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종(Sherwood et al. 2003). HTS 샘플의 275bp RT-PCR 앰플리콘과 3개의 추가 양성
샘플은 양방향 Sanger 시퀀싱(CAF)으로 검증되었으며 KY250488 가입과 96% 동일성
을 가졌습니다. 앰플리콘 간의 쌍별 nt 동일성은 98.55~100%였습니다. 이것은 남아프
리카에서 토마토반점위조바이러스 Tomato Spotted Wilt Orthotospovirus가 아가판투스
식물을 감염시키는 첫 번째 보고입니다. 이 연구는 토마토반점위조바이러스 Tomato
Spotted Wilt Orthotospovirus의 분포 및 발생에 대한 정보를 제공하고 번식 전에 식물
재료를 선별하는 묘목장의 필요성을 강조합니다.
3. 원 문
First Report of Tomato Spotted Wilt Orthotospovirus Infecting Agapanthus (Agapanthus
praecox) in South Africa
R. Bester, S. U. Demas, and H. J. Maree
Agapanthus praecox Willd. is an ornamental flowering plant that is indigenous to southern
Africa and was reported to be a host of tomato spotted wilt orthotospovirus (TSWV) in
Australia in 2000 (Wilson et al. 2000). Tomato spotted wilt orthotospovirus, belonging to the
genus Orthotospovirus of the family Tospoviridae, is a single-stranded negative-sense RNA
virus known to cause disease in many crops and ornamental plant species. This virus is in the
top 10 of the most economically important plant viruses worldwide (Rybicki 2015; Scholthof et
al. 2011). In May 2021, leaf material from three agapanthus (Agapanthus praecox) plants
displaying chlorotic mottling and yellow lesions was collected in Mbombela, South Africa. One
gram of symptomatic leaf material was used for total RNA extraction from each of the three
samples using a CTAB extraction protocol (Ruiz-Garcia et al. 2019). The three RNA extracts
were pooled, and a sequencing library was constructed using the Ion Total RNA-Seq Kit v2.0
and RiboMinus Plant Kit for RNA-Seq (ThermoFisher Scientific) at the Central Analytical
Facilities (CAF), Stellenbosch University. The RNA library was sequenced on an Ion Torrent
Proton Instrument (CAF). A total of 34,392,939 single-end reads were obtained. Data were
trimmed for quality with Trimmomatic (CROP:250, MINLEN:50). De novo assembly was
performed on the remaining 32,281,645 trimmed reads (average read length: 100 nucleotides
[nt], range: 50 to 250 nt) using SPAdes 3.13.0 and resulted in 4,788 contigs. BLASTn analysis
identified viral contigs longer than 1,000 nt with high nt identity to TSWV (six contigs), as well
as to the newly discovered viruses Agapanthus tungro virus (AgTV; one contig) and
Agapanthus velarivirus (AgVV; four contigs; Read et al. 2021). Read mapping was performed
against the relevant reference sequence with the highest nt identity to the contigs. For TSWV,
4,995, 21,221, and 14,574 reads mapped to segments L (KY250488), M (KY250489), and S
(KY250490) of the isolate LK-1, respectively, resulting in 99.97, 100.00, and 99.97% genome
coverage of the reference accessions. The nt identity between the reference accessions and
the consensus sequences generated (OP921761 to OP921763) was 97.26, 97.64, and 97.82% for
- 665 -
샘플은 양방향 Sanger 시퀀싱(CAF)으로 검증되었으며 KY250488 가입과 96% 동일성
을 가졌습니다. 앰플리콘 간의 쌍별 nt 동일성은 98.55~100%였습니다. 이것은 남아프
리카에서 토마토반점위조바이러스 Tomato Spotted Wilt Orthotospovirus가 아가판투스
식물을 감염시키는 첫 번째 보고입니다. 이 연구는 토마토반점위조바이러스 Tomato
Spotted Wilt Orthotospovirus의 분포 및 발생에 대한 정보를 제공하고 번식 전에 식물
재료를 선별하는 묘목장의 필요성을 강조합니다.
3. 원 문
First Report of Tomato Spotted Wilt Orthotospovirus Infecting Agapanthus (Agapanthus
praecox) in South Africa
R. Bester, S. U. Demas, and H. J. Maree
Agapanthus praecox Willd. is an ornamental flowering plant that is indigenous to southern
Africa and was reported to be a host of tomato spotted wilt orthotospovirus (TSWV) in
Australia in 2000 (Wilson et al. 2000). Tomato spotted wilt orthotospovirus, belonging to the
genus Orthotospovirus of the family Tospoviridae, is a single-stranded negative-sense RNA
virus known to cause disease in many crops and ornamental plant species. This virus is in the
top 10 of the most economically important plant viruses worldwide (Rybicki 2015; Scholthof et
al. 2011). In May 2021, leaf material from three agapanthus (Agapanthus praecox) plants
displaying chlorotic mottling and yellow lesions was collected in Mbombela, South Africa. One
gram of symptomatic leaf material was used for total RNA extraction from each of the three
samples using a CTAB extraction protocol (Ruiz-Garcia et al. 2019). The three RNA extracts
were pooled, and a sequencing library was constructed using the Ion Total RNA-Seq Kit v2.0
and RiboMinus Plant Kit for RNA-Seq (ThermoFisher Scientific) at the Central Analytical
Facilities (CAF), Stellenbosch University. The RNA library was sequenced on an Ion Torrent
Proton Instrument (CAF). A total of 34,392,939 single-end reads were obtained. Data were
trimmed for quality with Trimmomatic (CROP:250, MINLEN:50). De novo assembly was
performed on the remaining 32,281,645 trimmed reads (average read length: 100 nucleotides
[nt], range: 50 to 250 nt) using SPAdes 3.13.0 and resulted in 4,788 contigs. BLASTn analysis
identified viral contigs longer than 1,000 nt with high nt identity to TSWV (six contigs), as well
as to the newly discovered viruses Agapanthus tungro virus (AgTV; one contig) and
Agapanthus velarivirus (AgVV; four contigs; Read et al. 2021). Read mapping was performed
against the relevant reference sequence with the highest nt identity to the contigs. For TSWV,
4,995, 21,221, and 14,574 reads mapped to segments L (KY250488), M (KY250489), and S
(KY250490) of the isolate LK-1, respectively, resulting in 99.97, 100.00, and 99.97% genome
coverage of the reference accessions. The nt identity between the reference accessions and
the consensus sequences generated (OP921761 to OP921763) was 97.26, 97.64, and 97.82% for
- 665 -
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