Sweet Potato Feathery Mottle Virus Research Articles

First Report of Sweet Potato Virus E(SPVE) Infecting Sweet Potato in China.

Sweet potato (Ipomoea batatas [L.] Lam.) is a versatile crop, cultivated in the subtropical and tropical areas, as food, fodder, and industrial raw material crop. In China, sweet potato has been used as a health-care food in recent years, as it contains a wide range of nutrients and xenobiotic phytochemicals. However, viral diseases are major constraint for the sweet potato yield and quality, especially the seed production and quality. Over 30 species of viruses infect sweet potato worldwide (Clark et al. 2012). More recently, a few new viruses infected sweet potato were identified, such as sweet potato virus E (SPVE), which was reported in Korea(Jo et al. 2020). In May 2022, a sweet potato sample (JSXZ1) with virus-like symptom, such as mosaic and vein clearing were collected from sweet potato germplasm Xuzhou resource nursery, Jiangsu Province, China (N34˚16', E117˚18') (Fig. S1A). To investigate the virus disease, the sample JSXZ1 showing the typical symptoms of disease was prepared for Small-RNA (sRNA) deep-sequencing. The sRNA library was constructed using TruSeq™ Small RNA Sample Prep Kits (Illumina, San Diego, USA) and sequenced using the Illumine Hiseq 2500 platform by LC-Bop Technologies (Hangzhou) CO., LTD. The sample was sequenced to obtain 26, 358, 439 raw reads and 22, 969, 139 clean reads after quality control trimming and analysis. The Velvet 1.0.5 software was used to de novo assemble the clean reads (18 to 28 nt) into larger contigs, which were then compared with the nucleotide sequences in the National Center for Biotechnology Information (NCBI) database using the BLASTn algorithm. Viruses found in the sample were sweet potato latent virus (SPLV), sweet potato feathery mottle virus (SPFMV), sweet potato chlorotic stunt virus (SPCSV), sweet potato badnavirus A (SPBV-A) and sweet potato badnavirus B (SPBV-B). Surprisingly, besides the viruses listed above, 28 contigs matched sequences of SPVE isolate GS (MH388502). To verify the result, total RNA was extracted from the sample JSXZ1 and from other leave samples (JSXZ2-JSXZ5) that contained SPFMV, SPVC, SPLV, SPVG respectively stored in lab using FastPure Universal Plant Total RNA Isolation Kit (Vazyme Biotech Co., LTD, Nanjing, China). cDNA was synthesized using random primer (hexadeoxyribonucleotide mixture; pd(N)6). The cDNA serves as template in PCR using a newly designed primer pairs based on SPVE p1 gene (SPVE-F: 5'- TCACCAAAAAGAATGCTACAAC-3'/SPVE-R: 5'-GAAATCCTCCCACTCTCCATA-3'). An expected ~500-bp PCR fragment was obtained in JSXZ1, while none of the fragment was obtained from JSXZ2-JSXZ5 (Fig. S1B). The PCR fragment was cloned into pMD18-T vector (Takara Bio Inc., Beijing, China) and plasmid DNA from transformed Escherichia coli DH5α cell (n=3) were commercially sequenced by Sangon Biotech (Shanghai) Co., Ltd. The sequences of the three fragment clones we obtained were 100% identical when compared. A BLASTN analysis of the sequences revealed that they are specific to SPVE and shared 98.62% nucleotide identity to SPVE GS isolate (MH388502) and one sequence was submitted to GenBank (Accession number OQ948331). To determine the occurrence of SPVE in infected sweet potato plants, a total of 37 leaves samples with viral symptom collected from Shandong Province (n=6) and Jiangsu Province (n=31) were indexed by RT-PCR as described before. Only 9 (24.3%) out of 37 from Shandong (n=1) and Jiangsu (n=8) were positive to SPVE respectively. In addition, five additional viruses (SPFMV, SPVC, SPVG, SPLV, SPCSV) were detected among these 37 samples and always in a mixed infection of two or more viruses. To our knowledge, this is the first report of SPVE infecting sweet potato in China. Sweet potato is an important crop in China and other countries (Zhang et al. 2023). China is the largest sweet potato producer all over the world. In addition, as sweet potato is produced through the vegetative propagation mode, thus, more attention should be paid to detection and monitoring of occurrence of SPVE in China.

