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La nutrition azotée des plantes :
      Exploration de la diversité naturelle et étude de
                      gènes candidats
                                  Françoise DANIEL-VEDELE
                                  Institut Jean-Pierre Bourgin
                                                  INRA
                                          Versailles, France




h"p://www‐ijpb.versailles.inra.fr/
                         h"p://www6.inra.fr/saclay‐plant‐sciences/


  16 novembre 2012          Françoise Vedele : La nutrition azotée des plantes          Montpellier
Why working on nitrogen metabolism?


     Vegeta;ve
stage
                                        Reproduc;ve
stage





                                                         High
              Low


                                                         Nitrogen
          Nitrogen


   10
mM
             3
mM






16 novembre 2012       Françoise Vedele : La nutrition azotée des plantes          Montpellier
Field experiments – measuring changes in soil
                                          nitrate at 30 cm under a wheat crop

                                                                                                                                         20
                                40
                                                       Low N Plot
Daily Mean Soil Water Nitrate




                                                       High N Plot
                                                       Temperature                                                                       15
                                30




                                                                                                                                              Temperature (°C)
            (mM)




                                                                                                                                         10
                                20



                                10                                                                                                       5




                                 0                                                                                                    0
                                21/10/03   20/11/03   20/12/03      19/01/04      18/02/04      19/03/04     18/04/04         18/05/04
                                                                             Time                          From
T
Miller,
Rothamsted
center,
UK


           16 novembre 2012                           Françoise Vedele : La nutrition azotée des plantes                    Montpellier
World yield trends                                  Costs of inputs




16 novembre 2012   Françoise Vedele : La nutrition azotée des plantes   Montpellier
Nitrogen use efficiency


               Capacity of the plant to produce a supplement
            of yield (dY) for each added unit of N fertiliser (dNf)



                                  Yield                NUE = dY/dN

                                Ymax



                                  Y0




                       N mineralization             N fertilization
                                          0
                                                  N total


16 novembre 2012       Françoise Vedele : La nutrition azotée des plantes   Montpellier
From the Green revolution                             to the Green Tide




16 novembre 2012   Françoise Vedele : La nutrition azotée des plantes   Montpellier
Nitrogen use efficiency (NUE = NupE+NutE)
                          Nitrogen Assimilation Pathway


                                                                              N SEED FILLING




               N TRANSPORT




                                                                            N REMOBILISATION
    NITRATE REDUCTION
    AA SYNTHESIS
                   N ABSORPTION                                                N TRANSPORT



16 novembre 2012       Françoise Vedele : La nutrition azotée des plantes             Montpellier
Nitrate fluxes:
molecular actors




    7 NRT2
    51 NRT1
    7 CLC
    2 NAR2




Dechorgnat
et
al,
JEB
2011)

                                                                                    8

  16 novembre 2012             Françoise Vedele : La nutrition azotée des plantes        Montpellier
Characterization of AtNRT2.7
           Expression
of
AtNRT2.7
(Q‐RT‐PCR)
                                                                  Sub‐cellular
Localisa;on
of
NRT2.7
protein

                         194,5

%EF1
A1





           200

           150

           100
                 60,7

            50
 2,1
                    0,3
            5,9

             0

                Silique
 Dry
 Imbibed


                                      Root
            Leaf

                         seed
 seed

                 NRT2.7
expression
is
seed
specific


                                                                           NRT2.7
protein
is
located
on
the

pNRT2.7::G
pNRT2.7::GFP
 FM4‐64
                               GFP
                   tonoplast

FP

                              Nitrate
content
in
seeds



                                                                               nrt2.7
mutants
contain
less
nitrate




                                                                                           Chopin
et
al,
Plant
Cell,
2007


       16 novembre 2012               Françoise Vedele : La nutrition azotée des plantes                    Montpellier
AtNRT2.4: polarized localization and impact on
          growth for triple mutant mutants

pNRT2.4::NRT2.4-GFP                                                 nrt2.1 nrt2.2

                                                                    nrt2.1 nrt2.2 nrt2.4-1

Lateral root (N starvation)
                                                                    nrt2.1 nrt2.2 nrt2.4-2





Polarized plasma                                         nrt2.1 nrt2.2
      nrt2.1 nrt2.2 nrt2.4-1

membrane localisation



                                                                             Kiba et al, Plant Cell 2012

16 novembre 2012        Françoise Vedele : La nutrition azotée des plantes                  Montpellier
16 novembre 2012   Françoise Vedele : La nutrition azotée des plantes   Montpellier
Developmental variation in Arabidopsis thaliana




