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.
The Sainsbury Laboratory, John Innes Centre, Colney Lane, Norwich NR4 7UH, UK.
Rui
R
Lu
Isabelle
I
Malcuit
Peter
P
Moffett
Maria T
MT
Ruiz
Jack
J
Peart
Ai-Jiuan
AJ
Wu
John P
JP
Rathjen
Abdelhafid
A
Bendahmane
Louise
L
Day
David C
DC
Baulcombe
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