Genetic dissection of disease susceptibility in Arabidopsis to powdery and downy mildew has identified multiple susceptibility (S) genes whose impairment results in disease resistance. Although several of these S-genes have been cloned and characterized in more detail it is unknown to which degree their function in disease susceptibility is conserved among different plant species. Moreover, it is unclear whether impairment of such genes has potential in disease resistance breeding due to possible fitness costs associated with impaired alleles. Here we show that the Arabidopsis PMR4 and DMR1, genes encoding a callose synthase and homoserine kinase respectively, have functional orthologs in tomato with respect to their S-gene function. Silencing of both genes using RNAi resulted in resistance to the tomato powdery mildew fungus Oidium neolycopersici. Resistance to O. neolycopersici by SlDMR1 silencing was associated with severely reduced plant growth whereas SlPMR4 silencing was not. SlPMR4 is therefore a suitable candidate gene as target for mutagenesis to obtain alleles that can be deployed in disease resistance breeding of tomato.
References
[1]
Bowling SA, Guo A, Cao H, Gordon AS, Klessig DF, et al. (1994) A mutation in Arabidopsis that leads to constitutive expression of systemic acquired resistance. Plant Cell 6: 1845–1857.
[2]
Rate DN, Greenberg JT (2001) The Arabidopsis aberrant growth and death2 mutant shows resistance to Pseudomonas syringae and reveals a role for NPR1 in suppressing hypersensitive cell death. Plant J 27: 203–211.
[3]
Greenberg JT, Ausubel FM (1993) Arabidopsis mutants compromised for the control of cellular damage during pathogenesis and aging. Plant J 4: 327–341.
[4]
Dietrich RA, Delaney TP, Uknes SJ, Ward ER, Ryals JA, et al. (1994) Arabidopsis mutants simulating disease resistance response. Cell 77: 565–577.
[5]
Yu IC, Parker J, Bent AF (1998) Gene-for-gene disease resistance without the hypersensitive response in Arabidopsis dnd1 mutant. Proc Natl Acad Sci U S A 95: 7819–7824.
[6]
Vogel J, Somerville S (2000) Isolation and characterization of powdery mildew-resistant Arabidopsis mutants. Proc Natl Acad Sci U S A. 97: 1897–1902.
[7]
Van Damme M, Andel A, Huibers RP, Panstruga R, Weisbeek PJ, et al. (2005) Identification of arabidopsis loci required for susceptibility to the downy mildew pathogen Hyaloperonospora parasitica. Mol Plant Microbe Interact. 18: 583–592.
[8]
Hernández-Blanco C, Feng DX, Hu J, Sánchez-Vallet A, Deslandes L, et al. (2007) Impairment of cellulose synthases required for Arabidopsis secondary cell wall formation enhances disease resistance. Plant Cell. 19: 890–903.
[9]
Pavan S, Jacobsen E, Visser RG, Bai Y (2010) Loss of susceptibility as a novel breeding strategy for durable and broad-spectrum resistance. Mol Breed. 25: 1–12.
[10]
Lyngkjaer MF, Newton A C, Atzema J L, Baker SJ (2000) The barley mlo-gene: An important powdery mildew resistance source. Agronomie. 20: 745–756.
[11]
Consonni C, Humphry ME, Hartmann HA, Livaja M, Durner J, et al. (2006) Conserved requirement for a plant host cell protein in powdery mildew pathogenesis. Nat Genet. 38: 716–720.
[12]
Bai Y, Pavan S, Zheng Z, Zappel NF, Reinst?dler A, et al. (2008) Naturally occurring broad-spectrum powdery mildew resistance in a Central American tomato accession is caused by loss of mlo function. Mol Plant Microbe Interact. 21: 30–39.
[13]
Pavan S, Schiavulli A, Appiano M, Marcotrigiano AR, Cillo F, et al. (2011) Pea powdery mildew er1 resistance is associated to loss-of-function mutations at a MLO homologous locus. Theor Appl Genet. 123: 1425–1431.
