SEED DEVELOPMENT

(MATURATION)

JULIO MARCOS-FILHO
DEPT. OF CROP SCIENCE
USP/ESALQ
 INTRODUCTION
Start Î flower induction and differentiation

Seed development : a sequence of events

controlled by the genotype

Maturation: set of successive stages in

preparation for successful seed germination

Flowering and pollination are not uniform in

the same plant or within a population 2

INTRODUCTION
Initial studies: aimed at determining phenological
differences among species and the optimum harvest
time

First proposed criteria to identify seed maturity:

- Time from planting or seedling emergence to
harvesting
- Seed moisture content and morphological
characteristics to identify seed maturity
3
 SEED DEVELOPMENT
Delouche (1971) :

Seed maturation is a process that comprises a

set of morphological, physical, physiological
and biochemical events that occur from ovule
fertilization to the moment in which seeds
become physiologically independent of the
parent plant

DESICCATION
RESERVE DEPOSITION (PHASE IV)
(PHASE III)
Embryo dry weight

CELL DIVISION AND

ELONGATION
(PHASES I and II)

HISTODIFFERENTIATION

Days after flowering

Dure III (1975) 5
 GENERAL PARAMETERS TO CHARACTERIZE SEED
MATURATION

1960s and 1970s: individual seeds x plant

population
Seed sampling at pre-defined intervals and
identification of parameters associated
with maturation progress
Definition of seed changes in seed technology
research
6

GENERAL PARAMETERS TO IDENTIFY SEED

MATURATION

SEED MOISTURE CONTENT

Ovule fertilization and evolution during
maturation
Dry fruits x Fleshy fruits
Final dehydration

7
 Fresh weight

Moisture
content
Dry weight

TIME

Variations in moisture content, fresh weight and dry

weight during maturation of seed produced in dry (-
(- - -)
or fleshy fruits ( ).
(Carvalho and Nakagawa, 2000) 8

GENERAL PARAMETERS TO IDENTIFY SEED

MATURATION
SEED SIZE

R5 R6

Soybean pod and seed development (Ritchie et al., 1994) 9

 GENERAL PARAMETERS TO IDENTIFY SEED
MATURATION
SEED SIZE

R6 R7 R8 10

Species Period (days) Reference

Cotton 21-28 Carvalho (1972)

Soybean 64 Jacinto and Carvalho (1974)

Sorghum 15-20 Nagai (1973)

Wheat 40 Carvalho and Yanai (1976)

Days after flowering necessary for seeds of different species to attain

maximum size during maturation (Carvalho
(Carvalho and Nakagawa, 2000)

11
 DRY WEIGHT
number of cells

dry weight
Number of cells/seed x 10-6
40

(Dry weight (mg/seed)

6

30
4
20

2
10

2 5 8 11
Egli Frut development period (Phase I) 12

GENERAL PARAMETERS TO IDENTIFY SEED

MATURATION
SEED MOISTURE CONTENT + DRY WEIGHT
Moisture content

Dry weight

13
Egli
 Moisture content
Species Reference
(%)

Cotton 50-60 Carvalho (1972)

Peanut 47-50 Carvalho et al. (1976)
Oat 45 Frey et al. (1958)
Field bean 38-44 Neubern and Carvalho (1976)
Maize 25-30 Hunter et al. (1991)
Soybean 50 Andrews (1966)
Sorghum 23-30 Kersting et al. (1961)
Wheat 40 Carvalho and Yanai, 1976

Seed moisture content of different species at the time they reach

reach
maximum dry weight during development
14

GENERAL PARAMETERS TO IDENTIFY SEED MATURATION

GERMINATION

Primary root protrusion x Normal seedlings

Germination x Dormancy during maturation

VIGOR
15
 Germination
Species (days after anthesis)
Rye 05
Wheat 05
Sorghum 06 to 10
Clover 10
Cotton 22
Soybean 38

