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Understanding the mode of action of the symbiosis-specific NF-YA1 transcription factor in Medicago truncatula Carolina Rpodas Laboratoire des Interactions Plantes-Microorganismes INRA - Toulouse Paris, 13 th -16 th February 2017 AgreenSkills

SLIDE 1

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 AgreenSkills 4th Annual meeting Paris, 13th-16th February 2017

SLIDE 2

Nitrogen fixing symbiosis

WT Mtnf-ya1

Rhizobial infection and nodule organogenesis are affected in the Mtnf-ya1 mutant

Nod factor signaling pathway

Flavonoids

Rhizobium

Nod factor

Low nitrogen availability in the soil

Medicago truncatula – Sinorhizobium meliloti

Model legume Medicago truncatula

SLIDE 3

Identification of direct target genes of NF-YA1 by Chromatin Immunoprecipitation (ChIP-seq)

Mtnf-ya1 complementation

3xHA-NF-YA1

α-HA

Ongoing experiments: Generation of more material for ChIP-seq.

3xHA Ubiquitin promoter NF-YA1 T-ocs

CCAAT boxes

ChiP-qPCR

NF-YA1 binds the CCAAT Box 3 of ERN1 promoter

% o f Im p u t

B o x 6 B o x 3 2 0 4 0 6 0 8 0 1 0 0

C o n tro l H A -N F -Y A 1

SLIDE 4

Identification of target genes of NF-YA1 using RNAseq

T1 T2 T3 T4

S. Meliloti

inoculation

A17 Mtnf-ya1 NF-YA1 OE K9 RNA extraction RT-qPCR

RNAseq

NF-YA1 ENOD11 LEP

Nodulation markers expression pattern

R e la tiv e e x p re s io n le v e ls A 1 7 T 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 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 K 9 T 1 K 9 T 2 K 9 T 3 K 9 T 4 2 0 0 4 0 0 6 0 0 8 0 0 1 0 0 0 R e la tiv e e x p re s io n le v e ls 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 1 0 0 0 2 0 0 0 3 0 0 0 R e la tiv e e x p re s s io n le v e ls 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 5 1 0 1 5 2 0 2 5

Ongoing experiments: Generation of RNA libraries + sequencing.

SLIDE 5

Andreas Niebel Farid Regad Agnès Lepage Carolina Rípodas Federico Ariel Eve Coutant