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Sophora yellow stunt virus is a species of the genus Nanovirus. Sophora yellow stunt virus (SYSV) has a wide geographical distribution in Iran. In the current study, SYSV was identified from Kerman province (south‐eastern Iran) in five symptomatic aromatic herbs that are commonly used in the Iranian diet: parsley, dill, fenugreek, mint and basil. These infections were detected using either a PCR assay or high‐throughput sequencing. Eight circular single‐stranded DNA genomic components of five SYSV isolates as well as a new sophora isolate from south‐western Iran were PCR amplified, sequenced and characterized. Sequence analysis of SYSV genomic components showed that the Iranian isolates shared nucleotide identities of 64%–100% with each other and 63%–93% with an isolate from China. DNA‐Rs and DNA‐Cs were found to be the most diverse components relative to the others. Furthermore, associated alphasatellite molecules with the basil sample shared nucleotide identities of 80.7%–99.9% with the counterpart alphasatellite molecules in GenBank. Infectious clones for the genomic DNAs of SYSV were constructed using head‐to‐tail dimeric or partial dimeric inserts. The pathogenesis of SYSV was demonstrated by inoculation with the infectious DNA clones that induced typical symptoms in five plant species. Given the wide distribution of SYSV and its common host (Sophora alopecuroides) throughout Iran, as well as detection of the virus in plants of four families (Fabaceae, Apiaceae, Lamiaceae and Nitrariaceae), this virus is of economic importance within leguminous crops and in production of aromatic herbs in Iran. [ABSTRACT FROM AUTHOR]

Banana bunchy top disease (BBTD), caused by a Banana Bunchy Top Virus (BBTV), is the most devastating viral disease of bananas. The genome of multipartite BBTV is about 1 Kb in size and comprises six circular single-stranded (ss) DNA components, namely DNA-R, C, S, M, U3, and N. Gene expression in multipartite DNA viruses is highly flexible due to variations in the copy number and transcript levels of individual genomic components. This enables them to adapt to changing conditions and maintain optimal fitness. In the current study, the copy number of each genetic component of BBTV and its transcript level were determined in the leaf, midrib, and root tissue of the infected banana plant. The results of the study showed that DNA copies (copies/ng) were higher in midrib (11,105 ± 10,704.42), followed by leaf (6902.91 ± 1651.14), and were lowest in root (23.49 ± 9.21) tissues. In contrast, the transcript level was highest in the root (312.45 ± 106.69) and lowest in the midrib (0.72 ± 0.16). This suggests that BBTV independently regulates the transcriptional level of each DNA component. The distribution of BBTV in different tissues assists the identification of significantly higher viral load tissues for early and sensitive disease diagnosis, which will be beneficial for better disease management. [ABSTRACT FROM AUTHOR]

Banana bunchy top disease (BBTD), caused by a Banana Bunchy Top Virus (BBTV), is the most devastating viral disease of bananas. The genome of multipartite BBTV is about 1 Kb in size and comprises six circular single-stranded (ss) DNA components, namely DNA-R, C, S, M, U3, and N. Gene expression in multipartite DNA viruses is highly flexible due to variations in the copy number and transcript levels of individual genomic components. This enables them to adapt to changing conditions and maintain optimal fitness. In the current study, the copy number of each genetic component of BBTV and its transcript level were determined in the leaf, midrib, and root tissue of the infected banana plant. The results of the study showed that DNA copies (copies/ng) were higher in midrib (11,105 ± 10,704.42), followed by leaf (6902.91 ± 1651.14), and were lowest in root (23.49 ± 9.21) tissues. In contrast, the transcript level was highest in the root (312.45 ± 106.69) and lowest in the midrib (0.72 ± 0.16). This suggests that BBTV independently regulates the transcriptional level of each DNA component. The distribution of BBTV in different tissues assists the identification of significantly higher viral load tissues for early and sensitive disease diagnosis, which will be beneficial for better disease management.

