ESF provides the COST Office

through a European Commission contractCOST is supported

by the EU Framework Programme

Action Title: Omics Technologies for Crop Improvement,

Traceability, Determination of Authenticity, Adulteration and

Origin in Saffron (SAFFRONOMICS)

Action number: FA1101

Start date: 24/11/2011

End date: 23/11/2015

Year: 3rd

Chair: Maria Z. Tsimidou

Professor, Aristotle University of Thessaloniki. Greece

Annual Progress Conference

Plovdiv, Bulgaria, 28-29-30 April 2014

2

Saffron:

The dehydrated red

stigmas of the

flower of the plant

Crocus sativus L.

Bright red colour

Bitter taste

Unique flavour

3

The plant Crocus sativus L.

is a member of the Iridaceae

family.

The major producing countries

are Iran, India, Greece and Spain.

It is a perennial, triploid, sterile

plant and is reproduced by corms.

It reaches a height of 10 to 25

cm.

Its purple flowers consist of 6

tepals.

The pistil is located in the

centre of its flower with one ovary

from which a style is emerged.

Style is ended to a three-branch

stigma

Crocus sativus L.

Family: Iridaceae

Subfamily: Crocoideae

Genus: Crocus

Species: Crocus Sativus Linnaeus

style

stigma

ovary

4

Annual cycle of the

plant Crocus sativus L.

White Book, Saffron in Europe

corms: 3 to 6 years in the

same field

5

Saffron is expensive (~3500 €/kg) because the production process

requires manual labor

is of low yield (1-2 kg/hectare)

Trade values are determined based on the standard specifications described in the

ISO 3632-1&2, mainly on

Colouring strength (E1% at 440 nm)

Bitterness (as picrocrocin content, E1% at 250 nm)

Aroma (as safranal content, E1% at 330 nm)

Trade values

Based on the ISO trade standard for saffron (3632-1)

Absorbance at 257, 330 and 440 nm are related with the presence of

Picrocrocin

Safranal

Crocins

R1OOR28

9

O

11 13

20

15'15

13' 11'

20' 19'

9'

19 O

Rxn

RxnR1O

OR2

8

9

19

11 13

20

15 15'

13'

11'

9'

19'

20'

8'

O

O

Rxn

O

O

HO

HO

OH

O

OH

O

R1, R2: 1-3 Glucose moieties or H atom

all trans 13-cis

6

Scientific context and objectives (1/3)

Saffron is:

• a profitable High Value Agricultural Product (HVAP), of European in origin,

cultural and historical background

• fits food-safety trends

• a crop that fixes population in the rural areas

• The European Saffron industry is in crisis. Little or null impact in the CAP.

• Adulteration and mislabelling are key problems.

• The scientific human power involved in Saffron RTD is petite.

• This crop requires global actions, overcoming private and national

interests –actions are needed beyond EU borders.

7

Background / Problem statement

Brief reminder of MoU objectives:

8

Objective 1:

To build up a network of collaborative research on the structural

organization of Saffron genome, DNA fingerprinting, chemical

fingerprinting, proteomics, transcriptomics, and metabolomics of this crop.

Objective 2:

To preserve Crocus biodiversity, to carry out genetic improvement and to

protect quality, sustainability, and safety of production of saffron in Europe.

Objective 3

The long term achievement expected: CONTROL OF ADULTERATION

WORLDWIDE.

Scientific context and objectives (2/3)

Scientific context and objectives (3/3)

Research directions:

• ORGANISING INTERNATIONAL COOPERATIVE R&D and INNOVATION

• ASSEMBLING PREEMINENT EXPERTS (MULTIDISCIPLINARY APPROACH)

• INVOLVING PRODUCERS, MANUFACTURERS, MARKET STAKEHOLDERS &

CONSUMERS

9

This integrated knowledge will be the basis for the development

of saffron genetic improvement, for reliable innovative

techniques to produce high value saffron and to combat

especially bio-adulteration and fraud.

