Virus-induced Gene Silencing of 14-3-3 Genes Abrogates Dark Repression of Nitrate Reductase Activity in Nicotiana Benthamiana
Overview
Affiliations
In order to study the effect of repression of 14-3-3 genes on actual activity of the nitrate reductase (NR) in Nicotiana benthamiana leaves, Nb14-3-3a gene was silenced by virus-induced gene silencing (VIGS) method using potato virus X (PVX). Expression of Nb14-3-3a as well as Nb14-3-3b genes was altogether repressed in the leaves of PVX-14-3a-infected plants. Furthermore, two-dimensional gel electrophoresis and immunoblot analysis with anti-14-3-3 antiserum suggested that the expressions of Nb14-3-3a and Nb14-3-3b proteins are accordingly repressed in PVX-14-3a-infected plants. It is well known that binding of 14-3-3 proteins to phosphorylated NR leads to substantial decrease in NR activity of leaves under darkness. Therefore, we studied the changes in NR activity in response to light/dark transitions in the leaves of PVX-14-3a-infected plants. NR activation state was kept at a high level under darkness in PVX-14-3a-infected plants, but not in PVX-green fluorescent protein (GFP)-infected and control plants. This result suggests that Nb14-3-3a and/or Nb14-3-3b proteins are indeed involved in the inactivation of NR activity under darkness in N. benthamiana.
Determining novel functions of Arabidopsis 14-3-3 proteins in central metabolic processes.
Diaz C, Kusano M, Sulpice R, Araki M, Redestig H, Saito K BMC Syst Biol. 2011; 5:192.
PMID: 22104211 PMC: 3253775. DOI: 10.1186/1752-0509-5-192.
Ahn C, Han J, Lee H, Lee S, Pai H Plant Cell. 2011; 23(1):185-209.
PMID: 21216945 PMC: 3051261. DOI: 10.1105/tpc.110.074005.