Flower Speciesspecies

13 June 2011 ISTA GERMINATION SEMINAR GERMINATIONGERMINATION CHARACTERISTICSCHARACTERISTICS OFOF FLOWERFLOWER SPECIESSPECIES

RITA ZECCHINELLI - ISTA FLOWER SEED TESTING COMMITTEE Luca Flower species in the ISTA Rules:

. 352 species . 192 genera . 55 families

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI DICOTYLEDON FAMILIES Family N°genera N°species Family N°genera N°species Acanthaceae 1 1 Hypericaceae 1 1 Aizoaceae 1 1 Iridaceae 1 1 Amaranthaceae 3 6 Lamiaceae 14 21 Apiaceae 3 4 Linaceae 1 4 Apocynaceae 1 1 Malvaceae 6 7 Araliaceae 2 2 Nyctaginaceae 1 1 Asclepiadeaceae 1 1 Onograceae 2 4 Asteraceae 49 86 Papaveraceae 3 7 MONOCOTYLEDON FAMILIES Balsaminaceae 1 2 Plumbaginaceae 4 8 Family N°genera N°species Begoniaceae 1 2 Polemoniaceae 3 5 Amaryllidaceae 1 1 Boraginaceae 6 10 Polygonaceae 1 1 Asparagaceae 1 2 Brassicaceae 11 22 Portulaceae 1 1 Asphodelaceae 1 1 Campanulaceae 3 13 Primulaceae 3 12 Liliaceae 1 1 Capparidaceae 1 1 Proteaceae 1 1 Poaceae 2 2 Caryophyllaceae 6 16 Ranuncolaceae 7 20 Chenopodiaceae 1 1 Resedaceae 1 1 Cistaceae 1 1 Rosaceae 2 3 Convolvulaceae 2 5 Rutaceae 1 1 Crassulaceae 1 3 Saxifragaceae 1 1 Dipsacaceae 1 2 Scrophulariaceae 10 22 Fabaceae 4 8 Solanaceae 9 15 Gentianaceae 1 1 Tropaeolaceae 1 3 Geraniaceae 2 2 Valerianceae 1 1 Gesneriaceae 2 2 Verbenaceae 2 4 Hydrophyllaceae 3 4 Violaceae 1 3

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI What flower species have in common?

. ornamental use . fast changing trends in the market

. for seed testing laboratories

- less experience in standardized methods - lower number of samples/species than for agricultural species and vegetable - accreditation: all flower species in one crop group

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI GERMINATION METHODS IN THE ISTA RULES

Only one temperature for germination:

Agricultural species: 172 species/326 (53%) Flower species: 35 species/352 (10%)

Three temperatures for germination:

Agricultural species: 18 species/326 (5,5%) Flower species: 54 species/352 (15%)

Flower species: higher variability within the species lower degree of standardization lower degree of knowledge and experience

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI SEEDLING EVALUATION ACCORDING TO GROUPS (ISTA HANDBOOK ON SEEDLING EVALUATION)

Seedling Type Seedling Group

A A-1-1-1-1 Liliaceae

B A-1-2-1-1 Asphodelaceae

C A-1-2-2-1 Asparagaceae

D A-1-2-3-1 Poaceae

A-1-1-1-1 Amaryllidaceae

Acanthaceae, Aizoaceae, Amaranthaceae, Apiaceae, Apocynaceae, Araliaceae, Asclepiadeaceae, Asteraceae, Begoniaceae, Boraginaceae, Brassicaceae, Campanulaceae, Capparidaceae, Caryophyllaceae, Chenopodiaceae, Cistaceae, Convolvulaceae, Crassulaceae, Dipsacaceae, Fabaceae (Galega, Mimosa), Gentianaceae, Geraniaceae, Gesneriaceae, E A-2-1-1-1 Hydrophyllaceae, Hypericaceae, Lamiaceae, Linaceae, Nyctaginaceae, Onograceae, Papaveraceae, Plumbaginaceae, Polemoniaceae, Polygonaceae, Portulaceae, Primulaceae (Anagallis, Primula), Proteaceae, Ranuncolaceae, Resedaceae, Rosaceae, Rutaceae, Saxifragaceae, Scrophulariaceae, Solanaceae, Verbenaceae, Valerianceae, Violaceae

A-2-1-1-2 Balsaminaceae, Malvaceae

A-2-1-4-3 Primulaceae (Cyclamen)

F A-2-1-2-2 Fabaceae (Lupinus)

G A-2-2-2-2 Fabaceae (Latyrus), Tropaeolaceae

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Type C A. densiflorus Seedling Group A-1-2-2-1 Monocotyledons Hypogeal germination Epicotyl elongation Primary root essential

The seedling part that grows towards the light and turns green is the epicotyl with the terminal bud

Asparagus spp. A. densiflorus (Asparagaceae)

