Identification of Barley Yellow Dwarf Viruses in Poland
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Keywords:
Barley yellow dwarf
Luteovirus
Triticale
Secale
Barley yellow dwarf virus (BYDV) is a widespread complex of virus species of cereal crops in the world, known to infect both wild and cultivated plants of the family Poaceae, having the effect of yield reductions and subsequently large economic losses to farmers. Barley yellow dwarf disease (BYD) is a result of infection from BYDV species (genus: Luteovirus) and cereal yellow dwarf virus (CYDV; genus: Polerovirus). The symptoms of infection include the yellowing of leaf tips, rolling of leaf margins, stunting of plants, reduction in seed set and fertility. Taxonomically the viruses causing disease are the luteovirus species BYDV-MAV, BYDV-PAV and BYDV-PAS as well as the polerovirus CYDV-RPV. These are positive sense single-stranded RNA (+ssRNA) viruses vectored by different aphid species, including Sitobion avenae, Rhopalosiphum padi, Schizaphis graminun and Metopolphium dirhodum in a circulative nonpropagative manner. In Sweden, seasonal field surveys are conducted along with monitoring of aphid species to determine seasonal outbreak scenarios. In spring 2015 and 2017, BYD was confirmed from preliminary field surveys, identifying BYDV-PAV, BYDV-MAV and CYDV-RPV. This study was carried out to determine the viral diversity of isolates collected from plant material in southern Sweden, as well as to investigate the possible presence of phenotypic mixing or transcapsidation during infection in winter barley (Hordeum vulgare), winter wheat (Triticum aestivum) and winter rye (Secale cereale). DAS-ELISA and PCR amplification of the BYDV coat protein (CP) genes was used to determine viral diversity and the presence of phenotypic mixing or transcapsidation. Through sequence analyses of cloned PCR products, BYDV-PAV and BYDV-PAS were identified as viruses causing disease during outbreaks in spring 2015 and 2017. The BYDV-PAV isolates were closely related to previously recorded isolates of BYDV-PAV in Sweden, Latvia and Germany. In a phylogenetic analysis, Swedish and global BYDV-PAS isolates formed a well-supported group indicating that BYDV-PAS is a widespread virus. In addition, an isolate of Barley virus G, a virus first identified in barley of South Korea was identified in Swedish barley from the disease outbreak of 2015 and represents the first record of the virus in Sweden. DAS-ELISA results identified BYDV-PAV, BYDV-MAV and CYDV-RPV as viruses causing BYD disease in the outbreaks of 2015 and 2017, differing from that of CP gene sequence analysis which did not identify BYDV-MAV or CYDV-RPV. The results suggest that antibodies for BYDV-PAV and BYDV-MAV detect the CP of BYDV-PAS, because samples were most often positive for both BYDV-PAV and BYDV-MAV in DAS-ELISA, but BYDV-PAV and BYDV-PAS were detected by sequence analyses of the CP gene. Antibodies for CYDV-RPV seem to detect Barley virus G because of unspecific detection by DAS-ELISA antibodies for BYDV-PAV and BYDV-MAV, unable to adequately differentiate the CPs of these species. Future work should focus on ELISA antibody development for discrete identification of BYD virus species.
Barley yellow dwarf
Rhopalosiphum padi
Luteovirus
Secale
Rhopalosiphum maidis
Sitobion avenae
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Development and characterization of a new barley yellow dwarf virus (BYDV)-resistant wheat germplasm
A barley yellow dwarf virus (BYDV)-resistant line HG295 was selected from a cross between cv. 77-5433 and Zhong 5 after extensive investigation in field, greenhouse and ELISA. Cytological analysis revealed that it was an euploid line and genetically stable. The existence of alien DNA in HG295 was identified by RAPD and Southern hybridization analyses showed that the alien DNAs came from Zhong 5 or Th. intermedium. The differences of BYDV resistance between L1 and HG295 are discussed.
