Diane Jofuku Okamuro
IOS Division of Integrative Organismal Systems
BIO Directorate for Biological Sciences
Start Date:
September 1, 2002
Expires:
August 31, 2007 (Estimated)
Awarded Amount to Date:
$1928581
Investigator(s):
Sophien Kamoun sophien.kamoun@tsl.ac.uk (Principal Investigator)
Kamal Gajendran (Co-Principal Investigator)
Mark Waugh (Former Co-Principal Investigator)
Sponsor:
Ohio State University Research Foundation
1960 KENNY RD
Columbus, OH 43210 614/292-3732
NSF Program(s):
AFRICA, NEAR EAST, & SO ASIA, PLANT GENOME RESEARCH PROJECT
Field Application(s):
Program Reference Code(s):
BIOT, 9232, 9184, 9109, 5991
Program Element Code(s):
5976, 1329
ABSTRACT
Plant pathogenic microbes have the remarkable ability to manipulate biochemical, physiologc al and morphological processes in their host plants. These manipulations are achieved through a diverse array of virulence and avirulence factors that an either promote infection or trigger defense responses. Oomycetes, such as Phytophthora, downy-mildews and Pythium, form a unique branch of eukaryotic plant pathogens with an independent evolutionary history. Among the oomy etes, Phytophthora spp. cause some of the most destructive plant diseases in the world, and are arguably the most devastating pathogens of dicot plants. or example, Phytophthora infestans, the Irish potato famine pathogen, causes late blight, a disease that results in multibillion-dollar losses in potato and tomato production and is considered a significant threat to global food security. Structural genomics studies of Phytophthora are well under way within the framework of the Phytophthora Genome Consortium. However, despite their economic impact and singular phylogeny, oomycetes remain relatively understudied at a molecular level and key molecular events modulating basc pathogenicity and plant response remain poorly understood. The overall obje tive of this project is to use the technology of virus-mediated gene expression to carry out high throughput fun tional screening of P. infestans genes in plants. The specific objectives are to: 1. Perform high throughput functional expression screening of P. infestans DNAs in tomato and tobacco using a potato virus X (PVX)-based vector to identify DNAs that indu e resistance responses, disease-like symptoms and/or altered disease susceptibility. 2. Characterize the selected P. infestans DNAs and the encoded proteins with regards to primary sequence, gene expression, biological activity, and cellular localization during infection. 3. Characterize the molecular and cellular responses of tomato and tobacco to the sele ted P. infestans DNAs. The transient nature of the PVX expression system and the use of a recently developed Agrobacterium binary PVX vector will allow functional analysis of pathogen genes at an unprecedented high throughput rate. The functional screening will unravel a battery of novel Phytophthora genes that trigger a variety of cellular and molecular responses in plant cells and will firmly establish functional connections between pathogen genes and plant processes. The knowledge gained from these studies will provide significant insight into key molecular processes regulating an economically important pathosystem and will provide novel tools for improvement of disease resistan e in crop plants. Information and material generated from this project will be made freely available to the s ientific community through an interactive website that will include an annotated database of the selected DNAs and corresponding phenotype and expression data. The project will also offer outstanding opportunities for interdis iplinary training and outreach activities in functional genomics and bioinformatics revolving around the use of spectacular functional assays, historically and economically important disease pathosystems, and challenging concepts on the interaction between plants and microbes.
Deliverables:
1. Libraries of P. infestans cDNAs in virus vectors, including collection of Phytophthora genes that trigger various cellular and molecular responses in plant cells
2. Phenotypic and expression analyses of tomato and tobacco treated with Phytophthora effectors.
Bhattacharjee, S., Hiller, L. N., Liolios, K., Win, J., Kanneganti, T. -D., Young, C., Kamoun, S., and Haldar, K.. "The malarial host-targeting signal is conserved in the Irish potato famine pathogen.," PLoS Pathogens, v.22, 2006, p. e50.
Birch, P. R. J., Rehmany, A. P., Pritchard, L., Kamoun, S., and Beynon, J. L.. "Trafficking arms: oomycete effectors enter host plant cells," Trends in Microbiology, v.14, 2006, p. 8.
Bos, J. I. B., Armstrong, M., Whisson, S. C., Torto, T., Ochwo, M., Birch, P. R. J., and Kamoun, S.. "Intraspecific comparative genomics to identify avirulence genes from Phytophthora.," New Phytologist, v.159, 2003, p. 63.
