Identification and characterization of the Non-race specific Disease Resistance 1 (NDR1) orthologous protein in coffee

<p>Abstract</p> <p>Background</p> <p>Leaf rust, which is caused by the fungus <it>Hemileia vastatrix </it>(Pucciniales), is a devastating disease that affects coffee plants (<it>Coffea arabica </it>L.). Disadvantages that are associated with currently developed phytoprotection approaches have recently led to the search for alternative strategies. These include genetic manipulations that constitutively activate disease resistance signaling pathways. However, molecular actors of such pathways still remain unknown in <it>C. arabica</it>. In this study, we have isolated and characterized the coffee <it>NDR1 </it>gene, whose <it>Arabidopsis </it>ortholog is a well-known master regulator of the hypersensitive response that is dependent on coiled-coil type R-proteins.</p> <p>Results</p> <p>Two highly homologous cDNAs coding for putative NDR1 proteins were identified and cloned from leaves of coffee plants. One of the candidate coding sequences was then expressed in the <it>Arabidopsis </it>knock-out null mutant <it>ndr1-1</it>. Upon a challenge with a specific strain of the bacterium <it>Pseudomonas syringae </it>(DC3000::<it>AvrRpt2</it>), analysis of both macroscopic symptoms and <it>in planta </it>microbial growth showed that the coffee cDNA was able to restore the resistance phenotype in the mutant genetic background. Thus, the cDNA was dubbed <it>CaNDR1a </it>(standing for <it>Coffea arabica Non-race specific Disease Resistance 1a</it>). Finally, biochemical and microscopy data were obtained that strongly suggest the mechanistic conservation of the <it>NDR1</it>-driven function within coffee and <it>Arabidopsis </it>plants. Using a transient expression system, it was indeed shown that the CaNDR1a protein, like its <it>Arabidopsis </it>counterpart, is localized to the plasma membrane, where it is possibly tethered by means of a GPI anchor.</p> <p>Conclusions</p> <p>Our data provide molecular and genetic evidence for the identification of a novel functional <it>NDR1 </it>homolog in plants. As a key regulator initiating hypersensitive signalling pathways, <it>CaNDR1 </it>gene(s) might be target(s) of choice for manipulating the coffee innate immune system and achieving broad spectrum resistance to pathogens. Given the potential conservation of <it>NDR1</it>-dependent defense mechanisms between <it>Arabidopsis </it>and coffee plants, our work also suggests new ways to isolate the as-yet-unidentified <it>R</it>-gene(s) responsible for resistance to <it>H. vastatrix</it>.</p>