Synergy between an emerging monopartite begomovirus and a DNA-B component

In recent decades, a legion of monopartite begomoviruses transmitted by the whitefly Bemisia tabaci has emerged as serious threats to vegetable crops in Africa. Recent studies in Burkina Faso (West Africa) reported the predominance of pepper yellow vein Mali virus (PepYVMLV) and its frequent association with a previously unknown DNA-B component. To understand the role of this DNA-B component in the emergence of PepYVMLV, we assessed biological traits related to virulence, virus accumulation, location in the tissue and transmission. We demonstrate that the DNA-B component is not required for systemic movement and symptom development of PepYVMLV (non-strict association), but that its association produces more severe symptoms including growth arrest and plant death. The increased virulence is associated with a higher viral DNA accumulation in plant tissues, an increase in the number of contaminated nuclei of the phloem parenchyma and in the transmission rate by B. tabaci. Our results suggest that the association of a DNA-B component with the otherwise monopartite PepYVMLV is a key factor of its emergence.

Complete genome sequence of a previously undescribed monopartite begomovirus and betasatellite infecting Malvastrum coromandelianum in Cambodia

A previously undescribed monopartite begomovirus was identified from Malvastrum coromandelianum plants exhibiting yellow vein symptoms characteristic of begomoviruses, in Kampot province, Cambodia. The apparently full-length viral component was cloned and sequenced following enrichment of circular DNA by rolling circle amplification and restriction enzyme digestion. The genome of the virus was 2,737 nucleotides in length (KP188831), and exhibited an organization like that of other monopartite begomoviruses, sharing the highest nt identities of 87.7% with Ageratum yellow vein virus (AM940137). A satellite molecule was amplified from total DNA by PCR amplification with the betasatellite-specific primer pair β01/β02. The satellite molecule (1,346 nt, KP188832) had a structure characteristic like other betasatellites associated with begomoviruses, and shared the highest nt identity of 84.8% with Malvastrum yellow vein betasatellite (MN205547). According to the criteria established for species demarcation for classification of begomoviruses ( Geminiviridae ) and betasatellites ( Tolecusatellitidae ), respectively, the virus isolate from M. coromandelianum in Cambodia is a previously undescribed novel monopartite begomovirus species，for which the name Malvastrum yellow vein Cambodia virus (MaYVCV) is proposed, whereas, the betasatellite is identified as an previously undescribed novel betasatellite species, for which the name Malvastrum yellow vein Cambodia batesatellite (MaYVKHB) is proposed.

Geminiviruses: Taxonomic Structure and Diversity in Genomic Organization

Background: Geminiviridae is one of the best-characterized and hence, one of the largest plant-virus families with the highest economic importance. Its members characteristically have a circular ssDNA genome within the encapsidation of twinned quasi-icosaheadral virions (18-38 nm size-range). Objective: Construction of a narrative review on geminiviruses, to have a clearer picture of their genomic structure and taxonomic status. Methods: A thorough search was conducted for papers and patents regarding geminiviruses, where relevant information was used to study their genomic organization, diversity and taxonomic structure. Results: Geminiviruses have been classified into nine genera (viz., genus Begomovirus, Mastrevirus, Curtovirus, Topocuvirus, Becurtovirus, Turncurtovirus, Capulavirus, Eragrovirus and Grablovirus) having distinct genomic organizations, host ranges and insect vectors. Genomic organization of all genera generally shows the presence of 4-6 ORFs encoding for various proteins. For now, Citrus chlorotic dwarf-associated virus (CCDaV), Camellia chlorotic dwarf-associated virus (CaCDaV) and few other geminiviruses are still unassigned to any genera. The monopartite begomoviruses (and few mastreviruses) have been found associated with aplhasatellites and betasatellites (viz., ~1.3 kb circular ssDNA satellites). Recent reports suggest that deltasatellites potentially reduce the accumulation of helper-Begomovirus species in host plants. Some patents have revealed the methods to generate transgenic plants resistant to geminiviruses. Conclusion: Geminiviruses rapidly evolve and are a highly diverse group of plant-viruses. However, research has shown new horizons in tackling the acute begomoviral diseases in plants by generating a novel bio-control methodology in which deltasatellites can be used as bio-control agents and generate transgenic plants resistant to geminiviruses.

