A simplified method of constructing infectious clones of begomovirus employing limited restriction enzyme digestion of products of rolling circle amplification

Wu, Chia‐Ying; Lai, Yi-Chin; Lin, Na-Sheng; Hsu, Yau-Heiu; Tsai, H J; Liao, J. Y.; Hu, Chung-Chi

doi:10.1016/j.jviromet.2007.10.002

Cited by 29 publications

(30 citation statements)

References 29 publications

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“…Although it has been demonstrated that a clone containing a single genome copy is sufficient to ensure infectivity for certain begomoviruses [32], [33], [34], [35], the use of clones with duplicate copies of the sequences involved in the initiation of replication generally increases infectivity [31], [33], [36], [37], [38], [39], [40], [41]. Recently, a simple method for the construction of infectious begomovirus clones has been developed by cloning dimeric forms of the genome in a binary vector following partial digestion of the DNA obtained after rolling circle amplification with phage φ29 DNA polymerase [28], [29]. Despite the availability of relatively simple methodology for generating infectious clones of begomoviruses, which has been successfully applied to a large number of species, infectious clones were not obtained for any of the monopartite begomoviruses that infect Ipomoea spp., known as sweepoviruses.…”

Section: Discussionmentioning

confidence: 99%

“…Obtaining infectious begomovirus clones is a relatively easy process that has been simplified in various ways following the development of the rolling circle amplification (RCA) methodology using φ29 DNA polymerase [27], [28], [29]. Nevertheless, the genomes of sweepoviruses have been recalcitrant to rendering infectious clones to date [10], [21].…”

Section: Introductionmentioning

confidence: 99%

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Sweepoviruses Cause Disease in Sweet Potato and Related Ipomoea spp.: Fulfilling Koch's Postulates for a Divergent Group in the Genus Begomovirus

et al. 2011

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Sweet potato (Ipomoea batatas) and related Ipomoea species are frequently infected by monopartite begomoviruses (genus Begomovirus, family Geminiviridae), known as sweepoviruses. Unlike other geminiviruses, the genomes of sweepoviruses have been recalcitrant to rendering infectious clones to date. Thus, Koch's postulates have not been fullfilled for any of the viruses in this group. Three novel species of sweepoviruses have recently been described in Spain: Sweet potato leaf curl Lanzarote virus (SPLCLaV), Sweet potato leaf curl Spain virus (SPLCSV) and Sweet potato leaf curl Canary virus (SPLCCaV). Here we describe the generation of the first infectious clone of an isolate (ES:MAL:BG30:06) of SPLCLaV. The clone consisted of a complete tandem dimeric viral genome in a binary vector. Successful infection by agroinoculation of several species of Ipomoea (including sweet potato) and Nicotiana benthamiana was confirmed by PCR, dot blot and Southern blot hybridization. Symptoms observed in infected plants consisted of leaf curl, yellowing, growth reduction and vein yellowing. Two varieties of sweet potato, ‘Beauregard’ and ‘Promesa’, were infected by agroinoculation, and symptoms of leaf curl and interveinal loss of purple colouration were observed, respectively. The virus present in agroinfected plants was readily transmitted by the whitefly Bemisia tabaci to I. setosa plants. The progeny virus population present in agroinfected I. setosa and sweet potato plants was isolated and identity to the original isolate was confirmed by sequencing. Therefore, Koch's postulates were fulfilled for the first time for a sweepovirus.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Sweepoviruses Cause Disease in Sweet Potato and Related Ipomoea spp.: Fulfilling Koch's Postulates for a Divergent Group in the Genus Begomovirus

et al. 2011

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“…This technique does not require sequence information for cloning and is less expensive and time-consuming than conventional PCR-based methods (Wu et al, 2008; Bang et al, 2014). The amplification products can be sequenced directly (Jeske et al, 2010) or used for the cloning of geminiviral genomes (Inoue-Nagata et al, 2004; Wu et al, 2008) and biolistic inoculation (Jeske et al, 2010; Aranha et al, 2011). …”

