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  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Speed of seed imbibition related to seedling evaluation

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 1

    seedling evaluation Wagner MH, Demilly D, Blouin V,

    Ducournau S, Léchappé J.

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Seed imbibition: first step before

    seedling development

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 2

    Sowing Germination Growth Seedling

    Development of oilseed rape seedling

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Seed imbibition: first step before

    seedling development Imbibition Germination Growth

    W at

    er u

    pt ak

    e

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 3

    Time

    W at

    er

    from Bewley & Black, 1994

    Aim: test the image analysis parameters obtained before germination

    Radicle protusion

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    • Medicago truncatula

    4 6 8

    10 12 14 16

    se ed

    w ei

    gh t(

    m g)

    seeds at sowing time seeds during

    Seed imbibition measured by image analysis:

    seed area or volume is correlated to weight

    Demilly et al., 2007

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013

    2 4

    4 6 8 10 12 14

    se ed

    seed area (mm²)

    seeds during imbibition

    4

    y = 0.6038x + 0.8853

    R² = 0.865

    0

    2

    4

    6

    8

    10

    12

    14

    16

    0 5 10 15 20 25

    D ry

    s e

    e d

    m as

    s (m

    g)

    Dry seed volume on image (mm3)

    Demilly et al., 2007

    0

    50

    100

    150

    0 2 4 6 8

    % fr

    om in

    iti al

    v al

    ue

    hours after sowing

    moisture content (% dry weight)

    Area

    Volume

    • Brassica napus

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Two examples of application on

    crop species • Hardseededness in lucerne

    Cuticle Palisadic layer Malpighi cells

    Nutritive cells

    hard seed section fresh seed section

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 5

    • Ageing in oilseed rape

    Longitudinal sections of Medicago sativa seed coat MO x 63, PTM IMAC, SFR QUASAV

    OSR Cultivar

    Standard germination (%)

    Germination rate (%)

    1 97 99

    2 99 100

    3 90 99

    4 92 98

    5 98 99

    hard seed section fresh seed section

    Abnormal seedlings usually obtained after ageing tests

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Power of digital imaging for

    studying germination • 15 samples of each species

    Range of quality (%) Lucerne Oilseed rape

    Standard germination [40-65] [84.5-99.5]

    Abnormal seedlings [3-10] [0.5-13]

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 6

    • Automated imbibition and germination time courses at 20°C on 1 x 100 seeds (TP) previously

    weighed

    • Single seedling evaluation after 5 days

    Abnormal seedlings [3-10] [0.5-13]

    Ungerminated seeds [31-50] [0-2.5]

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Single seed analysis linked to

    seedling evaluation

    Area T0 Mass Initial

    Area

    increase Germination Seedling

    Single seed germination database obtained after image analysis (extract)

    Sowing time Image acquisition Image analysis

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 7

    0

    10

    20

    30

    40

    50

    60

    70

    80

    90

    100

    0 24 48 72 96 120

    G e

    rm in

    at io

    n (

    % )

    Time after sowing at 20°C(hours)

    11

    12

    13

    21

    22

    23

    31

    35

    36

    41

    42

    43

    51

    52

    53

    Sample date Camera zone seed

    Area T0

    mm²

    Mass

    (mg)

    Initial

    length (mm)

    increase

    (%)

    Germination time (hours)

