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Stripe rust Haustorial cDNAs Chuntao Yin, Xianming Chen, Scot Hulbert

Stripe rust Haustorial cDNAs Chuntao Yin, Xianming Chen, Scot Hulbert. Puccinia striiformis. Chuntao Yin. General Approach:. Isolate haustoria (pure as possible) Make cDNA library Sequence sample of library Sequence more or make new preparation Assemble unigenes/predict secreted proteins

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Stripe rust Haustorial cDNAs Chuntao Yin, Xianming Chen, Scot Hulbert

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  1. Stripe rust Haustorial cDNAs Chuntao Yin, Xianming Chen, Scot Hulbert Puccinia striiformis Chuntao Yin

  2. General Approach: • Isolate haustoria (pure as possible) • Make cDNA library • Sequence sample of library • Sequence more or make new preparation • Assemble unigenes/predict secreted proteins • Make expression constructs for in planta examination • of HR or ???

  3. One plate (384 clones) sequenced from first library Fungal ribosomal associated 25 No homology (fungal?) 19 Unknown fungal 11 U3 small nucleolar ribonucleoprotein 1 Nucleoside diphosphate kinase 1 Translation initiation factor eIF-1A subunit 1 Ubiquinone/menaquinone biosynthesis 1 Ubiquitin conjugating enzyme 1 Proteins with possible signal peptides 2 Unknown fungal with predicted signal anchors 2 Arabinose efflux permease (signal anchor) 1 65

  4. P. striiformis haustoria

  5. Maize line B73 (Rxo1) Mo17 (rxo1) Plasmids GFP only GFP and AvrRxo1 Bingyu Zhao Biolistic transient assay for HR-inducing genes

  6. Transient expression vector for HR assays

  7. Delivery of effectors by host and nonhost bacteria Rxo1 rxo1 Rxo1 rxo1 Burkholdaria andropogonis (maize pathogen) Xanthomonas oryzae pv. oryzicola (non pathogen)

  8. avr Pto fusions with Cya protein delivered into media by P. syringae DC3000: Schechter et al. (2004) J. Bact 164 aa 135 aa 100 aa Alan Collmer, Cornell Delivery of rust effectors as bacterial effector fusions? • Need to test DC3000 or DC3000 mutants for HR in cereals • Need to make and test constructs

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