Gene/Protein Disease Symptom Drug Enzyme Compound
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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.
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PMID:First Report of Sugarcane yellow leaf virus Infecting Barley in Tunisia. 3070 56

Barley yellow dwarf disease, a ubiquitous virus disease of cereal crops worldwide, is caused by a group of related, single-stranded RNA viruses assigned to Luteovirus (Barley yellow dwarf virus [BYDV] spp. PAV, PAS, MAV, and GAV) or Polerovirus (Cereal yellow dwarf virus-RPV) genera or unassigned to a genera (BYDV-SGV, BYDV-RMV, and BYDV-GPV) in the family Luteoviridae (1). Incidence of BYDV in cereal crops (e.g., barley, wheat, and oats) was high, and in recent years, reached epidemic levels in many regions of the Czech Republic. BYDV-PAV and BYDV-PAS have been identified in Czech cereal crops (2,4). Surveys of the incidence of BYDV were carried out using ELISA (SEDIAG SAS, Longvic, France) and one-step reverse transcription (RT)-PCR (Qiagen, Hilden, Germany) (2) during 2007 and 2008. Samples (125) were collected from different fields around the Czech Republic and 96 were BYDV positive. Three of the field isolates, CZ-6815, CZ-1561, and CZ-10844, from oat (Avena sativa; cv. Auron), winter wheat (Triticum aestivum; cv. Apache), and winter barley (Hordeum vulgare; cv. Merlot), respectively, were identified as BYDV-MAV by sequencing of the RT-PCR product (641-bp fragment) used to identify BYDV, which spanned 2839-3479 of the BYDV genome (GenBank Accession Nos. EF043235 and NC_002160) (2). The partial coat protein gene sequence of 483 nt was compared with the available sequences of 12 BYDV-PAV isolates (PAV-JP, PAV-NY, PAV-ILL, PAV-AUS, PAV-WG2, PAV-whG4y3, PAV-on21-4, Tahoe1, CA-PAV, HB3, FH3, and MA9501); nine BYDV-PAS isolates (PAS-129, PAS-64, WS6603, WG13, PAS-Tcb4-1, PASwaw5-9, FL2, PAS-Vd29, and PAS-MA9516); and six BYDV-MAV isolates (MAV-CA, MAV-PS1X1, MAV-Alameds268, LMB2a, SI-o4, and MAV-CN) by MEGA4 (3). Nucleotide and amino acid sequence identities for the three isolates ranged from 92.9 to 99.4% and 88.0 to 95.8%, respectively, for available BYDV-MAV isolates; 76.8 to 78.2% and 62.7 to 67.6%, respectively, for available BYDV-PAS isolates; and 77.6 to 79.3% and 65.5 to 70.4%, respectively, for available PAV isolates. The sequence data indicates that these isolates (CZ-6815, CZ-1561, and CZ10844; GenBank Accession Nos. FJ645747, FJ645758, and FJ645746, respectively) are BYDV-MAV. To my knowledge, this is the first record of BYDV-MAV in the Czech Republic. 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, NY, 2005. (2) J. K. Kundu. Plant Dis. 92:1587, 2008. (3) K. Tamura et al. Mol. Biol. Evol. 24:1596, 2007. (4) J. Vacke. Page 100 in: Sbornik Referatu z Odborneho Seminare, Aktualni Problemy Ochrany Polnich Plodin, Praha, 1991.
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PMID:First Report of Barley yellow dwarf virus-MAV in Oat, Wheat, and Barley Grown in the Czech Republic. 3075 62

