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A study conducted in Montecillo, Texcoco, Mexico, identified two viruses associated with symptoms of viral diseases in bean crops. The detected viruses were Orthotospovirus impatiensnecromaculae and two isolates of Potyvirus phaseoluteum. These viruses present significant differences from other viruses reported in different plant species. This finding highlights the importance of accurate identification for the management and control of these viral infections in bean crops in Mexico. [Extracted from the article]

East Asian Passiflora virus (EAPV) causes passionfruit woodiness disease, a major threat limiting passionfruit production in eastern Asia, including Taiwan and Vietnam. In this study, an infectious cDNA clone of a Taiwanese severe isolate EAPV-TW was tagged with a green fluorescent protein (GFP) reporter to monitor the virus in plants. Nicotiana benthamiana and yellow passionfruit plants inoculated with the construct showed typical symptoms of EAPV-TW. Based on our previous studies on pathogenicity determinants of potyviral HC-Pros, a deletion of six amino acids (d6) alone and its association with a point mutation (F8I, simplified as I8) were conducted in the N-terminal region of the HC-Pro gene of EAPV-TW to generate mutants of EAPV-d6 and EAPV-d6I8, respectively. The mutant EAPV-d6I8 caused infection without conspicuous symptoms in N. benthamiana and yellow passionfruit plants, while EAPV-d6 still induced slight leaf mottling. EAPV-d6I8 was stable after six passages under greenhouse conditions and displayed a zigzag pattern of virus accumulation, typical of a beneficial protective virus. The cross-protection effectiveness of EAPV-d6I8 was evaluated in both N. benthamiana and yellow passionfruit plants under greenhouse conditions. EAPV-d6I8 conferred complete cross-protection (100%) against the wild-type EAPV-TW-GFP in both N. benthamiana and yellow passionfruit plants, as verified by no severe symptoms, no fluorescent signals, and PCR-negative status for GFP. Furthermore, EAPV-d6I8 also provided complete protection against Vietnam's severe strain EAPV-GL1 in yellow passionfruit plants. Our results indicate that the attenuated mutant EAPV-d6I8 has great potential to control EAPV in Taiwan and Vietnam via cross-protection. [ABSTRACT FROM AUTHOR]

In Western Transdanubia in 2018 and in Central-Hungary in 2022 spiderwort plants showed flower breaking symptoms and mild mosaic on the foliage, which indicated a potential virus infection. One gram of symptomatic leaf sample was collected at both locations. Potyvirus-specific ELISA tests demonstrated potyvirus infection. To identify the virus species, potyvirus-specific RT-PCR was carried out on the samples. In both samples specific PCR products were detected and cloned into pGEM®-T Easy vector. The nucleotide sequences of the inserts were determined by Sanger sequencing. BLASTn searches on the complete coat protein region of both isolates demonstrated more than 99.87% identity with Tradescantia mild mosaic virus (TraMMV; accession number OL584375). Koch postulates were fulfilled by sap inoculating seed grown spiderwort plants. Phylogenetic analyses of the TraMMV coat protein sequences revealed two distinct evolutionary lineages: a tropical subgroup with at least 97.84% identity within the group and temperate subgroup with at least 98.97% identity within the group. One major difference between the subgroups was in the triplet responsible for vector transmission. The isolates belonging to the tropical subgroup had DAG triplets, while the temperate subgroup had NAG triplets. The difference in the triplets could be caused by natural diversification, directional selection or disruptive selection. License: CC BY-NC-ND [ABSTRACT FROM AUTHOR]

White yam (Dioscorea rotundata) plants collected from farmers' fields and planted at the Areka Agricultural Research Center, Southern Ethiopia, displayed mosaic, mottling, and chlorosis symptoms. To determine the presence of viral pathogens, an investigation for virome characterization was conducted by Illumina high-throughput sequencing. The bioinformatics analysis allowed the assembly of five viral genomes, which according to the ICTV criteria were assigned to a novel potyvirus (3 genome sequences) and a novel crinivirus (2 genome sequences). The potyvirus showed ~ 66% nucleotide (nt) identity in the polyprotein sequence to yam mosaic virus (NC004752), clearly below the demarcation criteria of 76% identity. For the crinivirus, the RNA 1 and RNA 2 shared the highest sequence identity to lettuce chlorosis virus, and alignment of the aa sequence of the RdRp, CP and HSP70h (~ 49%, 45% and 76% identity), considered for the demarcation criteria, revealed the finding of a novel virus species. The names Ethiopian yam virus (EYV) and Yam virus 1 (YV-1) are proposed for the two tentative new virus species. [ABSTRACT FROM AUTHOR]

Background: Pepper veinal mottle virus (PVMV) belongs to the genus Potyvirus within the family Potyviridae and is a major threat to pepper production, causing reduction in yield and fruit quality; however, efficient pesticides and chemical treatments for plant protection against viral infections are lacking. Hence, there is a critical need to discover highly active and environment-friendly antiviral agents derived from natural sources. Bacillus spp. are widely utilized as biocontrol agents to manage fungal, bacterial, and viral plant diseases. Particularly, Bacillus velezensis HN-2 exhibits a strong antibiotic activity against plant pathogens and can also induce plant resistance. Methods: The experimental subjects employed in this study were Bacillus velezensis HN-2, benzothiadiazole, and dufulin, aiming to evaluate their impact on antioxidant activity, levels of reactive oxygen species, activity of defense enzymes, and expression of defense-related genes in Nicotiana benthamiana. Furthermore, the colonization ability of Bacillus velezensis HN-2 in Capsicum chinense was investigated. Results: The results of bioassays revealed the robust colonization capability of Bacillus velezensis HN-2, particularly in intercellular spaces, leading to delayed infection and enhanced protection against PVMV through multiple plant defense mechanisms, thereby promoting plant growth. Furthermore, Bacillus velezensis HN-2 increased the activities of antioxidant enzymes, thereby mitigating the PVMV-induced ROS production in Nicotiana benthamiana. Moreover, the application of Bacillus velezensis HN-2 at 5 dpi significantly increased the expression of JA-responsive genes, whereas the expression of salicylic acidresponsive genes remained unchanged, implying the activation of the JA signaling pathway as a crucial mechanism underlying Bacillus velezensis HN-2-induced anti-PVMV activity. Immunoblot analysis revealed that HN-2 treatment delayed PVMV infection at 15 dpi, further highlighting its role in inducing plant resistance and promoting growth and development. Conclusions: These findings underscore the potential of Bacillus velezensis HN-2 for field application in managing viral plant diseases effectively. [ABSTRACT FROM AUTHOR]

Plant viruses cause severe losses in agricultural production. Parasite-derived resistance (PDR) offers a promising avenue for developing disease-resistant varieties independent of resistance genes. However, for potyviruses with great agricultural importance, such as soybean mosaic virus (SMV), systematic research on viral genes that can be used for PDR has not been conducted. In this study, we transiently expressed the untranslated region (UTR) or each protein-coding cistron of SMV in Nicotiana benthamiana to evaluate their potential role in conferring PDR. A viral suppressor of RNA silencing (VSR) was also applied to investigate the possible mechanisms of the PDR. The results showed that the transient overexpression of UTR and each cistron of SMV could inhibit SMV infection. The expression of VSR in N. benthamiana leaves could compromise UTR and most of the SMV cistron-mediated inhibition of SMV infection, indicating the involvement of RNA silencing in PDR. In comparison, the expression of VSR could not compromise the PDR conferred by coat protein (CP), P3N-PIPO, cylindrical inclusion (CI), and NIa-Pro, suggesting that these viral cistrons may play roles in PDR at the protein level. These results reveal diverse mechanisms in PDR conferred by different viral cistrons and provide potential gene candidates that can be used for transgenic approaches against SMV. [ABSTRACT FROM AUTHOR]