Loop‐mediated isothermal amplification assay: A novel disease diagnostics tool in sweetpotato seed quality assurance

AbstractSweetpotato is an important food crop in global production. However, sweetoptato viruses pose a threat to sustainable agriculture and cause significant economic loss. More than 30 viruses have been reported to date, with sweet potato feathery mottle virus (SPFMV) and sweet potato chlorotic stunt virus (SPCSV) occurring frequently and in combination, causing sweetpotato virus disease. The detection of viruses directly from sweetpotato is still a challenge, and efforts are being made toward this front. Therefore, rapid detection of viruses is critical for effective control. Current diagnostic tests are not sufficiently sensitive to reliably detect viruses directly from sweetpotato and require expensive laboratory equipment and a high level of skilled experience. Loop‐mediated isothermal amplification (LAMP) is sensitive and specific for both DNA/RNA amplification; it is affordable and its characteristics make it potentially suitable for on‐site testing. LAMP assays have been developed for several sweetpotato viruses: SPCSV, SPFMV, and sweet potato leaf curl virus (and related sweepoviruses). Laboratory validation showed a 100% diagnostic sensitivity for all three viruses. The LAMP on‐site testing results were comparable to those of polymerase chain reaction and reverse transcriptase polymerase chain reaction confirmatory laboratory tests. Interlab validation and packaging into affordable kits will ensure high adoption in sub‐Saharan Africa. The LAMP assay can be used for field surveys and monitoring the phytosanitary status of pre‐basic seed production in quarantine or certification program. This guarantees pathogen‐free planting material enters the seed system. We discuss opportunities for LAMP as a diagnostic assay for the rapid detection of sweetpotato viruses and its adoption as a quality control system for planting materials.

Diseases Incidence and Severity of Sweet potato feathery mottle virus in Sweetpotato in Gianyar Regency, Bali

Sweet potato feathery mottle virus (SPFMV) (Potyvirus) was first discovered in Indonesia in 2018. No one has reported information about the incidence and severity of SPFMV disease. Therefore, this study aims to determine the incidence and severity of SPFMV in Gianyar, Bali. The research was carried out directly in the field with observational parameters such as symptoms, incidence and disease severity. Observations were made every week when the plants were 63 to 91 days after planting (DAP). The results showed that SPFMV causes purple ringspot symptoms. The incidence of the disease did not increase when the plants were aged 63 to 91 DAP, while the severity of the disease increased every week. The highest incidence and severity of the disease were 70% and 20.7%. The main cause of SPFMV infection in the field is probably due to the continuous use of stem cuttings from previous crops.

Degeneration of cleaned-up, virus-tested sweetpotato seed vines in Tanzania

Viruses pose a major challenge to sweetpotato production in Tanzania. Use of cleaned-up, virus-tested seed vines distributed through a formal seed system is among the proposed strategies to address this challenge. However, virus-tested seed vines can get infected once in the field and it is not known how they will perform following several seasons of on farm propagation. We assessed the performance of virus-tested seed vines and farmer-sourced seed vines of a susceptible variety, Ejumula, and a relatively tolerant variety, Kabode, over five seasons to understand the trend in root yields, vine yields and virus incidences. The experiments were done in high and low virus pressure areas. The most prevalent viruses were sweet potato chlorotic stunt virus (SPCSV) followed by sweet potato feathery mottle virus (SPFMV) and sweet potato leaf curl virus (SPLCV), respectively. Both farmer-sourced and cleaned-up, virus-tested seed of cv. Ejumula were rapidly infected with SPCSV. The incidence of this virus on Ejumula's farmer-sourced material at the high-virus-pressure area reached 100% by the second season. The incidences for all three viruses remained stable for cv. Kabode across the five seasons. Plants generated from cleaned-up, virus-tested seed had lower incidences for all viruses compared to those from farmer-sourced planting material. Virus-tested seed produced significantly higher root yields for cv. Ejumula in the high-virus-pressure site, with a gradual drop across the seasons. The findings show that regular replenishment of clean, virus-tested seed is more economical in high-virus-pressure areas and for more susceptible varieties like cv. Ejumula. They also indicate that farmers may be reluctant to invest in cleaned-up, virus-tested seed in cases where they have virus-tolerant varieties such as cv. Kabode due to lack of obvious virus effect on yields.

