First report of Streptocarpus flower break virus in Streptocarpus hybrids in Aotearoa New Zealand.

Abstract

Streptocarpus (Cape primrose, family Gesneriaceae) is a genus of plants native to Southern Africa commonly grown indoors for their foliage and trumpet-shaped flowers. In Aoteroa New Zealand (NZ) to date, no viruses have been reported to infect plants of the Gesneriaceae (Veerakone et al. 2015). In September 2022, a plant of Streptocarpus hybrid exhibiting necrotic rings was observed in a hobbyist's greenhouse in Auckland, NZ. High-Throughput Sequencing (HTS) using MinIONTM (Oxford Nanopore Technologies), was applied as a first screen (Liefting et al. 2021). Phylogenetic analysis was performed using Geneious Prime 2021 (Biomatters Ltd, NZ). A BLASTn search with 622,847 obtained reads resulted in 3,260 and 4,340 matches to the sequences of Streptocarpus flower break tobamovirus (SFBV) and Impatiens necrotic spot orthotospovirus (INSV), respectively. A near-complete (98.5%) genome sequence of SFBV was obtained (GenBank accession No. OQ970154), which shared 99.52% nucleotide identity to a SFBV type isolate from Germany (GenBank accession No. NC_008365). A phylogenetic tree was also generated (e-Xtra). To confirm the presence of both viruses, leaf tissue was rub inoculated onto herbaceous indicator plants as described by Tang et al. (2013). Chenopodium amaranticolor and C. quinoa plants developed local lesions while Nicotiana occidentalis plants showed local necrosis followed by systemic leaf puckering by 14 days post inoculation (dpi). Nicotiana benthamiana and N. clevelandii plants showed systemic chlorosis but N. tabacum plants did not exhibit any symptoms by 28 dpi. Samples from indicators and Streptocarpus were tested by RT-PCR (SFBV) or RT-qPCR (INSV), using in-house designed primers: SFBV-forward (5'-GTCATCAGCCGGAGAGGTTC-3'), SFBV-reverse (5'-AGGGCGAGTCTCTTCCTCTG-3'), INSV-forward (5'-CAATCAGAGGGTGACTTGGAA-3'), INSV-reverse (5'-GACTTTCCGAAGACTTGATGC-3') and INSV-probe (5'-CCATTGTCCTTTATCATTCCAACAAG-3'). RT-PCR products (across MP and CP regions) of the expected size (357 bp) were amplified from the Streptocarpus sample and symptomatic indicators. All amplicons were sequenced in both directions and found to be identical to the obtained HTS sequence. The presence of INSV was confirmed in all Streptocarpus and inoculated indicators except N. tabacum by INSV-specific RT-qPCR. A further 77 Streptocarpus plants were collected from a greenhouse in Auckland that holds a collection of multiple Streptocarpus cultivars from across NZ and overseas. Twenty-five plants, either displaying flower colour-break (only one plant) or asymptomatic (24 plants), tested positive for SFBV by RT-PCR. All amplicons were sequenced and found to be identical. SFBV was first described from naturally infected Streptocarpus plants in 1995 in the Netherlands (Verhoeven et al. 1995), and then in Germany (Heinze et al. 2006) and the United States (Pappu & Druffel 2007). While INSV has been found in NZ in several plant genera (Veerakone et al., 2015), to our knowledge, this is the first report of SFBV in NZ. SFBV was thought to be associated with colour breaking of Streptocarpus flowers (Verhoeven et al. 1995) but the virus was detected in asymptomatic Streptocarpus plants in this study and in California (Pappu & Druffel 2007). Given SFBV-infected plants were purchased from several sources, and leaf cuttings for propagation are shared among hobbyists, SFBV is likely to have spread throughout NZ. How this will affect production is unclear at this stage.