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Genetics and plant breeding in Australia. CSIRO Vitis breeding: Dr. Rob Walker Power of genomic approaches: Dr. Chris Ford. A u s t r a l i a ’ s G r o w i n g F u t u r e. Grapevine Breeding . Viticultural Seminar Argentina May 2007

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slide1

Genetics and plant breeding in Australia

CSIRO Vitis breeding: Dr. Rob Walker

Power of genomic approaches: Dr. Chris Ford

slide2

A u s t r a l i a ’ s G r o w i n g F u t u r e

Grapevine Breeding

Viticultural Seminar

Argentina May 2007

Dr. Rob Walker, CSIRO Plant Industry

grapevine breeding in australia
Table grapes

Dried grapes

Wine grapes

Rootstocks

Grapevine breeding in Australia

CSIRO is working with Australia’s grape industries to develop new and improved grapevine varieties

tablegrape breeding and evaluation
Tablegrape breeding and evaluation

Table grapes

  • National program
  • Breeding by CSIRO
  • Evaluation in WA, Qld and NT by state agencies
  • Aiming for seedlessness, excellent taste, ripe and uniform maturity, large and uniform berry size and good production and postharvest characteristics.

M1 vines from

irradiated buds

Top-working

new selections

Range of colours, sizes, textures and flavours

Red

seedless

Bright red early

seedless

White muscat

seedless

Crisp white

seedless

Late white

seedless

slide5

Millennium Muscat™

Seeded

Very early

Muscat flavour

Domestic market

slide6

Magic Seedless™

Black

Early

Seedless

dried grapes
Dried grapes
  • Emphasis is on rain tolerant, low browning sultana types which are disease resistant and show consistent fruitfulness
  • Involves:
    • In-ovulo embryo rescue
    • Top-working for accelerated evaluation
    • Evaluation under mechanised systems (world’s best practice)
    • Comparison with comparator varieties
  • Range of selections under evaluation
rootstocks
Rootstocks

Screening techniques, selection & inheritance

Assessment of selections as grafted vines

Regional semi-commercial evaluation

Germplasm – Vitis spp.

Crosses  Interspecific families

Creation and recombination of genetic variability

Ease of propagation

Graft compatibility

Nematode tolerance

Mineral discrimination

1000s

  • Viticultural performance
    • Grafted vine vigour
    • Phylloxera tolerance
    • Yield
    • WUE & drought tolerance
    • Carbohydrate partitioning
  • Fruit quality
  • Wine quality

75

Commercial performance

G x E

4

Industry adoption

rootstocks for quality wine
Rootstocks for quality wine

Current Focus

Stock-scion compatibility

  • New rootstocksPBR recently released
  • to industry for evaluation
  • Selected for:
  • low to moderate vigour
  • ‘reduced’ potassium uptake
  • enhanced wine quality
  • tolerance of phylloxera and nematodes

Merbein 5489

Merbein 5512

Merbein 6262

Salt tolerance and water use efficiency

SALT SUSCEPTIBLE

SALT TOLERANT

winegrapes
Winegrapes
  • Emphasis on varieties suited to Australian conditions, particularly warm-climate regions.
  • Selection for yield performance, improved grape juice composition and wine quality.
  • January 2000 releases:

TyrianPBR, CiennaPBRandRubiennePBR

Compared with (parent) Cabernet Sauvignon, all 3 have improved yields, higher juice titratable acidity, higher wine colour density and total anthocyanins and have higher colour hue (brighter wine).

pre breeding
Pre-breeding
  • Genes and traits (CSIRO Plant Industry, Adelaide)
    • Colour and tannins
    • Grape berry development and ripening
    • Fruit flavour and aroma
    • Fungal pathology
    • Carotenoids, hormones and flavour
  • Rapid flowering genotypes and breeding efficiency

... to make this wine...

Growing these grapes...

... to respond to this consumer

gene expression and vine performance
Gene expression and vine performance

Microarray mapping of gene expression

(Mark Thomas and Chris Davies CSIRO Plant Industry Adelaide)

Future – microarrays will enable :-

Comparison of gene expression in:

- high versus low quality

- warm versus cool climate

- full versus restricted irrigation

International Grapevine Genome Program

Multinational research initiative to use a genomics approach to discover and determine the function of all grapevine genes.

key people and inputs
Key people and inputs
  • Table grapes (Peter Clingeleffer and Steve Sykes)
  • Dried grapes (Peter Clingeleffer, Steve Sykes and Steve Swain)
  • Rootstocks (Peter Clingeleffer, Steve Sykes, Rob Walker and Tim Jones)
slide15

