2019

GERMINATION
John Hampton
Seed Research Centre
Bio-Protection Research Centre
Lincoln University
New Zealand
Germination
• “the events up to visible radicle
emergence”
- a physiologist’s definition

• “all the processes up to and including the

production of a normal seedling”
- an agronomist’s definition

• germination begins with the intake of

water, and for the latter definition ends
when the young seedling can manufacture
its own food.
Successful germination therefore requires the
growth of the embryo into a small plant with
• a root system capable of absorbing water and
dissolving mineral nutrients
• a green leaf surface capable of photosynthesis

Initially food is provided from within the embryo

Starch, protein, enzymes sugars, amino
oil acids, fatty acids

Germination ends when photosynthesis begins

(seedling independent of storage material)
Cotyledons
= The leaf of an embryo functioning as a
storage or absorptive organ

One cotyledon = monocotyledon

e.g. cereals (where the cotyledon is called
the scutellum)

Two cotyledons = dicotyledon

e.g peas/beans
Germination may be Hypogeal or Epigeal

Hypogeal  cotyledons remain below the soil

with the seed coat

Epigeal  cotyledons carried above the soil

surface
Hypogeal Germination – Pisum sativum
Epigeal Germination – Phaseolus vulgaris
Requirements for germination

• Water – to raise the moisture content of the

embryonic tissue to 80 – 90%
• Oxygen – to allow the increased respiration
that accompanies the reactivation of the
embryo
• Favourable temperature – species dependent.
Range may be wide – many species germinate
best at around 25°C
Water
• Quiescent (resting) seeds are:
i) low in moisture
ii) inactive metabolically

• They therefore need water for:

i) enzyme activation
ii) breakdown, translocation and use of reserve
storage materials

• The amount needed varies with species and their

adaptation to the environment
Oxygen

• Required for the germination of most species

• Respiration increases quickly during
germination and  adequate supply of oxygen
is needed
• If insufficient oxygen, respiration changes from
aerobic to anaerobic, with the production of
ethanol and other toxins
Temperature

• All stages of germination are affected by

temperature:
i) minimum = lowest temperature at which
germination will proceed
ii) optimum = temperature giving the greatest
germination in the shortest time
(between 15 and 30°C)
iii) maximum = temperature just before that
which begins to break down
proteins (between 30 and 40°C)
Approximate temperature range and thermal optimum
of germination for various species
Range Thermal optimum
1. Seeds germinating well only at low (oC) (oC)
temperatures
leek 5-25 15-20
celery 5-25 10-15
cyclamen 10-20 15-18

2. Seeds germinating well only at relatively high

temperatures
cucumber 15-40 30-35
tomato 10-35 25
sorghum 10-40 35

3. Seeds germinating in a large temperature range

cabbage 5-40 25-30

carrot 5-40 25-30
barley 0-35 25-30
Time in days needed to obtain maximum germination at
the thermal optimum

Species Thermal optimum Time

(⁰C) (days)

Onion 25-30 4-5

Oat 25-30 2-3
Cabbage 25-30 2
Cyclamen 15-18 15-20
Doulas fir 25 50-70
• Seeds of many species need daily
fluctuating temperatures for optimum
germination:

i) dormancy breaking

ii) shift in inhibitor/promoter balance?

(it is known that ABA levels in seed are
lower at low temperatures)
Light

• A small number of species also require light

for germination
• Both light intensity and quality (wave length)
are important
• Light intensity required is 1100 – 2500 lux
(sunny day approx. 108,000 lux; overcast day
approx. 16,200 lux)
• Light quality: far-red light (>700 nm) inhibits
germination while red light (660 – 700 nm
promotes germination
• Phototreversible germination in seeds
involves phytochrome (a protein pigment)

Red (660 nm)

dormancy PR PFR germination
Far-Red (730 nm)
Inactive Active
Form Form

Dark
• Exposure to red light converts phytochrome
to the physiologically active far-red
absorbing form  germination proceeds

• Exposure to far-red light converts

phytochrome to the physiological inactive
red-absorbing form  germination blocked
• Phytochrome is synthesised in the seed as
PR (the physiologically inactive form), so
that most seeds required some light to
convert PR to PFR

• For many species a brief flash of light is all

that is required for this process
Pattern of Seed Germination

• Imbibition
• Enzyme activation
• Breakdown of storage tissues
• Initiation of embryo growth
• Protrusion of the radicle
• Seedling establishment
Imbibition

• = the uptake of moisture by absorption from

the germination media and hydration of the
seed tissues

• the first key event that moves the seed from

its quiescent state to the resumption of
embryo growth
Phase I = imbibition phase
•The seeds take up water, allowing the dry cells to
reconstitute themselves, and particularly their cell
membranes
Phase II = lag phase
•Water uptake stops
•The seed switches to germination mode and begins
metabolic activity
•Food reserves are mobilised
Phase III = germination phase
•Further water uptake for growth
•Cell elongation and division
•Enzymes activated and synthesised
•Radicle emergence
Enzyme activation