Initial germination (days) after ovule fertilization in some cultivated

cultivated
species (Delouche
(Delouche,, 1971) 16

Moisture Seed dry Vigor

Days after Germination
content weight (mg dry weight /
sowing (%)
(%) (mg/seed) seedling)
78 55.3 20.40 05 5.0
84 50.6 25.34 28 4.07
90 48.1 28.80 77 7.47
96 39.8 34.10 37 7.69
102 27.2 36.44 11 4.86
108 14.1 34.10 09 4.88
111 11.0 37.24 06 5.53
117 8.8 37.54 81 9.57
123 9.9 37.90 83 10.72
126 10.7 37.84 65 9.34

Variation of seed moisture content, dry weight, germination and vigor

during wheat maturation, cv. IAS -54. (Carvalho and Yanai, 1976) 17
 DETERMINATION OF PHYSIOLOGICAL MATURITY

CONCEPTS
a) Seed maturity is identified by the maximum
dry matter accumulation
b) Seed physiological maturity is reached
when there are no significant increases in
seed dry weight
c) Seed physiological maturity occurs when
seeds reach maximum dry weight,
germination, and vigor 18

DETERMINATION OF PHYSIOLOGICAL MATURITY

PREVAILING IDEA
Seed physiological maturity = maximum dry weight

“Relative Maturity”, “Morphological Maturity”, “Mass

Maturity”, “Harvest Maturity”, “Agronomic
Maturity”, “Time of Harvest”

Determination of physiological maturity in individuals

or in plant communities
19
 MOISTURE CONTENT SIZE

VIGOR

DRY WEIGHT

GERMINATION

20

Moisture Seed dry weight

Days after sowing 21
content (%) (mg/seed)
64 72.9 61.4
70 60.5 158.4
76 56.8 171.0
79(†) 43.9 202.8
82 38.2 203.8
88 21.8 194.8
94 41.0 208.6
100 30.0 206.4
106 18.1 194.8
Mean values of moisture content and dry weight during maturation of
bean seeds, cv Carioca (Neubern and Carvalho, 1976)
 DETERMINATION OF PHYSIOLOGICAL MATURITY

Difficult to identify the exact point of seed

physiological maturity (maximum dry weight)
NEED TO INCREASE PRECISION:
- Number of statistical replicates
- Reduce harvesting intervals
- Additional care in weighing

Use of 14 C to monitor reserve accumulation

22

DETERMINATION OF PHYSIOLOGICAL MATURITY

Visual indicators of seed physiological maturity

MAIZE:
“Black Layer”
“Milk Line”
SOYBEAN:
Pod and Seed Color

23
 PM

100

80

Percentage
60
Green
40 Yellow
Yellow, typical of cultivar

20

49 63 77 90

Days after R3

Marcos-Filho (1979)

24

25

“Black Layer”

(Mississippi State University extension service, http://msucares.com/crops/corn/corn2.html)

 Mature

Maturation of
tomato

Physiological potential

“Overripe”
26
Contreras

Mean period†
Species
(days)
Lettuce 0
Onion -4
Tobacco 0
Maize
Inbred 0
Single hybrid -5
Double hybrid -10
Soybean -7
Tomato +10
Wheat -6
Average and range in days before or after physiological maturity
maturity and seed vigor
of different cultivated species (TeKrony and Egli, 1997)
(†) (-
(-): days before seed maximum dry weight 27
(+): days after seed maximum dry weight
 Days after Germination(‡) Soluble Sugars(‡) Proteins(‡)
flowering FD† SD FD SD FD SD

26 00 68 1.1 0.1 0.57 0.09

32 00 68 1.5 0.2 0.51 0.18

42 65 -- 2.2 0.2 0.62 0.25

48 54 93 1.6 0.4 0.48 0.19

Percentage germination, leakage of sugar and protein from soybean

soybean seeds, cv
Chippewea, submitted to drying at different maturation stages
(Adams and Rinne, 1983)
(†) FD – seed threshing and fast drying immediately after harvest;
SL – slow seed drying still attached to pods.
(‡) Germination (%); Seed leachate (mg/seed) 28