CryoEM and stability analysis of virus-like particles of potyvirus and ipomovirus infecting a common host

Sweet potato feathery mottle virus (SPFMV) and Sweet potato mild mottle virus (SPMMV) are members of the genera Potyvirus and Ipomovirus, family Potyviridae, sharing Ipomoea batatas as common host, but transmitted, respectively, by aphids and whiteflies. Virions of family members consist of flexuous rods with multiple copies of a single coat protein (CP) surrounding the RNA genome. Here we report the generation of virus-like particles (VLPs) by transient expression of the CPs of SPFMV and SPMMV in the presence of a replicating RNA in Nicotiana benthamiana. Analysis of the purified VLPs by cryo-electron microscopy, gave structures with resolutions of 2.6 and 3.0 Å, respectively, showing a similar left-handed helical arrangement of 8.8 CP subunits per turn with the C-terminus at the inner surface and a binding pocket for the encapsidated ssRNA. Despite their similar architecture, thermal stability studies reveal that SPMMV VLPs are more stable than those of SPFMV.

Advances in the etiology of sweet potato (Ipomoea batatas (L.) Lam) yellow curling disease in Argentina

Sweet potato yellow curling (YC), the most severe disease of sweet potato detected in Argentina, causes symptoms and damage to sweet potato crops in all cultivated regions. Since 2010/11, the presence of four viruses has been detected in symptomatic cv. Arapey INIA: two potyviruses non-persistently transmitted by Myzus persicae (sweet potato feathery mottle virus, SPFMV and sweet potato virus G, SPVG); a closterovirus, sweet potato chlorotic stunt virus (SPCSV) and a geminivirus, sweet potato leaf curl virus (SPLCV), both transmitted by Bemisia tabaci in a semi-persistent and persistent manner, respectively. All the plants were collected from fields in Colonia Caroya, Córdoba province, Argentina. The objectives of the present work are to isolate and identify the virus or viruses involved in YC disease of sweet potato, and to elucidate the viral combination that reproduces YC symptoms. The most severe YC symptoms for this genotype in the field were only reproduced by a combination of the four viruses. The symptoms include chlorosis, stunting, mosaic, blistering, leaf curling, chlorotic spots, chlorotic patterns, leaf area reduction and distortion, and upward curling of leaf edges. The presence of each virus was detected by serological (DAS, NCM and TAS-ELISA) and molecular (PCR) tests. It is concluded that the interaction of SPFMV, SPVG, SPCSV and SPLCV is needed for the development of YC symptoms. Highlights Four viruses involved in yellow curling disease, SPFMV, SPVG, SPCSV and SPLCV were isolated and identified. The first antiserum produced in Argentina allows the fast, safe and efficient diagnosis of SPVG. The interaction of SPFMV, SPVG, SPCSV and SPLCV is needed for the manifestation of yellow curling in Argentina.

Elucidating the sweet potato virome in Greece with the aid of high‐throughput sequencing technology

