New approach for the construction of infectious clones of a circular DNA plant virus using Gibson Assembly

Ferro, Mayra Machado de Medeiros; Ramos-Sobrinho, Roberto; Xavier, César Augusto Diniz; Zerbini, F. Murilo; Lima, Gaus Silvestre de Andrade; Nagata, Tatsuya; Assunção, Iraíldes Pereira

doi:10.1016/j.jviromet.2018.10.017

Cited by 14 publications

(13 citation statements)

References 14 publications

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“…In contrast to In‐Fusion, Gibson assembly yields closed circular DNA molecules. Applied for reverse genetic studies of potyvirus clones (Bordat et al ., ; Pasin et al ., ), Gibson assembly has been used to generate binary infectious clones of members of genera Tymovirus (Blawid and Nagata, ), Carlavirus (Carvalho et al ., ), Comovirus (Bijora et al ., ), Potyvirus (Rose and Maiss, ), Polerovirus (Wetzel et al ., ), Benyvirus (Laufer et al ., ), Tobamovirus (Vasques et al ., ) and Begomovirus (Ferro et al ., ). Gibson assembly versions with improved fidelity are commercially available (e.g.…”

Section: Advanced Methods For Binary Infectious Clone Assemblymentioning

confidence: 97%

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Pasin

Menzel

Daròs

2019

Plant Biotechnology Journal

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Summary Recent metagenomic studies have provided an unprecedented wealth of data, which are revolutionizing our understanding of virus diversity. A redrawn landscape highlights viruses as active players in the phytobiome, and surveys have uncovered their positive roles in environmental stress tolerance of plants. Viral infectious clones are key tools for functional characterization of known and newly identified viruses. Knowledge of viruses and their components has been instrumental for the development of modern plant molecular biology and biotechnology. In this review, we provide extensive guidelines built on current synthetic biology advances that streamline infectious clone assembly, thus lessening a major technical constraint of plant virology. The focus is on generation of infectious clones in binary T‐ DNA vectors, which are delivered efficiently to plants by Agrobacterium . We then summarize recent applications of plant viruses and explore emerging trends in microbiology, bacterial and human virology that, once translated to plant virology, could lead to the development of virus‐based gene therapies for ad hoc engineering of plant traits. The systematic characterization of plant virus roles in the phytobiome and next‐generation virus‐based tools will be indispensable landmarks in the synthetic biology roadmap to better crops.

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Section: Advanced Methods For Binary Infectious Clone Assemblymentioning

confidence: 97%

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Pasin

Menzel

Daròs

2019

Plant Biotechnology Journal

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“…Therefore, PCR-based cloning by using overlapping primers to develop dimeric agro-infectious clones for the respective virus will be an easier approach. Ferro et al [174] have also developed the agro-infectious clones of a circular plant DNA virus by overlapping PCR-based amplification, and they have successfully validated this method on the mungbean.…”

Section: Concluding Remarks and Future Prospectsmentioning

confidence: 99%

Breeding for Enhancing Legumovirus Resistance in Mungbean: Current Understanding and Future Directions

Singh¹,

Singh

Pratap

et al. 2019

Agronomy

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Yellow mosaic disease (YMD) affects several types of leguminous crops, including the Vigna species, which comprises a number of commercially important pulse crops. YMD is characterized by the formation of a bright yellow mosaic pattern on the leaves; in severe forms, this pattern can also be seen on stems and pods. This disease leads to tremendous yield losses, even up to 100%, in addition to deterioration in seed quality. Symptoms of this disease are similar among affected plants; YMD is not limited to mungbean (Vigna radiata L. Wilczek) and also affects other collateral and alternate hosts. In the last decade, rapid advancements in molecular detection techniques have been made, leading to an improved understanding of YMD-causing viruses. Three distinct bipartite begomoviruses, namely, Mungbean Yellow Mosaic India Virus (MYMIV), Mungbean Yellow Mosaic Virus (MYMV), and Horsegram Yellow Mosaic Virus (HgYMV), are known to cause YMD in Vigna spp. Vigna crops serve as an excellent protein source for vegetarians worldwide; moreover, they aid in improving soil health by fixing atmospheric nitrogen through a symbiotic association with Rhizobium bacteria. The loss in the yield of these short-duration crops due to YMD, thus, needs to be checked. This review highlights the discoveries that have been made regarding various aspects of YMD affecting mungbean, including the determination of YMD-causing viruses and strategies used to develop high-yielding YMD-resistant mungbean varieties that harness the potential of related Vigna species through the use of different omics approaches.

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“…These DNA-A and DNA-B components were recovered from cactus samples ( Table 1 ). Specific primers were designed to amplify two copies of each cactus-derived begomoviruses component that were cloned in tandem to the binary vector pJL-89 [ 29 ], with the 35S promoter excluded as previously described by Ferro et al [ 30 ], using Gibson assembly [ 31 ] (New England Biolabs, Ipswich, MA, USA). Each clone was transformed into competent Escherichia coli XL1 Blue cells.…”

Section: Methodsmentioning

confidence: 99%

Identification of the Begomoviruses Squash Leaf Curl Virus and Watermelon Chlorotic Stunt Virus in Various Plant Samples in North America

Fontenele

Bhaskara

Cobb

et al. 2021

Viruses

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Geminiviruses are a group of plant-infecting viruses with single-stranded DNA genomes. Within this family, viruses in the genus Begomovirus are known to have a worldwide distribution causing a range of severe diseases in a multitude of dicotyledonous plant species. Begomoviruses are transmitted by the whitefly Bemisia tabaci, and their ssDNA genomes can be either monopartite or bipartite. As part of a viral survey, various plants including those in the families Alliaceae, Amaranthaceae, Apiaceae, Asteraceae, Brassicaceae, Cactaceae, Cucurbitaceae, Lamiaceae, Lauraceae, Malvaceae, Oleaceae and Solanaceae were sampled and screened for begomoviruses using both a high-throughput sequencing and a begomovirus-specific primer pair approach. Based on the sequences derived using these approaches, the full-length genome of various begomoviruses were amplified from plants using abutting primers. Squash leaf curl virus (SLCV) and watermelon chlorotic stunt virus (WCSV) were identified in Cactaceae (n = 25), Solanaceae (n = 7), Cucurbitaceae (n = 2) and Lamiaceae (n = 1) samples. WCSV is an Old World bipartite begomovirus that has only recently been discovered infecting watermelons in the Americas. Our discovery of WCSV in the USA is the first indication that it has reached this country and indicates that this virus might be widespread throughout North America. Phylogenetic analysis suggests WCSV was introduced to the New World twice. The detection of begomoviruses in cactus plants suggests possible spillover events from agricultural areas into native vegetation. Since WCSV and SLCV have previously been found in mixed infections, pseudo-recombination infection experiments were conducted. We demonstrate that WCSV DNA-B is successfully trans-replicated by SLCV DNA-A despite very low degree of similarity between the replication-associated iterative sequences present in their common region, an essential feature for binding of the replication associated protein. This study highlights the importance of viral surveys for the detection of spillover events into native vegetation, but also suggests the need for more surveillance of WCSV in the USA, as this virus is a serious threat to watermelon cultivation in the Middle East.

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New approach for the construction of infectious clones of a circular DNA plant virus using Gibson Assembly

Cited by 14 publications

References 14 publications

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Breeding for Enhancing Legumovirus Resistance in Mungbean: Current Understanding and Future Directions

Identification of the Begomoviruses Squash Leaf Curl Virus and Watermelon Chlorotic Stunt Virus in Various Plant Samples in North America

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