rootstock genetic background No Cochetel, Antoine Gautier, Elose - - PowerPoint PPT Presentation

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rootstock genetic background No Cochetel, Antoine Gautier, Elose - - PowerPoint PPT Presentation

Apr 27, 2023 40 likes •292 views

In grafted grapevines, physiological, transcriptional and hormonal responses to nutrient availability are strongly influenced by the rootstock genetic background No Cochetel, Antoine Gautier, Elose Mtier, Isabelle Merlin, Cyril Hvin,

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SLIDE 1

In grafted grapevines, physiological, transcriptional and hormonal responses to nutrient availability are strongly influenced by the rootstock genetic background

Noé Cochetel, Antoine Gautier, Eloïse Météier, Isabelle Merlin, Cyril Hévin, Jean-Bernard Pouvreau, Pierre Coutos-Thévenot, Michel Hernould, Zhanwu Dai, Philippe Vivin, Pierre-François Bert, Frédéric Escudié, Mindy Stephania Muñoz, Christophe Klopp, Serge Delrot, Sarah Jane Cookson, Nathalie Ollat, and Virginie Lauvergeat

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CULTIVATED GRAPEVINE, A GRAFTED PLANT

Phenology Yield Berry quality Tolerance to disease and abiotic stress

Rootstock Scion

Smith, 2004 Pouget, 2015

Berry Quality

Conferred vigour

  • Conferred vigor
  • A rootstock adapted to the environment
  • A vegetative growth compatible with the production of

qualitative berries

Rives, 1971 Champagnol, 1984 Ollat et al., 2003 Jones et al., 2009 Howell, 2001 Skinkis and Vance, 2013 Keller, 2015

How rootstocks control scion growth?

INT INTRODUCT CTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 3

HOW THE ROOTSTOCK CONTROLS THE SCION DEVELOPMENT?

  • Nitrate is the main source of nitrogen

N

N

  • Nitrate sensing and uptake are regulated by the rootstock to ensure an
  • ptimal absorption of this nutrient
  • Root and shoot growth are regulated in response to nitrate availability,

especially through root to shoot long distance hormone signalling Sensing of the availability and nitrate uptake Shoot and root growth regulation

Are the rootstocks different in terms of nitrate sensing and uptake?

Hormones

Rootstock Scion

  • Nitrogen is an essential nutrient crucial for grapevine growth, berry

yield and quality

  • Nitrate availability is heterogeneous in the soil

INT INTRODUCT CTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 4

HORMONAL REGULATION OF SCION GROWTH

  • Strigolactones:
  • Mainly synthesized in the roots
  • Regulated by nitrogen availability
  • Inhibit axillary bud outgrowth
  • Involved in arbuscular mycorrhizal fungi symbiosis in the rhizosphere

Do strigolactones contribute to the rootstock-dependent regulation of scion growth?

Waldie et al., 2014 Al-babili et al., 2015 Lopez-Obando et al., 2015 Pandey et al., 2016

N

N

Sensing of the availability and nitrate uptake Shoot and root growth regulation Strigolactone

Rootstock Scion

INT INTRODUCT CTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 5

PLANT MATERIAL

  • Choice of two rootstock genotypes known to differently control the growth of the same scion

1103P RGM CS CS

INT INTRODUCT CTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 6

N

N

  • Profiling of the short term transcriptomic responses in the roots
  • Heterogeneous nitrate availability

ARE THE ROOTSTOCKS DIFFERENT IN TERMS OF NITRATE SENSING AND UPTAKE?

Cochetel et al., JXB, 2017

1103P RGM CS CS N

N

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 7

ARE THE ROOTSTOCKS DIFFERENT IN TERMS OF NITRATE SENSING AND UPTAKE?

LN CS RGM 1103P CS LN LN LN HN HN LN HN 0.8 mM 5 mM Root tips harvesting 3 & 24 hpt LN HN

[NO3

  • ]

Cochetel et al., JXB, 2017

  • Split root system

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 8

ARE THE ROOTSTOCKS DIFFERENT IN TERMS OF NITRATE SENSING AND UPTAKE?