Frequent occurrence of Mungbean yellow mosaic India virus in tomato leaf curl disease affected tomato in Oman

Next generation sequencing (NGS) of DNAs amplified by rolling circle amplification from 6 tomato (Solanum lycopersicum) plants with leaf curl symptoms identified a number of monopartite begomoviruses, including Tomato yellow leaf curl virus (TYLCV), and a betasatellite (Tomato leaf curl betasatellite [ToLCB]). Both TYLCV and ToLCB have previously been identified infecting tomato in Oman. Surprisingly the NGS results also suggested the presence of the bipartite, legume-adapted begomovirus Mungbean yellow mosaic Indian virus (MYMIV). The presence of MYMIV was confirmed by cloning and Sanger sequencing from four of the six plants. A wider analysis by PCR showed MYMIV infection of tomato in Oman to be widespread. Inoculation of plants with full-length clones showed the host range of MYMIV not to extend to Nicotiana benthamiana or tomato. Inoculation to N. benthamiana showed TYLCV to be capable of maintaining MYMIV in both the presence and absence of the betasatellite. In tomato MYMIV was only maintained by TYLCV in the presence of the betasatellite and then only at low titre and efficiency. This is the first identification of TYLCV with ToLCB and the legume adapted bipartite begomovirus MYMIV co-infecting tomato. This finding has far reaching implications. TYLCV has spread around the World from its origins in the Mediterranean/Middle East, in some instances, in live tomato planting material. The results here may suggest that begomoviruses which do not commonly infect tomato, such as MYMIV, could be spread as a passenger of TYLCV in tomato.

Molecular and biological characterization of Chilli leaf curl virus and associated Tomato leaf curl betasatellite infecting tobacco in Oman

Background In Oman tobacco (Nicotiana tabacum; family Solanaceae) is a minor crop, which is produced only for local consumption. In 2015, tobacco plants exhibiting severe downward leaf curling, leaf thickening, vein swelling, yellowing and stunting were identified in fields of tobacco in Suhar Al-Batina region, Oman. These symptoms are suggestive of begomovirus (genus Begomovirus, family Geminiviridae) infection. Methods Circular DNA molecules were amplified from total DNA extracted from tobacco plants by rolling circle amplification (RCA). Viral genomes were cloned from RCA products by restriction digestion and betasatellites were cloned by PCR amplification from RCA product, using universal primers. The sequences of full-length clones were obtained by Sanger sequencing and primer walking. Constructs for the infectivity of virus and betasatellite were produced and introduced into plants by Agrobacterium-mediated inoculation. Results The full-length sequences of 3 begomovirus and 3 betasatellite clones, isolated from 3 plants, were obtained. Analysis of the full-length sequences determined showed the virus to be a variant of Chilli leaf curl virus (ChiLCV) and the betasatellite to be a variant of Tomato leaf curl betasatellite (ToLCB). Both the virus and the betasatellite isolated from tobacco show the greatest levels of sequence identity to isolates of ChiLCV and ToLCB identified in other hosts in Oman. Additionally clones of ChiLCV and ToLCB were shown, by Agrobacterium-mediated inoculation, to be infectious to 3 Nicotiana species, including N. tabacum. In N. benthamiana the betasatellite was shown to change the upward leaf rolling symptoms to a severe downward leaf curl, as is typical for many monopartite begomoviruses with betasatellites. Conclusions The leaf curl disease of tobacco in Oman was shown to be caused by ChiLCV and ToLCB. This is the first identification of ChiLCV with ToLCB infecting tobacco. The study shows that, despite the low diversity of begomoviruses and betasatellites in Oman, the extant viruses/betasatellites are able to fill the niches that present themselves.

