A systematic study to determine the extent of gene silencing in Nicotiana benthamiana and other Solanaceae species when heterologous gene sequences are used for virus-induced gene silencing.
Virus-induced gene silencing (VIGS) is a rapid and robust method for determining and studying the function of plant genes or expressed sequence tags (ESTs). However, only a few plant species are amenable to VIGS. There is a need for a systematic study to identify VIGS-efficient plant species and to determine the extent of homology required between the heterologous genes and their endogenous orthologs for silencing. Two approaches were used. First, the extent of phytoene desaturase (PDS) gene silencing was studied in various Solanaceous plant species using Nicotiana benthamiana NbPDS sequences. In the second approach, PDS sequences from a wide range of plant species were used to silence the PDS gene in N. benthamiana. The results showed that tobacco rattle virus (TRV)-mediated VIGS can be performed in a wide range of Solanaceous plant species and that heterologous gene sequences from far-related plant species can be used to silence their respective orthologs in the VIGS-efficient plant N. benthamiana. A correlation was not always found between gene silencing efficiency and percentage homology of the heterologous gene sequence with the endogenous gene sequence. It was concluded that a 21-nucleotide stretch of 100% identity between the heterologous and endogenous gene sequences is not absolutely required for gene silencing.
Senthil-Kumar
M
M
Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Pky, Ardmore, OK 73401, USA.|Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India.
Hema
R
R
Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India.
Anand
Ajith
A
Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Pky, Ardmore, OK 73401, USA.
Kang
Li
L
Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Pky, Ardmore, OK 73401, USA.
Udayakumar
M
M
Department of Crop Physiology, University of Agricultural Sciences, GKVK, Bangalore 560 065, India.
Mysore
Kirankumar S
KS
Plant Biology Division, The Samuel Roberts Noble Foundation, 2510 Sam Noble Pky, Ardmore, OK 73401, USA.
D001483:Base Sequence; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D017343:Genes, Plant / Q000235:genetics; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010942:Plant Viruses / Q000502:physiology*; D021122:Protein Subunits; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics / Q000378:metabolism; D019657:Solanaceae / Q000235:genetics*; D014026:Nicotiana / Q000235:genetics*
D021122:Protein Subunits; D010088:Oxidoreductases; C022398:phytoene dehydrogenase; D012273:Ribulose-Bisphosphate Carboxylase