AbstractIn 2018, typical symptoms of sweet potato virus disease (SPVD) such as purple ringspots, distortion and crinkling of the leaves appeared on a sweet potato crop located in southern Greece. High‐throughput sequencing (HTS) analysis revealed the presence of seven viruses, sweet potato feathery mottle virus (SPFMV), sweet potato chlorotic stunt virus (SPCSV), sweet potato virus C (SPVC), sweet potato virus G (SPVG), sweet potato virus 2 (SPV2), sweet potato leaf curl virus (SPLCV) and sweet potato pakakuy virus (SPPV). In silico analysis of the HTS data in combination with Sanger sequencing succeeded in obtaining the complete genome sequences of all seven viruses. Nucleotide sequence comparisons of complete viral genomes showed high similarity of all detected potyviruses and SPLCV with homologous sequences retrieved from GenΒank, whereas a new variant of SPCSV RNA1 was characterized and successfully transmitted onto Nicotiana benthamiana plants with the whitefly species Bemisia tabaci MED and MEAM1. Moreover, the Greek strain of SPPV showed higher similarity with SPPV strain B, even though it possesses a variable RT‐RNase H domain. Further phylogenetic analysis clustered Greek sweet potato virus isolates into various phylogenetic groups, whereas few new clades were formed in the cases of SPFMV, SPVG and SPCSV RNA1. Small‐scale surveys on three different sweet potato crops exhibiting typical, mild or no symptoms also indicated the close association of SPCSV with SPVD. Taken together, this study characterizes for the first time the virome of sweet potato crops in Greece and provides a glimpse into the evolutionary relationships of the identified viruses.

Development of a dual RT-RPA detection for Sweet potato feathery mottle virus and Sweet potato chlorotic stuntvirus

The disease co-infected by Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) is devastating in sweet potato, as it would give rise to the serious losses in both production and quality. Consequently, it is conducive for preventing and controlling this disease to detect these two viruses accurately and timely. Here we developed and optimized a dual reverse transcription recombinase polymerase amplification (RT-RPA) for rapid and accurate detection of SPFMV and SPCSV. Four special primers were designed based on the conserved sequences of SPFMV and SPCSV, respectively. The sensitivity of dual RT-RPA for SPFMV and SPCSV was 10−4 ng/μL at the optimal conditions in which the primer ratio between SPFMV and SPCSV was 2:1, and the reaction incubated for 25 min at a temperature of 39 °C. Both 61 sweet potato samples and 5 morning glory samples collected from China were tested using the dual RT-RPA successfully. Therefore, the dual RT-RPA is a reliable, rapid, sensitive method to detect these two viruses in sweet potato. It is the RT-RPA that was used for detection of SPFMV and SPCSV simultaneously firstly. This dual RT-RPA, as a convenient and powerful tool, will be useful to diagnose SPFMV and SPCSV.

Screening of sweet potato feathery mottle virus resistant sweet potato (Ipomoea batatas L., Lam.) cultivars in Kebbi State, Nigeria

Sweet potato is a food security crop because of its ability to withstand adverse climatic conditions. This security, however, is being threaten by viral diseases and use of resistant cultivars remain the best management. This research was conducted to screen cultivars of sweet potato against sweet potato feathery mottle virus (SPFMV) infection. The treatments consisted of five cultivars coded as CV1, CV2, CV3, CV4 and CV5, respectively. The asymptomatic experimental plants were established, maintained under screen house conditions and graft-inoculated using infected vines which were tested SPFMV positive using both Double Antibody Sandwich Enzyme-Linked Immunosorbent Assay (DAS-ELISA) and Polymerase Chain Reaction (PCR). The experiment was laid out in Completely Randomized Design (CRD) and replicated three times. Results obtained from disease incidence and symptom severity indicated that there was significant difference (P <0.05) among cultivars in their reaction to SPFMV infection. CV2 had the highest mean disease incidence (60.67%) while, CV3 had the lowest mean disease incidence (36.67%). CV1, CV3, and CV5 have the same lowest mean symptom severity score of 2.00 while, CV2 had the highest mean severity score of 4.00. Based on the reaction of the cultivars after inoculation, it could be concluded that, all the cultivars screened were susceptible to SPFMV but CV1, CV3 and CV4 cultivars have some degree of resistance to SPFMV infection and were therefore recommended for use by the farmers in the study area. This is the first research that screened sweet potato cultivars for resistance to SPFMV in Kebbi State, Nigeria.