Bos, J..I.B., Kanneganti, T. -D., Young, C., Cakir, C., Huitema, E., Win, J., Armstrong, M., Birch, P.R.J., and Kamoun, S.. "The C-terminal half of Phytophthora infestans RXLR effector AVR3a is sufficient to trigger R3a-mediated hypersensitivity and suppress INF1-induced cell death.," Plant Journal, v.4, 2006, p. 165.
Gajendran K, Gonzales MD, Farmer A, Archuleta E, Win J, Waugh ME, Kamoun S.. "Phytophthora functional genomics database (PFGD): functional genomics of Phytophthora-
plant interactions.," Nucleic Acids Research, v.34, 2006, p. D465.
Haldar, K; Kamoun, S; Hiller, NL; Bhattacharje, S; van Ooij, C. "Common infection strategies of pathogenic eukaryotes," NATURE REVIEWS MICROBIOLOGY, v.4, 2006, p. 922-931.
Huitema, E., Bos, J.I.B., Tian, M., Win, J., Waugh, M.E., and Kamoun, S.. "Linking sequence to phenotype in Phytophthora-plant interactions," Trends in Microbiology, v.12, 2004, p. 193.
Huitema, E., Vleeshouwers, V.G.A.A., Cakir, C., Kamoun, S., and Govers, F.. "Differences in intensity and specificity of hypersensitive response induction in
Nicotiana spp. by INF1, INF2A and INF2B of Phytophthora infestans," Molecular Plant-Microbe Interactions, v.18, 2005, p. 183.
Kamoun, S. "Groovy times: filamentous pathogen effectors revealed," CURRENT OPINION IN PLANT BIOLOGY, v.10, 2007, p. 358-365.
Kamoun, S.. "Molecular genetics of pathogenic oomycetes.," Eukaryotic Cell, v.2, 2003, p. 191.
Kanneganti, T. -D., Huitema, E., Vleeshouwers, V.G.A.A., Cakir, C., and Kamoun, S.. "Synergistic interactions of the plant cell death pathways induced by Phytophthora infestans Nep1-like protein PiNPP1.1 and INF1 elicitin.," Molecular Plant-Microbe Interactions, v.19, 2006, p. 854.
Kanneganti, TD; Bai, XD; Tsai, CW; Win, J; Meulia, T; Goodin, M; Kamoun, S; Hogenhout, SA. "A functional genetic assay for nuclear trafficking in plants," PLANT JOURNAL, v.50, 2007, p. 149-158.
Lamour, KH; Win, J; Kamoun, S. "Oomycete genomics: new insights and future directions," FEMS MICROBIOLOGY LETTERS, v.274, 2007, p. 1-8.
Liu, Z., Bos, J.I.B., Armstrong, M., Whisson, S.C., da Cunha, L., Torto-Alalibo, T., Win,
J., Avrova, A.O., Wright, F., Birch P.R.J., and Kamoun, S.. "Patterns of diversifying selection in the phytotoxin-like scr74 gene family of
Phytophthora infestans," Molecular Biology and Evolution, v.22, 2005, p. 659.
Rehmany, A. P., Gordon, A., Rose, L.E., Allen, R.L., Armstrong, M.R., Whisson, S.C.,
Kamoun, S., Tyler, B.M., Birch, P.R.J., and Beynon, J.L.. "Differential recognition of highly divergent downy mildew avirulence gene alleles by
RPP1 genes from two Arabidopsis lines.," Plant Cell, v.17, 2005, p. 1839.
Tian, MY; Win, J; Song, J; van der Hoorn, R; van der Knaap, E; Kamoun, S. "A Phytophthora infestans cystatin-like protein targets a novel tomato papain-like apoplastic protease," PLANT PHYSIOLOGY, v.143, 2007, p. 364-377.
Torto, T., Li, S., Styer, A., Huitema, E., Testa, A., Gow, N.A.R., van West, P., and Kamoun, S.. "EST mining and functional expression assays identify extracellular effector proteins from Phytophthora," Genome Research, v.13, 2003, p. 1675.
Win, J, Morgan, W., Bos, J., Krasileva, K.V., Cano, L.M., Chaparro-Garcia, A., Ammar, R., Staskawicz, B.J., and Kamoun, S.. "Adaptive evolution has targeted the C-terminal domain of the RXLR effectors of plant pathogenic oomycetes," Plant Cell, v.19, 2007, p. 2349.
Win, J., Kanneganti, T. -D., Torto-Alalibo, T., and Kamoun, S.. "Computational and comparative analyses of 150 full-length cDNA sequences from the
oomycete plant pathogen Phytophthora infestans.," Fungal Genetics and Biology, v.43, 2006, p. 20.