Working groups

1. Genetics, Genomics & Transcriptomics (coordinated research on these

fields). Leader: Silvia Fluch (AT); Co-Leader: A. Mozzarelli (IT)

2. Phytochemistry & Metabolomics (coordinated research on these fields).

Leader: M. Polissiou (GR); Co-leader: M. Carmona (ES)

3. Molecular and Phytochemical fingerprinting for breeding, traceability, and

authenticity (development of technological tools) Leader: P. Tarantilis

(GR); Co-Leader: S. Ordoudi (GR)

4. Dissemination, Project Management & Coordination. Leader: O. Santana

(ES); Co-Leader: JM. Thiercelin (FR)

10

0

5

10

15

20

YR 1YR 2

YR 3YR 4

18 21

8 8 Parties

Non-COST Countries

Grant Holder:

Universidad de Castilla-La Mancha (UCLM), Spain

Dr. Marta Roldan-Medina

Grant Holder manager

6

Action Parties

0

50

100

150

YR 1YR 2

YR 3YR 4

95

136

25 34 35 46

Total no. of indiv. Participants

ESRs

Female

7

Action participants

ISRAEL:1

MOROCCO:2

EGYPT: 1

TURKEY:2

IRAN:2

INDIA:1

AJERBAIJAN:1

USA:2

NEW Zealand: 2

SAUDI ARABIA:1

1

20 [6]

1

11 [4]

21 [7]

2

5 [2]

4

2

4 [2]

1

1

3

1

4

1

1

1

1

Use of COST Instruments

14

Activity (No.) Year 1 Year 2 Year 3 (plan) Year 4

MC/WG Meetings 1 MC meeting/

2 WG1-4 meetings

1 MC meeting

2 WG1-4 meetings

1 WG1 expert meeting

(on saffron genome sequencing)

1 MC meeting

1 WG1-4 meetings

2 expert meetings] (White Paper on saffron extraction

procedures regarding ISO quality

requirements)

(protocol on spectroscopic

techniques)

STSMs 5 STSMs 9 STSMs 7 max STSMs

Training Schools - - 1 Training School (on spectroscopic and

chemometric approaches)

1 Training School (on saffron proteomics)

Workshops or

Conferences

1 EU Seminar 1 Workshop

(on wild crocuses)

1 Annual Conference

1 Workshop

(on saffron genome)

1 Annual Conference

Joint

Publications*

*In scientific journals

- 2 3-5 expected

In FA1101 meetings 54 55

15

Progress in joint research has been achieved mainly through

exchange visits of young scientists from laboratories of

established experience in saffron quality and authenticity

aspects (Spain, Italy, Greece) to laboratories of established

experience in new analytical techniques (Greece, Spain,

Italy, the Netherlands) and between laboratories involved in

Plant Genomics (Spain, Italy, Germany).

•STSM leader Dr. Giovanni Giuliano, ENEA, IT (1st-2nd year)

•STSM leader Dr. Roberto Consonni, ISMAC, IT (3rd year -)

STSMs Applicant: Dr. Nikolaos Nenadis

Affiliation: Aristotle University of Thessaloniki (AUTH, Greece)

Subject: Ageing of saffron determined by PTR-MS (2012)

Applicant: Dr. Mateo Busconi

Affiliation: Università Cattolica del Sacro Cuore, Milano (Italy)

Subject: Development of molecular markers (AFLPs, SNPs) and analysis of data to ascertain genetic variability in the saffron 'CrocusBank' collection (2012)

Applicant: PhD. Student Eleni Naziri

Affiliation: Aristotle University of Thessaloniki (AUTH, Greece)

Subject: NMR based metabolomic characterization of saffron extracts from different geographical origins (2012)

STSMs Applicant: Sarah Frusciante

Affiliation: ENEA-Green Biotechnology Unit (ENEA, Rome (Italy)

Subject: Biochemical characterization of novel CCDs from Saffron (2013)

Applicant: Dr. Mahmoud SHARAF-ELDIN

Affiliation: Salman bin Abdulaziz University (SAU) / National Research Centre (NRC, Egypt)

Subject: Detection of saffron bio-adulteration (2013)

Applicant: PhD. Student Sofia Lalou

Affiliation: Aristotle University of Thessaloniki (AUTH, Greece)

Subject: NMR metabolite profiling of aged saffron (2013)

Applicant: Dr Olivia Costantina Demurtas

Affiliation: ENEA, Italian National Agency for New Technologies, Energy and Sustainable Development (ENEA, Italy)

Subject: Functional characterization of candidate transporters of saffron stigma

secondary metabolites (2013)

Applicant: Dr. Stella Ordoudi

Affiliation: Aristotle University of Thessaloniki (AUTH, Greece)

Subject: Meta-analysis of NMR data using chemometrics for assessment of age-related biomarkers in saffron (2013)

