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- Title
Construction of Infectious cDNA Clone of Brassica Yellows Virus Isolated from Strawberry and Establishment of TaqMan RT-qPCR.
- Authors
Zhao, Xiaoli; He, Chengyong; Gao, Dehang; Xu, Tengfei; Li, Xiaofeng; Liu, Junjie; Li, Shifang; Wang, Hongqing
- Abstract
The natural host range for brassica yellows virus (BrYV) is generally limited to Cruciferae. However, we found that BrYV can naturally infect strawberry. The full-length genome sequences of BrYV-MB (accession No. MZ666129) and BrYV-HY (accession No. ON060762) identified in strawberry from Yantai and Beijing, China, were obtained by high-throughput sequencing (HTS) combined with the RT-PCR and RACE techniques. The complete genome sequences of BrYV-MB and BrYV-HY are 5666 nt and contain six open reading frames (ORFs). The two isolates have the highest nucleotide (nt) sequence identity of 99.0%. The infectious cDNA clone of BrYV-HY was constructed through homologous recombination and used to agroinfiltrate Nicotiana benthamiana and Arabidopsis thaliana. The inoculated leaves of N. benthamiana showed necrotic symptoms after 4 days of inoculation (dpi), and the systematic leaves of A. thaliana exhibited purple symptoms at 14 dpi. To develop a rapid and high-sensitive method for the detection of BrYV, a TaqMan real-time fluorescence quantitative RT-PCR method (TaqMan RT-qPCR) was established. Under optimum reaction conditions, the sensitivity of the detection was as low as 100 fg and approximately 100-fold more sensitive than the conventional RT-PCR, so it can be used in large-scale testing.
- Subjects
YANTAI (Shandong Sheng, China); BEIJING (China); VIRUS cloning; PHYTOPLASMAS; STRAWBERRIES; WHOLE genome sequencing; BRASSICA; NICOTIANA benthamiana
- Publication
Plants (2223-7747), 2022, Vol 11, Issue 23, p3380
- ISSN
2223-7747
- Publication type
Article
- DOI
10.3390/plants11233380