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Cucumber mosaic virus-induced gene silencing in banana
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Cucumber mosaic virus-induced gene silencing in banana
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Journal Article
Cucumber mosaic virus-induced gene silencing in banana
2019
Overview
Banana (
Musa
spp.) is one of the world’s most important staple and cash crops. Despite accumulating genetic and transcriptomic data, low transformation efficiency in agronomically important
Musa
spp. render translational researches in banana difficult by using conventional knockout approaches. To develop tools for translational research in bananas, we developed a virus induced-gene silencing (VIGS) system based on a banana-infecting cucumber mosaic virus (CMV) isolate, CMV 20. CMV 20 genomic RNA 1, 2, and 3, were separately cloned in
Agrobacterium
pJL89 binary vectors, and a cloning site was introduced on RNA 2 immediately after the 2a open reading frame to insert the gene targeted for silencing. An efficient
Agrobacterium
inoculation method was developed for banana, which enabled the CMV 20 VIGS vector infection rate to reach 95% in our experiments. CMV 20-based silencing of
Musa acuminata
cv. Cavendish (AAA group)
glutamate 1-semialdehyde aminotransferase
(
MaGSA
) produced a typical chlorotic phenotype and silencing of
M. acuminata phytoene desaturase
(
MaPDS
) produced a photobleachnig phenotype. We show this approach efficiently reduced
GSA
and
PDS
transcripts to 10% and 18% of the control, respectively. The high infection rate and extended silencing of this VIGS system will provide an invaluable tool to accelerate functional genomic studies in banana.