Parsley severe stunt-associated virus (PSSaV) is a recently identified nanovirus first reported in Germany. During a survey for identification of nanoviruses infecting apiaceous plants in south-eastern Iran, PSSaV was identified and characterized using a combination of rolling circle amplification (RCA) and high-throughput sequencing. Parsley plant samples were collected from vegetable production farms in Kerman province. From two symptomatic samples (39Ba and 40Ba), seven PSSaV components (DNA-C, -S, -M, -R, -N, -U1 and -U2) with two phylogenetically distinct variants of DNA-R (R1 and R2) were identified. In common with the German isolate of PSSaV, no DNA-U4 component was identified. In addition, associated alphasatellite molecules were identified in samples 39Ba [n = 6] and 40Ba [n = 5]. Sequence analyses showed that concatenated component sequences of the two Iranian PSSaVs share 97.2% nucleotide identity with each other and 82% to the German isolate. The coat proteins (CPs) of the PSSaV Iranian sequences share 97.2% amino acid identity and ~ 84% identity with that of the German isolate. Sequence and phylogenetic analyses of a total of 11 recovered alphasatellites from the two samples can be classified into the genera Fabenesatellite [n = 2], Milvetsatellite [n = 1], Mivedwarsatellite [n = 2], Subclovsatellite [n = 2], Sophoyesatellite [n = 4] in the family Alphasatellitidae. Identification of PSSaV and other nanoviruses in wild and cultivated plants in Iran reveals that nanoviruses could be causing yield reduction in crops plants in this country. [ABSTRACT FROM AUTHOR]

Faba bean necrotic stunt virus (FBNSV) (genus Nanovirus; family Nanoviridae) is a multipartite virus comprising eight circular single-stranded DNA components, each about 1 kb in length. Also, two alphasatellites have often been reported to be associated with FBNSV infection. The only full-length genome sequence of an Iranian FBNSV isolate, FBNSV-[IR;Ka:FB], so far reported is one infecting faba bean (Vicia faba L.). To obtain more information about Iranian isolates of FBNSV (FBNSV-IRs) and their diversity, the full-length genome of four FBNSV isolates and associated alphasatellites obtained from faba bean (two isolates), common bean (Phaseolus vulgaris L., one isolate) and chickpea (Cicer arietinum L., one isolate) were characterized. All genomic components except U4 had the expected size in accordance with the FBNSV genomic sequences published in the GenBank. The component U4 had an additional stretch of 16 nucleotides (nts 765–780) in its non-coding region (NCR). Alphasatellites of the aforementioned isolates showed 98–100% sequence identity. Intra-component sequence analyses of FBNSV-IRs indicated a range of 98 to 100% nt identity between each single FBNSV-IRs component or between their associated alphasatellites. DNA-M and DNA-U2 exhibited the highest value of diversity, sharing 98–99% identity. Phylogenetic analyses indicated that FBNSV-IRs were clustered with a Moroccan isolate and some Azerbaijani isolates of FBNSV forming a clade distinct from Ethiopian isolates. Inter-component sequence analyses of NCRs revealed the DNA-S and DNA-U4 subgrouped together in all FBNSV-IRs isolates except the chickpea isolate in which NCRs of DNA-S and DNA–U4 formed separate subgroups. [ABSTRACT FROM AUTHOR]

Nanoviruses and geminiviruses are circular, single stranded DNA viruses that infect many plant species around the world. Nanoviruses and certain geminiviruses that belong to the Begomovirus and Mastrevirus genera are associated with additional circular, single stranded DNA molecules (~ 1-1.4 kb) that encode a replication-associated protein (Rep). These Rep-encoding satellite molecules are commonly referred to as alphasatellites and here we communicate the establishment of the family Alphasatellitidae to which these have been assigned. Within the Alphasatellitidae family two subfamilies, Geminialphasatellitinae and Nanoalphasatellitinae, have been established to respectively accommodate the geminivirus- and nanovirus-associated alphasatellites. Whereas the pairwise nucleotide sequence identity distribution of all the known geminialphasatellites (n = 628) displayed a troughs at ~ 70% and 88% pairwise identity, that of the known nanoalphasatellites (n = 54) had a troughs at ~ 67% and ~ 80% pairwise identity. We use these pairwise identity values as thresholds together with phylogenetic analyses to establish four genera and 43 species of geminialphasatellites and seven genera and 19 species of nanoalphasatellites. Furthermore, a divergent alphasatellite associated with coconut foliar decay disease is assigned to a species but not a subfamily as it likely represents a new alphasatellite subfamily that could be established once other closely related molecules are discovered. [ABSTRACT FROM AUTHOR]