Barley yellow dwarf
Germ plasm
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Trakya Region of Turkey has been one of the important cereal growing areas in Turkey. Previously sporadic and temporary infections of Yellow dwarf viruses (YDVs) have been reported in some parts of Turkey. YDV diseases on cereals however have been prevailing and causing yellowing, dwarfing, reddening and the reduction of grain yield on cultivated cereals since 1999 in the Trakya Region. YDV have been identified and their incidence and the rate of infections were investigated. Barley yellow dwarf virus-PAV (BYDV-PAV) was diagnosed as the most virulent and dominant one as Cereal yellow dwarf virus-RPV (CYDV-RPV) was also identified as another important virus in the area. In order to determine sources of YDVs and their over summering and overwintering hosts among the Poaceae weed species 326 symptomatic weed leaf samples and 82 intact weed plants were collected from road sides and hedge grows of cereal fields in 2010. In second year 357 weed leaf samples, 13 voluntary cereal leaves and 50 intact weed plants were also collected from same sites. Separately 7 aphid species were identified and 5 of them were used for vector transmission tests of YDVs from potted intact weeds to indicator barley (cv. Barbaros) seedlings. As a result of aphid transmissions from 15 weed species, 156 symptomatic barley leaf samples and from 6 weed species, 50 symptomatic barley samples were obtained in 2010 and 2011 respectively. So, totally 902 leaf samples were obtained from 42 weed, 3 voluntaries and 1 indicator barley species. DAS-ELISA and RT-PCR tests on 326 weed samples revealed the corresponding incidence rates were 54.60% for BYDV-PAV, 7.05% for CYDV-RPV, 5.52% for PAV+RPV, 14.41% for the other YDVs and being 81.59% total rate of virus incidence in weed samples in 2010. Test results on 370 leaf samples also revealed the incidences of BYDV-PAV as 14.86%, CYDV-RPV as 10.81%, PAV+RPV as 7.56% and the other YDVs as 48.91% totally being 82.16% rate of virus incidence from weed and voluntary cereal samples in 2011. Aphid transmitted barley samples revealed the similar incidences of viruses too. For molecular characterization the genomic region containing coat protein (CP) regions of BYDV-PAV and CYDV-RPV were amplified from selected weed species and samples by RT-PCR method. Specific DNA fragments in the sizes of 531 bp and 400 bp were amplified from 45 BYDV-PAV isolates from 24 weed species and 34 CYDV-RPV isolates from 15 weed species respectively. The selected DNA fragments of BYDV-PAV and CYDV-RPV were purified and sequenced for the determination of nucleotide sequences of CP genes of both virus isolates. Partial nucleotide sequences of 20 Turkish PAV weed isolates were determined and compared with other nine BYDV-PAV isolates in databases. Phylogenetic analysis of obtained and published nucleotide and amino acid sequences revealed the identity ranged from 86.67 - 99.80% and 70.05 - 99.40% respectively. Partial nucleotide sequences of 6 CYDV-RPV isolates were also compared with seven isolates of CYDV-RPV isolates in GenBank⁄EMBL. The nucleotide and amino acid sequences revealed the identity ranged from 80.44 - 95.86% and 62.50 - 93.33% identities respectively. To our knowledge, this is the first report of YDV’s in Poacea weed hosts in Turkey.
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Avena
Barley yellow dwarf
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The Matanuska-Susitna Valley is one of the most fertile regions in Alaska for growing cool-season vegetables. Barley (Hordeum vulgare) and oat (Avena sativa) crops are also sown for animal feed and green manure. The most damaging and widely distributed viral disease of small grains worldwide is barley yellow dwarf (BYD), caused by several species from two genera in the family Luteoviridae: luteovirus (Barley yellow dwarf virus [BYDV-MAV and BYDV-PAV]) and polerovirus (Cereal yellow dwarf virus [CYDV-RPV, formerly BYDV-RPV]) and three unassigned species (BYDV-RMV, BYDV-SGV, and BYDV-GPV) (2,4). Even though barley and oat have been grown in Alaska for more than 50 years, BYD has not been documented in small grains in this region. During September 2001, barley plants with bright yellow leaves were collected from five barley fields near Palmer. Three plants from each field were assayed using a reverse transcription-polymerase chain reaction (RT-PCR) protocol targeting members of the luteoviridae (3). The resulting ≈530-bp PCR product and its restriction fragment length polymorphism (RFLP) produced by digestion with NdeII implied that plants were infected with BYDV-PAV. In September 2002, three of the five sites were surveyed again for BYDV. Two of the fields (BF-1 and BF-2) had been replanted with barley and