Cucumber mosaic virus-induced gene silencing in banana

Abstract

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.

Using Genetic Engineering Techniques to Develop Banana Cultivars With Fusarium Wilt Resistance and Ideal Plant Architecture

“…Virus-induced gene silencing (VIGS) can be used to analyze gene function via the transient silencing of RNA expression; VIGS thus compensates for the time-consuming and inefficient nature of transgenic banana production ( Baulcombe, 1999 ). For example, Tzean et al (2019) developed a VIGS system based on a banana-infecting cucumber mosaic virus (CMV) to efficiently silence target genes. This system might be useful for gene functional analysis in bananas before stable genetic transformation.…”

“…Because bananas are nutritious and starchy, they are considered the fourth most important food crop after rice, wheat, and maize for the alleviation of human starvation in Africa by the United Nations Food and Agriculture Organization ( Song et al, 2018 ). In 2019, worldwide banana production was over 113 million tons; the two largest producers of bananas are India (36.7%) and China (13.8%) ( Tzean et al, 2019 ).…”

Application of cucumber mosaic virus to efficient induction and long-term maintenance of virus-induced gene silencing in spinach

“…CMV has been developed as a useful VIGS vector for many plants including Nicotiana benthamiana, tomato, pepper, Arabidopsis thaliana, lily, soybean, banana and maize (Hong et al 2012;Liu et al 2010;Otagaki et al 2006;Tzean et al 2019;Wang et al 2016). Recently, it has been demonstrated that we could silence even a gene of mycorrhizal fungi that colonized the root of N. benthamiana (Kikuchi et al 2016).…”

Simultaneous silencing of two different Arabidopsis genes with a novel virus-induced gene silencing vector

“…Given these features, VIGS is an attractive reverse genetic tool for functional genomics in plants [ 4 – 8 ]. In the past decade, a number of viral genomes has been modified as a powerful reverse genetic tool for the functional characterization of genes in plants, such as Tobacco rattle virus [ 9 ], Apple latent spherical virus [ 10 , 11 ], African cassava mosaic virus [ 12 ], and Cucumber mosaic virus [ 13 ]. However, most of the reported VIGS vectors only silence a single gene, and the VIGS vectors with visualizable indicator to evaluate the early penetrance of genes for reverse genetics are lacking.…”