HC-Pro and coal protein (CP) genes of a potyvirus facilitate cell-to-cell movement and are involved in the systemic movement of the viruses. The interaction between HC-Pro and CP is mandatory for aphid transmission. Two turnip mosaic virus (TuMV) isolates, RC4 and YC5, were collected from calla lily plants in Taiwan. The virus derived from the infectious clone pYC5 cannot move systemically in Chenopodium quinoa plants and loses aphid transmissibility in Nicotiana benthamiana plants, like the initially isolated virus. Sequence analysis revealed that two amino acids, P5 and A206, of YC5 CP uniquely differ from RC4 and other TuMV strains. Recombination assay and site-directed mutagenesis revealed that the fifth residue of leucine (L) at the N-terminal region of the CP (TuMV-RC4), rather than proline (P) (TuM V-YC5), is critical to permit the systemic spread in C. quinoa plants. Moreover, the single substitution mutant YC5-CPP5L became aphid transmissible, similar to RC4. Fluorescence microscopy revealed that YC5-GFP was restricted in the petioles of inoculated leaves, whereas YC5-CPp5L-GFP translocated through the petioles of inoculated leaves, the main stem, and the petioles of the upper uninoculated leaves of C. quinoa plants. In addition, YC5-GUS was blocked at the basal part of the petiole connecting to the main stem of the inoculated C. quinoa plants, whereas YC5-CPp51-GFP translocated to the upper leaves. Thus, a single amino acid, the residue Ls at the N-terminal region right before the f'DAGs motif, is critical for the systemic translocation ability of TuMV in a local lesion host and for aphid transmissibility in a systemic host. [ABSTRACT FROM AUTHOR]

Viruses employ a series of diverse translational strategies to expand their coding capacity, which produces viral proteins with common domains and entangles virus-host interactions. P3N-PIPO, which is a transcriptional slippage product from the P3 cistron, is a potyviral protein dedicated to intercellular movement. Here, we show that P3N-PIPO from watermelon mosaic virus (WMV) triggers cell death when transiently expressed in Cucumis melo accession PI 414723 carrying the Wmr resistance gene. Surprisingly, expression of the P3N domain, shared by both P3N-PIPO and P3, can alone induce cell death, whereas expression of P3 fails to activate cell death in PI 414723. Confocal microscopy analysis revealed that P3N-PIPO targets plasmodesmata (PD) and P3N associates with PD, while P3 localizes in endoplasmic reticulum in melon cells. We also found that mutations in residues L35, L38, P41, and I43 of the P3N domain individually disrupt the cell death induced by P3N-PIPO, but do not affect the PD localization of P3N-PIPO. Furthermore, WMV mutants with L35A or I43A can systemically infect PI 414723 plants. These key residues guide us to discover some WMV isolates potentially breaking the Wmr resistance. Through searching the NCBI database, we discovered some WMV isolates with variations in these key sites, and one naturally occurring I43V variation enables WMV to systemically infect PI 414723 plants. Taken together, these results demonstrate that P3N-PIPO, but not P3, is the avirulence determinant recognized by Wmr, although the shared N terminal P3N domain can alone trigger cell death. This work reveals a novel viral avirulence (Avr) gene recognized by a resistance (R) gene. This novel viral Avr gene is special because it is a transcriptional slippage product from another virus gene, which means that their encoding proteins share the common N-terminal domain but have distinct C-terminal domains. Amazingly, we found that it is the common N-terminal domain that determines the Avr-R recognition, but only one of the viral proteins can be recognized by the R protein to induce cell death. Next, we found that these two viral proteins target different subcellular compartments. In addition, we discovered some virus isolates with variations in the common N-terminal domain and one naturally occurring variation that enables the virus to overcome the resistance. These results show how viral proteins with common domains interact with a host resistance protein and provide new evidence for the arms race between plants and viruses. [ABSTRACT FROM AUTHOR]

RNA silencing is an innate immune mechanism of plants against invasion by viral pathogens. Artificial microRNA (amiRNA) can be engineered to specifically induce RNA silencing against viruses in transgenic plants and has great potential for disease control. Here, we describe the development and application of amiRNA-based technology to induce resistance to soybean mosaic virus (SMV), a plant virus with a positive-sense single-stranded RNA genome. We have shown that the amiRNA targeting the SMV P1 coding region has the highest antiviral activity than those targeting other SMV genes in a transient amiRNA expression assay. We transformed the gene encoding the P1-targeting amiRNA and obtained stable transgenic Nicotiana benthamiana lines (amiR-P1-3-1-2-1 and amiR-P1-4-1-2-1). Our results have demonstrated the efficient suppression of SMV infection in the P1-targeting amiRNA transgenic plants in an expression level-dependent manner. In particular, the amiR-P1-3-1-2-1 transgenic plant showed high expression of amiR-P1 and low SMV accumulation after being challenged with SMV. Thus, a transgenic approach utilizing the amiRNA technology appears to be effective in generating resistance to SMV. [ABSTRACT FROM AUTHOR]

The movement of potyviruses, the largest genus of single‐stranded, positive‐sense RNA viruses responsible for serious diseases in crops, is very complex. As potyviruses developed strategies to hijack the host secretory pathway and plasmodesmata (PD) for their transport, the goal of this study was to identify membrane and/or PD‐proteins that interact with the 6K2 protein, a potyviral protein involved in replication and cell‐to‐cell movement of turnip mosaic virus (TuMV). Using split‐ubiquitin membrane yeast two‐hybrid assays, we screened an Arabidopsis cDNA library for interactors of TuMV6K2. We isolated AtHVA22a (Hordeum vulgare abscisic acid responsive gene 22), which belongs to a multigenic family of transmembrane proteins, homologous to Receptor expression‐enhancing protein (Reep)/Deleted in polyposis (DP1)/Yop1 family proteins in animal and yeast. HVA22/DP1/Yop1 family genes are widely distributed in eukaryotes, but the role of HVA22 proteins in plants is still not well known, although proteomics analysis of PD fractions purified from Arabidopsis suspension cells showed that AtHVA22a is highly enriched in a PD proteome. We confirmed the interaction between TuMV6K2 and AtHVA22a in yeast, as well as in planta by using bimolecular fluorescence complementation and showed that TuMV6K2/AtHVA22a interaction occurs at the level of the viral replication compartment during TuMV infection. Finally, we showed that the propagation of TuMV is increased when AtHVA22a is overexpressed in planta but slowed down upon mutagenesis of AtHVA22a by CRISPR‐Cas9. Altogether, our results indicate that AtHVA22a plays an agonistic effect on TuMV propagation and that the C‐terminal tail of the protein is important in this process. [ABSTRACT FROM AUTHOR]

Tomato (Solanum lycopersicum) is the most important vegetable and fruit crop in the family Solanaceae worldwide. Numerous pests and pathogens, especially viruses, severely affect tomato production, causing immeasurable market losses. In Taiwan, the cultivation of tomato crops is mainly threatened by insect-borne viruses, among which pepper veinal mottle virus (PVMV) is one of the most prevalent. PVMV is a member of the genus Potyvirus of the family Potyviridae and is non-persistently transmitted by aphids. Its infection significantly reduces tomato fruit yield and quality. So far, no PVMV-resistant tomato lines are available. In this study, we performed nitrite-induced mutagenesis of the PVMV tomato isolate Tn to generate attenuated PVMV mutants. PVMV Tn causes necrotic lesions in Chenopodium quinoa leaves and severe mosaic and wilting in Nicotiana benthamiana plants. After nitrite treatment, three attenuated PVMV mutants, m4-8, m10-1, and m10-11, were selected while inducing milder responses to C. quinoa and N. benthamiana with lower accumulation in tomato plants. In greenhouse tests, the three mutants showed different degrees of cross-protection against wild-type PVMV Tn. m4-8 showed the highest protective efficacy against PVMV Tn in N. benthamiana and tomato plants, 100% and 97.9%, respectively. A whole-genome sequence comparison of PVMV Tn and m4-8 revealed that 20 nucleotide substitutions occurred in the m4-8 genome, resulting in 18 amino acid changes. Our results suggest that m4-8 has excellent potential to protect tomato crops from PVMV. The application of m4-8 in protecting other Solanaceae crops, such as peppers, will be studied in the future. [ABSTRACT FROM AUTHOR]