Combination Description Positive Negative Total CS/1103P HN3 vs LN3 132 73 205 HN24 vs LN24 5 3 8 CS/RGM HN3 vs LN3 449 103 552 HN24 vs LN24 221 710 931

  • 172 common differentially expressed genes (DEGs) in both combinations related to nitrogen
  • A more pronounced response in CS/RGM:
  • More DEGs
  • Common DEGs with a higher LFC

WGCNA revealed an interesting gene cluster related to strigolactone pathway.

Cochetel et al., JXB, 2017

  • De novo assembly of RNA sequencing results

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 9

STRIGOLACTONE GENE CLUSTER

Cochetel et al., JXB, 2017 Cochetel et al., JXB, 2018

Do strigolactones contribute to the rootstock- dependent regulation of scion growth?

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 10

FUNCTIONAL CHARACTERIZATION OF GRAPEVINE STRIGOLACTONE GENES

Cochetel et al., JXB, 2018

  • Genes involved in SL biosynthesis mainly expressed in the roots
  • Over-expression of grape genes in Arabidopsis restored hyper-branched mutants to the WT branching phenotype

Pouvreau et al., 2013 Phelipanche ramosa seeds

  • Overexpressing grapevine cells secreted SL-like compounds able to induce the germination of P. ramosa seeds

Grapevines produce SL-like compounds with similar roles as in other plant species.

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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IS SL PRODUCTION INDUCED BY N LIMITING CONDITIONS AND IMPACTED BY THE ROOTSTOCK IN MICROGRAFTS?

  • Phenotypic and molecular responses to nitrate - Hydroponic experiments

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 12

Pre-cultured in perlite Full nutrient solution 0N solution Treatment H2O HN 0N HN H2O HN 0N 0N 9 16 23 28 35 7 14 21 (Days) Gene expression profiling Exudates collection Week 1 Week 2

Cochetel et al., JXB, 2018

  • Hydroponics – Experimental design

IS SL PRODUCTION INDUCED BY N LIMITING CONDITIONS AND IMPACTED BY THE ROOTSTOCK IN MICROGRAFTS?

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 13

Pre-cultured in perlite Full nutrient solution 0N solution Treatment H2O HN 0N HN H2O HN 0N 0N 9 16 23 28 35 7 14 21 (Days) Exudates collection Week 1 Week 2

Cochetel et al., JXB, 2018

Gene expression profiling

  • Hydroponics – Gene expression profiling

DO N LIMITING CONDITIONS INDUCE EXPRESSION OF SL BIOSYNTHESIS GENES?

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 14

Cochetel et al., JXB, 2018

Combination Condition CS/1103P CS/RGM 0N HN

  • Expression of genes involved in SL

biosynthesis is regulated by N availability in grafted grapevines.

  • This

induction was higher in CS/RGM roots.

DO N LIMITING CONDITIONS INDUCE EXPRESSION OF SL BIOSYNTHESIS GENES?

0N solution Treatment

0N HN 0N 0N

Week 1 Week 2

  • Hydroponics – Gene expression profiling

MAX1

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 15

IS THE GENE EXPRESSION INDUCTION ASSOCIATED WITH SL PRODUCTION?

Pre-cultured in perlite Full nutrient solution 0N solution Treatment H2O HN 0N HN H2O HN 0N 0N 9 16 23 28 35 7 14 21 (Days) Transcript abundance quantification Exudates collection Week 1 Week 2

Cochetel et al., JXB, 2018

  • Hydroponics – Bio-assays

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 16

IS THE GENE EXPRESSION INDUCTION ASSOCIATED WITH SL PRODUCTION?

Cochetel et al., JXB, 2018

CS/1103P – 0N CS/1103P – HN CS/RGM – 0N CS/RGM – HN

  • Hydroponics – Bio-assays

Dilution 1/10

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 17

IS THE GENE EXPRESSION INDUCTION ASSOCIATED WITH SL PRODUCTION?

Cochetel et al., JXB, 2018

CS/1103P – 0N CS/1103P – HN CS/RGM – 0N CS/RGM – HN

  • Hydroponics – Bio-assays
  • In N limiting conditions, grafted grapevines

produced SL-like compounds.

  • SL production induction is earlier and stronger

in CS/RGM plants.

PDR1

Combination Condition CS/1103P CS/RGM 0N HN

Week 1 Week 2

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 18

IS THE GENE EXPRESSION INDUCTION ASSOCIATED WITH SL PRODUCTION?

