Towards Establishment of a Rice Stress Response Interactome

Rice (Oryza sativa) is a staple food for more than half the world and a model for studies of monocotyledonous species, which include cereal crops and candidate bioenergy grasses. A major limitation of crop production is imposed by a suite of abiotic and biotic stresses resulting in 30%–60% yield losses globally each year. To elucidate stress response signaling networks, we constructed an interactome of 100 proteins by yeast two-hybrid (Y2H) assays around key regulators of the rice biotic and abiotic stress responses. We validated the interactome using protein–protein interaction (PPI) assays, co-expression of transcripts, and phenotypic analyses. Using this interactome-guided prediction and phenotype validation, we identified ten novel regulators of stress tolerance, including two from protein classes not previously known to function in stress responses. Several lines of evidence support cross-talk between biotic and abiotic stress responses. The combination of focused interactome and systems analyses described here represents significant progress toward elucidating the molecular basis of traits of agronomic importance

Plant homeodomain protein involved in transcriptional regulation of a pathogen defense-related gene.

Transcription of the parsley pr2 gene, encoding pathogenesis-related protein 2 (PR2), is rapidly stimulated by fungal or bacterial elicitors. Previous work has revealed a 125-bp region within the pr2 promoter; this region encompasses all important cis-regulatory elements required for fungal elicitor-mediated expression. We now report the identification of a functionally relevant 11-bp DNA motif (CTAATTGTTTA) contained within this region; it specifically binds to factors present in both parsley and Arabidopsis nuclear protein extracts. From both plant species, full-length cDNA clones were isolated that encode proteins with high affinity fo this DNA motif. The proteins from both species contain stretches of 61 amino acids that are characteristic of homeodomain (HD) proteins. Binding studies and use of a polyclonal antiserum raised against a fusion polypeptide of glutathione S-transferase with the HD portion of the parsley protein indicated that the 11-bp DNA motif is a potential in vivo target site and that the HD protein is contained within the observed complex formed between the DNA motif and nuclear protein extracts. Transient expression studies using the authentic and a mutated target site suggested a functional role of the HD-DNA interaction in the regulation of the pr2 gene expression