Understanding the mode of action of the symbiosis-specific NF-YA1 transcription factor in Medicago truncatula Carolina Rípodas Laboratoire des Interactions Plantes-Microorganismes INRA - Toulouse Paris, 13 th -16 th February 2017 AgreenSkills 4th Annual meeting
Medicago truncatula – Sinorhizobium meliloti Nitrogen fixing symbiosis Nod factor signaling pathway WT Mtnf-ya1 Rhizobial infection and nodule organogenesis are affected in the Mtnf-ya1 mutant Flavonoids Rhizobium Nod factor Model legume Low nitrogen availability in Medicago truncatula the soil
Identification of direct target genes of NF-YA1 by Chromatin Immunoprecipitation (ChIP-seq) Mtnf-ya1 complementation Ubiquitin promoter NF-YA1 T-ocs 3xHA 3xHA-NF-YA1 α -HA ChiP-qPCR CCAAT boxes 1 0 0 C o n tro l H A -N F -Y A 1 8 0 % o f Im p u t 6 0 NF-YA1 binds the CCAAT Box 3 of ERN1 promoter 4 0 2 0 0 B o x 6 B o x 3 Ongoing experiments: Generation of more material for ChIP-seq.
Identification of target genes of NF-YA1 using RNAseq T 0 T 1 T 2 T 3 T 4 A17 S. Meliloti RNA Mtnf-ya1 RT-qPCR inoculation extraction NF-YA1 OE K9 RNAseq Nodulation markers expression pattern NF-YA1 ENOD11 LEP 3 0 0 0 2 5 1 0 0 0 R e la tiv e e x p re s io n le v e ls R e la tiv e e x p re s s io n le v e ls R e la tiv e e x p re s io n le v e ls 2 0 8 0 0 2 0 0 0 1 5 6 0 0 1 0 4 0 0 1 0 0 0 5 2 0 0 0 0 0 A 1 7 T 0 A 1 7 T 1 A 1 7 T 2 A 1 7 T 3 A 1 7 T 4 6 3 8 7 T 0 6 3 8 7 T 1 6 3 8 7 T 2 6 3 8 7 T 3 6 3 8 7 T 4 0 1 2 3 4 K 9 T 0 K 9 T 1 K 9 T 2 K 9 T 3 K 9 T 4 0 1 2 3 4 0 1 2 3 4 A 1 7 T 0 A 1 7 T 1 A 1 7 T 2 A 1 7 T 3 A 1 7 T 4 6 3 8 7 T 0 6 3 8 7 T 1 6 3 8 7 T 2 6 3 8 7 T 3 6 3 8 7 T 4 K 9 T 0 K 9 T 1 K 9 T 2 K 9 T 3 K 9 T 4 T T T T T T T T T T T T T T T 7 7 7 7 7 7 7 7 7 7 9 9 9 9 9 1 1 1 1 1 8 8 8 8 8 K K K K K A A A A A 3 3 3 3 3 6 6 6 6 6 Ongoing experiments: Generation of RNA libraries + sequencing.
Acknowledgments Eve Coutant Andreas Niebel Agnès Lepage Federico Ariel Carolina Rípodas Farid Regad
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