Floral Fragrances

P. Hugueney, ENSL

What is flower scent ?

pump

Flower scent analysis :"dynamic headspace chromatography"

flowerattachedto plant

containerfilter

trap

1 : sampling

Flower scent analysis using "headspace chromatography"

2 :analysis

Gas ChromatographyMass Spectroscopy

trap

More than 100 different volatile compounds are detected in the scent of a single rose flower

OH

OH

OH

OH

OCH3

COOCH3

COOCH3

OH

H3CO OCH3

CH3

COOCH3

H3CO OCH3

CH3

H3CO OCH3

CH3

COOCH3

OH

COOCH3

OH

COOCH3

OH

COOCH3

OH

OH

OCH3

OH

The reason why flowers are scented ...

Olfactory organ of insects : the antenna

V

Antenna : containsolfactory neurons

electroantennography :detection of neuron

membranedepolarization

V GC-EADGas Chromatography

with ElecroAntennographicDetection

Sex pheromone mimicry in the orchid Ophrys sphegodes

Schiestl et al. (2000), J. Comp. Physiol. A 186 : 567-574

Flower scent analysis by " nose detection"

A "pleasant" scent is relative….

OH

OH

OH

OH

OCH3

COOCH3

COOCH3

OH

H3CO OCH3

CH3

COOCH3

H3CO OCH3

CH3

H3CO OCH3

CH3

COOCH3

OH

COOCH3

OH

COOCH3

OH

COOCH3

OH

OH

OCH3

OHUnderstanding scent compounds biosynthesis

Major classes of flower scent compounds :

- Terpenes

- Aromatic compounds

- Fatty acid derivatives

= Organizing scent compounds into metabolic pathways

Understanding scent compounds biosynthesis

Terpenes (25000 natural compounds)

OPPsterols

carotenoids

chlorophyll

OH

β-pinene

OH

geraniol

H

H

OHH(-) menthol

linalool

limonene

Isopentenyl pyrophosphate

Primary metabolism

Most advanced models :

- Peppermint (Mentha x piperata) : Rodney Croteau

- Clarkia breweri : Eran Pichersky (about 10 scent genes cloned ...)

Photo : J.C. Caissard

Photo : J. D'Auria

Rosa x centifolia ( Rose de Mai )

- grown in the south of France (Grasses)

- 1982 : 350 tons of flowers

- used in some perfumes

Rosa x damascena essential oil : 15 - 20 tons oil / year

: 1500 - 10000 €€€€/kg

: 4 tons petals (800h) for 1kg oil

A more recent but very promising model : the rose

- Rosa x hybrida : initial crosses involving less than

10 wild rose species

- more than 2 centuries of breeding and selection

- about 25000 registered varieties

- most economically important ornamental plant

What is a rose ?

R. odorata

Rosa chinensiscv. “Old Blush”

R. moschata

Rosa gallica Rosa phoenicia

R. damascena

R. biferaRosa moschata

NoisetteRoses

Yellow TeaRoses

Pink TeaRoses

Modern Roses

R. chinensis R. gigantea

R. odorataR. gallica

R. gallicaR. bifera

PortlandRoses

BourbonRoses

Chinese Hybrids

BourbonRoses

Hybrid Tea Roses

Hybrid Perpetual RosesTea Scented Roses

Modern roses genealogy

Interest of crossing "European" and "Chinese" Roses

"European" Roses : ex : Rosa gallica, Rosa x damascena - temperate climate - yearly flowering in May

"Chinese" Roses : ex : Rosa chinensis - subtropical climate- recurrent flowering

Modern hybrid roses : - temperate climate- recurrent flowering- selection of aesthetic traits

wild rose modern rose

Secretory epidermis

Terpenes

Phenylethanol

Aromatic compounds

Fatty acid derivatives

Rose ketones

etc...

Rose scent

GC analysis of R. x damascena scent

pA

min

phen

ylet

hano

l Ger

anio

lC

itro

nello

lne

rol

2 main types of scent in "wild" roses

"European" Type : ex : Rosa x damascena - phenylethanol - monoterpenes

Geraniol

Citronellol

OHnerol

OH

2-phenylethanol

"Chinese" Type : ex : Rosa chinensis, Rosa gigantea

- monoterpenes

- fatty acid derivatives

- phenolic compounds

OH

cis-3-hexenol

geraniol

CH3O OCH3

OCH3

trimethoxybenzene

2 main types of scent in "wild" roses

"Combined" Type :

Geraniol

OH

2-phenylethanol

Citronellol

OHnerol

H3CO OCH3

CH3

3,5-dimethoxytoluene

modern roses scent...

pA

2-ph

nylt

hano

l

Royal Red

min

modern roses scent...

"Scentless" Type :

Why are many modern roses scentless ?

This trait was lost in recent breeding processes, with selection programs based on shape, colour, disease resistance, longevity...

Characterization of genes and enzymes involved in scent production in roses

- 2 years ago : 20 rose sequences in gene databases

- Genomic approach : random sequencing of genes

expressed in rose petals

Channelière et al., 2002Guterman et al., 2002

Protein synthesis (5.1%)

Protein and lipid turnover (2.4%)

Cell wall (1.9%)

Secreted proteins(2.8%)

Membraneassociated

(10.9%)

Hormoneregulated

(1.9%)

Defense /stress related

(15.1%)

Signal transduction (4.0%)Transcription

(1.2%)Other (8.6%)

Unknown function (9.2%)

No blast hit (26.6%)

Primary metabolism (6.6%)

Secondary metabolism (2.6%)

RNA binding (0.9%)

Functional classification of the rose petal ESTs

ß-ionone ß-damascenone

Linalool

Geraniol

Detection thresholds (ppb) (human)

ß-ionone 0.007

ß-damascenone 0.009

Linalool 6

Citronellol 40

Geraniol 75Citronellol

(1 µl / 100 m3)

Alister Stella Gray (Noisette)

Rose ketones biosynthesis

O

carotenoid- dioxygenase

O

Rose ketones :

oxidativeclivage

Carotenoids

- ß-damascenone

- ß-ionone

β- carotene

retinal

OH

retinoic acid

β- apocarotenal

β- ionone

symetric cleavage asymetric cleavage

Carotenoid cleavage in mammals

The "Rose" lab

The "Rose" lab in St Etienne

S. BaudinoF. JullienJ.L. MagnardV. Bergougnoux

S. Channelière P. Vergne C. Dolle

G. Scalliet

J. Szecsi

P. Hugueney M. Cock M. Bendahmane

Plant Reproduction and Development (C.Dumas), CNRS,INRA,ENSL,UCBL