Tomato Twisted Leaf Virus: A Novel Indigenous New World Monopartite Begomovirus Infecting Tomato in Venezuela

Begomoviruses are one of the major groups of plant viruses with an important economic impact on crop production in tropical and subtropical regions. The global spread of its polyphagous vector, the whitefly Bemisia tabaci, has contributed to the emergence and diversification of species within this genus. In this study, we found a putative novel begomovirus infecting tomato plants in Venezuela without a cognate DNA-B component. This begomovirus was genetically characterized and compared with related species. Furthermore, its infectivity was demonstrated by agroinoculation of infectious clones in tomato (Solanum lycopersicum) and Nicotiana benthamiana plants. The name Tomato twisted leaf virus (ToTLV) is proposed. ToTLV showed the typical genome organization of the DNA-A component of New World bipartite begomoviruses. However, the single DNA component of ToTLV was able to develop systemic infection in tomato and N. benthamiana plants, suggesting a monopartite nature of its genome. Interestingly, an additional open reading frame ORF was observed in ToTLV encompassing the intergenic region and the coat protein gene, which is not present in other closely related begomoviruses. A putative transcript from this region was amplified by strand-specific reverse transcription-PCR. Along with recent studies, our results showed that the diversity of monopartite begomoviruses from the New World is greater than previously thought.

Tomato leaf curl Patna virus causing tomato leaf curl disease in Bangladesh

Tomato leaf curl virus (ToLCV) has appeared as a potential threat to the tomato production in the world. ToLCV, a member of the family Geminiviridae may contain either bipartite or monopartite genome. The genetic nature of a monopartite ToLCV isolate characterized from the tomato leaf curl diseased samples of Jamalpur district, Bangladesh (ToLCV-JB) has been reported. The products of rolling circle amplification (RCA) were digested, cloned and sequenced. Sequence analysis revealed the features of begomovirus genome organization in the ToLCV-JB isolate, containing six open reading frames. BLAST analysis showed 100% sequence similarity with tomato leaf curl Patna virus (EU862323.1) and more than 80% similarity with other reported monopartite begomoviruses. Hence, the virus isolate was registered as Tomato leaf Curl Patna virus-[Bangladesh:Jamalpur:2014] isolate ToLCV-JB (Genebank Accession: KU933675.1) according to the suggestion of NCBI. Recombination analysis also did not show any genetic exchange between ToLCV-JB and ToLCV-Patna virus. Moreover, they belong to the same cluster as observed in phylogenetic analysis. The present work suggests the possibility of cross-border spread of ToLCV-Patna viruses without mutation and this could pose a threat to tomato production in Bangladesh as well as in the Asian continent.

In-Silico Investigation of Betasatellite Complexity in Papaya Leaf Curl Disease Complex

Papaya leaf curl disease complex (PLCD) impose heavy losses to papaya farmers across India. It is mainly reported to be caused by Papaya leaf curl virus and associated begomoviruses carrying a DNA-A and betasatellite molecules. Both components are required for infliction of severe symptoms. The investigation into molecular diversity is necessary to devise intervention techniques against these begomoviruses. Based on the outcome of diversity, preferred resistance strategy against a disease complex should be generic in nature, which provides an advantage of a broad range resistance against a variety of plant virus causing same disease symptoms. For this purpose, we studied the molecular diversity through investigation of genetic complexity of the betasatellite genome using various computational methods and probed genetic complexity of betasatellite component. Multiple sequence alignment and recombination analysis were performed to understand the evolutionary relationship of betasatellite components with a DNA-A present as a helper genome in case of monopartite begomoviruses. The overall results indicate that the betasatellite molecules have evolved independently of DNA-A component and their association is a result of co-infection and due to inter- and intraspecific interaction with various forms of virus infection in plants. The information generated through this study has potential application in designing intervention strategies against PLCD, which is widespread in Indian sub-continent region that includes countries like India, Pakistan, Nepal and Bangladesh.