Detection and elimination of viruses infecting sweet potatoes in Hungary

AbstractSweet potato has been grown in Hungary for the last three decades, and its popularity is increasing among farmers and consumers. Its production is hampered by pests and diseases due to poor agricultural practices, such as the use of virus‐infected propagation materials. We tested the presence of 15 viruses by PCR and quantitative PCR in 110 sweet potato plants collected from seven regions in Hungary. Seven viruses in single or multiple infections associated with a wide range of foliar symptoms were detected: sweet potato chlorotic stunt virus (SPCSV), sweet potato virus G (SPVG), sweet potato virus C (SPVC), sweet potato feathery mottle virus (SPFMV), sweet potato virus 2 (SPV2), sweet potato leaf curl virus (SPLCV), and sweet potato pakakuy virus (SPPV). This is the first report on the occurrence of the begomovirus SPLCV in sweet potatoes in Hungary. The infectivity and identity of these viruses were confirmed through bioassays (grafting to Ipomoea setosa) and sequencing of the PCR‐amplified sections of their genomes, respectively. Due to the necessity for virus‐free sweet potato propagation material in Hungary, virus elimination was carried out successfully in five out of six genotypes important for Hungarian farmers using heat treatment and meristem tip culture. All five viruses detected in the plants before heat treatment were removed except SPPV, which persists after heat treatment. Production and strict regulation of virus‐free sweet potato propagation materials are recommended to avoid exacerbating the virus situation and protect Hungarian farmers from further losses.

Sweet potato feathery mottle virus (internal cork disease of sweet potato)

Sweet potato feathery mottle virus (internal cork disease of sweet potato)

Cloning and Characterization of Eukaryotic Translation Initiation Factor 4E (eIF4E) Gene Family in <i>Ipomoea batatas</i> L. (Lam) for Understanding Hexaploid Sweetpotato-Virus Interactions

Characterization of genes related to sweetpotato to viral disease resistance is critical for understanding plant-pathogen interactions, especially with feathery mottle virus infection. For example, genes encoding eukaryotic translation initiation factor (eIF)4E, its isoforms, eIF(iso)4E, and the cap-binding protein (CBP) in plants, have been implicated in viral infections aside from their importance in protein synthesis. Full-length cDNA encoding these putative eIF targets from susceptible/resistant and unknown hexaploid sweetpotato (Ipomoea batatas L. Lam) were amplified based on primers designed from the diploid wild-type relative Ipomoea trifida consensus sequences, and designated IbeIF4E, IbeIF(iso)4E and IbCBP. Comparative analyses following direct-sequencing of PCR-amplified cDNAs versus the cloned cDNA sequences identified multiple homeoalleles: one to four IbeIF4E, two to three IbeIF(iso)4E, and two IbCBP within all cultivars tested. Open reading frames were in the length of 696 bp IbeIF4E, 606 bp IbeIF(iso)4E, and 675 bp IbCBP. The encoded single polypeptide lengths were 232, 202, and 225 amino acids for IbeIF4E, IbeIF(iso)4E, and IbCBP, with a calculated protein molecular mass of 26 kDa, 22.8 kDa, and 25.8 kDa, while their theoretical isoelectric points were 5.1, 5.57, and 6.6, respectively. Although the homeoalleles had similar sequence lengths, single nucleotide polymorphisms and multi-allelic variations were detected within the coding sequences. The multi-sequence alignment performed revealed a 66.9% - 96.7% sequence similarity between the predicted amino acid sequences obtained from the homeoalleles and closely related species. Furthermore, phylogenetic analysis revealed ancestral relationships between the eIF4E homeoalleles and other species. The outcome herein on the eIF4E superfamily and its correlation in sequence variations suggest opportunities to decipher the role of eIF4E in hexaploid sweetpotato feathery mottle virus infection.

Targeting of SPCSV-RNase3 via CRISPR-Cas13 confers resistance against sweet potato virus disease.