Introduction/Objective. Ayocote bean (Phaseolus coccineus) has potential as a source of resistance in breeding programs because it exhibits greater tolerance to plant pathogens than P. vulgaris. However, its sanitary characterization is insipient; therefore, the purpose of this work was to carry out an etiological-epidemiological diagnosis, with emphasis on presumptive symptoms of viral and phytoplasmic organisms, and a typical fungal signs of powdery mildew. Materials and Methods: A plot (50 x 62 m) of flowering Ayocote bean was selected. It was divided into 80 (8 x 10) quadrats (6 x 6 m) and 720 subquadrats (2 x 2 m). From 25 plants with powdery-mildew-type leaf symptoms, mycelium was collected with adhesive tape for light microscopy observation and taxonomic identification. Length-width measurements were made on 60 conidia. Pure mycelium collected in situ and ex situ from 1-5 leaflets/plant was used for genomic analysis by PCR with universal primers ITS1 and ITS4. Samples were sequenced in Macrogen Inc. Korea. A total of 63 plants and 121 trifoliate leaves with viral and phytoplasmic symptoms were collected by direct sampling. In 88/121 samples, genomic analysis was performed by PCR with universal primers for Potyvirus (1), Begomovirus (2), and Phytoplasmas (1). Sequence editing and analysis were performed in SeqAssem and BLASTn/GenBank. Phylogenetic constructions were developed in Mega 11 with MUSCLE, Maximum Likelihood (ML), and HKY substitution model (1000-Bootstrap). Putative powdery mildew severity (%), flower damage (%), Macrodactylus sp. adult density, and plant vigor (%) were evaluated in 80 quadrats (3subquadrats/quadrat) with App-Monitor®v1.1 configured with a 5-class scale. In GoldenSurfer® v10, Kriging geostatistical analysis was performed to determine the spatial interrelationship between these variables. Results: Erysiphe vignae was identified as associated with powdery mildew of P. coccineus. The fungus, with hyaline, ovoid to ellipsoid conidia measuring 31.74 ± 0.3419 µm x 15.11 ± 0.1579 µm, without the presence of fibrosin bodies, had 100% genomic homology. This is the first report in Mexico. With average July-August temperature and relative humidity of 16.3 °C (±5.8) and 92.8 % (±10.7), respectively, powdery mildew leaf incidence and severity were 65.3 and 22.7 % (±16.9, range: 0 - 66.5 %), respectively. The most inductive focus (60-80 % severity) had an aggregate e 4-quadrat pattern (96 m2, lag = 4 and s2-s = 450). Inoculum dispersal was significantly associated with dominant North-South winds and plant vigor (lag = 4 and s2-s = 470). Flower damage was inconclusive in its spatial association with powdery mildew and Macrodactylus sp. suggesting uncorrelated events. No Potyvirus, Begomovirus, or Phytoplasmas were detected associated with yellowing, leaf distortion, mosaic, internode shortening, and other symptoms observed in situ. This confirms the relative tolerance/resistance reported for P. coccineus. Conclusion: E. vignae (Erysiphales: Erysiphaceae) associated with P. coccineus is reported for the first time in Mexico with moderate to intense epidemic level, which indicates its susceptible condition to this fungus. However, negative results for Potyvirus, Begomovirus, and Phytoplasmas, validate the apparent tolerance/resistance of P. coccineus to these organisms. [ABSTRACT FROM AUTHOR]

The best-characterized functional motifs of the potyviral Helper-Component protease (HC-Pro) responding for aphid transmission, RNA silencing suppression, movement, symptom development, and replication are gathered in this review. The potential cellular protein targets of plant virus proteases remain largely unknown despite their multifunctionality. The HC-Pro catalytic domain, as a cysteine protease, autoproteolytically cleaves the potyviral polyproteins in the sequence motif YXVG/G and is not expected to act on host targets; however, 146 plant proteins in the Viridiplantae clade containing this motif were searched in the UniProtKB database and are discussed. On the other hand, more than 20 interactions within the entire HC-Pro structure are known. Most of these interactions with host targets (such as the 20S proteasome, methyltransferase, transcription factor eIF4E, and microtubule-associated protein HIP2) modulate the cellular environments for the benefit of virus accumulation or contribute to symptom severity (interactions with MinD, Rubisco, ferredoxin) or participate in the suppression of RNA silencing (host protein VARICOSE, calmodulin-like protein). On the contrary, the interaction of HC-Pro with triacylglycerol lipase, calreticulin, and violaxanthin deepoxidase seems to be beneficial for the host plant. The strength of these interactions between HC-Pro and the corresponding host protein vary with the plant species. Therefore, these interactions may explain the species-specific sensitivity to potyviruses. [ABSTRACT FROM AUTHOR]

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IntroductionCanola (Brassica napus L.) is one of the most important crops worldwide. Turnip yellows virus (TuYV), transmitted by aphids, is one of the most damaging viruses affecting canola crops and is challenging to control. With the prediction of more intense and prolonged drought events due to future climate change, an additional factor may extensively impact the epidemiology of plant diseases. This study aimed to understand the impact of drought on canola plants infected with TuYV and to explore the relationship between virus infection and drought.MethodsTwo glasshouse experiments were conducted: 1. Competition: Four plants (two infected, two non-infected) were grown in the same pot. 2. No Competition: One plant was grown per pot. In both experiments, infected and non-infected canola plants were exposed to well-watered conditions, water stress (simulated drought), and terminal drought. Various plant traits were recorded, including biomass, leaf area, height, number of leaves, chlorophyll content, water use efficiency, and virus symptom expression.ResultsBoth virus infection and water stress reduced dry biomass, leaf area, and height. Virus infection alone reduced canola biomass by up to 49% compared to non-infected, well-watered controls. Under water stress or terminal drought, the biomass of TuYV-infected plants was further reduced by up to 71% and 65%, respectively. Virus infection also reduced the number of leaves, although water treatment alone did not. Chlorophyll content was higher in water-stressed and terminal drought plants compared to well-watered ones, while virus infection reduced chlorophyll content. The impact of drought and virus infection was more pronounced when plants were under competition.DiscussionGiven the expected increase in prolonged and frequent droughts in many canola-growing regions due to climate change, a significant detrimental effect on canola production due to the combined influence of drought and TuYV is anticipated. This study underscores the need for developing mitigation strategies to protect canola production in a changing climate.

Papaver somniferum L. produces therapeutically-useful alkaloids like morphine, codeine, papaverine, and thebaine that accumulate in the latex of its capsule. Apart from its alkaloids, the edible seeds of the plant have high nutritional value and culinary use, with the seed oil being rich in the health-promoting unsaturated fatty acids. Sujata is low alkaloid-producing latex-less culinary variety of P. somniferum developed from an alkaloid-rich gum harvest parent (Sampada) for curtailing the narcotic menace of morphine and opium (dried latex). Earlier, it has been shown that the expression of stearoyl-acyl-carrier protein desaturase (PsSAD) in the young capsule and the proportion of unsaturated fatty acids in the seeds are higher in Sujata as compared to Sampada. Here, we studied bacterial endophytes isolated from Sujata tissues for their role in defining the fatty acid (saturated stearic versus unsaturated oleic) profile in its leaves and seeds. Besides, a potyvirus infecting P. somniferum was characterized through its coat protein gene sequencing and transmission electron microscopy and the two genotypes (Sujata and Sampada) contrasting in their phenotypes (oil/fatty acid, alkaloid and latex profiles) were compared for their susceptibility towards it through indicator plant- and qRT-PCR-based virus infectivity assays. In both the assays, Sujata was found to have higher susceptibility to the potyvirus, as compared to Sampada. The most plausible reason for this could be the higher PsSAD expression in the leaves of Sujata, which resulted in higher relative levels of oleic acid and lower relative levels of stearic acid in its leaves as compared to that of Sampada.