Pea necrotic yellow dwarf virus (PNYDV) is a multipartite, circular, single-stranded DNA plant virus. PNYDV encodes eight proteins and the function of three of which remains unknown-U1, U2, and U4. PNYDV proteins cellular localization was analyzed by GFP tagging and bimolecular fluorescence complementation (BiFC) studies. The interactions of all eight PNYDV proteins were tested pairwise in planta (36 combinations in total). Seven interactions were identified and two (M-Rep with CP and MP with U4) were characterized further. MP and U4 complexes appeared as vesicle-like spots and were localized at the nuclear envelope and cell periphery. These vesicle-like spots were associated with the endoplasmatic reticulum. In addition, a nuclear localization signal (NLS) was mapped for U1, and a mutated U1 with NLS disrupted localized at plasmodesmata and therefore might also have a role in movement. Taken together, this study provides evidence for previously undescribed nanovirus protein-protein interactions and their cellular localization with novel findings not only for those proteins with unknown function, but also for characterized proteins such as the CP. [ABSTRACT FROM AUTHOR]

Viruses in the genus Nanovirus of the family Nanoviridae generally have eight individually encapsidated circular genome components and have been predominantly found infecting Fabaceae plants in Europe, Australia, Africa and Asia. For over a decade Sophora alopecuroides L. (Fabaceae) plants have been observed across Iran displaying dwarfing, yellowing, stunted leaves and yellow vein banding. Using a high-throughput sequencing approach, sequences were identified within one such plant that had similarities to nanovirus genome components. From this plant, the nanovirus-like molecules DNA-R ( n = 4), DNA-C ( n = 2), DNA-S ( n = 1), DNA-M ( n = 1), DNA-N ( n = 1), DNA-U1 ( n = 1), DNA-U2 ( n = 1) and DNA-U4 ( n = 1) were amplified, cloned and sequenced. Other than for the DNA-R, these components share less than 71% identity with those of other known nanoviruses. The four DNA-R molecules were highly diverse, sharing only 65–71% identity with each other and 64–86% identity with those of other nanoviruses. In the S. alopecuroides plant 14 molecules sharing 57.7–84.6% identity with previously determined sequences of nanovirus-associated alphasatellites were also identified. Given the research activity in the nanovirus field during the last five years coupled with high-throughput sequence technologies, many more diverse nanoviruses and nanovirus-associated satellites are likely to be identified. [ABSTRACT FROM AUTHOR]

Banana bunchy top virus (BBTV), belonging to the genus Babuvirus, is the most devastating and widespread banana virus. Banana and plantain are major crops in terms of household income and food security in Democratic Republic of Congo (DRC). Despite the large area under banana and plantain cultivation in the country, before this study, the genetic characterization of BBTV isolates had only been undertaken for two provinces. In the study presented here, genetic variation in BBTV was assessed from 52 BBTV isolates collected in five out of 11 provinces in DRC (Bandundu, Bas-Congo, Katanga, Kinshasa and Kasaï Oriental) and in two provinces using sequences previously described in databases. Full genome sequencing of DNA-R components was performed, revealing low genetic variation (98-100 % nucleotide identity) among the BBTV isolates detected. The phylogenetic analyses showed that all the DRC isolates were clustered in the South Pacific clade of BBTV. Based on the coding region for the replication initiator protein, haplotype diversity was estimated to be 0.944 ± 0.013, with 30 haplotypes from 68 isolates in DRC. Such diversity shows a haplotype distribution mainly at the sub-regional level in DRC. In addition, the sequence determination from the whole genome of selected isolates confirmed low genetic variation among isolates from seven DRC provinces (97-100 % nucleotide identity). This study strengthened the hypothesis of a single BBTV introduction some time ago, followed by the spread of the virus in the country. [ABSTRACT FROM AUTHOR]

