means that economic sustainability of cereal cultivation in several European countries including France and United Kingdom is now again threatened by this aphid‐vectored viral disease. In this review, we summarize the current knowledge on the YDV pathosystem, describe management options against YDD, analyse the impacts of the neonicotinoid ban in Europe, and consider future strategies to control YDV. © 2020 Society of Chemical Industry     

Analysis of Accumulation Patterns of <Emphasis Type="Italic">Barley yellow dwarf virus-PAV</Emphasis> (BYDV-PAV) in Two Resistant Wheat Lines

Barley yellow dwarf (BYD) is one of the main viral diseases of small-grain cereals. This disease, reported on numerous plant species of the Poaceae family, is caused by a complex of eight viral species including the species Barley Yellow Dwarf Virus-PAV (BYDV-PAV), frequently found in western Europe. Resistance sources against BYDV-PAV are scarce and only identified in perennial Triticineae. Some BYDV-resistant wheat lines have been obtained by introgressing these resistances into bread wheat germplasms. Genetic and biological characterization of the resulting lines has been undertaken. However, little information on the resistant behaviour of these lines during the early stages of the infection process is available. To evaluate the resistance of two genetically distinct resistant lines (Zhong ZH and TC14), 1740 young plantlets, belonging to susceptible reference hosts (barley cv. Express and wheat cv. Sunstar), Zhong ZH or TC14 wheat lines, were inoculated in controlled conditions with French BYDV-PAV isolates. The infection process was monitored during the first 21 days after inoculation (DAI) using a semi-quantitative ELISA. A standardized protocol including five successive samplings of leaves from all inoculated plants and the collection of plant roots at the end of the monitored period was carried out. This protocol enabled an assessment of the infection percentage and the evolution of the viral load in plants from the 7th DAI to the 21st DAI. Statistical analyses of the BYDV infection kinetics using raw ELISA data, a model of the time-dependent variation of the percentage of infected plants and the area under concentration progress curves (AUCPC) demonstrated that Zhong ZH and TC14 lines (1) reduce the development rate of the BYD disease during the first days of infection, (2) decrease the infection efficiency of BYDV-PAV isolates, in the leaves, from 98.7% for susceptible plant genotypes to 81.9% and 71.7% for Zhong ZH and TC14, respectively, (3) reduce the virus load in the leaves of infected plants and (4) are not spared from BYDV infection, as 95.1% of Zhong ZH and 90.2% of TC14 inoculated plants accumulated viral particles in roots and/or in leaves at 21 DAI. These results confirm the BYDV-partial resistant behaviour of both Zhong ZH and TC14 lines. The development rate of the disease was the single parameter that allowed the distinction between the two resistant sources present in the tested lines.     

Common reed (Phragmites communis) is a natural host of important cereal viruses in the Trakya region of Turkey

Common reed (Phragmites communis Trin.), a perennial grass, is a widespread weed in the Trakya region of Turkey. Reed leaf samples were collected in 2004 and 2005, and tested for the presence of theMaize dwarf mosaic virus (MDMV),Sugarcane mosaic virus (SCMV),Barley yellow dwarf virus-PAV (BYDV-PAV),Cereal yellow dwarf virus-RPV (CYDV-RPV) andWheat dwarf virus (WDV) by DAS-ELISA, PTA-ELISA and Western blot analysis. MDMV was identified in five out of sixP. communis samples that exhibited characteristic virus-like symptoms in 2004. The remaining sample was co-infected with MDMV and BYDV-PAV. Transmission electron microscopy confirmed the presence of flexuous rod-shaped virus particles in four samples that reacted positively for MDMV in ELISA. In 2005, ELISA revealed that nine out of 234 samples that were collected in two different locations were infected with MDMV, nine with SCMV, and three with BYDV-PAV. No sample contained CYDV-RPV, JGMV and WDV. Our results confirm that the common reed is a host of BYDV-PAV and indicate, for the first time, that it is also a natural host of MDMV and SCMV.P. communis most likely acts as a reservoir of these three viruses in the Trakya region in Turkey. http//www.phytoparasitica.org posting Sept. 13, 2006.     