the other (OF-3) was planted with oats. Leaf samples from 36 symptomatic barley plants from each field and 60 symptomatic oat plants were randomly collected and stored at -80°C. In 2002, in addition to RT-PCR and RFLP analyses, enzyme-linked immunosorbent assays (ELISA) using Agdia kits (Agdia, Elkhart, IN) for BYDV-PAV, CYDV-RPV, and BYDV-SGV were also performed (1). First, RT-PCR and RFLP were completed on all samples using 0.5 g of tissue. Of samples from BF-1, BF-2, and OF-3, 61, 100, and 70%, respectively, generated luteoviridae-specific fragments. The RFLP profiles from barley were all PAV-like, whereas 71% of oat samples were PAV-like, and 29% were of an unknown pattern. No bands were observed from apparently healthy field plants. ELISA (0.2 g of tissue) was performed on all PCR-positive samples, resulting in 22, 97, and 33% detection for BYDV-PAV from BF-1, BF-2, and OF-3, respectively. An additional 29% of oat samples (OF-3) tested positive for CYDV-RPV, whereas none of the barley plants tested positive. One oat plant had a mixed infection with both PAV and RPV profiles, and all oat plants with the unidentified RFLP pattern were serologically positive for RPV. No BYDV-SGV was detected in either barley or oats. The PCR assay was clearly more sensitive than ELISA, especially for plants that had mature and necrotic tissue, which were predominately found in BF-1 and OF-3. Based on these direct tests on the coat protein's nucleic acid (PCR) and serology (ELISA), it is concluded that two distinct viruses, BYDV-PAV and CYDV-RPV, were found in oats, whereas only the former was found in barley. To my knowledge, this is the first report of luteovirus and polerovirus infection in small grains in Alaska. References: (1) M. F. Clark and A. N. Adams. J. Gen. Virol. 34, 475, 1977. (2) C. J. D'Arcy and P. A. Burnett. Barley Yellow Dwarf: 40 Years of Progress. The American Phytopathological Society, St. Paul, MN, 1995. (3) N. L. Robertson and R. French. J. Gen. Virol. 72,1473, 1991. (4) M. H. V. van Regenamortel et al. Virus Taxonomy. Seventh Report of the International Committee on Taxonomy of Viruses. Academic Press, NY, 2000.
Barley yellow dwarf
Luteovirus
Avena
Avena fatua
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Luteovirus
Barley yellow dwarf
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A field survey was conducted in Tunisia in the North-Eastern regions (Bizerte, CapBon and Zaghouan), the North-Western region (Kef) and the Central-Eastern region (Kairouan) during the 2011/2012 growing season, in order to determine the incidence and the geographic distribution of Barley yellow dwarf virus (BYDVs) in barley fields. Tissue blot immunoassays (TBIA) showed that BYDV was most common in Zaghouan (incidence 14%), Cap Bon (14%) and Bizerte (35%), in randomly collected samples from these three locations.Among the different BYDVs identified, BYDV-PAV (64%) was the most common followed by BYDV-MAV (16%) and CYDV-RPV (3%). The coat protein gene sequences of six isolates collected from different regions shared >98% pairwise similarity. In comparisons with other BYDV sequences from around the world, the Tunisian sequences shared greatest homology with isolates 109 and ASL1 from the United States of America and Germany (≈97%), and <90% with all other isolate sequences available in public databases.
Barley yellow dwarf
Sequence homology
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Sugarcane yellow leaf virus (ScYLV) causes severe leaf symptoms in sugarcane (Saccharum spp.). It is a single-stranded RNA virus assigned to the genus Polerovirus, family Luteoviridae (1). ScYLV is transmitted by two aphid species, Melanaphis sacchari and Rhopalosiphum maidis. Although barley (Hordeum vulgare), oats (Avena sativa), and wheat (Triticum spp.) are susceptible to ScYLV when experimentally inoculated (3), this virus, related serologically to Barley yellow dwarf virus (BYDV)-RPV (4), has never been detected naturally in these cereals. In this study, 240 barley leaves were randomly collected from six fields in Tunisia following a north-south trend during the high infestation periods (March/April) in the 2013 growing season. Samples were tested by DAS-ELISA, using three antibodies (Bioreba AG, Switzerland), two of them, BYDV-B and BYDV-F, specific to luteoviruses corresponding to BYDV-PAV and BYDV-MAV, respectively, and the third one, BYDV-RPV, specific to the polerovirus synonymous to Cereal yellow dwarf virus (CYDV)-RPV. Based on DAS-ELISA, 30 samples were found positive for B/CYDV infection; 17 out of the 30 infected samples contained a single serotype, BYDV-PAV, and 13 out of the 30 infected samples contained two serotypes, PAV and RPV. Total RNA was extracted from all positive samples, and RT-PCR of the viral CP gene was performed with