Plant virus species of the family Nanoviridae have segmented genomes with the highest known number of segments encapsidated individually. They thus likely represent the most extreme case of the so-called multipartite, or multicomponent, viruses. All species of the family are believed to be transmitted in a circulative nonpropagative manner by aphid vectors, meaning that the virus simply crosses cellular barriers within the aphid body, from the gut to the salivary glands, without replicating or even expressing any of its genes. However, this assumption is largely based on analogy with the transmission of other plant viruses, such as geminiviruses or luteoviruses, and the details of the molecular and cellular interactions between aphids and nanoviruses are poorly investigated. When comparing the relative frequencies of the eight genome segments in populations of the species Faba bean necrotic stunt virus (FBNSV) (genus Nanovirus) within host plants and within aphid vectors fed on these plants, we unexpectedly found evidence of reproducible changes in the frequencies of some specific segments. We further show that these changes occur within the gut during early stages of the virus cycle in the aphid and not later, when the virus is translocated into the salivary glands. This peculiar observation, which was similarly confirmed in three aphid vector species, Acyrthosiphon pisum, Aphis craccivora, and Myzus persicae, calls for revisiting of the mechanisms of nanovirus transmission. It reveals an unexpected intimate interaction that may not fit the canonical circulative nonpropagative transmission. [ABSTRACT FROM AUTHOR]

Banana bunchy top disease is caused by the Banana bunchy top virus (BBTV), which is a species of the genus Babuvirus in the family Nanoviridae. In this study, we collected 61 isolates of BBTV from Sumatra Island. The sequences of DNA-R and DNA-S of 61 samples and DNA-U3 of 37 samples were determined. Although the Sumatra population has different sequences in the stem-loop region in DNA-U3, it is generally very similar, with little or no variation according to planting area or the varieties and genome types of the host. The comparison of sequences between Sumatra isolates and those from other areas showed that the sequence identity with the Asian group is higher than with the South Pacific group. The genetic analysis showed that the Sumatra population has very low diversity compared with other areas. Recombination analysis revealed that there was no evidence of recombination in the Sumatra population. The selection pressure analysis indicated that all Sumatra populations received negative selection. The selection pressure among the viruses infecting the banana varieties having the genotypes of AAB or ABB received a slightly weaker negative selection than that of the viruses infecting the varieties with AA or AAA genotypes. A neutrality test suggested the sudden expansion of the West Sumatra population. This is the first report to survey the BBTV on Sumatra Island. [ABSTRACT FROM AUTHOR]

Abstract: RNA silencing refers to processes that depend on small (s)RNAs to regulate the expression of eukaryotic genomes. In plants, these processes play critical roles in development, in responses to a wide array of stresses, in maintaining genome integrity and in defense against viral and bacterial pathogens. We provide here an updated view on the array of endogenous sRNA pathways, including microRNAs (miRNAs), discovered in the model plant Arabidopsis, which are also the basis for antiviral silencing. We emphasize the current knowledge as well as the recent advances made on understanding the defense and counter-defense strategies evolved in the arms race between plants and DNA viruses on both the nuclear and the cytoplasmic front. This article is part of a Special Issue entitled: MicroRNA''s in viral gene regulation. [Copyright &y& Elsevier]

Banana bunchy top disease (BBTD) caused by Banana bunchy top virus (BBTV) is one of the most devastating diseases of banana and poses a serious threat for cultivars like Hill Banana (Syn: Virupakshi) and Grand Naine in India. In this study, we have cloned and sequenced the complete genome comprised of six DNA components of BBTV infecting Hill Banana grown in lower Pulney hills, Tamil Nadu State, India. The complete genome sequence of this hill banana isolate showed high degree of similarity with the corresponding sequences of BBTV isolates originating from Lucknow, Uttar Pradesh State, India, and from Fiji, Egypt, Pakistan, and Australia. In addition, sixteen coat protein (CP) and thirteen replicase genes (Rep) sequences of BBTV isolates collected from different banana growing states of India were cloned and sequenced. The replicase sequences of 13 isolates showed high degree of similarity with that of South Pacific group of BBTV isolates. However, the CP gene of BBTV isolates from Shervroy and Kodaikanal hills of Tamil Nadu showed higher amino acid sequence variability compared to other isolates. Another hill banana isolate from Meghalaya state had 23 nucleotide substitutions in the CP gene but the amino acid sequence was conserved. This is the first report of the characterization of a complete genome of BBTV occurring in the high altitudes of India. Our study revealed that the Indian BBTV isolates with distinct geographical origins belongs to the South Pacific group, except Shervroy and Kodaikanal hill isolates which neither belong to the South Pacific nor the Asian group. [ABSTRACT FROM AUTHOR]