A. setaceus ISTA Germination methods Counts Temperature Breaking Substrate (°C) First Final dormancy (d) (d)

A. setaceus TP; BP; S 2030; 20 7-14 35 Soak in water for 24 h

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Asparagus: development of the seedling during the test •Start of germination: the primary root pierces the seed coat, followed by a short hypocotyl and the basal part of the cotyledons (the upper part remains in the seed coat and absorbs reserves from the endosperm) - A. setaceus: has a very stout primary root; terminal secondary roots don’t normally develop (A) bud small - A. densiflorus: secondary roots normally develop; scale-leaves one of them serves as a water storage organ (B) epicotyl •The epicotyl emerges from the basal part of the cotyledon; basal part it bears several small scale-leaves, of cotyledon the top ones form the terminal bud hypocotyl

secondary roots (one serving as a A water storage organ) primary root B B 1 2 3 4

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Asparagus densiflorus: abnormal seedlings

Normal seedling Abnormal seedlings

1 2 3 4 5 6 7 8 9 10

1 - 12/01 Secondary root serving as water storage organ weak or missing 2 - 3 - 11/04 Primary root missing 4 - 5 - 21/05 Epicotyl missing 6 - 12/01; 21/05 Secondary root and epicotyl missing 7- 9 - 11/03; 11/04 Primary root retarded or missing 10 - 00/09 Seedling as a whole decayed as a result of primary infection

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Asteraceae Type E Seedling Group A-2-1-1-1 Dycotiledons Epigeal germination Dahlia pinnata Without epicotyl elongation Primary root essential

The seedling part that grows towards the light is the hypocotyl with two cotyledons attached that turn green

ISTA Germination methods

Counts Temperature Breaking Substrate (°C) First Final dormancy (d) (d)

TP; BP 2030; 20; 15 4-7 21 Prechill

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Dahlia: development of the seedling during the test

•Start of germination: the primary root pierces the seed coat (1) •The primary root elongates rapidly (2) •The hypocotyl elongates and raises the cotyledons (3) •The cotyledons become green and start photosynthesis (4).

During the test period, the seedling rarely develops secondary roots. The epicotyl doesn’t develop and the terminal bud is hardly visible between the cotyledons.

1 2 3 4

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Dahlia pinnata: abnormal seedlings

Normal seedling

1 2

3 4 5 1 - 11/04 Primary root missing 2 - 11/03 Primary root is retarded 3 - 31/05 Cotyledons are discoloured 4 - 31/05 Cotyledons are necrotic 5 - 00/01 The seedling as a whole is deformed

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Lamiaceae Type E Seedling Group A-2-1-1-1 Dycotiledons Epigeal germination Without epicotyl elongation Primary root essential

The seedling part that grows towards the light is the hypocotyl with two cotyledons attached that turn green

ISTA Germination methods Counts Temperature Breaking Substrate (°C) First Final dormancy (d) (d)

Coleus blumei TP; BP 2030; 20 5-7 21 Light

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Coleus: development of the seedling during the test

During the test period, the seedling rarely develops secondary roots. The epicotyl doesn’t develop and the terminal bud is hardly visible between the cotyledons.

1 2 3 4

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Coleus blumei: abnormal seedlings

1 2 3

Normal seedling Abnormal seedling

1 - 11/03 Primary root retarded 2 - 11/12 Primary root decayed as a result of primary infection 3 - 11/04 Primary root missing

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Primulaceae Type E Seedling Group A-2-1-4-3 Dycotiledons Epigeal germination Tuberous hypocotyl Several seminal roots, instead of a primary root

The seedling part that grows towards the light is the hypocotyl with two cotyledons attached that turn green Cyclamen persicum

ISTA Germination methods Counts Temperature Breaking Substrate (°C) First Final dormancy (d) (d) KNO ; soak TP; BP; S 20; 15 14-21 35 3 13 June 2011 ISTA GERMINATION SEMINAR in water 24 h RITA ZECCHINELLI (germination in the dark is suggested) Cyclamen: development of the seedling during the test The seed of Cyclamen contains a small, straight embryo embedded in endosperm. •Start of germination: the seed coat is pierced (1) •The hypocotyl forms a tuber (2) •Several seminal roots develop at the base of the hypocotyl (3) •When the hypocotyl has completely emerged, it is followed by the petiole of one cotyledon (4) The blade of the cotyledon (A) seldom occurs during the test period. The terminal bud takes place on the top of the tuber, at the base of the cotyledon (B). The first foliage leaf needs time to start the development.

A B

1 2 3 4

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI NORMAL SEEDLINGS: examples

Unusual, but normal

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI ABNORMAL SEEDLINGS: examples

Normal seedling 00/01 – Seedling deformed

Normal seedling 11/02 Primary root stubby

11/08 Primary root shows negative 21/02 Hypocotyl not Primarygeotropism root with negative forming a tuber geotropism

Normal seedling Normal seedling

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI ABNORMAL SEEDLINGS: examples

31/07 Cotyledon decayed as a result of primary infection

Attention must be paid to sign of decay at the point where the cotyledon petiole leaves the seed coat

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Tropaeolaceae Tropaeolum Tropaeolum majus peregrinum

Tropaeolum peltophorum

Tropaeolum spp.

