First Report of a Rhabdovirus Infecting Passion Fruit ( <i>Passiflora edulis</i> ) in Yunnan Province, China

Passion fruit (Passiflora edulis) is a fruit crop with high economic and nutritional value that is widely cultivated in tropical and subtropical regions around the globe. Diseases caused by viruses pose a serious threat to the production of passion fruit worldwide (Wu et al. 2024). Recently, several isolates of rhabdoviruses, such as citrus-associated rhabdovirus (CiaRV), passiflora cytorhabdovirus (PFCV), and passiflora nucleorhabdovirus (PaNV), have been identified and characterized in Passiflora spp. collected from China and Brazil (Zhang et al. 2021; Vidal et al. 2025). In November 2020, a total of ten P. edulis plants displaying chlorosis and leaf curling symptoms were collected in Wenshan City, Yunnan Province, China. To identify viruses potentially associated with this disease, leaf samples were collected from a P. edulis plant exhibiting typical disease symptoms and subjected to lncRNA sequencing using an Illumina NovaSeq 6000 platform. As a result, a total of 77,384,252 clean reads were generated following the removal of adapters and low-quality sequences using Trimmomatic software (v. 0.39) with default settings (Bolger et al. 2014). After the removal of host-derived reads, the resulting reads were assembled de novo into 105,632 contigs. The assembled contigs were subsequently searched against the NCBI non-redundant protein database to detect potential viral pathogens using BLASTx with a cutoff e-value of 10−5 (Wang et al. 2024). As a result, three contigs were aligned to three known passion fruit viruses: Passion fruit severe mottle-associated virus (PFSMoAV, accession no. MK449340), Telosma mosaic virus (TeMV, accession no. MH623078), and East Asian Passiflora virus (EAPV, accession no. OL956953 ) (Xie et al. 2019; Fu et al. 2023). Notably, a contig of 13,490 nucleotides (nt) exhibited high sequence similarity (≥ 86.0%) to members of the Cytorhabdovirus caricae species within the genus Cytorhabdovirus, suggesting that this virus might be a new member of the family Rhabdoviridae, for which the name “Passion fruit-associated rhabdovirus” (PaRV) is tentatively proposed. Subsequently, the complete genome sequence of PaRV was obtained using RT-PCR and 5’- and 3’-RACE (rapid amplification of cDNA ends) approaches. The entire genomic sequence of PaRV is 13,546 nt in length and has been submitted to the GenBank database under the accession number MZ272468. BLASTn analysis revealed that PaRV exhibited sequence identities of 98.9% (MT302544), 98.3% (MT302543), and 97.8% (PQ721446) identical to different CiaRV isolates deposited in the GenBank database. Phylogenetic analysis based on complete genome sequences indicated that PaRV was most closely related to CiaRV-PB1 (accession no. MT302544), previously identified in the paper bush (Zhang et al. 2021). To assess the infectivity of PaRV, symptomatic passion fruit leaf sap was mechanically inoculated onto six healthy Nicotiana benthamiana plants, with healthy passion fruit leaf sap serving as a mock control. After 2 weeks, plants inoculated with symptomatic sap exhibited leaf curling and chlorosis symptoms, whereas the control plants remained asymptomatic. All symptomatic leaves tested positive for PaRV using RT-PCR with specific primers (PaRV-F1: 5’-ATACTTGTAATTTGTGGCCCTC-3’ and PaRV-R1: 5’-CGCTACCCTCTTGATGATACTC-3’) (6/6). For comparison, PaRV was not detected in the control sample. These findings are consistent with symptoms previously reported in Nicotiana species infected with other rhabdoviruses (Higgins et al., 2016; Fránová et al., 2019). To our knowledge, this is the first report of the natural occurrence of PaRV in the agroecosystem in Yunnan, China. Our study provides an important update on the prevalent PaRV field strains currently circulating in China, providing valuable insights into viral evolution and epidemiological characteristics.