Dry weight Physiological Potential

Days after anthesis

Contreras
Maturation of tomato 29
 SEED MATURITY X HARVEST TIME

- Determination of harvest time

- Delayed harvest: problems

30

Date Yield Date Yield

(L1) (kg/ha) (L2) (kg/ha)
09/03/81 5.255 11/03/81 5.104

19/03/81 4.493 01/04/81 4.797

08/04/81 4.398 22/04/81 4.695

05/05/81 3.904 12/05/81 4.440

18/05/81 3.888 05/06/81 4.236

Effect of harvest time on maize seed yield in two locations of

Parana state, Brazil
(Hadlich, 1983) 31
 ⇑ RH
⇓ RH

⇑ RH

⇑ RH

⇓ RH
32

FIELD WEATHERING

33
 YEAR PM HT 14* 28*

1973 94 88 58 55

1974 85 83 85 41

1975 89 85 66 57

1976 83 28 13 07

1977 71 62 40 05

Vigor (accelerated aging) of ‘Kent’ soybean seeds as

influenced by harvest time for five experimental years
(TeKrony et al., 1980).

PM = physiological maturity; HT = adequate

harvest time; 14 and 28 = days after HT 34

SEED MATURITY X HARVEST TIME

- Harvesting at physiological maturity?

- Variations in seed moisture content and

in plant characteristics

- Visual parameters: black layer, milk line,

seed and/or fruit color, glumes and pedicel

35
 SEED MATURITY X HARVEST TIME
- Uniformity of maturation

Soybean Tomato

36
Cotton

II

III

Carrot
37
 Umbel Umbel / Proportion Germination First Count 1000 seeds
Order Plant (%) (%) (%) (g)
Primary 1 11 75 54 2.28
Secondary 11 58 65 43 2.15
Tertiary 24 31 54 37 1.99

Number of umbels per plant, contribution to plant seed production

(%), germination (%), vigor (germination first count - %) and weight of
1000 seeds associated with umbel order in carrot (Nascimento, 1991)

38

RESERVE ACCUMULATION DURING SEED

DEVELOPMENT

- Sequence of genetically programmed events

- Monocots Î endosperm; Dicots Î cotyledons

- Final yield: seed number + seed growth rate + duration

of seed fill

39
 RESERVE ACCUMULATION DURING SEED
DEVELOPMENT
1. NUTRIENT TRANSLOCATION FROM THE PARENT PLANT

Photoassimilates (sugars, amino acids and other

solutes) are transported via phloem

Plant reserve accumulation in endospermic seeds

Nutrient translocation to seeds

Short distance transport

40

NUTRIENT TRANSLOCATION – short distance

starchy endosperm

transfer cells embryo

chalazal region basal endosperm

pedicel
phloem
41
 RESERVE ACCUMULATION DURING SEED
DEVELOPMENT

1. NUTRIENT TRANSLOCATION FROM THE PARENT PLANT

Plant reserve accumulation in non-endospermic seeds

42

NUTRIENT TRANSLOCATION – short distance

f f

ee
h
ct

43
 RESERVE ACCUMULATION DURING SEED
DEVELOPMENT
2. NUTRIENT ASSIMILATION

Mature seeds Î two or three main types of reserves

Synthesis is parallel during seed development

Synthesis occurs in different cellular compartments:

starch, in amyloplasts
lipids, in spherosomes
proteins, in cytosol and endoplasmic reticulum

44

2. NUTRIENT ASSIMILATION

Carbohydrates

Lipids

Proteins

Nucleic acids

45
MAIZE
Dry Weight DNA
300 0,06

mg
200 0,04
mg

100 0,02

30 0,5
Proteins RNA
20 0,3
mg

mg
10 0,1

15 30 45 15 30 45
Days after fertilization 46
whole seed endosperm embryo

2. NUTRIENT ASSIMILATION

LEGUMES

Accumulation of carbohydrates precedes those of

lipids and proteins

47
 50 Proteins

Percentage 40

30 Lipids

20

Carbohydrates
10

30 50 70
Days after flowering

Changes in carbohydrate, lipid and protein contents during

soybean seed development (Adapted from Konno, 1979)
48

cell division cell elongation

140 1400 7

DNA
proteins (mg/cotyledon)
DNA (µg/cotyledon)