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Tropaeolum Type G spp. Seedling Group A-2-2-2-2 Dycotiledons Epigeal germination Tropaeolum majus Epicotyl elongation The primary root may be replaced by secondary roots

The seedling part that grows towards the light and turns green is the epicotyl with the primary leaves

Tropaeolum peltophorum ISTA Germination methods

Temperature Counts Substrate ( °

C) Breaking First Final dormancy (d) (d)

TP; 20; 15 4-7 21 Prechill BP; S (and 2030 T. majus) Tropaeolum peregrinum

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Tropaeolum majus: development of the seedling during the test

•Start of germination: the primary root pierces the seed coat (1) •The primary root elongates rapidly (2) •Secondary roots soon develop (3) •The hypocotyl is hardly discernible, the epicotyl elongates rapidly (4)

The buds in the axils of the cotyledons are usually dormant except if the terminal bud is damaged.

1 2 3 4

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Tropaeolum majus: abnormal seedlings

1 2 3

Normal seedling 4 5 6

1 - 00/01 Seedlings deformed 2 - 00/09 Seedling decayed as a result of primary infection 3 - 11/03 Primary root retarded 4 - 11/04 Primary root missing 5 - 21/01 Epicotyl short and thick 6 - 21/03 Epicotyl deeply cracked 7 - 21/12 Epicotyl decayed as a result of primary infection 7

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Fabaceae Lupinus spp.

Lupinus nanus

Lupinus polyphyllus Lupinus hartwegii

Lupinus hybrids

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Type F Seedling Group A-2-1-2-2 Lupinus hartwegii Dycotiledons Epigeal germination Lupinus spp. Epicotyl elongation (flower species) ? The primary root may be replaced by secondary roots

The seedling part that grows towards the light and turns green is the hypocotyl with two cotyledons And the epicotyl with the primary leaves

ISTA Germination methods

Temperature Counts Substrate ( °

C) Breaking First Final dormancy (d) (d)

L. hartwegii: pierce TP; 2030; seed; or file 4-7 21 fragment off testa at BP; S 20 Lupinus nanus Lupinus polyphyllus cotyledon end; prechill

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Lupinus polyphyllus: development of the seedling during the test •Start of germination: the primary root breaks through the seed coat (1) •The primary root elongates rapidly (2) and soon produces numerous secondary roots •The hypocotyl elongates and the cotyledons are freed from the seed coat (3) ? •The cotyledons become green and start photosynthesis (4).

1 2 3

Substrate: paper

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Lupinus polyphyllus vs Lupinus albus Type F - Seedling Group A-2-1-2-2: secondary roots soon produced? Lupinus polyphyllus (flower) Lupinus albus (vegetable)

NO any secondary roots produced YES

(or only a few and very late) secondary roots produced

Substrate: sand

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Lupinus polyphyllus

Type F - Seedling Group A-2-1-2-2 Dycotiledons → YES Epigeal germination - Epicotyl elongation → YES The primary root may be replaced by secondary roots → ?

The seedling part that grows towards the light and turns green is the hypocotyl with two cotyledons and the epicotyl with the primary leaves

Change to:

Type F - Seedling Group A-2-1-2-1 (new) ?

Change only the description of the

Development of the seedling during the test?

The other flower species of Lupinus should be also investigated. Looking for seeds!

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Lupinus poliphyllus: abnormal seedlings

1 2 3

1 - 11/04 Primary root missing 2 - 42/03 Primary leaf shredded or otherwise deformed 3 - 00/09 Seedling decayed as a result of primary infection

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI ISTA Flower Seed Committee

Flower species: there is some work to do

WORKING GROUPS

1) ISTA Flower Seed Testing Handbook 2) Validation study for Felicia heterophylla (germination method/s) 3) Validation study for Alcea rosea (germination method in sand) 4) Sample size for flower species (Tab. 2A part 3) 5) Identification of Helianthus species by seed … Seedling evaluation in flower species of Lupinus (NEW! To be still discussed)

Anyone willing to co-operate is welcome!

13 June 2011 ISTA GERMINATION SEMINAR RITA ZECCHINELLI Photos of seeds and seedlings Sylvie Ducournau Zita Ripka Digital Atlas of Economic Plants (http://www.plantatlas.eu/) USDA Plant Database (http://plants.usda.gov/java/) Digital collection of INRAN-ENSE Laboratorio Analisi Sementi, Italy ISTA Handbook on Flower Seed Testing 1st Edition 2008 ISTA Handbook on Seedling Evaluation 3rd Edition 2003 ISTA Handbook on Pure Seed Definitions 3rd Edition 2010

Acknowledgements Fabio Ferrari, Alessandra Arioli and colleagues Sylvie Doucournau Joël Léchappé

and

Luca Frigerio, the young artist of the drawing ‘Germination of a seed of violet’

AndAnd thankthank youyou forfor youryour attention!attention!