ABSTRACT Potato virus Y (PVY) is recognized as one of the most common and destructive pathogens seriously affecting potato producing areas worldwide. More recently PVYNTN and PVYN:O/N-Wi, have emerged as the main strains present in the PVY infected plants detected in Brazilian potato fields. In this study, samples of potato collected in south part of Minas Gerais - Brazil were first tested by DAS-ELISA and then by RT-PCR multiplex in order to discriminate the PVY strains. Afterward, part of them was tested by RT-qPCR to confirm and quantify the viruses in infected tissues. The sensitivity of the techniques for detecting PVY isolates present in the sampled locations was investigated, as well as the occurrence of mixed infections, aiming to understand the general epidemiological picture of this pathogen in potato producing fields. In the multiplex RT-PCR test, the samples with O and N serotypes were identified as infected with PVYNTN and PVYN:O/N-Wi strains. When tested by RT-PCR for amplification of PVYE, 41 samples (67,2%) were positive, having a characteristic electrophoretic profile for this recombinant strain, and 9 isolates were also observed with atypical patterns for recombinant PVYE. The best technique to detect mixed infection was RT-qPCR, with the concentration of PVYNTN being much higher than that of PVYN:O/N-Wi. These results show the importance of using the most suitable method for the diagnosis and surveying of PVY strains in crop fields and reveal, for the first time, the dissemination of PVYE recombinants to several Brazilian potato fields.

Abstract Background Plant responses to a wide range of stresses are known to be regulated by epigenetic mechanisms. Pathogen-related investigations, particularly against RNA viruses, are however scarce. It has been demonstrated that Arabidopsis thaliana plants defective in some members of the RNA-directed DNA methylation (RdDM) or histone modification pathways presented differential susceptibility to the turnip mosaic virus. In order to identify genes directly targeted by the RdDM-related RNA Polymerase V (POLV) complex and the histone demethylase protein JUMONJI14 (JMJ14) during infection, the transcriptomes of infected mutant and control plants were obtained and integrated with available chromatin occupancy data for various epigenetic proteins and marks. Results A comprehensive list of virus-responsive gene candidates to be regulated by the two proteins was obtained. Twelve genes were selected for further characterization, confirming their dynamic regulation during the course of infection. Several epigenetic marks on their promoter sequences were found using in silico data, raising confidence that the identified genes are actually regulated by epigenetic mechanisms. The altered expression of six of these genes in mutants of the methyltransferase gene CURLY LEAF and the histone deacetylase gene HISTONE DEACETYLASE 19 suggests that some virus-responsive genes may be regulated by multiple coordinated epigenetic complexes. A temporally separated multiple plant virus infection experiment in which plants were transiently infected with one virus and then infected by a second one was designed to investigate the possible roles of the identified POLV- and JMJ14-regulated genes in wild-type (WT) plants. Plants that had previously been stimulated with viruses were found to be more resistant to subsequent virus challenge than control plants. Several POLV- and JMJ14-regulated genes were found to be regulated in virus induced resistance in WT plants, with some of them poisoned to be expressed in early infection stages. Conclusions A set of confident candidate genes directly regulated by the POLV and JMJ14 proteins during virus infection was identified, with indications that some of them may be regulated by multiple epigenetic modules. A subset of these genes may also play a role in the tolerance of WT plants to repeated, intermittent virus infections.

Sugarcane mosaic virus (SCMV) (genus, Potyvirus; family, Potyviridae) is widespread, deleterious, and the most damaging pathogen of sugarcane (Saccharum officinarum L. and Saccharum spp.) that causes a substantial barrier to producing high sugarcane earnings. Sugarcane mosaic disease (SCMD) is caused by a single or compound infection of SCMV disseminated by several aphid vectors in a non-persistent manner. SCMV has flexuous filamentous particle of 700–750 nm long, which encapsidated in a positive-sense, single-stranded RNA molecule of 9575 nucleotides. RNA interference (RNAi)-mediated antiviral innate immunity is an evolutionarily conserved key biological process in eukaryotes and has evolved as an antiviral defense system to interfere with viral genomes for controlling infections in plants. The current study aims to analyze sugarcane (Saccharum officinarum L. and Saccharum spp.) locus-derived microRNAs (sof-miRNAs/ssp-miRNAs) with predicted potential for targeting the SCMV +ssRNA-encoded mRNAs, using a predictive approach that involves five algorithms. The ultimate goal of this research is to mobilize the in silico- predicted endogenous sof-miRNAs/ssp-miRNAs to experimentally trigger the catalytic RNAi pathway and generate sugarcane cultivars to evaluate the potential antiviral resistance surveillance ability and capacity for SCMV. Experimentally validated mature sugarcane (S. officinarum, 2n = 8X = 80) and (S. spp., 2n = 100–120) sof-miRNA/ssp-miRNA sequences (n = 28) were downloaded from the miRBase database and aligned with the SCMV genome (KY548506). Among the 28 targeted mature locus-derived sof-miRNAs/ssp-miRNAs evaluated, one sugarcane miRNA homolog, sof-miR159c, was identified to have a predicted miRNA binding site, at nucleotide position 3847 of the SCMV genome targeting CI ORF. To verify the accuracy of the target prediction accuracy and to determine whether the sugarcane sof-miRNA/ssp-miRNA could bind the predicted SCMV mRNA target(s), we constructed an integrated Circos plot. A genome-wide in silico-predicted miRNA-mediated target gene regulatory network was implicated to validate interactions necessary to warrant in vivo analysis. The current work provides valuable computational evidence for the generation of SCMV-resistant sugarcane cultivars.

Background: Plant responses to a wide range of stresses are known to be regulated by epigenetic mechanisms. Pathogen-related investigations, particularly against RNA viruses, are however scarce. It has been demonstrated that Arabidopsis thaliana plants defective in some members of the RNA-directed DNA methylation (RdDM) or histone modification pathways presented differential susceptibility to the turnip mosaic virus. In order to identify genes directly targeted by the RdDM-related RNA Polymerase V (POLV) complex and the histone demethylase protein JUMONJI14 (JMJ14) during infection, the transcriptomes of infected mutant and control plants were obtained and integrated with available chromatin occupancy data for various epigenetic proteins and marks. Results: A comprehensive list of virus-responsive gene candidates to be regulated by the two proteins was obtained. Twelve genes were selected for further characterization, confirming their dynamic regulation during the course of infection. Several epigenetic marks on their promoter sequences were found using in silico data, raising confidence that the identified genes are actually regulated by epigenetic mechanisms. The altered expression of six of these genes in mutants of the methyltransferase gene CURLY LEAF and the histone deacetylase gene HISTONE DEACETYLASE 19 suggests that some virus-responsive genes may be regulated by multiple coordinated epigenetic complexes. A temporally separated multiple plant virus infection experiment in which plants were transiently infected with one virus and then infected by a second one was designed to investigate the possible roles of the identified POLV- and JMJ14-regulated genes in wild-type (WT) plants. Plants that had previously been stimulated with viruses were found to be more resistant to subsequent virus challenge than control plants. Several POLV- and JMJ14-regulated genes were found to be regulated in virus induced resistance in WT plants, with some of them poisoned to be expressed in early infection stages. Conclusions: A set of confident candidate genes directly regulated by the POLV and JMJ14 proteins during virus infection was identified, with indications that some of them may be regulated by multiple epigenetic modules. A subset of these genes may also play a role in the tolerance of WT plants to repeated, intermittent virus infections. [ABSTRACT FROM AUTHOR]

