Grapevine adaptation to abiotic stress: an overview N. Ollat, E Marguerit, F. Lecourieux, A. Destrac-Irvine, S. Cookson, V. Lauvergeat, F. Barrieu, Z. Dai, E. Duchêne, G. Gambetta, E. Gomes, D. Lecourieux, C. van Leeuwen, T. Simonneau, L. Torregrosa, P. Vivin, S. Delrot

A big thank to a great staff

Pollution, climate change and reduction of inputs

Context

http://www.globalcarbonproject.org/

High incertainty More differences between seasons and dry and humid regions

Increase of soil and air pollution : N2O, radiation, salinity, nutrient availability

CO2 and temperature rise Precipitations and drought risks

Any environmental conditions that reduce growth and yield

below optimum levels (Cramer et al., 2011)

Abiotic stress : • Water, temperature, light,

chemical • Duration, intensity, time of

occurrence • Multistress Responses of plants: • Dynamic • Complex (reversible or not) • Organ specific

Abiotic stress

Cramer et al., 2011

• Cell wall metabolism • Water potential

gradients • Inhibition of cell growth • Inhibition of protein

synthesis/modification of regulation

• Energy metabolism • Sugar transport and

storage

General plant response to abiotic stress

epigenetic control

Adaptation to abiotic stress

For a crop : to maintain yield and quality under adverse conditions For a perenial crop: to survive over years to extreme adverse conditions

Adaptation means both a « process » and a « status » (Cooper and Hammer, 1996)

How to define adaptation ?

escape, avoidance, tolerance, resistance

A process « to adapt » A status « to be adapted »

Genotype (or population) : a new combination of favorable alleles (or changes in the allele frequency

within a population)

Genotype : a given combination of favorable alleles

Escape, avoidance, tolerance, resistance

Across generations Short to life cycle of the individual

Adaptation sensu stricto Constitutive Regulation = Acclimation (Plasticity of traits)

Short term Long term

Existing diversity

Selective value Functional Developmental

Genetic architecture

Heritability Reversible Less reversible

High WUE Gs = f (ψ) Stomatal density

• Identification of mechanisms underlying acclimation and adaptation

• Abiotic stress : drought, temperature, mineral deficiencies…. when/where/how ?

• Traits of interest for adaptation : final (yield, quality) or intermediate (WUE, K/tartrate, developmental traits as phenology and root system)

Which targets ?

Some examples

-0.3 -0.2 -0.1 0.0 0.1 0.2 0.3

-0.3

-0.2

-0.1

0.0

0.1

0.2

0.3

PC1

PC

2

S1_20a

G1_Ta

SG_60a

S1_40a S1_Ta

GG_TaSG_Ta

SG_40a

SG_40b

SG_20a

G1_40a

GG_60a

S1_Tb

S1_40b

G1_40b

GG_60b

GG_Tb

S1_60a

SG_Tb

S1_40c

S1_60b

S1_20b

GG_20a

GG_60c

G1_20a

G1_Tb

S1_60c

GG_40a

G1_40c

SG_20b

GG_20bG1_60a

SG_40

GG_Tc

G1_20b

S1_Tc

S1_20c

SG_Tc

SG_60b

G1_60b

GG_40b

G1_60c

G1_20c

G1_TcSG_20c

SG_60c

GG_20c

GG_40c

-20 -10 0 10

-20

-10

010

PCA for gene expression

Principal component 1 (33%)

Pri

nci

pal

co

mp

on

en

t 2

(2

5%

)

110R

RGM

C

MWD

LWD

Drought responses in roots

HWD

Days after treatment

-3 0 3 6 9 12 15 18 21 24

SW

C (

kg H

2O

/kg s

oil)

