SEED DEVELOPMENT (MATURATION)

JULIO MARCOS-FILHO DEPT. OF CROP SCIENCE USP/ESALQ

INTRODUCTION
Start 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

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
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RESERVE DEPOSITION (PHASE III)

DESICCATION (PHASE IV)

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
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GENERAL PARAMETERS TO IDENTIFY SEED MATURATION

SEED MOISTURE CONTENT Ovule fertilization and evolution during maturation Dry fruits x Fleshy fruits
Final dehydration
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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 ( ). 8 (Carvalho and Nakagawa, 2000)

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 Cotton Soybean Sorghum Wheat

Period (days) 21-28 64 15-20 40

Reference Carvalho (1972) Jacinto and Carvalho (1974) Nagai (1973) Carvalho and Yanai (1976)

Days after flowering necessary for seeds of different species to attain maximum size during maturation (Carvalho and Nakagawa, 2000) (Carvalho 11

DRY WEIGHT
number of cells Number of cells/seed x 10-6 dry weight
6 30 4 20 2 10 (Dry weight (mg/seed) 40

Egli

11

Frut development period (Phase I)

12

SEED MOISTURE CONTENT + DRY WEIGHT

Moisture content

GENERAL PARAMETERS TO IDENTIFY SEED MATURATION

Dry weight

Egli

13

Species Cotton Peanut Oat Field bean Maize Soybean Sorghum Wheat

Moisture content (%) 50-60 47-50 45 38-44 25-30 50 23-30 40

Reference Carvalho (1972) Carvalho et al. (1976) Frey et al. (1958) Neubern and Carvalho (1976) Hunter et al. (1991) Andrews (1966) Kersting et al. (1961) 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
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Species Rye Wheat Sorghum Clover Cotton Soybean

Germination (days after anthesis) 05 05 06 to 10 10 22 38

Initial germination (days) after ovule fertilization in some cultivated cultivated species (Delouche, 1971) (Delouche, 16

Days after sowing 78 84 90 96 102 108 111 117 123 126

Moisture content (%) 55.3 50.6 48.1 39.8 27.2 14.1 11.0 8.8 9.9 10.7

Seed dry weight (mg/seed) 20.40 25.34 28.80 34.10 36.44 34.10 37.24 37.54 37.90 37.84

Germination (%) 05 28 77 37 11 09 06 81 83 65

Vigor (mg dry weight / seedling) 5.0 4.07 7.47 7.69 4.86 4.88 5.53 9.57 10.72 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
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MOISTURE CONTENT

SIZE

VIGOR

DRY WEIGHT GERMINATION

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Days after sowing 64 70 76 79() 82 88 94 100 106

Moisture content (%) 72.9 60.5 56.8 43.9 38.2 21.8 41.0 30.0 18.1

Seed dry weight (mg/seed) 61.4 158.4 171.0 202.8 203.8 194.8 208.6 206.4 194.8

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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
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DETERMINATION OF PHYSIOLOGICAL MATURITY

Visual indicators of seed physiological maturity MAIZE:

Black Layer Milk Line

SOYBEAN: Pod and Seed Color

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PM
100 80

Percentage

60 40 Green Yellow Yellow, typical of cultivar 20

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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
Contreras

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Species Lettuce Onion Tobacco Maize Inbred Single hybrid Double hybrid Soybean Tomato Wheat

Mean period (days) 0 -4 0 0 -5 -10 -7 +10 -6

Average and range in days before or after physiological maturity and seed vigor maturity of different cultivated species (TeKrony and Egli, 1997) () (-): days before seed maximum dry weight ((+): days after seed maximum dry weight

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Days after flowering 26 32 42 48

Germination() FD 00 00 65 54 SD 68 68 -93

Soluble Sugars() FD 1.1 1.5 2.2 1.6 SD 0.1 0.2 0.2 0.4

Proteins() FD 0.57 0.51 0.62 0.48 SD 0.09 0.18 0.25 0.19

Percentage germination, leakage of sugar and protein from soybean seeds, cv soybean 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)

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Dry weight

Physiological Potential

Days after anthesis

Contreras

Maturation of tomato

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SEED MATURITY X HARVEST TIME