During the surveys conducted from green space in Shiraz City, Iran, in the summer of 2022, leaf chlorosis and mosaic symptoms were observed on the leaves of black-eyed-susans (Rudbeckia hirta), a Dahlia sp., and Mexican Petunia (Ruellia brittoniana) plants. Total genomic RNA was separately extracted from 10 symptomatic and one symptomless (negative control) leaf samples and subjected to Reverse Transcription-Polymerase Chain Reaction (RT-PCR) using a potyvirus degenerate primer pair (NIb3R, NIb2F). RT-PCR resulted in the amplification of an approximately 350 bp DNA fragment in all symptomatic samples, while no such fragment was amplified from the symptomless plant. The amplified DNA fragment was subjected to Sanger sequencing, and its size was determined to be exactly 350 bp, and confirmed that it belongs to the Nib gene of potato virus Y (PVY). The nucleotide (nt) sequence of the amplicons was compared with the nt sequence of the same region of some other PVY isolates that were available in the GenBank. The sequence analysis revealed that the R. brittoniana isolate exhibited the highest (98.8%) similarity to a PVY isolate from the USA with the Acc. No. of KY_848029.1. Similarly, the R. hirta and the Dahlia isolates showed the highest (98.3%, 98.4 %, respectively ) similarity to a PVY isolate from Kazakhstan with the Acc. No. of ON_583980.1. [ABSTRACT FROM AUTHOR]

Sugarcane mosaic virus (SCMV) (genus, Potyvirus; family, Potyviridae) is widespread, deleterious, and the most damaging pathogen of sugarcane (Saccharum officinarum L. and Saccharum spp.) that causes a substantial barrier to producing high sugarcane earnings. Sugarcane mosaic disease (SCMD) is caused by a single or compound infection of SCMV disseminated by several aphid vectors in a non-persistent manner. SCMV has flexuous filamentous particle of 700–750 nm long, which encapsidated in a positive-sense, single-stranded RNA molecule of 9575 nucleotides. RNA interference (RNAi)-mediated antiviral innate immunity is an evolutionarily conserved key biological process in eukaryotes and has evolved as an antiviral defense system to interfere with viral genomes for controlling infections in plants. The current study aims to analyze sugarcane (Saccharum officinarum L. and Saccharum spp.) locus-derived microRNAs (sof-miRNAs/ssp-miRNAs) with predicted potential for targeting the SCMV +ssRNA-encoded mRNAs, using a predictive approach that involves five algorithms. The ultimate goal of this research is to mobilize the in silico- predicted endogenous sof-miRNAs/ssp-miRNAs to experimentally trigger the catalytic RNAi pathway and generate sugarcane cultivars to evaluate the potential antiviral resistance surveillance ability and capacity for SCMV. Experimentally validated mature sugarcane (S. officinarum, 2n = 8X = 80) and (S. spp., 2n = 100–120) sof-miRNA/ssp-miRNA sequences (n = 28) were downloaded from the miRBase database and aligned with the SCMV genome (KY548506). Among the 28 targeted mature locus-derived sof-miRNAs/ssp-miRNAs evaluated, one sugarcane miRNA homolog, sof-miR159c, was identified to have a predicted miRNA binding site, at nucleotide position 3847 of the SCMV genome targeting CI ORF. To verify the accuracy of the target prediction accuracy and to determine whether the sugarcane sof-miRNA/ssp-miRNA could bind the predicted SCMV mRNA target(s), we constructed an integrated Circos plot. A genome-wide in silico-predicted miRNA-mediated target gene regulatory network was implicated to validate interactions necessary to warrant in vivo analysis. The current work provides valuable computational evidence for the generation of SCMV-resistant sugarcane cultivars. [ABSTRACT FROM AUTHOR]

Copyright of Jurnal Fitopatologi Indonesia is the property of IPB University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Analysis of the viromes of three symptomatic Fabaceae plants, i.e., red clover (Trifolium pratense L.), pea (Pisum sativum L.), and common bean (Phaseolus vulgaris L.), using high-throughput sequencing revealed complex infections and enabled the acquisition of complete genomes of a potyvirus, bean yellow mosaic virus (BYMV). Based on phylogenetic analysis, the Slovak BYMV isolates belong to two distinct molecular groups, i.e., VI (isolate FA40) and XI (isolates DAT, PS2). Five commercial pea genotypes (Alderman, Ambrosia, Gloriosa, Herkules, Senator) were successfully infected with the BYMV-PS2 inoculum and displayed similar systemic chlorotic mottling symptoms. Relative comparison of optical density values using semi-quantitative DAS-ELISA revealed significant differences among virus titers in one of the infected pea genotypes (Ambrosia) when upper fully developed leaves were tested. Immunoblot analysis of systemically infected Alderman plants showed rather uneven virus accumulation in different plant parts. The lowest virus accumulation was repeatedly detected in the roots, while the highest was in the upper part of the plant stem. [ABSTRACT FROM AUTHOR]

Impatiens walleriana was introduced into Brazil and is now widely cultivated as an ornamental plant. Impatiens plants from a public garden showing leaf mosaic and colour break symptoms were submitted to total RNA extraction and high throughput sequencing. Identity of 99% with Amaranthus leaf mottle virus (AmLMV, Potyvirus) was observed. This is the first report of AmLMV in both Impatiens walleriana and the Americas. [ABSTRACT FROM AUTHOR]

Viruses rely completely on host translational machinery to produce the proteins encoded by their genes. Controlling translation initiation is important for gaining translational advantage in conflicts between the host and virus. The eukaryotic translation initiation factor 4E (eIF4E) has been reported to be hijacked by potyviruses for virus multiplication. The role of translation regulation in defence and anti‐defence between plants and viruses is not well understood. We report that the transcript level of eIF6 was markedly increased in turnip mosaic virus (TuMV)‐infected Nicotiana benthamiana. TuMV infection was impaired by overexpression of N. benthamiana eIF6 (NbeIF6) either transiently expressed in leaves or stably expressed in transgenic plants. Polysome profile assays showed that overexpression of NbeIF6 caused the accumulation of 40S and 60S ribosomal subunits, the reduction of polysomes, and also compromised TuMV UTR‐mediated translation, indicating a defence role for upregulated NbeIF6 during TuMV infection. However, the polysome profile in TuMV‐infected leaves was not identical to that in leaves overexpressing NbeIF6. Further analysis showed that TuMV NIb protein, the RNA‐dependent RNA polymerase, interacted with NbeIF6 and interfered with its effect on the ribosomal subunits, suggesting that NIb might have a counterdefence role. The results propose a possible regulatory mechanism at the translation level during plant–virus interaction. [ABSTRACT FROM AUTHOR]

Maize lethal necrosis (MLN) is a viral disease caused by host co-infection by maize chlorotic mottle virus (MCMV) and a potyvirus, such as sugarcane mosaic virus (SCMV). The disease is most effectively managed by growing MLN-resistant varieties. However, the relative importance of MCMV and potyvirus resistance in managing this synergistic disease is poorly characterized. In this study, we evaluated the effects of SCMV and/or MCMV resistance on disease, virus titers, and synergism and explored expression patterns of known potyvirus resistance genes TrxH and ABP1. MLN disease was significantly lower in both the MCMV-resistant and SCMV-resistant inbred lines compared with the susceptible control Oh28. Prior to 14 days postinoculation (dpi), MCMV titers in resistant lines N211 and KS23-6 were more than 100,000-fold lower than found in the susceptible Oh28. However, despite no visible symptoms, titer differences between MCMV-resistant and -susceptible lines were negligible by 14 dpi. In contrast, systemic SCMV titers in the potyvirus-resistant line, Pa405, ranged from 130,000-fold to 2 million-fold lower than susceptible Oh28 as disease progressed. Initial TrxH expression was up to 49,000-fold lower in Oh28 compared with other genotypes, whereas expression of ABP1 was up to 4.5-fold lower. Measures of virus synergy indicate that whereas MCMV resistance is effective in early infection, strong potyvirus resistance is critical for reducing synergist effects of co-infection on MCMV titer. These results emphasize the importance of both potyvirus resistance and MCMV resistance in an effective breeding program for MLN management. [ABSTRACT FROM AUTHOR]

