Abstract
Quinoa (Chenopodium quinoa), native to the Andean region of South America, has been recognized as a potentially important crop in terms of global food and nutrition security since it can thrive in harsh environments and has an excellent nutritional profile. Even though challenges of analyzing the complex and heterogeneous allotetraploid genome of quinoa have recently been overcome, with the whole genome-sequencing of quinoa and the creation of genotyped inbred lines, the lack of technology to analyze gene function in planta is a major limiting factor in quinoa research. Here, we demonstrate that two virus-mediated transient expression techniques, virus-induced gene silencing (VIGS) and virus-mediated overexpression (VOX), can be used in quinoa. We show that apple latent spherical virus (ALSV) can induce gene silencing of quinoa phytoene desaturase (CqPDS1) in a broad range of quinoa inbred lines derived from the northern and southern highland and lowland sub-populations. In addition, we show that ALSV can be used as a VOX vector in roots. Our data also indicate that silencing a quinoa 3,4-dihydroxyphenylalanine 4,5-dioxygenase gene (CqDODA1) or a cytochrome P450 enzyme gene (CqCYP76AD1) inhibits betalain production and that knockdown of a reduced-height gene homolog (CqRHT1) causes an overgrowth phenotype in quinoa. Moreover, we show that ALSV can be transmitted to the progeny of quinoa plants. Thus, our findings enable functional genomics in quinoa, ushering in a new era of quinoa research.
Highlights
Rapid and large-scale changes in environmental and social conditions require rapid and fundamental changes in crop productivity and diversity (Eshed and Lippman, 2019)
We obtained inocula for virus-induced gene silencing (VIGS) analyses, including the packaged viruses of apple latent spherical virus (ALSV)-CqPDSN, ALSV-CqPDSC, and ALSVWT, from the uninoculated upper leaves with chlorotic spots of quinoa plants inoculated with ALSV-RNA1 complementary DNA (cDNA) plasmid combined with ALSV-RNA2 VIGS vectors either harboring or not a specific 300-bp trigger sequence required for VIGS (Supplementary Figure 1)
We obtained inocula for VIGS analyses, including the packaged viruses of ALSV-CqRHT1, from the uninoculated upper leaves with chlorotic spots of quinoa plants (Iw line) inoculated with ALSV-RNA1 cDNA plasmid combined with ALSV-RNA2 VIGS vectors either harboring or not a specific 300-bp trigger sequence required for VIGS
Summary
Rapid and large-scale changes in environmental and social conditions require rapid and fundamental changes in crop productivity and diversity (Eshed and Lippman, 2019). ALSV VIGS and VOX in Quinoa production will require that people shift from a livestock-based diet to a plant-based diet (Shepon et al, 2018). This shift will need to be accompanied by an increase in the production of proteinrich crops (Eshed and Lippman, 2019). Diversifying staple crops is another requirement for achieving a food supply that is sustainable, resilient, and suited to local environments (Massawe et al, 2016)
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