Tomato black ring virus (TBRV) is the only member of the Nepovirus genus that is known to form defective RNA particles (D RNAs) during replication. Here, de novo generation of D RNAs was observed during prolonged passages of TBRV isolates originated from Solanum lycopersicum and Lactuca sativa in Chenopodium quinoa plants. D RNAs of about 500 nt derived by a single deletion in the RNA1 molecule and contained a portion of the 5' untranslated region and viral replicase, and almost the entire 3' non-coding region. Short regions of sequence complementarity were found at the 5' and 3' junction borders, which can facilitate formation of the D RNAs. Moreover, in this study we analyzed the effects of D RNAs on TBRV replication and symptoms development of infected plants. C. quinoa, S. lycopersicum, Nicotiana tabacum, and L. sativa were infected with the original TBRV isolates (TBRV-D RNA) and those containing additional D RNA particles (TBRV + D RNA). The viral accumulation in particular hosts was measured up to 28 days post inoculation by RT-qPCR. Statistical analyses revealed that D RNAs interfere with TBRV replication and thus should be referred to as defective interfering particles. The magnitude of the interference effect depends on the interplay between TBRV isolate and host species.

Viruses cause epidemics on all major crops of agronomic importance, and a timely and accurate identification is essential for control. High throughput sequencing (HTS) is a technology that allows the identification of all viruses without prior knowledge on the targeted pathogens. In this paper, we used HTS technique for the detection and identification of different viral species occurring in single and mixed infections in plants in Poland. We analysed various host plants representing different families. Within the 20 tested samples, we identified a total of 13 different virus species, including those whose presence has not been reported in Poland before: clover yellow mosaic virus (ClYMV) and melandrium yellow fleck virus (MYFV). Due to this new finding, the obtained sequences were compared with others retrieved from GenBank. In addition, cucurbit aphid-borne yellows virus (CABYV) was also detected, and due to the recent occurrence of this virus in Poland, a phylogenetic analysis of these new isolates was performed. The analysis revealed that CABYV population is highly diverse and the Polish isolates of CABYV belong to two different phylogenetic groups. Our results showed that HTS-based technology is a valuable diagnostic tool for the identification of different virus species originating from variable hosts, and can provide rapid information about the spectrum of plant viruses previously not detected in a region.

Tomato black ring virus (TBRV), a member of the Nepovirus genus, is a serious plant pathogen distributed worldwide. It causes significant damage to several economically important crops, such as artichoke or strawberry. The TBRV bipartite genome consists of two polyadenylated single‐stranded positive‐sense RNA molecules, which may be accompanied by subviral particles such as defective interfering RNAs (DI RNAs) and satellite RNAs (satRNAs). In this study, we obtained the complete genome sequence of six TBRV isolates originating from different hosts and determined the presence of eight TBRV satRNAs. Subsequently, genetic variability, recombination, phylogenetic and selection pressure analyses were performed. The results revealed that the TBRV population is genetically diverse. The occurrence of potential recombination events, evidence of positive selection pressure acting on particular codons and the diversification of satRNAs within the TBRV population indicated that the virus mutates and can rapidly adapt to new environmental conditions or hosts. The presented data shed some light on TBRV evolutionary dynamics and epidemiology.

In recent years, the occurrence of cucumber mosaic virus (CMV) has been noted in zucchini crops in Poland. Beside characteristic isolates, which displayed mosaics and chlorosis on infected plants, new necrotic isolates have also been identified. Here, we analysed the molecular variability of 27 isolates of CMV collected from zucchini in various regions of the country. Sequence and phylogenetic analysis based on the genes encoding the coat (CP) and movement (MP) proteins revealed that the Polish isolates belong to two subgroups: IA and II, with the prevalence of subgroup II. New recombinant variants with an IA-MP/II-CP pattern for RNA3 were also detected.

Tomato black ring virus (TBRV) is a worldwide‐distributed RNA virus infecting a wide range of different host plants, including crop species, trees, shrubs, and weeds. Here, we investigated the molecular evolution of TBRV and its adaptability to different plant species. The TBRV‐Pi isolate was used to generate five independent evolution lineages serially passaged in either quinoa, tobacco, or tomato plants. After 15 passages, the genetic variability present in all the lineages was characterized for the movement (MP) and coat (CP) coding cistrons. We addressed two main questions: to what extent does the amount of genetic variability in the TBRV genome depend on the host species, and are there host species‐specific adaptive mutations? Overall, 201 different nucleotide substitutions emerged during the evolution experiment, with some of them appearing multiple times in different lineages; two of them (one in CP and one in MP) were unique for a particular host plant. We have shown that the degree of genetic differentiation depends on the host species in which the virus evolved, and that positive selection is operating upon certain residues, particularly in CP. Moreover, we have characterized new types of defective RNAs that arose during the TBRV‐Pi evolution in tobacco. Furthermore, this is the first report of a defective RNA from the RNA2 of TBRV.