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Virus‐induced multiple gene silencing to study redundant metabolic pathways in plants: Silencing the starch degradation pathway in Nicotiana benthamiana
Author(s) -
George Gavin M.,
Bauer Rolene,
Blennow Andreas,
Kossmann Jens,
Lloyd James R.
Publication year - 2012
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201100469
Subject(s) - nicotiana benthamiana , gene silencing , biology , gene , microbiology and biotechnology , gene expression , enzyme , biochemistry , genetics
Virus‐induced gene silencing (VIGS) is a rapid technique that allows for specific and reproducible post‐transcriptional degradation of targeted mRNA. The method has been proven efficient for suppression of expression of many single enzymes. The metabolic networks of plants, however, often contain isoenzymes and gene families that are able to compensate for a mutation and mask the development of a silencing phenotype. Here, we show the application of multiple gene VIGS repression for the study of these redundant biological pathways. Several genes in the starch degradation pathway [disproportionating enzyme 1; ( DPE1 ), disproportionating enzyme 2 ( DPE2 ), and GWD ] were silenced. The functionally distinct DPE enzymes are present in alternate routes for sugar export to the cytoplasm and result in an increase in starch production when silenced individually. Simultaneous silencing of DPE1 and DPE2 in Nicotiana benthamiana resulted in a near complete suppression in starch and accumulation of malto‐oligosaccharides.