0.08

0.12

0.16

0.20

0.24

0.28

0.32

CTL

LWD

MWD

HWD

Peccoux, 2011; Barrieu, unpublished

3 scion-rootstocks combinations CS/CS, CS/RGM, CS/110R

Bordo platform

4 levels of soil water content during 2 weeks

Root tips Microarrays NimbleGen

Intensity of water deficit

o CS and 110R: more genes related to oxidative stress response and carbon metabolism

o RGM : more regulated genes, modification of cell wall properties

o Differences in term of responses / intensity of stress

PIP1.1-L

PIP1.3/5-L

PIP2.1-L

ABF2-L

SnRK2.6-L

SnRK2.6-R

-1

-0.75

-0.5

-0.25

0

0.25

0.5

0.75

1

-1 -0.75 -0.5 -0.25 0 0.25 0.5 0.75 1

F2

(2

8 %

)

F1 (35 %)

ABA metabolism and regulatory pathway

Aquaporins

Discriminant responses among genotypes

101-14

110R

140Ru

161-49

41B Mgt

Grenache

RGM

SO4

Syrah

-12

-8

-4

0

4

8

-14 -10 -6 -2 2 6 10 14 F

2 (

28 %

)

F1 (35 %)

V. vinifera

V. berlandieri x V. rupestris

V. riparia & V. riparia x V. rupestris

o Genotypes are grouped according to their background

o VviABF2, VviSnRK2.6, VviPIP1.1, VviPIP2.1 and Vvi PIP1-3/5 in leaves are discriminant among genotypes