[14]
Humphry M, Reinst?dler A, Ivanov S, Bisseling T, Panstruga R (2011) Durable broad-spectrum powdery mildew resistance in pea er1 plants is conferred by natural loss-of-function mutations in PsMLO1. Mol Plant Pathol. 12: 866–878.
[15]
Nishimura MT, Stein M, Hou BH, Vogel JP, Edwards H, et al. (2003) Loss of a callose synthase results in salicylic acid-dependent disease resistance. Science. 301: 969–972.
[16]
Van Damme M, Zeilmaker T, Elberse J, Andel A, de Sain-van der Velden M, et al. (2009) Downy mildew resistance in Arabidopsis by mutation of HOMOSERINE KINASE. Plant Cell 21: 2179–2189.
[17]
Stuttmann J, Hubberten HM, Rietz S, Kaur J, Muskett P, et al. (2011) Perturbation of Arabidopsis amino acid metabolism causes incompatibility with the adapted biotrophic pathogen Hyaloperonospora arabidopsidis. Plant Cell. 23: 2788–2803.
[18]
G?llner K, Schweizer P, Bai Y, Panstruga R (2008) Natural genetic resources of Arabidopsis thaliana reveal a high prevalence and unexpected phenotypic plasticity of RPW8-mediated powdery mildew resistance. New Phytol. 177: 725–742.
[19]
Zhang F, Maeder ML, Unger-Wallace E, Hoshaw JP, Reyon D, et al. (2010) High frequency targeted mutagenesis in Arabidopsis thaliana using zinc finger nucleases. Proc Natl Acad Sci U S A. 107: 12028–12033.
[20]
Shukla VK, Doyon Y, Miller JC, DeKelver RC, Moehle EA, et al. (2009) Precise genome modification in the crop species Zea mays using zinc-finger nucleases. Nature. 459: 437–441.
[21]
Townsend JA, Wright DA, Winfrey RJ, Fu F, Maeder ML, et al. (2009) High-frequency modification of plant genes using engineered zinc-finger nucleases. Nature. 459: 442–445.
[22]
Lyznik LA, Djukanovic V, Yang M, Jones S (2012) Double-strand break-induced targeted mutagenesis in plants. Methods Mol Biol. 847: 399–416.
[23]
Kurowska M, Daszkowska-Golec A, Gruszka D, Marzec M, Szurman M, et al. (2011) TILLING: a shortcut in functional genomics. J Appl Genet. 52: 371–390.
[24]
Bai Y, van der Hulst R, Bonnema G, Marcel TC, Meijer-Dekens F, et al. (2005) Tomato defense to Oidium neolycopersici: dominant Ol genes confer isolate-dependent resistance via a different mechanism then recessive ol-2. Mol Plant Microbe Interact. 18: 354–362.
[25]
Dereeper A, Guignon V, Blanc G, Audic S, Buffet S, et al. (2008) Phylogeny.fr: robust phylogenetic analysis for the non-specialist. Nucl Acids Res. 36: W465–W469.
[26]
Wesley SV, Helliwell CA, Smith NA, Wang MB, Rouse DT, et al. (2001) Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 27: 581–590.
[27]
Truett GE, Heeger P, Mynatt RL, Truett AA, Walker JA, et al. (2000) Preparation of PCR-quality mouse genomic DNA with hot sodium hydroxide and Tris (HotSHOT). BioTechniques 29: 52–54.
[28]
Heilersig HJB, Loonen A, Bergervoet M, Wolters AMA, Visser RGF (2006) Post-transcriptional gene silencing of GBSSI in potato: effects of size and sequence of the inverted repeats. Plant Mol Biol. 60: 647–662.
[29]
Ellinger D, Naumann M, Falter C, Zwikowics C, Jamrow T, et al. (2013) Elevated early callose deposition results in complete penetration resistance to powdery mildew in Arabidopsis. Plant Physiology 161: 1433–1444.