                   
Natural
varia;on
in
Arabidopsis
thaliana:

                   
Seed
colour
and
seed
size
varia;on

                   
Inflorescence
architecture
varia;on

                   
Leaf
shape
and
rose"e
size
varia;on

16 novembre 2012      Françoise Vedele : La nutrition azotée des plantes   Montpellier
Bay 0 x Shahdara: Summary of N related QTLs
              I                           II                              III                            IV                                V
                                                                                                                              DM10.7 +
                                                              NO10.6 -          + DM3.2      AA10.7 -                        AA10.9 +
                                                           NP10.3 -
                                                            AA10.5 -
                                                                         L3
    - NP3.4
                                                                                - AA3.3       NP10.5 +            + NP3.6     NO10.8 -
                                                                                                                              NP10.6 -
                              DM10.3 -           + AA3.2
   AA10.1 -
                                          L2
    + NP3.1
                                                                                             DM10.6 -             - DM3.4

  NP10.1 -                    NO10.4 -                                                                            + NP3.7
                                                                                                                   + AA3.5
 DM10.1 +                     NO10.5 -
                              NP10.2 -          - NP3.2                                      NO10.7 -
  DM10.2 +                    DM10.4 +
  NO10.1 -

AA10.2 +                                                                 L4
       - AA3.4
                                                                                 - NP3.5                                     DM10.8 +
 NO10.2 -     L1
 + DM3.1
                              DM10.5 -
                                                + NP3.3
                                                             NP10.4 +
                                                                                + DM3.3      AA10.8 +                         NP10.7 -

                              AA10.4 -                       AA10.6 +


 NO10.3 +
                  + AA3.1
 AA10.3 +




                                                                                                              DM: dry matter
                                                                                                              NO: nitrate content
                                                                                                              NP: Total N content
                                                                                                              AA: amino acid conten
      Loudet
et
al,
Plant
Physiol
2003
                                                                       10 mM / 3 mM
    16 novembre 2012                     Françoise Vedele : La nutrition azotée des plantes                   
              Montpellier
An Ideal Plant for NUE




                           BASF
                                         14

 16 novembre 2012   Françoise Vedele : La nutrition azotée des plantes         Montpellier
Deciphering the NUE mechanisms in Arabidopsis




16 novembre 2012   Françoise Vedele : La nutrition azotée des plantes   Montpellier
Uptake and remobilization in Arabidopsis




16 novembre 2012          Françoise Vedele : La nutrition azotée des plantes   Montpellier
Variability in biomass and N% gains between N+ and N-
                    1.2
               Biomass
Gain


                    1.0


                    0.8


                    0.6


                    0.4


                    0.2





                    0.7

                                            N%
Gain

                    0.6

                    0.5

                    0.4

                    0.3

                    0.2

                    0.1





 16 novembre 2012          Françoise Vedele : La nutrition azotée des plantes   Montpellier
16 novembre 2012   Françoise Vedele : La nutrition azotée des plantes   Montpellier
Plant response to nitrogen availability
         Arabidopsis Response to Nitrogen Availability
                                    AReNA

   Nitrate transport           Regulation by N                       Natural variation




Phenotypes, QTL Physiological analysis Reverse Genetics Transcriptome

                              Key metabolic processes

16 novembre 2012       Candidate Genes for increasing NUE
                    Françoise Vedele : La nutrition azotée des plantes           Montpellier
16 novembre 2012   Françoise Vedele : La nutrition azotée des plantes   Montpellier

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La nutrition azotée des plantes : Exploration de la diversité naturelle et études des gènes candidats

  • 1. La nutrition azotée des plantes : Exploration de la diversité naturelle et étude de gènes candidats Françoise DANIEL-VEDELE Institut Jean-Pierre Bourgin INRA Versailles, France h"p://www‐ijpb.versailles.inra.fr/
 h"p://www6.inra.fr/saclay‐plant‐sciences/
 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 2. Why working on nitrogen metabolism? Vegeta;ve
stage
 Reproduc;ve
stage
 High
 Low