Lu1/Lu4 primers (2). A product of 531 bp was cloned and sequenced. The identities among the sequences determined varied between 80 to 100%, and from the 17 samples containing BYDV-PAV, six distinct BYDV-PAV sequences were revealed and named PAV-TN1 to PAV-TN6 (GenBank Accession No. JX402453 to JX402457 and KF271792). Fortuitously, all 13 positive samples corresponding to the serotypes PAV-RPV exhibited 98.7 to 99.3% identity with ScYLV isolates. These 13 samples contained three distinct sequences that were named ScYLV-Tun1 to ScYLV-Tun3 (GenBank Accession No. KF836888 to KF836890). Of the 17 PAV-positive samples collected, six were infected with PAV-TN1, four with PAV-TN2, four with PAV-TN3, one with PAV-TN4, one with PAV-TN5, and the last one with PAV-TN6. Of the 13 ScYLV-positive samples, seven were infected with ScYLV-Tun1, four with ScYLV-Tun2, and two with ScYLV-Tun3. Phylogenetic analysis showed that PAV-TN sequences formed a very tight cluster (>98%) corresponding to BYDV subspecies PAV-II, whereas all three Tunisian ScYLV sequences were clustered together. This study provides the first report of ScYLV isolates infecting barley crops in Tunisia, and confirms serological cross-reactivity between ScYLV and BYDV-RPV when commercial antibodies against BYDV-RPV are used. References: (1) C. J. D'Arcy and L. L. Domier. Page 891 in: Virus Taxonomy, 8th Report of the ICTV. C. M. Fauquet et al., eds. Springer-Verlag, New York, 2005. (2) N. L. Robertson and R. French. J. Gen. Virol. 72:1473, 1991. (3) S. Schenck and A. T. Lehrer. Plant Dis. 84:1085, 2000. (4) J. Vega et al. Plant Dis. 81:21, 1997.
Barley yellow dwarf
Rhopalosiphum maidis
Luteovirus
Avena
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Wheat (Triticum aestivum L.) is a natural host of many viruses. Yellow dwarf viruses belonging to the Luteoviridae family are important virus species that cause economic loss by restricting wheat production worldwide. Surveys were conducted in 2017 to determine Yellow dwarf viruses (BYDV-PAV, BYDV-MAV, BYDV-SGV, BYDV-RMV, and CYDV-RPV) and their infection rates in wheat production areas in Mardin province. 400 fresh leaf samples collected were tested by Multiplex reverse transcription-polymerase chain reaction (m-RT-PCR). The overall infection rate was found to be 3%. BYDV-PAV has been identified as the most widespread virus with a 2.5% presence rate. It was found out that BYDV-SGV, CYDV-RPV, and BYDV-RMV infections were lower, with rates of 1.75%, 0.5% and 0.25% respectively. In the current study, double infections were detected in 8 samples. The overall infection rate of the detected viruses (BYDV-PAV, BYDV-SGV, CYDV-RPV, BYDV-RMV) was found to be lower than the records reported in previous similar studies. No BYDV-MAV infection was found in any of the wheat samples tested. The cDNA of the coat protein (CP) gene of a BYDV-PAV isolate randomly selected from virus-positive samples was cloned, bidirectionally sequenced, and the phylogenetic relationship revealed. According to the phylogenetic analysis with 19 different isolates in the NCBI database of BYDV-PAV Mardin isolate, it showed the highest genetic similarity by 95.52% with the Germany isolate (KY634926) while the lowest similarity rate was 89.22% with the Germany and Pakistan isolates (KY634886 and JQ811489). The presence of BYDV-PAV, BYDV-SGV, CYDV-RPV, and BYDV-RMV were reported for the first time with this survey study conducted in Mardin.
Barley yellow dwarf
Infection rate
Luteovirus
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Barley yellow dwarf disease is a worldwide ubiquitous virus disease of cereal crops.In order to characterize the B/CYDV isolates occurring in Tunisia, 240 barley leaves were randomly sampled from 6 fields following a North-South trend and analyzed by serological and molecular tests.DAS-ELISA results showed 40 positive samples with a prevalence of barley yellow dwarf virus (BYDV)-PAV (77.5%), followed by cereal yellow dwarf virus (CYDV)-RPV (25%) and BYDV-MAV (15%).Studies of the geographic distribution showed a high incidence of B/CYDV in the Tunisian Southern provinces.RT-PCR assays were performed to amplify the viral coat protein gene (CP) and sequence analyses revealed six BYDV-PAV haplotypes named PAV-TN1 to PAV-TN6.Phylogenetic analysis showed that the six Tunisian haplotypes were close to BYDV-PAV-II subspecies and had a strong similarity with Moroccan, Czech, French and German haplotypes.Although PAV-TN2 and PAV-TN5 showed up to 10% divergence from BYDV-PAV-II at the amino acid level, it seems to belong to the same subspecies but in a separated cluster.Our results will be important in developing appropriate control measures against BYDV disease in Tunisia.
Barley yellow dwarf
Subspecies
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