Detection and sequencing of Barley yellow dwarf virus (BYDV) in cereal fields of Hamedan province
Paper ID : 1878-24IPPC (R3)
Authors
1Plant protection department, Agriculture faculty, Buali sina university,Hamedan
2Plant protection department, Agriculture faculty, Buali sina university, Hamedan, Iran
Abstract
Barley yellow dwarf viruses are the most widespread and important viruses of cereals. These viruses belong to family luteoviridae. Barley yellow dwarf virus (BYDV) can result in a total yield reduction of 80% in wheat. Studies revealed that BYDV is present in most parts of Iran. Due to the importance and rank of the crop cultivation in Hamedan province, the spread of this virus in the country, climate conditions and vector activity provide the suitable situations for the development of the virus in Hamedan province. For detection and study on the distribution of BYDV, samples were collected during the Spring of 2020-2021 from Hamedan wheat and barley fields including Malayer, Nahavand, Tuyserkan, Hamedan, Asadabad, Bahar, Kabudarahang, Famenin and Razan. 40 plant samples with yellowing and dwarfing symptoms were collected from each region. The samples tested for BYDV infection by indirect ELISA using specific BYDV antibody (Agdia) according to the Converse and Martin method. The results showed the presence of BYDV in most collected samples. Positive reactions to BYDV antibody in %80, %90, %80, %80, %50, %50, %90, %90 and %90 of the Malayer, Nahavand, Tuyserkan, Hamedan, Asadabad, Bahar, Kabudarahang, Famenin and Razan samples were recorded, respectively. To confirm the infections, total RNA extraction were performed from the positive samples in ELISA, using the Column RNA isolation kit (Denazist) according to the manufacturer's protocol. RT-PCR were performed with specific BYDV-PAV primer pair that is the prevalent strain of the virus in most parts of the country. These primers amplify a 900bp fragment of the genome (a part of polymerase gene). RT-PCR results, confirmed the results of indirect ELISA. Then, a sample of Tuyserkan selected and sequenced. Nucleotide sequence of the Tuyserkan isolate confirmed the BYDV-PAV infection and showed that this isolate has maximum homology of more than 97% with Kahnouj, Karaj, Yazd and Arak isolates. While it revealed the least similarity (84-85% similarity) with other Iranian isolates including Bushehr, Esfandaghe, Kermanshah Saveh, Abasabad and Rostagh isolates. The group which include Tuyserkan isolate, have more than 10 percent diversity with other isolates at amino acid level in polymerase gene and were proposed as a distinct species previously. The results showed the diversity of this virus in the country.
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