Sweet potato (Ipomoea batatas) is one of the most important crops in the world, and its production rate is mainly decreased by the sweet potato virus disease (SPVD) caused by the co‐infection of sweet potato chlorotic stunt virus (SPCSV) and sweet potato feathery mottle virus. However, methods for improving SPVD resistance have not been established. Thus, this study aimed to enhance SPVD resistance by targeting one of its important pathogenesis‐related factors (i.e., SPCSV‐RNase3) by using the CRISPR‐Cas13 technique. First, the RNA targeting activity of four CRISPR‐Cas13 variants were compared using a transient expression system in Nicotiana benthamiana. LwaCas13a and RfxCas13d had more efficient RNA and RNA virus targeting activity than PspCas13b and LshCas13a. Driven by the pCmYLCV promoter for the expression of gRNAs, RfxCas13d exhibited higher RNA targeting activity than that driven by the pAtU6 promoter. Furthermore, the targeting of SPCSV‐RNase3 using the LwaCas13a system inhibited its RNA silencing suppressor activity and recovered the RNA silencing activity in N. benthamiana leaf cells. Compared with the wild type, transgenic N. benthamiana plants carrying an RNase3‐targeted LwaCas13a system exhibited enhanced resistance against turnip mosaic virus TuMV‐GFP and cucumber mosaic virus CMV‐RNase3 co‐infection. Moreover, transgenic sweet potato plants carrying an RNase3‐targeted RfxCas13d system exhibited substantially improved SPVD resistance. This method may contribute to the development of SPVD immune germplasm and the enhancement of sweet potato production in SPVD‐prevalent regions.

First report of Sweet potato virus G in sweet potato in Italy

Sweet potato (Ipomoea batatas) is the sixth most important food crop in terms of production globally. In Italy, sweet potato is mainly cultivated in Veneto province (northeast Italy), and to a lesser extent in Lazio and Apulia provinces, on a total area of about 400 hectares. Among the sweet potato pests, viruses are the second most important constraint next to sweet potato weevil (Barkessa, 2018). In 2020, tubers of six different genotypes, with uncertain geographical origins, were sent by a grower from Veneto province to IPSP-CNR, with the aim of ascertaining their phytosanitary status. Two tubers of each genotype were grown in an insect-proof greenhouse (26/22°C, day/night) under natural light conditions and transplanted in 25 cm diameter pots containing peat bog-moss as a substrate. After about one month, leaves of only one genotype (identified as DP/578) showed light diffuse chlorotic symptoms (Figures 1, 2). Sap extracts from both plants of each of the six genotypes were tested with ELISA using broad-spectrum potyvirus antibodies (Bioreba, Switzerland). Only the symptomatic genotype DP/578 tested positive to potyvirus antibodies. Total RNA was extracted from the leaves of the two plants of DP/578 and from a healthy plant, with the Plant RNA Kit (Omega Bio-Tek, USA), and subjected to RT-PCR using the universal Potyviridae forward primer (5'-GGBAAYAAYAGYGGDCARCC-3'; Gibbs & Mackenzie, 1997) and the reverse primer SPFMV-Rv (5'-TTGCACACCCCTCATTCCYAAG-3'; Parrella et al., 2006). An amplification product of approximately 1700 bp was produced only from samples with symptoms but not from the healthy control. Sequences obtained from the two symptomatic plants were identical (GenBank Accession No. MW456564) and showed highest identity (98.2%) with the CH2-32-1 isolate of Sweet potato virus G (SPVG) from China (MK778832). SPVG alone is not considered a real threat to sweet potato production since infected plants rarely display symptoms (Rännäli et al., 2008). Significant yield losses have been recorded only when SPVG co-infects sweet potato with other potyviruses such as Sweet potato feathery mottle virus (SPFMV) (Clark & Hoy, 2006). Previously, outbreaks of SPFMV were reported in sweet potato cultivation in central Italy (Parrella et al., 2006). In the current study, attempts to amplify other sweet potato closely related potyviruses were unsuccessful using RT-PCR developed by Li et al. (2012). Based on our observation and results (symptoms recorded and serological detection of SPVG), the yield effects of SPVG may have been overlooked in sweet potato. Considering the number of existing SPVG strains and of sweet potato landraces/varieties, further investigation would be desirable to understand the real incidence of SPVG in individual sweet potato crops and its effects on production.

Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses

Globally, Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) occur frequently and in combination cause sweetpotato virus disease (SPVD). Many viral diseases are economically important and negatively impact the production and movement of germplasm across regions. Rapid detection of viruses is critical for effective control. Detection and quantification of viruses directly from sweetpotato remains a challenge. Current diagnostic tests are not sensitive enough to reliably detect viruses directly from the plant or require expensive laboratory equipment and expertise to perform. We developed a simple and rapid loop‐mediated isothermal amplification (LAMP) assay for the detection of SPFMV, SPCSV and begomoviruses related to sweet potato leaf curl virus (SPLCV). Laboratory validation recorded 100 % diagnostic sensitivity for all the three viruses. The LAMP assays were customized for field testing using a lyophilized thermostable isothermal master mix in a ready-to-use form that required no cold chain. The average time to positivity (TTP) was: SPFMV 5−30 min, SPCSV 15–43 min s and begomoviruses 28−45 mins. LAMP on-site testing results were comparable to PCR and RT-PCR confirmatory laboratory tests. The LAMP assay is a powerful tool for rapid sweetpotato virus detection at a reasonable cost and thus could serve as quality control systems for planting materials.

Species and genetic variability of sweet potato viruses in China

China is the world’s largest producer of sweet potato (Ipomoea batatas (L.) Lam.). Considering that there are numerous sweet potato-producing regions in China and sweet potato is a vegetatively propagated crop, the genetic diversity of sweet potato viruses could be high in the country. However, studies on species and genetic variabilities of sweet potato viruses in China are limited, making it difficult to prevent and control viral diseases in this crop. During 2014–2019, sweet potato samples with viral disease-like symptoms were randomly collected from sweet potato fields in 25 provinces in China. Twenty-one virus species, including 12 DNA and 9 RNA viruses, were identified in the samples using next-generation sequencing, polymerase chain reaction and rolling-circle amplification methods. One novel sweepovirus species, Sweet potato leaf curl Hubei virus (SPLCHbV), was identified. Two species, Sweet potato collusive virus and Tobacco mosaic virus, were identified for the first time in sweet potato in China. Full-length or nearly full-length genomic sequences of 111 isolates belonging to 18 viral species were obtained. Genome sequence comparisons of potyvirus isolates obtained in this study indicate that the genome of sweet potato virus 2 is highly conserved, whereas the other four potyviruses, sweet potato feathery mottle virus, sweet potato virus G, sweet potato latent virus and sweet potato virus C, exhibited a high genetic variability. The similarities among the 40 sweepovirus genomic sequences obtained from eight sweepovirus species are 67.0–99.8%. The eight sweepoviruses include 14 strains, of which 4 novel strains were identified from SPLCHbV and 1 from sweet potato leaf curl Guangxi virus. Five sweet potato chlorotic stunt virus (SPCSV) isolates obtained belong to the WA strain, and the genome sequences of SPCSV are highly conserved. Together, this study for the first time comprehensively reports the variability of sweet potato viruses in China.

In Vitro Identification and In Vivo Confirmation of Inhibitors for Sweet Potato Chlorotic Stunt Virus RNA Silencing Suppressor, a Viral RNase III.

ABSTRACTSweet potato virus disease (SPVD), caused by synergistic infection of Sweet potato chlorotic stunt virus (SPCSV) and Sweet potato feathery mottle virus (SPFMV), is responsible for substantial yield losses all over the world. However, there are currently no approved treatments for this severe disease. The crucial role played by RNase III of SPCSV (CSR3) as an RNA silencing suppressor during the viruses’ synergistic interaction in sweetpotato makes it an ideal drug target for developing antiviral treatment. In this study, high-throughput screening (HTS) of small molecular libraries targeting CSR3 was initiated by a virtual screen using Glide docking, allowing the selection of 6,400 compounds out of 136,353. We subsequently developed and carried out kinetic-based HTS using fluorescence resonance energy transfer technology, which isolated 112 compounds. These compounds were validated with dose-response assays including kinetic-based HTS and binding affinity assays using surface plasmon resonance and microscale thermophoresis. Finally, the interference of the selected compounds with viral accumulation was verified in planta. In summary, we identified five compounds belonging to two structural classes that inhibited CSR3 activity and reduced viral accumulation in plants. These results provide the foundation for developing antiviral agents targeting CSR3 to provide new strategies for controlling sweetpotato virus diseases.IMPORTANCE We report here a high-throughput inhibitor identification method that targets a severe sweetpotato virus disease caused by coinfection with two viruses (SPCSV and SPFMV). The disease is responsible for up to 90% yield losses. Specifically, we targeted the RNase III enzyme encoded by SPCSV, which plays an important role in suppressing the RNA silencing defense system of sweetpotato plants. Based on virtual screening, laboratory assays, and confirmation in planta, we identified five compounds that could be used to develop antiviral drugs to combat the most severe sweetpotato virus disease.