RNA (µg/cotyledon)

RNA
100 1000 5
proteins

60 600 3

20 200 1

10 14 18 22 26
Days after flowering 49
Stage Dry weight M.C. Proteins Oil Sugars
(mg/seed) (%) (mg/seed) (mg/seed) (mg/seed)
R4 0.2 78.4 ---- ---- ----
R5 5.9 83.1 2.5 0.1 1.3
R6 123.6 62.4 42.0 26.7 19.3
R7 194.2 51.9 71.9 31.7 30.8
R8 188.3 9.7 73.4 36.2 32.4

Variation in soybean composition during maturation

(Dornbos and McDonald, 1986)
50

HORMONES

Cytokinins

Gibberellins

Auxins

Abscisic acid

51
 ENVIRONMENTAL FACTORS AFFECTING SEED
DEVELOPMENT

- Soil Fertility
Adequate supply Î seed size and weight

- Water
Effects of water deficits
Water excess
Time of occurrence
Association with temperature stress
52

Association of water deficit and thermal stress during soybean

seed filling (Franç
(França Neto and Krzyzanowski)
53
 alternate dry
and humid
periods

water deficit
water deficit

APROSMAT, 2006 54

ENVIRONMENTAL FACTORS AFFECTING SEED

DEVELOPMENT

- Temperature

Critical value: 35oC during seed filling

“Forced maturation” and greenish seeds

55
 56

Tetrazolium 1-3
Germination (%) Emergence (%)
(%)
Color
3 3 3
Initial months
Initial Initial months
months

Yellow 88 87 75 73 87 88

Green 60 19 52 20 58 38

Soybean: presence of green seeds and effects on

germination and vigor
(Scheren and Tolentino Jr, 2005)

57
ENVIRONMENTAL FACTORS AFFECTING SEED
DEVELOPMENT

- Light
Photosynthesis x plant and seed performance
Retention of flowers and fruits

- Seed position on the plant

58

ENVIRONMENTAL FACTORS AFFECTING SEED

DEVELOPMENT

Seed 4-stage developmental pattern: cell division,

cell elongation, reserve accumulation, desiccation

Seeds do not germinate while attached to the

parent plant

During most of seed development:

Formation and activity of enzymes involved in
synthesis processes, managed by m-RNA
59
 METABOLISM REVERSION FROM DEVELOPMENT TO
GERMINATION

Metabolic reversion from synthesis to hydrolysis

Synthesis and activity of ABA
High concentration during embryogenesis

Synthesis and activity of ABA

Decrease during late maturation desiccation

Management by m-RNA
60

HORMONE INFLUENCE

cyt gib aux

Seed dry
ABA weight

Fertilization Maturity

61
 Desiccation is the “trigger” to switch to metabolism
reversion

synthesis mobilization

Does drying play a role in this switch?

Henk Hilhorst
62

DESICCATION TOLERANCE DURING SEED

MATURATION
Desiccation during seed maturation:
developmental pattern, driving seed embryo to a
quiescent stage in preparation for germination

Desiccation Tolerance:
The ability to recover biological functions after
drying to equilibrium with moderately dry air and
then resume normal function when rehydrated
This ability depends on the ability to maintain
membrane structure and prevent protein denaturation
63
 DESICCATION TOLERANCE DURING SEED
MATURATION
Desiccation Tolerance:
Intolerant Phase:
Cell division and elongation + part of reserve
accumulation period
Tolerant Phase:
Final phase of reserve accumulation
Premature and rapid desiccation:
Damage to enzyme and protein synthesis
Loss of cell turgidity, damage to cell membranes,
enzymes, proteins and nucleic acid structure
64

DESICCATION TOLERANCE DURING SEED MATURATION

PROTECTIVE SUBSTANCES OR MECHANISMS

- LEA (late embryogenesis abundant) Proteins

- Antioxidant systems

- Soluble sugars: raffinose, stachyose, sucrose

- “Heat shock” proteins

- Slow drying
65