A control group of plants received an equal volume of 0.05% Tween 80 buffer spray. Subsequent to fifteen days of inoculation, the plants that received the treatment manifested similar symptoms to the originally diseased specimens, whereas the controls exhibited no signs of illness. The infected leaves yielded C. karstii, which was re-isolated and subsequently characterized using morphological traits and a multi-gene phylogenetic tree analysis. The pathogenicity test, conducted three times, yielded similar results, thereby confirming Koch's postulates. Biomedical science This report, to our knowledge, details the inaugural occurrence of Banana Shrub leaf blight in China, specifically caused by C. karstii. The disease reduces the aesthetic and financial worth of Banana Shrub, and this research forms a crucial basis for future strategies in disease prevention and cure.
As a key food crop in some developing countries, the banana (Musa spp.) holds an important place in tropical and subtropical regions as a fruit. China's extensive history in cultivating bananas has positioned it as the second-largest banana producer on a global scale. The area devoted to banana planting exceeds 11 million hectares, according to FAOSTAT 2023. Banana mild mosaic virus (BanMMV), a flexuous filamentous virus, infects bananas and is classified as a banmivirus within the Betaflexiviridae family. Infection of Musa spp. is often asymptomatic, and the virus's worldwide distribution likely contributes to its high prevalence, as indicated in the study by Kumar et al. (2015). Transitory symptoms, including mild chlorotic streaks and mosaics, frequently manifest on young leaves as a consequence of BanMMV infection (Thomas, 2015). The presence of banana streak viruses (BSV) and cucumber mosaic virus (CMV) alongside BanMMV can intensify the mosaic patterns associated with BanMMV, according to Fidan et al. (2019). In October 2021, throughout eight cities encompassing four in Guangdong (Huizhou, Qingyuan, Zhanjiang, Yangjiang), two in Yunnan (Hekou and Jinghong), and two in Guangxi (Yulin and Wuming), a total of twenty-six leaf samples were procured, each exhibiting possible banana viral disease symptoms. Following thorough mixing of the contaminated samples, we partitioned them into two distinct batches and dispatched them to Shanghai Biotechnology Corporation (China) for metatranscriptomic sequencing. Every sample included a quantity of leaves equivalent to about 5 grams. For the purpose of ribosomal RNA depletion and library preparation, the Zymo-Seq RiboFree Total RNA Library Prep Kit (Zymo Research, USA) was selected. Shanghai Biotechnology Corporation (China) performed Illumina sequencing (Illumina NovaSeq 6000). Paired-end (150 bp) sequencing of the RNA library was carried out on an Illumina HiSeq 2000/2500 sequencer. Employing CLC Genomics Workbench (version 60.4), clean reads were assembled from metagenomic data via a de novo assembly. The National Center for Biotechnology Information (NCBI)'s non-redundant protein database facilitated the BLASTx annotation procedure. From the de novo assembly of the 68,878,162 clean reads, 79,528 contigs were ultimately generated. A 7265-nucleotide contig displayed the highest nucleotide sequence similarity (90.08%) with the BanMMV isolate EM4-2 genome; its GenBank accession number is [number]. Return OL8267451, it is imperative. Following the design of primers specific to the BanMMV CP gene (Table S1), leaf samples from eight cities (n=26) underwent testing. The results indicated only one Musa ABB Pisang Awak sample, originating from Guangzhou’s Fenjiao region, demonstrated infection. Anaerobic biodegradation BanMMV-infected banana leaves exhibited subtle chlorosis and yellowing at the leaf margins (Fig. S1). Our investigation into the BanMMV-infected banana leaves yielded no detection of additional banana viruses, like BSV, CMV, and banana bunchy top virus (BBTV). Staurosporine RNA, harvested from the infected plant leaves, was sequenced and the resulting contig's integrity across the complete sequence was affirmed using overlapping PCR amplification (Table S1). All ambiguous regions were subjected to PCR and RACE amplification, and Sanger sequencing was performed on the amplified products. Excluding the poly(A) tail, the complete genome of the candidate virus measured 7310 nucleotides. GenBank's accession number ON227268 contains the sequence from the Guangzhou isolate, BanMMV-GZ. Supplementary Figure 2 demonstrates the schematic organization of the genome sequence in BanMMV-GZ. The genome of this virus possesses five open reading frames (ORFs), including one encoding RNA-dependent RNA polymerase (RdRp), three triple gene block proteins (TGBp1 to TGBp3) critical for cell-to-cell transmission, and a coat protein (CP), akin to other BanMMV isolates (Kondo et al., 2021). The neighbor-joining phylogenetic method, applied to the full genome's complete nucleotide sequence and the RdRp gene's sequence, unambiguously located the BanMMV-GZ isolate within the collection of all BanMMV isolates (Figure S3). Based on our present knowledge, this report signifies the first observation of BanMMV's infection of bananas in China, thereby expanding the global expanse of this viral disease. Hence, a more comprehensive examination of BanMMV's presence and frequency throughout China is imperative.