o VviSnRK2.6 is the only root discriminant variable

P94, P114, P 164, P180, P181

Rossdeutsch, 2015; Rossdeutsch et al., 2016

TT

SW

_07

TT

SW

_08

TT

SW

_09

Tr_

WD

_0809

TT

SW

_070809

Coefb

_070809

RGM3

Tr_

C_0809

--

CS1

VMC6E10

VMC16D4

VMC9B5

VMC2E9

VMC4C6

Coefa

_09

Coefb

_09

NT

RF

TS

W6

0%

_ 09

NT

RF

TS

W4

0%

_ 09

Coefb

_070809

CS5

NT

RF

TS

W4

0%

_ 07

NT

RF

TS

W2

0%

_ 07

NT

RF

TS

W4

0%

_ 070809

NT

RF

TS

W2

0%

_ 070809

RGM13

QTLs mapping for drought responses

VVMD7 0.0

VVIb22 16.8

VVMD6 25.9

VVIq06 45.8

VMC8D11 57.5 vvc10 62.1 VMC1A12 69.5

VVIq17 79.3

VVIv04 87.6

CR7

VMC2f12 0.0 VVC20 3.1

VVIp04 14.4 VVIv15a 20.6 VVIm07 24.8 VMC9F4x 28.2 VVIh02a 32.1

VMC1B11 52.1

VVIb66 64.9

VMC2H10 76.3

CR8

VMC2G2 VMC2H9

VMC4H5 VMC4G6 VMC5G1

IRT1f 0.0 VMC4f8 5.2

VVC19 16.5 VVIQ57 22.1 VVIb94 25.9

VVIn61 45.9

VVIs21 54.6

VMC9F2 72.6

VVIf52 81.6 VMC9D3 88.2

CR1

VVIB01 0.0

Male 13.0 Fem 13.1 VVIb23 15.4

VVIo55 26.3 VMC2C10 31.7

VMC5G7 43.5

VVIu20a 52.6 VVIU20 56.5

VMC7G3 64.3

CR2

VMC2E7 0.0

UDV021 16.1 VVIh02e 23.2 VMC3F3 24.3 VVMD36 25.6 VVIB59 25.7 VMC9F4cs 27.6 VVIn54 31.7 IRT1d 37.1

CR3

VMCNG1F1 0.0 IRT1a IRT1h2 4.0 VVIr46 6.7 VMC4D4 14.5 VMC7H3 16.2

VMC2b5a 33.6 VMC2b5c 34.8 VrZAG21 38.1 VVIn75 39.1 VMC2b5b 43.4

VRZAG83 61.0

CR4

VVC06 0.0

VVC22 9.3 VVII52 12.8

VVIt68 24.2

VVIv21 33.9 VVIn33 35.6 VMC6E10 37.7 VVC71 38.6 VMC16D4 43.8 VMC9B5 50.9

VMC2E9 62.4 VVIn40 65.4 VMC4c6 68.6

CR5

FRD3a 0.0 IRT1i 0.2

16.0 17.5 IRT1c 19.3

VVIc50 22.6 29.5 33.9 35.6

VVIp28 40.2 VVIn31 43.7 VVIp37 46.6 VVIm43 51.0 VVIs62 56.8

CR6

Marguerit et al., 2009; 2012

V. vinifera x V. riparia progeny as rootstocks

Bordo platform

Progesterone 5-beta reductase (POR) Predicted protein

D4H, NCED Glutathione S transferase Alkanal reductase Class IV Chitinase Unnamed protein

Lipoxygenase (LOX)

Microsatellite linkage map

• Transpiration • Water use efficiency • Responses to SWC • TTSW

• Transpiration • Plant conductance • Δ water potential • Water use efficiency

QTLs mapping for gas exchanges regulation under drought

PHENOARCH platform

Syrah x Grenache progeny

Coupel-Ledru et al., 2014; 2016; 2017

PHENOARCH platform

Transpiration Conductance

Water potential gradients

http://us.123rf.com/400wm/400/400/marigranula/marigranula0907/marigranula090700265/5245339-grapevine-leaf-isolated-on-white-background.jpg

Developmental rate is related to temperature > thermal time Heat Sum = Σ (Tmaxi-Tmin i)/2 From i = 60 to n. ( GFV model, Parker et al., 2011)

Heat Sum = Σ (Tmaxi-Tbase )

From i = 45 to n. Tbase = 2, 10, 6°C (Duchêne et al., 2010)

Temperature and phenology

Van Leeuwen and Destrac, 2017

Vitadapt

262 degree-days 21 days

Developmental rate is related to temperature > thermal time Heat Sum = Σ (Tmaxi-Tmin i)/2 From i = 60 to n. ( GFV model, Parker et al., 2011)

Heat Sum = Σ (Tmaxi-Tbase )

From i = 45 to n. Tbase = 2, 10, 6°C (Duchêne et al., 2010)

Temperature and phenology

Van Leeuwen and Destrac, 2017

Vitadapt

229 degree-days 14 days

CA

Developmental rate is related to temperature > thermal time Heat Sum = Σ (Tmaxi-Tmin i)/2 From i = 60 to n. ( GFV model, Parker et al., 2011)

Heat Sum = Σ (Tmaxi-Tbase )

From i = 45 to n. Tbase = 2, 10, 6°C (Duchêne et al., 2010)