Cochetel et al., JXB, 2018

CS/1103P – 0N CS/1103P – HN CS/RGM – 0N CS/RGM – HN

  • Hydroponics – Bio-assays
  • In N limiting conditions, grafted grapevines

produced SL-like compounds.

  • SL production induction is earlier and stronger

in CS/RGM plants.

PDR1

Combination Condition CS/1103P CS/RGM 0N HN

Week 1 Week 2

PDR1 gene expression is induced in N limiting conditions and more strongly in CS/RGM.

CS/RGM CS/1103P CS/RGM CS/1103P CS/1103P – 0N CS/1103P – HN CS/RGM – 0N CS/RGM – HN

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 19

IS THE GENE EXPRESSION INDUCTION ASSOCIATED WITH SL PRODUCTION?

Cochetel et al., JXB, 2018

CS/1103P – 0N CS/1103P – HN CS/RGM – 0N CS/RGM – HN

  • Hydroponics – Bio-assays
  • In N limiting conditions, grafted grapevines

produced SL-like compounds.

  • SL production induction is earlier and stronger

in CS/RGM plants.

PDR1

Combination Condition CS/1103P CS/RGM 0N HN

Week 1 Week 2

PDR1 gene expression is induced in N limiting conditions and more strongly in CS/RGM.

CS/RGM CS/1103P CS/RGM CS/1103P CS/1103P – 0N CS/1103P – HN CS/RGM – 0N CS/RGM – HN

Scion branching inhibition by N limiting conditions was more pronounced in CS/RGM.

Are these results reproducible with older plants grown in greenhouse?

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 20

IN GREENHOUSE CONDITIONS, 1103P CUTTINGS ARE MORE BRANCHED THAN RGM.

Cochetel et al., JXB, 2018

  • Woody cuttings of 1103P and RGM were grown and watered

with LN (0.8 mM N) or HN (5 mM N) solutions during 45 days

  • HN: main stem growth was promoted in RGM compared to 1103P
  • LN:
  • Reduction of LB outgrowth for RGM
  • Reduction of stem development in 1103P

Greenhouse experiments confirmed that N limiting conditions induce a stronger inhibition

  • f bud outgrowth for RGM.

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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IN GREENHOUSE CONDITIONS, CS/1103P IS MORE BRANCHED THAN CS/RGM.

Cochetel et al., JXB, 2018

  • CS/1103P and CS/RGM plants were grown and watered with

HN (5 mM N) solution during 45 days

Bud outgrowth was reduced when RGM is the rootstock.

  • CS/1103P plants are more branched than CS/RGM plants
  • Differences even more pronounced after one year of culture

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 22

IN SUMMARY

  • In response to a heterogeneous nitrogen availability, CS/RGM transcriptome was more impacted than CS/1103P
  • Gene co-expression network analysis revealed a strigolactone-specific gene cluster
  • Putative genes involved in SL pathway seem to play similar role as in other species
  • In N limiting conditions, RGM seems to produce more SLs
  • SL production is correlated to a higher LB outgrowth inhibition in N limiting conditions for RGM or CS/RGM

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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NITROGEN, THE OTHER MINERALS AND THE ROOT GROWTH

  • Interaction between N and P
  • Transcriptional and post-transcriptional regulation of the same protein, HSR1
  • Nitrate drives the phosphate starvation response through IDR1
  • Root growth regulation

Medici et al., 2015 Medici et al., 2016 Kellermeier et al., 2014

  • How do grapevine rootstocks modify phosphorus concentration in scion?

Gautier et al., 2018

P availability regulates SL biosynthesis and has a synergetic action with N for association with arbuscular mycorrhizal fungi.

Poster P99

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 24

GRAPEVINE AND ITS RHIZOSPHERE

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

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SLIDE 25

ACKNOWLEDGEMENTS

  • UMR Biologie du Fruit et Pathologie, Bordeaux, France (Arabidopsis transformation)
  • Pierre Coutos-Thévenot, Poitiers, France (Embryogenic cell transformation)
  • Frédéric Escudié and Christophe Klopp, Toulouse, France (RNAseq analysis)
  • Jean-Bernard Pouvreau, Nantes, France (Bio-assays)
  • Sandrine Ruffel, Montpellier, France (Split-root experiment)

INTRODUCTION RESULTS CONCLUSION PERSPECTIVES

ncochetel@unr.edu @NCochetel