High throughput virus-induced gene silencing implicates heat shock protein 90 in plant disease resistance.
Virus-induced gene silencing was used to assess the function of random Nicotiana benthamiana cDNAs in disease resistance. Out of 4992 cDNAs tested from a normalized library, there were 79 that suppressed a hypersensitive response (HR) associated with Pto-mediated resistance against Pseudomonas syringae. However, only six of these clones blocked the Pto-mediated suppression of P.syringae growth. The three clones giving the strongest loss of Pto resistance had inserts corresponding to HSP90 and also caused loss of Rx-mediated resistance against potato virus X and N-mediated tobacco mosaic virus resistance. The role of HSP90 as a cofactor of disease resistance is associated with stabilization of Rx protein levels and could be accounted for in part by SGT1 and other cofactors of disease resistance acting as co-chaperones. This approach illustrates the potential benefits and limitations of RNA silencing in forward screens of gene function in plants.
Lu
Rui
R
The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK.
Malcuit
Isabelle
I
Moffett
Peter
P
Ruiz
Maria T
MT
Peart
Jack
J
Wu
Ai-Jiuan
AJ
Rathjen
John P
JP
Bendahmane
Abdelhafid
A
Day
Louise
L
Baulcombe
David C
DC
D000595:Amino Acid Sequence; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D020868:Gene Silencing; D017343:Genes, Plant; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000235:genetics* / Q000382:microbiology / Q000821:virology; D010940:Plant Proteins / Q000235:genetics*; D017863:Potexvirus / Q000472:pathogenicity; D017346:Protein Serine-Threonine Kinases / Q000502:physiology; D044224:Pseudomonas syringae / Q000472:pathogenicity; D017386:Sequence Homology, Amino Acid; D014026:Nicotiana / Q000235:genetics / Q000382:microbiology / Q000821:virology; D014027:Tobacco Mosaic Virus / Q000472:pathogenicity
D018076:DNA, Complementary; D018744:DNA, Plant; D018841:HSP90 Heat-Shock Proteins; D010940:Plant Proteins; C084354:Pto protein, Lycopersicon; D017346:Protein Serine-Threonine Kinases