Temperature and phenology

Van Leeuwen and Destrac, 2017

Vitadapt

506 degree-days 21 days

P59, P83, P91, P154

V V I r 4 6 0 . 0

V M C 4 d 4 1 2 . 6 V M C 7 h 3 1 4 . 6

V r Z A G 2 1 3 7 . 2

V V I n 7 5 4 3 . 4

V V M D 3 2 5 3 . 3 V V I p 3 7 5 4 . 4

V M C 6 g 1 0 6 7 . 8

R I G W 0 4

GST1

GST2

V V I p 1 7 a 0 . 0

V M C 5 h 1 1 4 . 3

V V I p 1 1 1 8 . 8

V V I v 7 0 2 5 . 2 V V I p 3 1 2 6 . 6 V V I m 0 3 2 7 . 1

V V I p 3 4 4 1 . 7 V V I v 3 3 4 2 . 8

V M C 7 b 1 5 1 . 2

R I G W 1 9

VvWRKY3

GST3 GST4

Budburst

V V M D 7 0 . 0 V r Z A G 6 2 3 . 2

V V M D 6 1 5 . 9 V M C 5 H 5 1 8 . 8

V V I v 3 6 . 2 3 0 . 0

V M C 9 a 3 . 1 4 0 . 3

VMC8d11 5 6 . 7

V V I p 7 5 7 8 . 8

U D V 0 1 6 . 2 8 7 . 7

V V I n 5 6 9 2 . 1

R I G W 0 7

SGR7/SR

LOBD39

VvFT

Id1

VvSVP1

V V I p 0 5 _ G W 0 . 0 V V I p 0 5 _ R I 0 . 8

V M C 9 c 1 7 . 7 V V I q 3 2 1 0 . 2

V V C 3 4 1 7 . 9

V M C 2 c 3 7 . 7

V V M D 2 4 0 . 8

V V I n 6 4 2 . 5 V V I p 2 6 5 . 5 V V I n 9 4 9 . 0

R I G W 1 4

VvFUL-L VvSEP1

VvCOL2 VvFLC2

V V I n 5 2 0 . 0

V V I t 6 5 1 2 . 1 V V C 0 5 1 2 . 7 V M C 3 g 1 1 1 8 . 3

U D V 0 5 2 2 4 . 3

V V M D 3 7 4 4 . 9 V V M D 5 4 6 . 3

V M C 4 b 7 - 2 5 3 . 7

R I G W 1 6

VvPYL

VvHB10

V M C 2 a 3 0 . 0

V V I v 1 6 9 . 4

V V I m 1 0 4 9 . 2

V V I u 0 4 5 6 . 6

V V M D 1 7 7 4 . 7

V V I n 1 6 8 3 . 1

V M C 7 f 2 9 1 . 2

V V I m 3 3 9 9 . 1

R I G W 1 8

VvSUT2-3

VvSUT2-2

VvMSA VvABF7

Flowering Véraison

Duchêne et al, 2012

Temperature and phenology

Riesling x Gewurtztraminer population

Length of periods in DD

O25, O39, O40, O42, P3, P147, P153

Gra

pe

vin

e G

row

th a

nd D

eve

lopm

enta

l P

att

ern

s

and th

eir R

espo

nse

s t

o E

leva

ted

Te

mp

era

ture

N L

uch

air

e, M

Rie

nth

, C R

om

ieu

, C H

ou

el, Y

G

ibo

n, O

Tu

rc, B

Mu

ller,

L T

orr

egro

sa, A

Pel

leg

rin

o

PI 10

PI 5

PI 25

VPD (2 kPa) PAR/14 h PP

(560 µmol.m-2.s-1)

15°-35° Photo/Nyctiperiod

High T° either at night or day degrades energy supply

Whole vine C balance

Torregrosa et al., 17th Meeting of ASEV-Japan, 10/6/2017, Kyoto, Japan

Temperature and development

Microvine Photosynthesis and respiration

O40, O59

Torregrosa et al., 2017, Lecourieux et al., 2017

Temperature and berry development

Microvines

Two temperature regimes (N/D) 22°C/12°C 30°C/20°C

Heat stress +8°C (12h) 1 to 14 days

Fruiting cuttings

Fruiting cuttings (Cabernet Sauvignon)

Lecourieux et al., 2017

Transcriptomic analysis (Grapevine Nimblegen Arrays) Proteomic analysis (Label-free LC-MS/MS) Metabolomic analysis (LC-MS/MS)

