TY - JOUR
T1 - Virus-induced multiple gene silencing to study redundant metabolic pathways in plants
T2 - silencing the starch degradation pathway in Nicotiana benthamiana
AU - George, Gavin M.
AU - Bauer, Rolene
AU - Blennow, Per Gunnar Andreas
AU - Kossmann, Jens
AU - Lloyd, James R.
N1 - Special Issue: Focus: Synthetic biology
PY - 2012/7
Y1 - 2012/7
N2 - 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.
AB - 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.
U2 - 10.1002/biot.201100469
DO - 10.1002/biot.201100469
M3 - Journal article
C2 - 22345045
SN - 1860-6768
VL - 7
SP - 884
EP - 890
JO - Biotechnology Journal
JF - Biotechnology Journal
ER -