South Korean passion fruit (Passiflora edulis) has been documented as a host for various viral diseases, including those attributable to the papaya leaf curl Guangdong virus, cucumber mosaic virus, East Asian Passiflora virus, and euphorbia leaf curl virus (Joa et al., 2018; Kim et al., 2018). Greenhouse-grown P. edulis plants in Iksan, South Korea, displayed virus-like symptoms, such as leaf and fruit mosaic patterns, curling, chlorosis, and deformation, in June 2021. This affected over 2% of the 300 plants (8 exhibiting symptoms and 292 without). RNA from symptomatic leaves of a single P. edulis plant, pooled together, was extracted using the RNeasy Plant Mini Kit (Qiagen, Germany) to produce a total RNA sample, and the TruSeq Stranded Total RNA LT Sample Prep Kit (Illumina, San Diego, CA) was subsequently used to construct a transcriptome library. NGS methodology, using the Illumina NovaSeq 6000 system from Macrogen Inc. (Korea), was employed. The software Trinity (Grabherr et al. 2011) was used to carry out a de novo assembly of the resulting 121154,740 reads. Using BLASTn, a total of 70,895 contigs, each longer than 200 base pairs, were assembled and annotated from the NCBI viral genome database. The numerical expression 212.0 holds a specific position. The 827 nucleotide contig sequence was determined to match milk vetch dwarf virus (MVDV), a member of the Nanoviridae family's nanovirus genus (Bangladesh isolate, accession number). This JSON schema contains a list of sentences, each uniquely structured. LC094159 presented a nucleotide identity of 960%, whereas the 3639-nucleotide contig indicated a correspondence with Passiflora latent virus (PLV), a Carlavirus member of Betaflexiviridae (Israel isolate, accession number). Sentences are to be returned in a list format within this JSON schema. A nucleotide identity of 900% was determined for sequence DQ455582. Verification of the NGS results involved isolating RNA from symptomatic leaves of the same P. edulis plant, using a viral gene spin kit (iNtRON Biotechnology, Seongnam, Korea). The RNA was then subjected to RT-PCR using primers specific to the viruses: PLV-F/R targeting the PLV coat protein, MVDV-M-F/R targeting the MVDV movement protein and MVDV-S-F/R targeting the MVDV coat protein. PLV, as indicated by a 518-base-pair PCR product, was detected, while no amplification of the MVDV product was observed. A nucleotide sequence was derived from the directly sequenced amplicon and deposited in GenBank (acc. number.). Transform these sentences ten times, generating distinct structural arrangements without reducing the original length. OK274270). Return this JSON schema. In a BLASTn analysis, the nucleotide sequence of the PCR product displayed 930% identity with PLV isolates from Israel (accession number MH379331) and 962% identity with PLV isolates from Germany (accession number MT723990), respectively. Out of eight plants in the Iksan greenhouse, six passion fruit leaves and two fruit samples exhibiting PLV-like symptoms were selected for RT-PCR analysis, with six of these samples testing positive for PLV. Notwithstanding the widespread detection of PLV, one leaf and one fruit from the collection did not show any trace of this compound. Extracts from systemic leaves of plants were used as inoculum for mechanical sap inoculation of P. edulis and indicator plants, including Chenopodium quinoa, Nicotiana benthamiana, N. glutinosa, and N. tabacum. P. edulis presented with vein chlorosis and yellowing on its systemic leaves at 20 days post inoculation. At 15 days post-inoculation, necrotic lesions were visually detected on the inoculated N. benthamiana and N. glutinosa leaves, and Plum pox virus (PLV) infection was verified using reverse transcription polymerase chain reaction (RT-PCR) on symptomatic leaf samples. This study's focus was on determining the infectability and potential for transmission of PLV within commercially grown passion fruit in the southern region of South Korea. Whereas persimmon (Diospyros kaki) in South Korea experienced no symptoms associated with PLV, no pathogenicity testing for passion fruit was reported in the literature (Cho et al., 2021). The natural infection of passion fruit with PLV in South Korea, for the first time observed, is accompanied by clear symptoms. The selection of healthy propagation materials and the evaluation of potential losses in passion fruit production are essential.
Capsicum chlorosis virus (CaCV), a member of the Orthotospovirus genus within the Tospoviridae family, was first observed infecting capsicum (Capsicum annuum) and tomato (Solanum lycopersicum) in Australia in 2002, as documented by McMichael et al. The infection's subsequent propagation was observed across a range of plants, encompassing waxflower (Hoya calycina Schlecter) in the United States (Melzer et al. 2014), peanut (Arachis hypogaea) in India (Vijayalakshmi et al. 2016), the spider lily (Hymenocallis americana) (Huang et al. 2017), chilli pepper (Capsicum annuum) (Zheng et al. 2020), and Feiji cao (Chromolaena odorata) (Chen et al. 2022) in China.