Applicant: Dr. Omar Santana Méridas

Affiliation: Parque Científico y Tecnológico (JCCM-PCYTA, Spain)

Subject: Sourcing and metabolomic study of new sources of Crocus-based bioactive extracts (2013)

STSMs

Applicant: Ms Soukaina Chaouqi

Affiliation: Institut National de la Recherche Agronomique (INRA, Morocco)

Subject: Evaluation of the effect of dehydration, drying and storage conditions on secondary compounds of Moroccan saffron (2013)

SUMMARY OF

COMPLETED

STSMs

2013: 9 completed STSMs 2014: 5 STSMs planned

More information available in

http://www.saffronomics.org/download-center/cat_view/5-completed-stsms.html

STSMs

• Progress was also achieved in dissemination about saffron properties, authenticity and consumer awareness as well as for expansion of saffron cultivation (Austria, FYROM, Israel, Egypt, Saudi Arabia) through mass media and public lectures.

• FACE BOOK 20

• WEB page (www.saffronomics.gr) and newsletter are useful platforms for communicating various types of information on “SaffronOmics” and other related activities. • Outreach activities for Krokos Kozanis was an outcome of an MSc thesis for spreading information about the properties of herbs and spices and in particular for authentic saffron to high school pupils. Castilla la Mancha prepares outreach activities for elementary school pupils.

Progress vs objectives (2/2)

• Results were the 54 communications by a total of 95 different authors presented in the 3 COST FA1101 scientific meetings taken place in the 1rst year (2012) and the 55 ones presented by a total of 116 different authors in the 2 scientific meetings in the 2nd year (2013) (http://www.saffronomics.org/download-center/).

• Publications: 1 book of Proceedings from 1rst year scientific meetings (ISBN -13:978-84-616-0964-2 and 2 joint research papers (Ordoudi, S.A., de los Mozos Pascual, M., Tsimidou, M.Z., Food Chemistry, 2014, 150, 414-421 / Sánchez-Vioque, R., Polissiou, M., Astraka, K., de los Mozos-Pascual, M., Tarantilis, P., Herraiz-Peñalver, D., Santana-Méridas, O. Industrial Crops and Products, 2013, 49, 150-159.

• Agreement to share reference samples of saffron dried stigmas to get comparable scientific results among laboratories (in process for a third harvesting period)

• Agreement to collaborate in ring tests for the improvement of analytical protocols (under reconsideration due to new literature available)

• Data bank for genomics, transcriptomics, proteomics (under construction) • Data bank for spectroscopic data (in process) • Workshop in wild crocuses: set up principles for species prioritization and study beyond Crocus

Bank data. • Expert group meeting on saffron genome sequencing: the output was a White Paper for project

plan outline. • Expression of interest of more research groups to participate in our Action • Institute of Natural Resource Sciences, Zurich University of Applied Sciences, Prof. Grunder, J. • Laboratoire de Chimie Analytique Bio-Inorganique et Environnement, IPREM, CNRS-UPPA,

Pau, France, Dr. Pedrero Zayas, Z. • Institute of Applied Biosciences CENTRE FOR RESEARCH AND TECHNOLOGY HELLAS –

CERTH, Dr. Madesis, P. • Dr. Galanakis,Ch. Research & Innovation Director, Galanakis Laboratories –Chania Greece

21

Progress vs objectives (3/2)

22

None of the above activities would have been

planned or achieved

without the Action’s research network.

23

Invited Speakers in the 2nd Year Meetings

Dr. Christina Fournaki,

taxonomist, Curator of the Botanic

garden of MaiCh (Crete, Greece)

Dr. Panagiotis Kefalas, expert in

secondary metabolites Omics

techniques, Director of Laboratory

of Chemistry of Natural Products

and Analytical Chemistry MaiCh

(Crete, Greece)

Dr. Anaya Sarpaki

Archaeologist–archaeobotanist,

independent Scholar in Santorini

excavations

WG2 Interim meeting on Phytochemistry and

Metabolomics and WG1/WG3/WG4 Looking for Synergy

and Dissemination Capacity Building (MaiCh, Crete,

Greece)

Dr. Apostolos Spyros

Assistant Professor, NMR

specialist Laboratory, Department

of Chemistry, University of Crete

(Heraklion Crete, Greece)

1st Annual Conference on Omics technologies for

crop improvement and traceability in saffron and allied

species (AIT, Tulln, Austria)