In Situ Localization of Barley Yellow Dwarf Virus-PAV 17-kDa Protein and Nucleic Acids in Oats

ABSTRACT Barley yellow dwarf virus strain PAV (BYDV-PAV) RNA and the 17-kDa protein were localized in BYDV-PAV-infected oat cells using in situ hybridization and in situ immunolocalization assays, respectively. The in situ hybridization assay showed labeling of filamentous material in the nucleus, cytoplasm, and virus-induced vesicles with both sense and antisense nucleic acid probes, suggesting that the filamentous material found in BYDV-PAV-infected cells contains viral RNA. BYDV-PAV negative-strand RNA was detected before virus particles were observed, which indicates that RNA replication is initiated before synthesis of viral coat protein in the cytoplasm. The 17-kDa protein was associated with filamentous material in the cytoplasm, nucleus, and virus-induced vesicles. The labeling densities observed using antibodies against the 17-kDa protein were similar in the nucleus and cytoplasm. No labeling of the 17-kDa protein was observed in plasmodesmata, but filaments in the nuclear pores occasionally were labeled. Since BYDV-PAV RNA and 17-kDa protein colocalized within infected cells, it is possible that single-stranded viral RNA is always associated with the 17-kDa protein in vivo. The 17-kDa protein may be required for viral nucleic acid filaments to traverse the nuclear membrane or other membrane systems.     

Improved method for assaying maize plant resistance to maize rough dwarf disease by artificial inoculation and real-time RT-PCR

The study of maize rough dwarf disease (MRDD) and breeding for resistance requires inoculation of maize plants by means of planthoppers. The plant age, insect density and inoculation duration are main factors in the success of maize rough dwarf disease inoculation. These parameters were tested using a susceptible maize inbred line Ye478. Using one or two-leaf plants, 15 planthoppers per plant and a five day inoculation duration, the line Ye478 was the most susceptible with 100% diseased plants; F112132 was moderately susceptible with 60% diseased plants and 90110 and F022411 were resistant without any disease. The results were consistent with those from six years of field studies. Using enzyme-linked immunosorbent assay (ELISA) and real-time quantitative RT-PCR, rice black-streaked dwarf virus was detected in severely diseased plants. The plants were rated from 0 to 3 according to their symptoms at the time of flowering. Plants scoring 0, 1 and 2 could not be distinguished by ELISA, only by real-time quantitative RT-PCR. All of the plants with a score of 3 were positive by ELISA and real-time quantitative RT-PCR. The significant differences in the average viral contents in plants with different symptom ratings could be distinguished by using real-time RT-PCR.     

Linear-motion tattoo machine and prefabricated needle sets for the delivery of plant viruses by vascular puncture inoculation

Vascular puncture inoculation (VPI) of plant viruses previously has been conducted either manually or by use of a commercial engraving tool and laboratory-fabricated needle arrays. In an effort to improve this technique, a linear-motion tattoo machine driving industry-standard needle arrays was tested as a means of delivering plant viruses into maize and small grain seed embryos. The new method was applied in the successful transmission of maize rayado fino virus (MRFV), the type member of the genus Marafivirus, from an archived sample to maize. Subsequent transfer of MRFV from the sap of an infected plant using the method produced an average infection rate in maize of 70% (range 39–93%). Maize, oat, and triticale were successfully infected with oat blue dwarf virus (OBDV) using the method; similar infection rates were observed between maize seeds inoculated with the tattoo machine and those inoculated with the engraving machine when using prefabricated needle arrays. No infection was obtained in repeated tests with barley yellow dwarf virus (BYDV-PAV) or cereal yellow dwarf virus (CYDV-RPV) using either sap or RNA from infectious cloned cDNA. Replacement of the original engraving-tool with a linear-motion tattoo machine in VPI provides greater flexibility and convenience in a quiet, readily-available instrument, while improving reproducibility through the use of prefabricated needle arrays.     

青海省小麦种质资源对大麦黄矮病毒的抗性鉴定

为明确青海省不同小麦种质资源对大麦黄矮病毒(Barley yellow dwarf virus,BYDV)的抗性差异,于2014—2015年采用堆测法人工接种鉴定了178份种质资源的抗病性。结果显示,不同小麦种质资源对黄矮病的抗性存在较大差异,甘A100、川766、陕1059、兰麦-2的病情指数依次为14.83、23.60、23.99和24.66,表现出较好的抗病性;病情指数在50.00以上的高感品种有69份,包括尕老汉、白板麦、兴热密穗等,其中白板麦病情指数高达72.35,高于感病对照阿勃;其余种质资源病情指数在25.00~50.00之间,表现为感病;甘A101、甘A99、藏515、藏519、木汉麦、群科大白麦、拉胎板麦、小红麦-2和朗县折达25初期表现感病,后期恢复健康,有一定的耐病性;抗病性不同的种质资源感染BYDV后,对小麦产量的影响差异很大,抗病对照中4产量损失8.96%,耐病品种产量损失在13.64%~19.74%之间,高病品种尕老汉产量损失达41.91%,表明小麦种质资源中抗BYDV的品种极少。     