Showy rattlepod (Crotalaria spectabilis Roth.) and phasey bean [Macroptilium lathyroides (L.) Urb.] plants exhibiting symptoms of a possible virus infection were found in Piracicaba municipality, São Paulo State, Brazil. Flexuous filamentous particles and cytoplasmatic inclusions typical of potyvirus infection were observed in leaf extracts and cells of symptomatic leaves, respectively, by transmission electron microscopy. Total RNA was extracted from the leaves of symptomatic plants, and RT-PCR followed by nucleotide sequencing of the amplicons showed that both plants were infected with bean common mosaic virus (BCMV). The potyvirus was mechanically transmitted to healthy plants of showy rattlepod, phasey bean, and sunn hemp (Crotalaria junceae L.). Myzus persicae transmitted the virus from both species of infected plants to healthy plants of common beans (Phaseolus vulgaris L.). This is the first report of BCMV infecting showy rattlepod and phasey bean plants in Brazil. [ABSTRACT FROM AUTHOR]

Background: In cellular organisms, inosine triphosphate pyrophosphatases (ITPases) prevent the incorporation of mutagenic deaminated purines into nucleic acids. These enzymes have also been detected in the genomes of several plant RNA viruses infecting two euphorbia species. In particular, two ipomoviruses produce replicase-associated ITPases to cope with high concentration of non-canonical nucleotides found in cassava tissues. Method: Using high-throughput RNA sequencing on the wild euphorbia species Mercurialis perennis, two new members of the families Potyviridae and Secoviridae were identified. Both viruses encode for a putative ITPase, and were found in mixed infection with a new partitivirid. Following biological and genomic characterization of these viruses, the origin and function of the phytoviral ITPases were investigated. Results: While the potyvirid was shown to be pathogenic, the secovirid and partitivirid could not be transmitted. The secovirid was found belonging to a proposed new Comovirinae genus tentatively named "Mercomovirus", which also accommodates other viruses identified through transcriptome mining, and for which an asymptomatic pollen-associated lifestyle is suspected. Homology and phylogenetic analyses inferred that the ITPases encoded by the potyvirid and secovirid were likely acquired through independent horizontal gene transfer events, forming lineages distinct from the enzymes found in cassava ipomoviruses. Possible origins from cellular organisms are discussed for these proteins. In parallel, the endogenous ITPase of M. perennis was predicted to encode for a C-terminal nuclear localization signal, which appears to be conserved among the ITPases of euphorbias but absent in other plant families. This subcellular localization is in line with the idea that nucleic acids remain protected in the nucleus, while deaminated nucleotides accumulate in the cytoplasm where they act as antiviral molecules. Conclusion: Three new RNA viruses infecting M. perennis are described, two of which encoding for ITPases. These enzymes have distinct origins, and are likely required by viruses to circumvent high level of cytoplasmic non-canonical nucleotides. This putative plant defense mechanism has emerged early in the evolution of euphorbias, and seems to specifically target certain groups of RNA viruses infecting perennial hosts. [ABSTRACT FROM AUTHOR]

Eukaryotic translation initiation factor 4E (eIF4E), which plays a pivotal role in initiating translation in eukaryotic organisms, is often hijacked by the viral genome‐linked protein to facilitate the infection of potyviruses. In this study, we found that the naturally occurring amino acid substitution D71G in eIF4E is widely present in potyvirus‐resistant watermelon accessions and disrupts the interaction between watermelon eIF4E and viral genome‐linked protein of papaya ringspot virus‐watermelon strain, zucchini yellow mosaic virus or watermelon mosaic virus. Multiple sequence alignment and protein modelling showed that the amino acid residue D71 located in the cap‐binding pocket of eIF4E is strictly conserved in many plant species. The mutation D71G in watermelon eIF4E conferred resistance against papaya ringspot virus‐watermelon strain and zucchini yellow mosaic virus, and the equivalent mutation D55G in tobacco eIF4E conferred resistance to potato virus Y. Therefore, our finding provides a potential precise target for breeding plants resistant to multiple potyviruses. [ABSTRACT FROM AUTHOR]

Pepper (Capsicum spp.) is an important solanaceous cash crop in Benin; however, productivity is limited due to several key constraints, especially diseases caused by viruses. We sought to understand farmers' perceptions of viral diseases, management strategies deployed, and to identify the virus population affecting pepper production in Benin. To assess farmers' perceptions and management of viral diseases, a survey was carried out in four agroecological zones of Benin. A total of 144 pepper farmers were interviewed using the snowball method. A total of 52 pepper leaf samples with virus-like symptoms were collected and diagnosed by reverse-transcriptase polymerase chain reaction (RT-PCR) or PCR. Pepper production systems varied across agroecological zones (P # 0.001) with a predominance of farms practicing monoculture (82%). The majority of farmers (89%) indicated that pests and diseases were the main constraints to increased production. Cucumber mosaic virus (92% of the total samples), Pepper vein yellow virus (52%), and Pepper veinal mottle virus (50%) were the major viruses detected in pepper fields in Benin. There were both single (29%) and mixed (71%) infections of the viruses, suggesting that mixed infections are common for pepper in Benin, confounding efforts to reduce virus infections. Nearly 100% of the farmers surveyed were not aware of these viral diseases. They also could not directly relate symptoms of virus infection to the presence of aphids, whiteflies, or thrips. Farmers relied primarily on synthetic insecticides (93%) to control virus vectors. Interestingly, some farmers applied commercial (12%) and homemade (17%) biopesticides, with neem-based preparations being the most widely used. A total of 15% of farmers used companion cropping with maize, mint or basil and 43% of farmers used crop rotation as a cultural management practice to control viral disease and vector pressure in pepper fields. The implications of this work include the importance of training farmers and extension agents on diagnosis of viruses and their vectors causing viral diseases. This study provides baseline information for the development of host-resistant cultivars and deployment of integrated pest management strategies for pepper in Benin to reduce farmer losses. [ABSTRACT FROM AUTHOR]

Symptoms such as severe mosaic, yellowing and necrosis of leaves have been observed in iris (Iris spp.) plants for many years. In this study, the presence of potyvirus species that can cause similar symptoms in iris plants was investigated. Field surveys were carried out in the province of Bilecik in the South Marmara region-Türkiye in the year 2022. In the study, a total of 41 iris plants were collected, including 34 symptomatic and 7 asymptomatic ones. Potyvirus infections were detected by conventional molecular methods using degenerate primers for amplification of the partial NIb (nuclear inclusion protein b) gene region specific to the genus Potyvirus. As a result of the molecular assays, potyvirus infection was detected in 6 plants showing symptoms caused by viruses. After nucleotide sequencing of the 6 Potyvirus fragments obtained, the infections were determined to be caused by iris severe mosaic virus (ISMV). These isolates showed over 94% nucleotide similarity among themselves and 82-99% similarity with the global isolate. Phylogenetic analyses showed that Potyvirus species were successfully separated among themselves according to the NIb gene region obtained with these degenerate primers. [ABSTRACT FROM AUTHOR]

Bean common mosaic virus (BCMV) was detected on common bean (Phaseolus vulgaris) plants showing wrinkled and/or narrow leaves, curling, shrinking and chlorosis of leaves, dwarfing of plants, and mottled pods in Inner Mongolia and named BCMV-22Huhe. Its genome has a size of 10,062 bp and was deposited in GenBank under the accession number OR778613. It is closely related to BCMV-Az (GenBank accession no. KP903372, in China) in the lineage of AzBMV. A recombination event was detected for BCMV-22Huhe among the 99 BCMV isolates published in the NCBI GenBank database, showing that BCMV-CJ25 (MK069986, found in Mexico) was a potential major parent, and the minor parent is unknown. This work is the first description of the occurrence of BCMV in Inner Mongolia, China. [ABSTRACT FROM AUTHOR]