- Determination of harvest time

- Delayed harvest: problems

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Date (L1) 09/03/81 19/03/81 08/04/81 05/05/81 18/05/81

Yield (kg/ha) 5.255 4.493 4.398 3.904 3.888

Date (L2) 11/03/81 01/04/81 22/04/81 12/05/81 05/06/81

Yield (kg/ha) 5.104 4.797 4.695 4.440 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

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FIELD WEATHERING

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YEAR 1973 1974 1975 1976 1977

PM 94 85 89 83 71

HT 88 83 85 28 62

14* 58 85 66 13 40

28* 55 41 57 07 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

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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
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SEED MATURITY X HARVEST TIME

- Uniformity of maturation

Soybean

Tomato

Cotton

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II

III

Carrot

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Umbel Order Primary Secondary Tertiary

Umbel / Plant 1 11 24

Proportion Germination First Count (%) (%) (%)

1000 seeds (g)

11 58 31

75 65 54

54 43 37

2.28 2.15 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

endosperm; Dicots

cotyledons

- Final yield: seed number + seed growth rate + duration of seed fill
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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
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NUTRIENT TRANSLOCATION short distance

starchy endosperm

transfer cells chalazal region

embryo basal endosperm pedicel phloem

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

Plant reserve accumulation in non-endospermic seeds

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NUTRIENT TRANSLOCATION short distance

f ee
h

ct

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RESERVE ACCUMULATION DURING SEED DEVELOPMENT 2. NUTRIENT ASSIMILATION

Mature seeds 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
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2. NUTRIENT ASSIMILATION

Carbohydrates Lipids Proteins Nucleic acids

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MAIZE
Dry Weight 300 mg 200 DNA 0,06 0,04 mg mg

100

0,02

30 Proteins mg 20 RNA

0,5 0,3

10

0,1

15

30

45 15 Days after fertilization

endosperm

30

45

46
embryo

whole seed

2. NUTRIENT ASSIMILATION

LEGUMES

Accumulation of carbohydrates precedes those of lipids and proteins

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50 40 30 20

Proteins

Percentage

Lipids

Carbohydrates 10

30

50

70

Days after flowering

Changes in carbohydrate, lipid and protein contents during soybean seed development (Adapted from Konno, 1979)

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cell division
140

cell elongation
1400 7

100

RNA proteins

1000

60

600

20

200

10

14 18 Days after flowering

22

26

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proteins (mg/cotyledon)

DNA (g/cotyledon)

DNA RNA (g/cotyledon)

Stage R4 R5 R6 R7 R8

Dry weight

(mg/seed) 0.2 5.9 123.6 194.2 188.3

M.C. (%) 78.4 83.1 62.4 51.9 9.7

Proteins Oil Sugars (mg/seed) (mg/seed) (mg/seed) ---------2.5 42.0 71.9 73.4 0.1 26.7 31.7 36.2 1.3 19.3 30.8 32.4

Variation in soybean composition during maturation (Dornbos and McDonald, 1986)

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HORMONES

Cytokinins Gibberellins Auxins Abscisic acid

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ENVIRONMENTAL FACTORS AFFECTING SEED DEVELOPMENT

- Soil Fertility Adequate supply - Water

seed size and weight

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

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Association of water deficit and thermal stress during soybean seed filling (Frana Neto and Krzyzanowski) (Fran 53

alternate dry and humid periods

water deficit water deficit

APROSMAT, 2006

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ENVIRONMENTAL FACTORS AFFECTING SEED DEVELOPMENT

- Temperature Critical value: 35oC during seed filling Forced maturation and greenish seeds

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Germination (%)

Color

Tetrazolium 1-3 (%)

Emergence (%)

Initial
Yellow Green

3
months

Initial 75 52

3
months

Initial 87 58

3
months

88 60

87 19

73 20

88 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

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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
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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
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HORMONE INFLUENCE

cyt

gib

aux

ABA

Seed dry weight

Fertilization

Maturity

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Desiccation is the trigger to switch to metabolism reversion

synthesis

mobilization

Does drying play a role in this switch?

Henk Hilhorst

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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
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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
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DESICCATION TOLERANCE DURING SEED MATURATION

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
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