感染大麦黄矮病毒的小麦中vsiRNA特征分析

 RNA沉默是植物中一种保守的抗病毒机制,其以大量病毒来源的小干扰RNA(virus-derived small interfering RNAs,vsiRNAs)的产生为标志,病毒在侵染寄主过程中可通过vsiRNAs靶向寄主转录本以对抗这种防御机制。BYDV-GAV引起的小麦黄矮病导致小麦黄化和矮化症状,对小麦生产构成严重威胁。通过深度测序技术,分析了感染BYDV-GAV的感病小麦品种‘小偃6号’中vsiRNAs的特征,共得到11 384个vsiRNAs,并预测到37 784个寄主靶基因。发现来源于BYDV-GAV基因组的正义和反义链的vsiRNAs的数量分布大致相等,在5’末端具有A和C偏好性,长度主要在21nt~22nt。靶基因的功能分析表明这些靶基因参与了广泛的生物学功能,尤其是在寄主-病原物互作中占的比重最大。选取25个参与寄主-病原互作的抗性相关基因进行定量验证,发现接种BYDV-GAV后有15个明显下调,6个上调,4个微弱下调,表明测序结果和靶基因预测可靠。推测BYDV-GAV可以通过vsiRNAs干扰寄主抗性基因表达和信号转导,从而实现对感病寄主的侵染。研究结果对揭示BYDV-GAV与小麦互作的分子机制具有重要意义。     

10种病毒抑制剂对小麦黄矮病的防治效果

在田间条件下研究了10种病毒抑制剂对小麦黄矮病的防治效果。结果表明,不同药剂处理对小麦黄矮病均有一定的抑制作用,其中6%寡糖·链蛋白WP、25%宁南·嘧菌酯AS和0.5%菇类蛋白多糖AS的防效分别为64.26%、58.44%和53.18%。不同病毒抑制剂处理的小麦产量有不同程度的增加,6%寡糖·链蛋白WP、20%吗胍·乙酸铜WP、0.5%葡聚烯糖SP和25%宁南·嘧菌酯AS增产率分别为21.5%、20.9%、20%和19.5%。说明6%寡糖·链蛋白WP是一种有前途的小麦黄矮病抑制剂,可在农业生产实践中进行推广应用。     

Plant community diversity influences vector behaviour and Barley yellow dwarf virus population structure

The species composition of a plant community can affect the distribution and abundance of other organisms including plant pathogens. The goal of this study was to understand the role of host diversity in the transmission of two Barley yellow dwarf virus (BYDV) species that share insect vectors and hosts. Greenhouse experiments measured the transmission rate of BYDV species PAV and PAS from infected oat plants to healthy agricultural and wild grasses and from these species back to healthy oat seedlings. In the field component of the study, the rate of spread of PAV and PAS was measured in monoculture plots planted with agricultural grasses. In greenhouse experiments, the aphid vector more readily transmitted PAV from agricultural grasses and more readily inoculated PAS to the wild grass species assayed. In the field experiment, disease prevalence was greater in wheat, but there was no difference in the rate of spread of PAV and PAS. These results indicate an interaction between vector and host genotype that selects for greater PAV transmission in grain crops, contributes to differences in disease prevalence between grass types, and maintains pathogen diversity within the larger plant community (i.e. agricultural and non‐agricultural hosts).     

小麦黄矮病及其抗性育种研究进展

由大麦黄矮病毒(barley yellow dwarf viruses, BYDVs)侵染引起的小麦黄矮病(wheat yellow dwarf disease)是世界小麦上的主要病害之一, 对小麦生产及粮食安全构成了严重威胁。本文综述了BYDVs的生物学特征、分类地位、致病机理及抗性育种等相关研究, 并基于相关研究进展, 提出了进一步扩大抗性基因挖掘、加快优异抗病基因克隆、加强抗病机理研究等方面的建议, 旨在为我国小麦黄矮病抗性育种及科学防治提供一定的参考。     

大麦黄矮病毒侵染小麦miRNA鉴定和特征分析

 大麦黄矮病毒(barley yellow dwarf viruses,BYDVs)引起的小麦黄矮病严重威胁我国麦类生产,造成严重经济损失。植物中的miRNA调控植物生长发育、信号转导及对外界压力的反应,通过调控植物抗性基因的表达影响植物与病原物的互作。本研究对感染BYDV-GAV 后3 d、7 d及健康对照的‘小偃6号'小麦样品进行miRNA测序,合并去冗余后分别得到99、96、95个已知的miRNA序列和806、809、1 024个新miRNA序列。对这些miRNA进行差异表达分析,BYDV-GAV侵染后3 d和7 d的小麦样品,与对照相比上调表达的miRNA数量分别为3个和7个,下调表达的为14个和12个。将差异表达的miRNA利用psRNATarget进行靶基因预测,共得到1 254个靶标基因。靶基因的KEGG和GO富集分析,进一步明确了其功能及作用通路。对14个病毒病症状相关的靶基因进行定量分析,结果表明随病毒侵染时间的延长,这些靶基因出现差异性表达,显示miRNA参与了寄主与病毒的互作。研究结果有助于揭示BYDV-GAV与寄主小麦的互作机理。     