Barley yellow dwarf disease, an important, ubiquitous virus disease of cereal crops worldwide, is caused by a group of related single-stranded RNA viruses assigned to luteovirus (Barley yellow dwarf virus (BYDV) spp. PAV, PAS, MAV, and GAV) or polerovirus (Cereal yellow dwarf virus-RPV) genera or unassigned to a genera (BYDV-SGV, BYDV-RMV, and BYDV-GPV) in the family Luteoviridae (2). Incidence of BYDV in cereal crops (e.g., barley, wheat, and oats) was high and reached epidemic levels in recent years in many regions of the Czech Republic. Previously, only PAV isolates have been identified here on the basis of serological detection (4), although antibodies to differentiate between PAV, PAS, and MAV are not widely available. Field samples of cereal crops were routinely tested in 2006 and 2007 and BYDVs were detected by ELISA. One-step-reverse transcription (RT)-PCR (Qiagen, Hilden, Germany) was adapted for BYDV detection using primer pairs BYcpF (5'-CCACTAGAGAGGTGGTGAATG-3') and BYcpR (5'-CCGGTGTTGAGGAGTCTACC-3') designed from conserved sequences identified by aligning multiple BYDV sequences available in public databases. These primers amplify a 641-bp fragment spanning nucleotides 2839-3479 from PAV (GenBank Accession No. EF043235) or PAS (GenBank Accession No. NC_002160) that includes a region of the coat protein gene and the intergenic region. RT-PCR amplicons were generated from two field isolates, PS-RuJK (spring wheat isolate, cv. Granny, collected in July 2007 from experimental plots at the CRI in Prague) and JE-120JK (winter barley isolate, cv. Merlot, collected in January 2008 from a barley field in Rychnov), both of which induced severe BYD symptoms. Amplicons were sequenced in both directions in a CEQ2000XL sequencer (Beckman Coulter, Fullerton, CA). The partial coat protein gene sequence of 483 nt of PS-RuJK and JE-120JK was analyzed and compared with available sequences of 26 PAV, 17 PAS, and 13 MAV isolates by MEGA4 (3). PS-RuJK (GenBank Accession No. EU863652) nucleotide and amino acid sequence identities ranged from 96.3 to 99.2% and 93.7 to 98.7%, respectively, for available PAS isolates, and 89.9 to 90.5% and 85.5 to 86.9%, respectively, for available PAV isolates, and 78.3 to 79.5% and 70.0 to 72.5%, respectively, for available MAV isolates. Similarly, nucleotide and amino acid sequence identities JE-120JK (GenBank Accession No. EU863653) ranged from 95.2 to 98.6% and 90.6 to 96.9%, respectively, for PAS isolates, 88.8 to 90.1% and 83.1 to 84.4%, respectively, for PAV isolates, and 77.6 to 78.7% and 67.5 to 70.0%, respectively, for MAV isolates. Also, both of these isolates have the conserved amino acid motif "SIPGS" that is usually present in a variable region of the coat protein gene on the surface of virion (1) at position 52 to 56 of amino acid sequences of all published PAS-like isolates, including Vd29:AY167109, FH1:AJ223588, MA9516:AJ007926, FL2:AJ223586, ASL-1:AJ810418, and WS6603:DQ285680, contrary to "PVFRP" or "LISGP" motif in PAV or MAV, respectively. Therefore, the sequence data clearly confirm that these two isolates belong to the PAS species. To our knowledge, this is the first record of PAS detected in the Czech Republic. References: (1) C. A. Chay et al. Phytopathology 86:370, 1996. (2) 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, NY, 2005. (3) K. Tamura et al. Mol. Biol. Evol. 24:1596, 2007. (4) J. Vacke. Page 100 in: Sbornik Referatu z Odborneho Seminare, Aktualni Problemy Ochrany Polnich Plodin. Praha, 1991.
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PMID:First Report of Barley yellow dwarf virus-PAS in Wheat and Barley Grown in the Czech Republic. 3076 55

The grasses Sorghastrum nutans (Indian grass), Schizachyrium scoparium (little bluestem), Panicum virgatum (switchgrass), and Andropogon gerardii (big bluestem) are four of the most common plant species present in a tallgrass prairie (1). Infection with barley yellow dwarf virus (BYDV, family Luteoviridae) is of interest in these species because of the potential effects of the virus on tallgrass prairie plant communities and the potential for tallgrass prairie to function as a reservoir of the virus for infection in wheat or barley fields. In a previous inoculation experiment, an unidentified strain of BYDV transmitted by the aphid species Rhopalosiphum padi was reported to infect S. scoparium but none of the other three grass species (2). We sampled for the presence of five virus strains in at least 50 blooming plants of each grass species in a natural tallgrass prairie stand in August 2000. Samples were collected in watersheds that were designated 1B, 1D, K1A, 20B, and 20C at Konza Prairie Biological Station in the Flint Hills near Manhattan, KS. To detect the virus, we used enzyme-linked immunosorbent assay (ELISA) with antibodies purchased from Agdia (Elkhart, IN). For the PAV, MAV, RMV, and SGV strains, we used double-antibody sandwich ELISA with alkaline phosphatase label. For Cereal yellow dwarf virus (RPV), we used compound direct ELISA with alkaline phosphatase label. The scoring of ELISA results was based on comparison with infected and uninfected control plants of the same species. Symptoms of infection in the field were difficult to interpret visually, since plants in this natural environment often showed multiple symptoms of stress. None of the five strains were detected in 51 individuals of S. nutans. For 50 individuals of S. scoparium, the incidence of infection by the different strains was 4% for MAV, 0% for PAV, 2% for RMV, 0% for RPV, and 58% for SGV. For 51 individuals of P. virgatum, the incidence of infection was 31% for MAV, 0% for PAV, 0% for RMV, 0% for RPV, and 4% for SGV. For 64 individuals of A. gerardii, the incidence of infection was 59% for MAV, 0% for PAV, 0% for RMV, 0% for RPV, and 3% for SGV. The impact of BYDV on these tallgrass prairie species remains to be determined. The PAV strain is the most commonly reported strain in wheat in Kansas but was not recovered from these grass species. References: (1) C. C. Freeman. The flora of Konza Prairie: A historical review and contemporary patterns. Pages 69-80 in: Grassland Dynamics. A. K. Knapp et al., eds. Oxford, 1998. (2) W. N. Stoner. Plant Dis. Rep. 60:593, 1976.
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PMID:Barley Yellow Dwarf Disease in Natural Populations of Dominant Tallgrass Prairie Species in Kansas. 3081 73