Field and laboratory experiments were carried out on the island of Oahu, HI, to compare the susceptibility of the two most commonly grown banana (Musa sp.) cultivars in the state ('Dwarf Brazilian' or Santa Catarina [locally known as dwarf apple I and 'Williams') to the aphid-borne Banana bunchy top virus (genus Babuvivus, family Nanouiridae, BBTV). Several morphological and physiological features of the two cultivars were monitored to determine whether the banana aphid, Pentalonia nigronervosa Coquerel (Hemiptera: Aphididae), transmits BBTV to both cultivars at a similar rate; and whether after successful inoculation, does each cultivar respond similarly to viral infection. Results from the laboratory experiment showed that a similar percentage of both cultivars were infected with BBTV by aphid vectors (>90% for both cultivars). However, field results showed a significantly lower percentage of dwarf apple (39%) infected with BBTV compared with Williams (79%). We also found that all physiological and morphological features measured (i.e., plant height, leaf area, canopy, chlorophyll level, and moisture content) for both cultivars were impacted similarly by BBTV. The incubation period, or the time between plant infection and initial appearance of disease symptoms, was similar for both cultivars. Results also showed that BBTV transmission efficiency was lower in the field than in the laboratory, despite that more aphids per plant were used for field than laboratory inoculation tests. The results highlight the potential use of less susceptible cultivars to help manage BBTV and the importance of screening banana varieties in the field to determine their response to vectors and associated diseases. [ABSTRACT FROM AUTHOR]

Field experiments were conducted in Oahu, Hawaii, to investigate the effects of banana bunchy top virus (BBTV) infection on growth and morphology of banana ( Musa acuminata). The time interval between aphid inoculation of BBTV and the initial appearance of disease symptoms (i.e. incubation period) was also determined. Plants infected with BBTV showed a significant reduction in petiole size (i.e. length and distance), plant canopy and height, leaf area, pseudostem diameter and chlorophyll content compared with control plants. Growth differences between virus-infected and control plants were not observed until 40–50 days after the plants were inoculated with viruliferous aphids. Other growth parameters such as petiole width and leaf production were not statistically different between infected and control plants. The incubation period of banana bunchy top disease or appearance of symptoms ranged from 25 to 85 days after aphid inoculation. However, PCR assays provided earlier detection of BBTV in banana plants compared with visual symptoms. [ABSTRACT FROM AUTHOR]

The study of the transmission biology of insect-borne plant viruses is important to develop disease control practices. We characterized the transmission of a nanovirus, Banana bunchy top virus (BBTV), by its aphid vector Pentalonia nigronervosa Coquerel (Hemiptera, Aphididae) with respect to temperature, vector life stage, and plant access time. Adult aphids transmitted BBTV more efficiently than third instar nymphs at all temperatures tested. Adult aphids transmitted the virus more efficiently at 25 and 30°C than at 20°C, but temperature had no impact on transmission efficiency by nymphs. By decoupling the relationship between temperature and aphid BBTV acquisition or inoculation, we determined that temperature affected inoculation events more strongly than acquisition. Longer plant access periods increased viral acquisition and inoculation efficiencies in a range of 60 mm to 24 h. Both BBTV acquisition and inoculation efficiencies peaked after 18 h of plant access period. We also show that BBTV transmission by P. nigronervosa requires a latent period. Our results demonstrate that vector transmission of BBTV is affected by temperature, vector life stage, and plant access period. [ABSTRACT FROM AUTHOR]