大麦黄矮病毒侵染小麦miRNA鉴定和特征分析

 大麦黄矮病毒(barley yellow dwarf viruses,BYDVs)引起的小麦黄矮病严重威胁我国麦类生产,造成严重经济损失。植物中的miRNA调控植物生长发育、信号转导及对外界压力的反应,通过调控植物抗性基因的表达影响植物与病原物的互作。本研究对感染BYDV-GAV 后3 d、7 d及健康对照的‘小偃6号'小麦样品进行miRNA测序,合并去冗余后分别得到99、96、95个已知的miRNA序列和806、809、1 024个新miRNA序列。对这些miRNA进行差异表达分析,BYDV-GAV侵染后3 d和7 d的小麦样品,与对照相比上调表达的miRNA数量分别为3个和7个,下调表达的为14个和12个。将差异表达的miRNA利用psRNATarget进行靶基因预测,共得到1 254个靶标基因。靶基因的KEGG和GO富集分析,进一步明确了其功能及作用通路。对14个病毒病症状相关的靶基因进行定量分析,结果表明随病毒侵染时间的延长,这些靶基因出现差异性表达,显示miRNA参与了寄主与病毒的互作。研究结果有助于揭示BYDV-GAV与寄主小麦的互作机理。     

Coat protein-mediated resistance to isolates of barley yellow dwarf in oats and barley

Tissue cultures of GAF30/Park oats were biolistically co-transformed with constructs containing the coat protein (CP) genes of the P-PAV, MAV-PS1 or NY-RPV isolates of barley yellow dwarf virus (BYDV), together with a construct containing the bar gene for herbicide resistance and the uidA reporter gene. Transformed, herbicide-resistant tissue cultures were screened by PCR for the presence of the CP genes. Fertile regenerated plants were recovered from some CP-transformed tissue cultures. T₁ progeny of these plants were screened for resistance to the BYDV isolate corresponding to the introduced gene by inoculation with viruliferous aphids followed by ELISA tests. Variation in ELISA values for GAF30/Park control plants made interpretation of the data difficult, but oat plants resistant to each of the three isolates of BYDV (ELISA values less than 0.3; virus titers equivalent to less than 25% of infected controls) were identified in T₁ generations. Further testing of MAV-PS1 CP-transformed lines to the T₂ generation, NY-RPV CP-transformed lines to the T₃ generation and P-PAV CP-transformed lines to the T₄ generation identified further resistant plants. Similarly, immature embryos and calli of the barley cultivar Golden Promise were biolistically bombarded with constructs containing the CP gene of the P-PAV isolate of BYDV and the bar and uidA reporter genes, lines of self-fertile P-PAV CP-transformed barley plants were developed, and T₁plants were screened for resistance to P-PAV. Eight plants from six lines showed moderate to high levels of resistance to P-PAV that correlated with the presence of the CP gene. Plants giving low ELISA values were also found in other lines, even though the CP gene was not detected in these plants. Some T₂ plants derived from resistant parents that contained the CP gene were themselves highly resistant.     

Occurrence of barley yellow dwarf virus in cereals and grasses of the low-rainfall wheatbelt of South Australia

South Australia is in the dry temperate zone where most cereal crops are grown in an area of low rainfall, with a crop-free season from December to April. The incidence of barley yellow dwarf virus (BYDV) was assessed by ELISA from 1989 to 1991 in wheat crops and irrigated pastures of South Australia. The incidence of BYDV was low in most wheat crops of the low-rainfall area in 1989 and 1990 (less than 1% of plants infected), but moderate levels of infection (1–10%) were observed in some early-sown crops. BYDV infection was more widespread in the high-rainfall area (south east of South Australia). A high incidence of BYDV was observed in the irrigated pastures of the three areas surveyed (4–86%). Of the five previously described strains, the Rhopalosiphum padi/Sitobion avenae strain (PAV) was the most common in wheat samples (> 90%). PAV and the R. padi-specific strain (RPV) were found in pasture grasses, alone or in mixed infection. Virus incidence was greater in Festuca spp. (56%) and Lolium perenne (30%) than in other species (2-–9%).