Copyright of Revista U.D.C.A Actualidad & Divulgación Científica is the property of Universidad de Ciencias Aplicadas y Ambientales U.D.C.A and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

El cultivo de maracuyá amarillo es afectado por múltiples problemas fitosanitarios. Entre los principales, están los virus de los géneros Potyvirus (soybean mosaic virus, SMV), Cucumovirus (cucumber mosaic virus, CMV), Tymovirus (passion fruit yellow mosaic virus, PFYMV) y Begomovirus (passionfruit leaf distortion virus, PLDV). Información acerca de la prevalencia e interacción entre estos virus es escasa. El objetivo del presente estudio fue verificar la prevalencia e identificar los tipos de infecciones virales mixtas de potyvirus, cucumovirus, tymovirus y begomovirus en cultivos de maracuyá en Valle del Cauca, Colombia. Muestras foliares de maracuyá con síntomas virales fueron recolectadas y se purificaron sus ácidos nucleicos. La identificación de los virus se realizó por PCR empleando cebadores universales por género viral, y cebadores específicos para los virus SMV, CMV, PFYMV y PLDV. Se analizó la incidencia de cada virus por municipio y se comparó la severidad de los síntomas con el diagnóstico realizado. En total se colectaron 66 muestras en los municipios de Toro, Dagua, Roldanillo, Bolívar, La Unión y El Cerrito. Se estableció que los virus con mayor prevalencia fueron PLDV, seguido por SMV y CMV. No se detectó la presencia de PFYMV en las muestras analizadas. Se identificó por primera vez la presencia de infecciones dobles y triples entre potyvirus, cucumovirus y begomovirus, encontrándose mayor severidad de síntomas a mayor número de virus identificado por muestra. La presente investigación provee resultados clave para el diseño de estrategias de control de enfermedades virales en maracuyá.

Celery mosaic virus (CeMV), member of genus Potyvirus, is reported for the first time in Indonesia, from celery plants (Apium graveolens) in a vegetable field at Berastagi, North Sumatera Province. The plants possessed mosaic and vein clearing symptoms on the leaves as typical of CeMV infection. Virus incidence was confirmed by RT-PCR using degenerate potyvirus primer which amplified partial coat protein and 3’-UTR of the viral genome. Phylogenetic tree analysis placed Indonesian CeMV isolates in one separated clade within CeMV group and shared 96.5%–96.7% nucleotide identity with exemplar isolate of CeMV.

Kitosan dan ekstrak kasar daun bugenvil diketahui mampu mengendalikan beberapa virus tanaman termasuk Bean common mosaic virus strain Blackeye Cowpea (BCMV-BlC). Untuk mengurangi penggunaan bahan baku dan peningkatan keefektifan juga stabilitasnya, kedua bahan perlu disintesis menggunakan teknologi nanpopartikel (NP). Penelitian bertujuan menyintesis nanopartikel kitosan, ekstrak daun bugenvil, dan kombinasinya menggunakan modifikasi metode gelasi ionik, mengarakterisasi NP dan mengevaluasi potensi NP untuk mengendalikan BCMV-BlC pada tanaman indikator Chenopodium amaranticolor. Nanopartikel kitosan (Kit-NP), ekstrak daun bugenvil (EDB-NP), dan kombinasinya (KEDB-NP) berhasil disintesis dengan rata-rata ukuran partikel berturut-turut sebesar 99.72, 163.68, dan 221.42 nm berdasarkan pengamatan di bawah transmission electron microscope (TEM) dan berbentuk bola (sferis) berdasarkan hasil analisis scanning electron microscope (SEM). Hasil analisis fourier transform infrared (FTIR), KEDB-NP menunjukkan gugus fungsi berturut-turut hidroksil, alkena, amina, cincin aromatik, dan senyawa alifatik organohalogen. Daun C. amaranticolor yang diberi perlakuan Kit-NP, EDB-NP, KEDB-NP dan juga non-NP pada konsentrasi 100-800 ppm sebelum inokulasi virus menunjukkan jumlah rata-rata lesio lokal nekrotik (LLN) nyata lebih rendah dibandingkan kontrol tanaman sakit tanpa perlakuan dan tidak berbeda nyata antarkonsentrasi. Perlakuan NP dan non-NP sebelum inokulasi virus secara nyata memperpanjang periode inkubasi dan menekan pembentukan LLN dengan keefektifan 67.5%–100% dibandingkan dengan aplikasi setelah inokulasi virus. Di antara konsentrasi yang diuji, konsentrasi NP antara 100 dan 300 ppm yang diaplikasikan sebelum penularan virus menunjukkan paling efektif dalam menghambat pembentukan LLN. Sintesis EDB-NP, Kit-NP, dan KEDB-NP menggunakan sangat sedikit bahan baku, namun keefektifannya sebanding dengan bentuk non-NPnya dalam mengendalikan infeksi BCMV.

East Asian Passiflora virus (EAPV) causes passionfruit woodiness disease, a major threat limiting passionfruit production in eastern Asia, including Taiwan and Vietnam. In this study, an infectious cDNA clone of a Taiwanese severe isolate EAPV-TW was tagged with a green fluorescent protein (GFP) reporter to monitor the virus in plants. Nicotiana benthamiana and yellow passionfruit plants inoculated with the construct showed typical symptoms of EAPV-TW. Based on our previous studies on pathogenicity determinants of potyviral HC-Pros, a deletion of six amino acids (d6) alone and its association with a point mutation (F8I, simplified as I8) were conducted in the N-terminal region of the HC-Pro gene of EAPV-TW to generate mutants of EAPV-d6 and EAPV-d6I8, respectively. The mutant EAPV-d6I8 caused infection without conspicuous symptoms in N. benthamiana and yellow passionfruit plants, while EAPV-d6 still induced slight leaf mottling. EAPV-d6I8 was stable after six passages under greenhouse conditions and displayed a zigzag pattern of virus accumulation, typical of a beneficial protective virus. The cross-protection effectiveness of EAPV-d6I8 was evaluated in both N. benthamiana and yellow passionfruit plants under greenhouse conditions. EAPV-d6I8 conferred complete cross-protection (100%) against the wild-type EAPV-TW-GFP in both N. benthamiana and yellow passionfruit plants, as verified by no severe symptoms, no fluorescent signals, and PCR-negative status for GFP. Furthermore, EAPV-d6I8 also provided complete protection against Vietnam’s severe strain EAPV-GL1 in yellow passionfruit plants. Our results indicate that the attenuated mutant EAPV-d6I8 has great potential to control EAPV in Taiwan and Vietnam via cross-protection.

Plant viruses cause severe losses in agricultural production. Parasite-derived resistance (PDR) offers a promising avenue for developing disease-resistant varieties independent of resistance genes. However, for potyviruses with great agricultural importance, such as soybean mosaic virus (SMV), systematic research on viral genes that can be used for PDR has not been conducted. In this study, we transiently expressed the untranslated region (UTR) or each protein-coding cistron of SMV in Nicotiana benthamiana to evaluate their potential role in conferring PDR. A viral suppressor of RNA silencing (VSR) was also applied to investigate the possible mechanisms of the PDR. The results showed that the transient overexpression of UTR and each cistron of SMV could inhibit SMV infection. The expression of VSR in N. benthamiana leaves could compromise UTR and most of the SMV cistron-mediated inhibition of SMV infection, indicating the involvement of RNA silencing in PDR. In comparison, the expression of VSR could not compromise the PDR conferred by coat protein (CP), P3N-PIPO, cylindrical inclusion (CI), and NIa-Pro, suggesting that these viral cistrons may play roles in PDR at the protein level. These results reveal diverse mechanisms in PDR conferred by different viral cistrons and provide potential gene candidates that can be used for transgenic approaches against SMV.