Section: Rolling Circle Amplification (Rca) and Infectious Clones As mentioning

confidence: 99%

Biotechnological Advancements and Begomovirus Management in Okra (Abelmoschus esculentus L.): Status and Perspectives

et al. 2017

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Despite the importance of okra, as one of the important vegetable crop, very little attention has been paid to its genetic improvement using advanced biotechnological tools. The exploitation of marker assisted breeding in okra is often limited due to the availability of a few molecular markers, the absence of molecular genetic-map(s), and other molecular tools. Chromosome linkage-groups were not yet constructed for this crop and reports on marker development are very scanty and mostly hovering around cultivar characterization. Besides, very little progress has been observed for transgenic development. However, high throughput biotechnological tools like chromosome engineering, RNA interference (RNAi), marker-assisted recurrent selection (MARS), genome-wide selection (GWS), targeted gene replacement, next generation sequencing (NGS), and nanobiotechnology can provide a rapid way for okra improvement. Further, the etiology of many deadly viral diseases like the yellow vein mosaic virus (YVMV) and okra enation leaf curl virus (OELCV) in okra is broadly indistinct and has been shown to be caused by various begomovirus species. These diseases cause systemic infections and have a very effective mode of transmission; thus, preventing their spread has been very complicated. Biotechnological interventions have the potential to enhance okra production even under different viral-stress conditions. In this background, this review deals with the biotechnological advancements in okra per se along with the begomoviruses infecting okra, and special emphasis has been laid on the exploitation of advanced genomic tools for the development of resistant varieties.

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“…Tandemly repeated infectious DNA clones of the SL-1 isolate were constructed by rolling circle amplification (RCA) as described by Wu et al (2008) with some modification. Total DNA was isolated from tissue of SL-1 infected melon and the circular genomic DNAs were amplified by RCA using the TempliPhi Amplification kit (Amersham Pharmacia).…”

Section: Construction Of Infectious Clones and Agroinfectionmentioning

confidence: 99%

Identification and characterization of a mechanical transmissible begomovirus causing leaf curl on oriental melon

Chang

Tsai

et al. 2010

Eur J Plant Pathol

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Oriental melon plants, Cucumis melo var. makuwa cv. Silver Light, showing virus-induced symptoms of mosaic, leaf curl and puckering were observed in the fields of eastern Taiwan in 2007. A virus culture, designated as SL-1, isolated from the diseased melon was established in systemic host plants, Nicotiana benthamiana and oriental melon, by mechanical inoculation. SL-1 did not react to the antisera against common cucurbit-infecting RNA viruses. Viral DNAs extracted from the diseased plant were amplified with the degenerate primers for begomoviruses. The full-length genomic DNA-A and DNA-B of SL-1 were sequenced and found to be closest, with 97.7% and 90.6% nucleotide identity, respectively, to Tomato leaf curl New Delhi begomovirus (ToLCNDV) cucumber isolate from a group of cucurbit-infecting begomoviruses. The virus SL-1 was designated as ToLCNDV oriental melon isolate (ToLCNDV-OM). The pathogenicity of ToLCNDV-OM was confirmed by agroinfection. Progeny virus from the agroinfected N. benthamiana plants was able to infect oriental melon by mechanical inoculation and caused symptoms similar to the original diseased melon in the field. The ToLCNDV-OM also infected five other species of cucurbitaceous plants by mechanical inoculation. This is the first report of a new ToLCNDV isolate causing severe disease on oriental melon in Taiwan

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A simplified method of constructing infectious clones of begomovirus employing limited restriction enzyme digestion of products of rolling circle amplification

Cited by 29 publications

References 29 publications

Sweepoviruses Cause Disease in Sweet Potato and Related Ipomoea spp.: Fulfilling Koch's Postulates for a Divergent Group in the Genus Begomovirus

Sweepoviruses Cause Disease in Sweet Potato and Related Ipomoea spp.: Fulfilling Koch's Postulates for a Divergent Group in the Genus Begomovirus

Biotechnological Advancements and Begomovirus Management in Okra (Abelmoschus esculentus L.): Status and Perspectives

Identification and characterization of a mechanical transmissible begomovirus causing leaf curl on oriental melon

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