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dc.contributor.author
Lentz, Ezequiel M.
dc.contributor.author
Kuon, Joel‑Elias
dc.contributor.author
Alder, Adrian
dc.contributor.author
Mangel, Nathalie
dc.contributor.author
Zainuddin, Ima M.
dc.contributor.author
McCallum, Emily J.
dc.contributor.author
Anjanappa, Ravi B.
dc.contributor.author
Gruissem, Wilhelm
dc.contributor.author
Vanderschuren, Hervé
dc.date.accessioned
2018-09-04T08:02:47Z
dc.date.available
2018-09-03T07:31:28Z
dc.date.available
2018-09-04T08:01:30Z
dc.date.available
2018-09-04T08:02:47Z
dc.date.issued
2018
dc.identifier.issn
1746-4811
dc.identifier.other
10.1186/s13007-018-0340-5
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/286434
dc.identifier.doi
10.3929/ethz-b-000286434
dc.description.abstract
Aim We report the construction of a Virus-Induced Gene Silencing (VIGS) vector and an agroinoculation protocol for gene silencing in cassava (Manihot esculenta Crantz) leaves and roots. The African cassava mosaic virus isolate from Nigeria (ACMV-[NOg]), which was initially cloned in a binary vector for agroinoculation assays, was modified for application as VIGS vector. The functionality of the VIGS vector was validated in Nicotiana benthamiana and subsequently applied in wild-type and transgenic cassava plants expressing the uidA gene under the control of the CaMV 35S promoter in order to facilitate the visualization of gene silencing in root tissues. VIGS vectors were targeted to the Mg2+-chelatase gene in wild type plants and both the coding and promoter sequences of the 35S::uidA transgene in transgenic plants to induce silencing. We established an efficient agro-inoculation method with the hyper-virulent Agrobacterium tumefaciens strain AGL1, which allows high virus infection rates. The method can be used as a low-cost and rapid high-throughput evaluation of gene function in cassava leaves, fibrous roots and storage roots. Background VIGS is a powerful tool to trigger transient sequence-specific gene silencing in planta. Gene silencing in different organs of cassava plants, including leaves, fibrous and storage roots, is useful for the analysis of gene function. Results We developed an African cassava mosaic virus—based VIGS vector as well as a rapid and efficient agro-inoculation protocol to inoculate cassava plants. The VIGS vector was validated by targeting endogenous genes from Nicotiana benthamiana and cassava as well as the uidA marker gene in transgenic cassava for visualization of gene silencing in cassava leaves and roots. Conclusions The African cassava mosaic virus—based VIGS vector allows efficient and cost-effective inoculation of cassava for high-throughput analysis of gene function in cassava leaves and roots.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
BioMed Central
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.subject
Cassava
en_US
dc.subject
VIGS
en_US
dc.subject
ACMV
en_US
dc.subject
Geminivirus
en_US
dc.subject
Agrobacterium tumefaciens
en_US
dc.subject
Gene silencing
en_US
dc.subject
Agroinoculation
en_US
dc.title
Cassava geminivirus agroclones for virus-induced gene silencing in cassava leaves and roots
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 4.0 International
dc.date.published
2018-08-27
ethz.journal.title
Plant Methods
ethz.journal.volume
14
en_US
ethz.journal.issue
1
en_US
ethz.journal.abbreviated
Plant methods
ethz.pages.start
73
en_US
ethz.size
9 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.scopus
ethz.publication.place
London
en_US
ethz.publication.status
published
en_US
ethz.date.deposited
2018-09-03T07:31:47Z
ethz.source
SCOPUS
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2018-09-04T08:01:32Z
ethz.rosetta.lastUpdated
2018-12-02T13:18:24Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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