Experimental design and sampling of heat stress exp

Temperature and berry acclimation

Heat stress +8°C (12h) 1 to 14 days

Nimblegen microarrays Vitis HX12K, (FC >2

Green Veraison

Ripening

7 500 DEGs 36

Putative key players of grape acclimation / adaptive responses to heat

1- HSFA2: master regulator of

thermotolerance in plants

2- GOLS1: protection against

abiotic stresses

1- Belong to key regulatory

hubs in hormone and stress

signalling in plants

2- No functional role assigned

HSPs

RLK

Enz. : VvGOLS1

FTs : VvHSFA2,

VvAP2/ERF, VvbHLH

Signalling Secondary Metabolism Transport Epigenetic

processes

HEAT TOLERANCE Poster 127 P16

Te

mp

era

ture

desyn

chro

niz

es s

ug

ar

and o

rga

nic

acid

me

tab

olis

m

in g

rape

s a

nd r

em

od

els

th

eir

tra

nscrip

tom

e

Mal

ate/

Tar

trat

e

Individual berries

M. R

ienth

, L

. T

orr

egro

sa, G

. S

ara

h,

M. A

rdis

so

n1

, J-M

Brillo

ue

t, C

. R

om

ieu

Torregrosa et al., 17th Meeting of ASEV-Japan, 10/6/2017, Kyoto, Japan

Temperature and berry composition

Two temperature regimes (N/D) 22°C/12°C 30°C/20°C

P49

Mapping fruit quality traits

New genotyping and phenotyping tools

Tartrate

Potassium

C H

oue

l, A

Do

lige

z, M

Rie

nth

, S

Foria

, N

Luch

aire, A

Pe

llegrin

o,

C R

om

ieu, L

Torr

egro

sa

Iden

tifica

tion

of sta

ble

QT

Ls fo

r ve

ge

tative

and

repro

du

ctive

tra

its in

th

e m

icro

vin

e (

Vitis

vin

ife

ra L

.)

Picovigne X Ugni blanc flb progeny

P61

RixGW progeny, 2014

[Lin

alo

ol] in

mic

rog/k

g

0500

1000

1500

2000

*

***

***

[Gera

nio

l] (

mic

rog/k

g)

020

00

6000

10000

16E

204E

209E

210E

40

71G

48E

4E

69E

GW

643

Ri4

9

*

Temperature and aroma profiles

15°C night/24°C day 21°C night/30°C day

Linalol

Geraniol

Duchêne et al., unpublished

Riesling x Gewurtztraminer progeny

Fruiting cuttings

Two temperature regimes

P2, P43, P144

LN

CS

LN HN

1103P

131 73

1 0

4 3

HN3 vs LN3 HN24 vs LN24

CS/1103P

CS/RGM 383 58

66 45

155 665

HN3 vs LN3 HN24 vs LN24

136 76

212

Up Down Total

604 768

1369

Up Down Total

LN

CS

RGM

LN HN

LN

0 hpt 3 & 24 hpt

LN HN

0,8 mM 5 mM

Root tips RNA-Seq

o 172 genes commonly and differentially expressed for the two combinations (G6PDH, GS, NR, NIR)

o For 1103P, a majority of DEG are related to nitrogen nutrition (81%)

o For RGM, more differentially expressed genes, and stronger effects (induction or repression) (NRT2.4a, BTB, GTL1, NTF6.3, strigolactone biosynthesis)

o Temporal differences between genotypes (ethylene)

Response to mineral nutrition

Cochetel et al., 2017; 2018 P75, P144, P175

V. berlandieri

V. rupestris

V. riparia

Other

Greffadapt

Cabernet-sauvignon

P content in petiole at veraison

Variability for mineral content

Gautier et al., in process

Poster P99

O56, P76, P137, Poster 173

Tandonnet et al., 2018 27

V. vinifera x V. riparia 138 individuals as rootstocks

Root system as a key parameter of adaptation Root system as a key parameter

Poster 182

Conclusions

o Grapevine fit into the general model for abiotic stress response mechanisms

o Original results (new components in regulatory pathways, genetic architecture of traits and analyses of diversity)

o Which responses are leading to acclimation and

adaptation ? o Interactions between abiotic and biotic stress

responses ? o Highly polygenic traits ? o Modelling for phenotypes (P67) and genotype

(genomic selection P82)

• To re-enforce the hidden half community • Phylloxera Symposium in Bordeaux 2013

• Root Symposium in Rauscedo 2014

• Next….

• To exchange about • Traits of interest

• Phenotyping procedures and facilities

• Genetic and genomic ressources

• Specific approaches to analyse interactions (GxG, GxGxEaxEb)

• To built common ressources • Genomic ressources dedicated to root and rootstock studies

• Data bases of phenotypic traits related to root and roostock performances

Proposal : a working group within IGGP

An International Root and Rootstock Initiative ?

Thank you for your attention