Genetics and plant breeding in Australia

CSIRO Vitis breeding: Dr. Rob Walker

Power of genomic approaches: Dr. Chris Ford

genomics and post genomics approaches to understanding grape berry composition

Genomics and post-genomics approaches to understanding grape berry composition

Christopher M Ford

School of Agriculture Food and Wine

The University of Adelaide

grapevine biotechnology
Grapevine biotechnology
  • Classical era – ‘genetic modification’
    • Technically difficult with grapevines
    • Widespread opposition to GM crops
  • Post-genomics – a ‘systems approach’
    • All genes identified ( + physical mapping)
    • Control of gene expression
    • All proteins and metabolites identified
      • varietal; developmental; stress-response
    • Control of metabolic pathways - composition
grape berry acidity
Grape berry acidity
  • 2 major acids – tartaric, malic
  • Malic acid – central metabolite
  • Tartaric acid – unique, unusual and important
  • Use of acids in winemaking
    • $$s costs
    • Future considerations – global warming…
  • Context – physiology of TA and acid metabolism
pathways to tartaric acid
Pathways to tartaric acid

O

COOH

1COOH

1

CO

OH

2

OH

COOH

COOH

COOH

A

HO

HO

2

3

HO

O

O

OH

OH

4

COOH

CHO

HO

HO

HO

HO

3

L-tartaric acid

+

OH

C4/C5 cleavage

OH

OH

4

5CHO

HO

HO

HO

5

CH2OH

CH2OH

CH2OH

CH2OH

6

6CH2OH

L-Ascorbic acid

2-keto-L-gulonic acid

L-idonic acid

5-keto-D-gluconic acid

Glycoaldehyde

C2/C3 cleavage

B

3COOH

OH

1COOH

4

HO

+

5

2

COOH

CH2OH

6

Oxalic acid

L-threonic acid

the search for ta synthesis components
The search for TA synthesis components
  • Classical approaches –
    • biochemical
    • homologous genes
    • grapevines as experimental systems…
  • Molecular resources –
    • analysis of transcripts – identity and abundance
    • predicted enzymatic activity of encoded sequences
slide21

Identification of candidate sequences

1. cDNA libraries are clustered based upon similarity of EST expression pattern

Leaf

Flower

Flower

Bud, root

& shoot

Root & leaf

veraison, pre-

veraison

Veraison

Ripe berry

Petiole, stem

Pre-veraison

Pre-veraison

Post-veraison

leaf

berry

petiole, flower, bud, root

2. Unigenes, grouped on similarity of expression across all 55 cDNA libraries

slide22

Cross referencing candidate genes for in-depth sequence analysis

Identification of differentially expressed cDNAs: 565 candidate genes

Limiting EST number to no less than 6 per TC narrowed the number of candidate genes:87 candidate genes

Domain and motif screening for oxidoreductase enzymes:8 candidate genes

slide23

Berries were sampled from 25 species of grapevine. Organic acid levels were compared for each species.

Tartaric acid

Oxalic acid

slide24

Correlating biosynthesis with gene expression

A. One species identified in this study makesNO tartaric acid

B

B

B. Gene expression

studies showed that

one candidate gene

correlated with tartaric

acid biosynthesis;

other candidates were

expressed in both TA

and non TA

accumulating

grapevines

slide25

Combined metabolic and transcriptional profiling to identify candidate tartrate biosynthetic genes

Prov. Patent 503479

Overexpression in E. coli of the protein encoded by candidate gene assays and tag-facilitated purification allowed determination of its catalytic activity

Enzymology revealed substrate specificity and catalytic activity for a key TA synthesis intermediate

outcomes and future directions
Outcomes, and future directions
  • Continuing to investigate the pathway and regulation of TA synthesis
  • Integrating malate and tartaric acid metabolism
  • Understanding the metabolism of ascorbic acid during grape berry development
the people
The people…
  • University of Adelaide:
    • Seth DeBolt (PhD candidate 2003-2006)
    • Vanessa Melino (PhD candidate 2005- )
    • Steve Tyerman, Matt Hayes
  • Flinders University of South Australia:
    • Crystal Sweetman (PhD candidate 2006- )
    • Crista Burbidge (PhD candidate 2007- )
    • Kathy Soole
  • University of California (Davis)
    • Doug Cook
the funding
The funding
  • This project was supported by the Australian Government’s Cooperative Research Centre Program and conducted by the Cooperative Research Centre for Viticulture.
  • This work was supported by Australia’s grape growers and winemakers through their investment body, the Grape and Wine Research and Development Corporation, with matching funds from the Australian Government.