    Seedling type

    1 101102 1 hg 11 5.1 2.3 3.7 0.0 56 PA

    1 101102 1 hg 12 5.4 2.1 3.9 5.4 SD

    1 101102 1 hg 13 5.6 3 3.7 28.3 20 PN

    1 101102 1 hg 14 5.4 2.4 3.8 1.4 PN

    1 101102 1 hg 15 3.5 1.3 3.1 3.2 36 PA

    1 101102 1 hg 16 4.6 1.4 3.7 5.6 SD

    1 101102 1 hg 17 4.6 2 3.4 6.5 35 PN

    1 101102 1 hg 18 5.2 2.3 3.4 -4.3 SD

    1 101102 1 hg 19 4.1 2.2 3.2 6.3 SD

    1 101102 1 hg 21 6.0 2.6 4.1 3.1 39 PN

    1 101102 1 hg 22 4.6 2.6 3.3 5.6 SD

    1 101102 1 hg 23 5.3 2.7 3.8 3.5 SD

    1 101102 1 hg 24 4.6 2.2 3.6 6.5 61 PN

    1 101102 1 hg 25 4.5 1.5 3.5 4.9 29 PN

    1 101102 1 hg 26 6.1 2.1 3.8 46.1 20 PN

    1 101102 1 hg 27 3.6 1.3 3.1 6.2 18 PN

    1 101102 1 hg 28 4.3 1.4 3.5 6.9 SD

    1 101102 1 hg 29 5.5 2.8 3.5 2.0 SD

    1 101102 1 hg 31 5.0 2.5 3.7 3.7 SD

    1 101102 1 hg 32 5.0 2.1 3.5 63.4 21 PN

    1 101102 1 hg 33 5.6 2.7 4.0 3.3 SD

    1 101102 1 hg 34 4.1 1.8 3.4 6.3 SD

    1 101102 1 hg 35 4.2 1.1 3.3 0.0 55 PN

    1 101102 1 hg 36 4.1 1.6 3.4 5.5 PN

    1 101102 1 hg 37 3.7 1.0 2.8 32.7 SNG

    Automated germination time courses

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Seed coat thickness and

    hardseededness in Lucerne • X-rays before sowing seeds to detect hard seeds on

    4 lots containing more than 40% of hard seeds

    Hard seeds sorted after 8 hours of

    HS HS HS

    HS HS

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 8

    0

    20

    40

    60

    80

    100

    1 2 3 4

    N o

    rm al

    g e

    rm in

    at io

    n (

    % )

    Seed lot

    No scarification Global scarification Hard seeds scarification

    after 8 hours of imbibition

    Hard seeds have all adherent testa

    HS

    HS HS HS HS

    HS

    HSHSHSHSHS

    HS

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Imbibition with or without seed

    scarification in lucerne

    Mean after mechanical scarification (25 seeds per lot)

    8.0

    9.0

    10.0

    Se e

    d a

    re a

    m e

    as u

    re d

    b y

    im ag

    e a

    n al

    ys is

    (m m

    ²)

    Imbibition of 15 seedlots at 20°C

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 9

    Mean before mechanical scarification (100 seeds per lot)

    3.0

    4.0

    5.0

    6.0

    7.0

    0 1 2 3 4 5 6

    Se e

    d a

    re a

    m e

    as u

    re d

    b y

    im ag

    e a

    n al

    ys is

    (m m

    ²)

    Time after sowing (hours)

  • G r o u p e d ’ É t u d e e t d e c o n t r ô l e d e s V a r i é t é s E t d e s S e m e n c e s

    Oilseed rape imbibition at 20°C is

    less important but also significant • 15 seed lots from 5 cultivars

    9.00

    10.00

    Ss e

    e d

    v o

    lu m

    e o

    b ta

    in e

    d b

    y im

    ag e

    a n

    al ys

    is (

    m m

    3 ) 11

    12

    13

    3030thth ISTA Symposium ISTA Symposium -- Antalya Antalya -- 14 14 JuneJune 20132013 10

    4.00

    5.00

    6.00

    7.00

    8.00

    0 2 4 6 8 10 12

    Ss e

    e d

    v o

    lu m

    e o

    b ta

    in e

    d b

    y im

    ag e

    a n

    al ys

    is (

    m m

    Time after sowing (hours)

    13

    21

    22

    23

    31

    35

    36

    41

    42

    43

    51

    52

    53

    Cultivar Thousend seed weight (g)

    1 5.0

    2 4.6

    3 4.8

    4 4.8

    5 6.2

  • G r o u p e d ’ É t u d e e t d e

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