Articles of Significant Interest in This Issue

Sweet potato virus disease (SPVD) caused by synergistic infection of Sweet potato chlorotic stunt virus (SPCSV) and Sweet potato feathery mottle virus (SPFMV) leads to dramatic yield loss. Wang et al. (e00107-21) report in vitro identification and in vivo confirmation of inhibitors targeting the SPCSV RNase III (CSR3), an RNA silencing suppressor. As a result, five compounds belonging to two structural classes which inhibited CSR3 activity and reduced viral accumulation in plants were identified. These results provide the foundation for developing antiviral agents targeting viral RNase III to provide new strategies for controlling plant viruses.

Investigations on the Prevalence of Two Sweet Potato Viruses and their Potential Weed Reservoirs in Egypt

GLOBALLY, sweet potato (Ipomoea batatas (L.) Lam., family Convolvulaceae), is an important vegetable crop, unluckily, the viral infection induces a significant yield loss. The current study aims to identify the viruses infecting sweet potato in Egypt and highlights the role of the interfacing weeds as viruses reservoir. The study covered both of transmission, host range, inclusion bodies, virus morphology, histopathology, serological diagnosis, genome identification using RT-PCR and multiplex RT-PCR. The results revealed that the potyviruses sweet potato feathery mottle virus (SPFMV) and sweet potato virus G (SPVG) are the most devastating viruses affecting sweet potato in Egypt. Coinfection with of both viruses was the common feature in both of sweet potato and its interfacing weeds. A wider host range in families Convolvulaceae, Chenopodiaceae and Euphorbiaceae, was detected. The infection symptoms on sweet potato were comparable to that detected in the interfaced weeds. The infection-incidence in perennial Convolvulaceae and Tiliaceae weed-species showed 100% virus incidence. The characteristic amorphous inclusion bodies for the detected viruses and the cytopathological alterations on the host cells were observed under Transmission Electron Microscopy (TEM). The nucleotide sequences of the partial coat protein gene of the detected viruses were compared to the isolate available in GenBank. This work recommends the importance of weed interfacing crops as virus reservoir, and in light, the importance of multidisciplinary research work to resolve the virus problems threats in potential crops. Identifying viruses and their incidence increase the efficiency of the management strategies to control the virus spread.

Transcriptome analysis provides insights into the responses of sweet potato to sweet potato virus disease (SPVD)

Sweet potato (Ipomoea batatas) ranks among the most important crops in the world and provides nutritional and economic sustainability for subsistence farmers in sub-Saharan Africa. Its production is mainly constrained by sweet potato virus disease (SPVD) caused by the coinfection of two positive-sense single-stranded RNA viruses, sweet potato chlorotic stunt virus (SPCSV) and sweet potato feathery mottle virus (SPFMV). Current understanding of sweet potato responses to SPCSV and SPFMV at the molecular level remains very limited. In this study, we performed deep sequencing of both messenger RNA (mRNA) and small RNA (sRNA) populations in an SPVD-susceptible cultivar ‘Beauregard’ upon viral infection, to identify biological pathways that contribute to both general and specific host responses to these important viral pathogens. We found that pathways related to stress response and signaling were significantly affected by viral infection. sRNA components of these pathways were predominantly affected in late stages of the coinfection by SPCSV and SPFMV. We identified several novel microRNAs that were responsive to viral infection, some of which were predicted to target nucleotide-binding site leucine-rich repeat (NBS-LRR) disease resistance genes. The downregulation of the salicylic acid-mediated defense response pathway in particular seems to be a result of the viral infection process, and can in part explain the susceptible nature of the ‘Beauregard’ cultivar.