Dr. Jaroslav Dolezel

Centre of the Region Hana for

Biotechnological and Agricultural

Research (Czech Republic)

Dr. Salim al-Babili

University of Science and

Technology (Kingdom of Saudi

Arabia)

Dr. Konstantinos Aliferis,

expert in

metabolomics,Department of

Plant Sciences, McGill University

(Quebec, Canada)

Dr. Thomas Prohaska, stable

isotopes expert, University of

Natural Resources and Life

Sciences

(Vienna, Austria)

24

MaiCh meeting promotion to

the local press and TV

channels

Highlights of the 2nd Year events

Round Table of “PHYTOBUISENESS”

One day meeting in Kozani (Greece)

organized by the Saffron

Cooperative and Crocus Kozanis

Products SA.

1st Annual Conference (Tulln, Austria)

WG1-4 Meeting (Chania, Greece)

Working with pupils of the high-school in Krokos

(Kozani, Greece) on various saffron aspects

Future Plan and Challenges (1/2)

Further advance in the genetic characterization, estimation of biodiversity and

development of molecular and chemical descriptors for saffron and Crocus spp. at

the CROCUSBANK World Saffron and Crocus Collection (www.crocusbank.org).

To evaluate the available genomic tools and modern genetic and breeding

approaches looking for crop improvement in saffron and ornamental crocuses.

The application of omics techniques to detect “new-generation” adulterants in

saffron.

To initiate genomic typing of saffron in PDOs and recognized areas, as tools for

traceability applications, determination of authenticity, and for fighting against fraud

of origin, labelling and marketing in this HVAP, the highest-priced European food

product.

25

Future Plan and Challenges for the 1st trimester of the 3rd Year

26

WG1

Writing a White Paper on saffron genome

sequencing with contributions from various

partners.

Prepare the workshop on saffron genome.

Prepare a core collection of wild saffron

samples within the network.

WG2

Data Bank for spectral data after collaboration

of WG2&3&4.

Joint publications between WG2&3&4 (from

collaborations established in the frame of COST

network).

Writing a White Paper on saffron extraction.

WG3 WG4

Maintenance and management of the

“Saffronomics” website.

Dissemination/Outreach activities about saffron

aspects for pupils of different ages.

Distribution of samples of saffron to get

complementary scientific results by different

Omics techniques in different laboratories for a

third harvesting period).

Organization of two training Schools to present

the advanced spectroscopic techniques applied

to the quality characterization of saffron. The 1st

one will be on spectroscopic techniques and

chemometrics and the 2nd one on saffron

proteomics.

Significant Highlights in Science or Networking

• One proposal to the Call: FP7-PEOPLE-2013-ITN.

• One proposal in Call: FP7‐KBBE‐2013‐7 ‐ single‐stage -

KBBE.2013.2.4‐01 “Assuring quality and authenticity in

the food chain”.

• One project funded in Call: FP9-KBBE.2013.3.1-01 “Plant

High Value Products-from discovery to final product”.

• Project: “From DISCOvery to products: A next generation

pipeline for the sustainable generation of high-value

plant products” (Grant agreement no: 613513. Start Date

November 2013).

27

Challenges

Deviations are mainly due to financial restrictions in terms of

national or European funding.

Collaboration between academics and industrial partners -

even when agreements are signed – is not funded easily.

More data are expected in the field of Genomics and

Transcriptomics. This work is tedious and expensive.

28

• Involvement of SAFFRONOMICS partners with saffron

producers and companies strengthen the links among all

interested parties (Italy, Spain, Greece).

• Expansion of saffron cultivation has become a strategic plan

(Austria).

• Familiarization of the young generation from producing areas

with saffron cultivation, authenticity and properties has become

a priority in the activities of certain partners (Greece).

29

According to template used by

IFT for outreach educational

activities

Objective:

To disseminate the knowledge about saffron

spice and increase its comsuption within Europe

through a program focused in primary schools (6

to 11years old), using an environmental program

adapted to each age.

Stages:

- Design and development of the teaching

methodology, activity proposals, and teacher´s

guide adapted to the current contents in the

European curriculum.

- Design and development of students working

books to students from 1st to 6th primary school.

- Design, ilustration, layout and printing.

Outcomes:

-Teacher guide (digital material, 90-100 pages)

- 6 Printed students books (400 , 8-10 pages

each)

-Languages: English, Spanish, Greek.

35

OUR GRATIDUTE to

Ioanna, Cassia, Christophe