 Nitrogen
 Nitrogen

 10
mM
 3
mM

 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 3. Field experiments – measuring changes in soil nitrate at 30 cm under a wheat crop 20 40 Low N Plot Daily Mean Soil Water Nitrate High N Plot Temperature 15 30 Temperature (°C) (mM) 10 20 10 5 0 0 21/10/03 20/11/03 20/12/03 19/01/04 18/02/04 19/03/04 18/04/04 18/05/04 Time From
T
Miller,
Rothamsted
center,
UK
 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 4. World yield trends Costs of inputs 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 5. Nitrogen use efficiency Capacity of the plant to produce a supplement of yield (dY) for each added unit of N fertiliser (dNf) Yield NUE = dY/dN Ymax Y0 N mineralization N fertilization 0 N total 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 6. From the Green revolution to the Green Tide 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 7. Nitrogen use efficiency (NUE = NupE+NutE) Nitrogen Assimilation Pathway N SEED FILLING N TRANSPORT N REMOBILISATION NITRATE REDUCTION AA SYNTHESIS N ABSORPTION N TRANSPORT 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 8. Nitrate fluxes: molecular actors 7 NRT2 51 NRT1 7 CLC 2 NAR2 Dechorgnat
et
al,
JEB
2011)
 8
 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 9. Characterization of AtNRT2.7 Expression
of
AtNRT2.7
(Q‐RT‐PCR) Sub‐cellular
Localisa;on
of
NRT2.7
protein
 194,5
 %EF1
A1
 200
 150
 100
 60,7
 50
 2,1
 0,3
 5,9
 0
 Silique
 Dry
 Imbibed

 Root
 Leaf
 seed
 seed
 NRT2.7
expression
is
seed
specific

 NRT2.7
protein
is
located
on
the
 pNRT2.7::G pNRT2.7::GFP
 FM4‐64
 GFP
 tonoplast
 FP
 Nitrate
content
in
seeds nrt2.7
mutants
contain
less
nitrate
 Chopin
et
al,
Plant
Cell,
2007
 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 10. AtNRT2.4: polarized localization and impact on growth for triple mutant mutants pNRT2.4::NRT2.4-GFP nrt2.1 nrt2.2
 nrt2.1 nrt2.2 nrt2.4-1
 Lateral root (N starvation) nrt2.1 nrt2.2 nrt2.4-2
 Polarized plasma nrt2.1 nrt2.2
 nrt2.1 nrt2.2 nrt2.4-1
 membrane localisation Kiba et al, Plant Cell 2012 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 11. 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 12. Developmental variation in Arabidopsis thaliana 
Natural
varia;on
in
Arabidopsis
thaliana:
 
Seed
colour
and
seed
size
varia;on
 
Inflorescence
architecture
varia;on
 
Leaf
shape
and
rose"e
size
varia;on
 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 13. Bay 0 x Shahdara: Summary of N related QTLs I II III IV V DM10.7 + NO10.6 - + DM3.2 AA10.7 - AA10.9 + NP10.3 - AA10.5 - L3
 - NP3.4 - AA3.3 NP10.5 + + NP3.6 NO10.8 - NP10.6 - DM10.3 - + AA3.2 AA10.1 - L2
 + NP3.1 DM10.6 - - DM3.4 NP10.1 - NO10.4 - + NP3.7 + AA3.5 DM10.1 + NO10.5 - NP10.2 - - NP3.2 NO10.7 - DM10.2 + DM10.4 + NO10.1 - AA10.2 + L4
 - AA3.4 - NP3.5 DM10.8 + NO10.2 - L1
 + DM3.1 DM10.5 - + NP3.3 NP10.4 + + DM3.3 AA10.8 + NP10.7 - AA10.4 - AA10.6 + NO10.3 + + AA3.1 AA10.3 + DM: dry matter NO: nitrate content NP: Total N content AA: amino acid conten Loudet
et
al,
Plant
Physiol
2003
 10 mM / 3 mM 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes 
 Montpellier
  • 14. An Ideal Plant for NUE BASF
 14
 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 15. Deciphering the NUE mechanisms in Arabidopsis 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 16. Uptake and remobilization in Arabidopsis 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 17. Variability in biomass and N% gains between N+ and N- 1.2
 Biomass
Gain
 1.0
 0.8
 0.6
 0.4
 0.2
 0.7
 N%
Gain
 0.6
 0.5
 0.4
 0.3
 0.2
 0.1
 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 18. 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 19. Plant response to nitrogen availability Arabidopsis Response to Nitrogen Availability AReNA Nitrate transport Regulation by N Natural variation Phenotypes, QTL Physiological analysis Reverse Genetics Transcriptome Key metabolic processes 16 novembre 2012 Candidate Genes for increasing NUE Françoise Vedele : La nutrition azotée des plantes Montpellier
  • 20. 16 novembre 2012 Françoise Vedele : La nutrition azotée des plantes Montpellier