Global food production is challenged by plant pathogens that cause significant crop losses. Fungi, bacteria, and viruses have long threatened sustainable and profitable agriculture. The danger is even higher in vegetatively propagated horticultural crops, such as garlic. Currently, quarantine, rouging infected plants, and control of natural vectors are used as the main means of disease and pest control in garlic crops. Agricultural biotechnology, meristem-tip culture, and cryotherapy offer solutions for virus eradication and for the multiplication of 'clean stocks', but at the same time, impact the symbiotic and beneficial components of the garlic microbiome. Our research involves the first metatranscriptomic analysis of the microbiome of garlic bulb tissue, PCR analyses, and a biological assay of endophytes and pathogens. We have demonstrated that in vitro sanitation methods, such as shoot tip culture or cryotherapy can alter the garlic microbiome. Shoot tip culture proved ineffective in virus elimination, but reduced bacterial load and eliminated fungal infections. Conversely, cryotherapy was efficient in virus eradication but demolished other components of the garlic microbiome. Garlic plants sanitized by cryotherapy exhibited a lower survival rate, and a longer in vitro regeneration period. The question arises whether total eradication of viruses, at the expense of other microflora, is necessary, or if a partial reduction in the pathogenic load would suffice for sanitized garlic production. We explore this question from both scientific and commercial perspectives. [ABSTRACT FROM AUTHOR]

The genus Potyvirus is considered as the largest among plant single‐stranded (positive‐sense) RNA viruses, causing considerable economic damage to vegetable and fruit crops worldwide. Through the coordinated action of four viral proteins and a few identified host factors, potyviruses exploit the endomembrane system of infected cells for their replication and for their intra‐ and intercellular movement to and through plasmodesmata (PDs). Although a significant amount of data concerning potyvirus movement has been published, no synthetic review compiling and integrating all information relevant to our current understanding of potyvirus transport is available. In this review, we highlight the complexity of potyvirus movement pathways and present three potential nonexclusive mechanisms based on (1) the use of the host endomembrane system to produce membranous replication vesicles that are targeted to PDs and move from cell to cell, (2) the movement of extracellular viral vesicles in the apoplasm, and (3) the transport of virion particles or ribonucleoprotein complexes through PDs. We also present and discuss experimental data supporting these different models as well as the aspects that still remain mostly speculative. [ABSTRACT FROM AUTHOR]

In April 2022, virus‐like symptoms were observed in a Cymbopogon spp. plant grown in the backyard of a house and in Java citronella (C. winterianus) plants in a flower shop in Piracicaba, São Paulo State, Brazil. In April 2023, ca. 700,000 Ceylon citronella (C. nardus) plants showing symptoms of mosaic were observed in a field crop in Dois Corrégos (SP). Evidence for a possible potyvirus infection was revealed by electron microscopy and confirmed by molecular assays. Nucleotide sequencing of amplicons from RT‐PCR identified the potyvirus infecting a Cymbopogon spp. plant, Java and Ceylon citronella as sugarcane mosaic virus (SCMV, genus Potyvirus). Phylogenetic analysis of the coat protein gene indicated that the SCMV isolates from Cymbopogon (C1), Java (JC1), and Ceylon citronella (CC) belong to the clade of SCMV from sugarcane. The C1 isolate, and an isolate from sugarcane (SCMV‐SU) (control) were mechanically transmitted to Java citronella, sorghum, sugarcane and maize plants. Aphis gossypii transmitted the C1 and SU isolates to Java citronella plants, while Melanaphis sacchari transmitted only the SU isolate to Java citronella plants. This is the first molecular characterization of SCMV from Cymbopogon spp. plants. [ABSTRACT FROM AUTHOR]

The garlic (Allium sativum L.) cultivated in the O'Higgins Region, the main garlic-producing area in Chile, is the commercial type known as pink garlic, which presents a low yield and a high prevalence of symptoms associated with viral infections. A survey was carried out in the three localities of the region that cultivate this vegetable to identify, by reverse transcription polymerase chain reaction (RT-PCR), the presence of Onion yellow dwarf virus (OYDV) and Leek yellow stripe virus (LYSV). The results indicated a prevalence of 99.5% and 54.0% respectively. In addition, a molecular and phylogenetic analysis of the amplified genome fragments for both viruses was performed. Using the nucleotide BLAST (BLASTn) tool, nucleotide sequences were compared with sequences available in the National Center for Biotechnology Information (NCBI) database. The Chilean sequences were aligned with complete sequences available at NCBI and a phylogenetic tree was constructed using the Neighbor-Joining method. The results showed genetic variability among Chilean isolates. [ABSTRACT FROM AUTHOR]

Kudzu (Pueraria montana var. lobata), a plant native to Southeastern Asia, has become a major noxious weed covering millions of hectares in the Southern United States. A kudzu patch displaying virus-like symptoms located in Ackerman, northeastern Mississippi (MS), was used as a source for virus isolation and characterization involving mechanical and vector transmission, ultrastructural observation, surveys, Sanger and high-throughput genome sequencing, and sequence analyses. The results revealed the presence of a new potyvirus in infected kudzu, closely related to wisteria vein mosaic virus (WVMV) and provisionally named kudzu chlorotic ring blotch virus (KudCRBV). Genome features and pairwise comparison with six WVMV genomes currently available in GenBank and three additional isolates from MS sequenced in this work suggest that KudCRBV is likely a member of a new species in the genus Potyvirus. Furthermore, under experimental conditions, KudCRBV was successfully transmitted by cotton and potato aphids and mechanically to soybean and beans. A state-wide survey revealed several kudzu patches infected by the virus in northern MS. [ABSTRACT FROM AUTHOR]

Background: Cowpea [Vigna unguiculata (L) Walp.] is one of the Paraguayan rural families' main crops, serving as an essential protein and carbohydrate source. Cowpea aphid borne mosaic virus (CABMV) and cowpea severe mosaic virus (CPSMV) were identified infecting cowpea plants. Disease control caused by both viruses is difficult because there is no information about local cowpea resistant cultivars and vector control is not practical. Methods: In the present work, sixteen cowpea genotypes/cultivars were mechanically inoculated with local isolates of CABMV and CPSMV to identify resistant genotypes/cultivars that can be used in breeding programs. Virus infections were determined by symptoms expression and confirmed by PTA-ELISA. Result: Genotypes Arroz rojo (V. angularis), TVu 379, TVu 382, TE94-256-2E and TE97-309G-9 were resistant to CABMV. Genotypes Arroz rojo (V. angularis), CNCX-698-128F, TVu 379, TVu 382, TVu-3961, TE97-309G-9 and TE97-309G-3 were resistant to CPSMV. Overall, this study showed that local cowpea cultivars do not offer any resistance to virus infection and the need for resistant germplasms for cowpea breeding programs in the country. [ABSTRACT FROM AUTHOR]

Regulation of protein synthesis is a strong determinant of potyviral pathogenicity. The Potyviridae family is the largest family of plant-infecting positive sense RNA viruses. Similar to the animal-infecting Picornaviridae family, the potyviral RNA genome lacks a 5′ cap, and instead has a viral protein (VPg) linked to its 5′ end. Potyviral genomes are mainly translated into one large polyprotein relying on a single translation event to express all their protein repertoire. In the absence of the 5′ cap, the Potyviridae family depends on cis -acting elements in their 5′ untranslated regions (UTR) to recruit the translation machinery. In this review, we summarize the diverse 5′UTR-driven, cap-independent translation mechanisms employed by the Potyviridae family including scanning-dependent mechanism, internal initiation, and the stimulatory role of the VPg. These mechanisms have direct implications on potyviral pathogenicity, including host range specificity and resistance. Finally, we discuss how these viral strategies could not only inform new avenues for engineering and/or breeding for crop resistance but would also provide opportunities for the development of biotechnological tools for large-scale protein production in plant systems. [ABSTRACT FROM AUTHOR]