• Begins during Phases I and II of imbibition

• Enzyme activation during Phase II allows:
i) breakdown of stored tissue
ii) transfer of nutrients to the growing point
iii) synthesis of new enzymes
iv) production of sucrose and other energy
products
The Germination Test

Objective
• To determine the maximum germination potential
of a seed lot
• To provide data for comparing the quality of
different lots and also estimating field planting
value
Conditions
• Controlled and standardised
• Optimum for the species to allow regular, rapid
and complete germination
e.g. from the ISTA Rules
Barley  20°C for 7 days after first
having broken dormancy
Perennial
ryegrass  20/30°C for 14 days after first
having broken dormancy
Pea  20°C for 8 days
Pine  20°C for 28 days
Pansy  20/30°C for 21 days after first
having broken dormancy
Germination

…the emergence and development of the

seedling to a stage where its essential
structures indicate whether or not it is able to
develop further into a satisfactory plant under
favourable conditions in soil.

(ISTA 2014)
Percentage Germination

…the proportion by number of seeds which have

produced seedlings classified as normal under
the condition of and within the timeframe of
the test.
Normal Seedlings

…show the potential for continued

development into satisfactory plants when
grown in good quality soil and under
favourable conditions of moisture,
temperature and light.
…have all the essential structures (root
system; shoot axis; cotyledons; terminal
buds; coleoptile) needed for continued
development.
1. Intact seedlings: seedlings with all their
essential structures well developed,
complete, in proportion, and healthy.
2. Seedlings with slight defects: seedlings
with certain slight defects (e.g. slight
retardation of the primary root; primary
leaf with limited damage), provided they
show an otherwise satisfactory and
balanced development.
3. Seedlings with secondary infection:
seedlings affected by micro-organisms
from sources other than the seed.
Abnormal Seedlings

…do not show the potential to develop into a

normal plant.
1. Damaged seedlings: essential structures
missing or severely damaged.
2. Deformed seedlings: weak
development/physiological disturbances.
3. Decayed seedlings: death/decay resulting
from seed infection.
Ungerminated Seeds

1. Hard seeds: seeds which have not

imbibed water but are viable.
2. Fresh seeds: seeds (other than hard
seeds) which remain clean and firm (but
probably dormant).
3. Dead seeds: seeds which are not fresh or
hard and have not produced any part of a
seedling.
General Principles

• Use seeds from the pure seed fraction of

the purity test (or pure seeds selected at
random from the working sample).
• Test 4 replicates of 100 seeds.
• Use the test method prescribed for the
species in the ISTA Rules.
Methods Using Paper

1. TP (top of paper)
• Seeds are geminated on top of one or more
layers of paper
• Transparent boxes with lids
• Petri dishes
• On trays in germination cabinet

• Must prevent papers drying out

2. BP (between paper)

• Seeds are germinated between two layers of

paper (e.g. rolled towels)

• Place rolls in upright position

• Cover with plastic bag to prevent drying out

Sand

• Seeds are placed on a level layer of moist

sand and covered with 10 – 20 mm
uncompressed sand depending on size of
the seed.
• Sand must not be too wet.
• Sand may be used instead of paper when
the evaluation of a diseased sample is
impracticable because of contamination of
the paper substrate.
Method requirements
1. Moisture
• Sufficient for germination but not excessive
• Water pH 6.0 – 7.5

2. Temperature
• Uniform throughout the germination apparatus
• Within  2°C of the specified temperature
• Where alternating temperatures specified (e.g.
20/30°C), maintain lower temperature for 16h
and higher for 8h.
3. Light
•Not required for most species, but better developed
seedlings are produced if some light available
•In some tropical and subtropical grasses, light may
promote germination of dormant samples (e.g.
Bothriochloa spp.; Chloris guyana)
4.Choice of method
•The ISTA Rules often provide alternate methods (e.g.
soybean: BP; S 20 – 30; 25°C)
•Choice of method depends on facilities available and
experience of the analysts
Seed Quality: Germination Standards

• Most countries have some form of legislation

designed to provide legal protection to both
buyers and sellers
• This legislation may contain minimum
standards for seed quality components (e.g.
germination)
• These may apply for domestic and also
imported seed lots
Examples of minimum germination standards for
imported seed lots

perennial ryegrass = 60% (South Africa);

80% (Canada)
red clover = 70% (Argentina);
85% (Chile)

Do these standards protect the buyer?

Table 1. Some possible germination test results for seed lots which
meet an 80% minimum germination standard

Seed lot Normal Abnormal Dead Seeds

Seedlings Seedlings (%)
(%) (%)
1 80 18 2

2 80 3 17

3 80 9 11

4 80 15 5
These minimum germination standards are saying:

“it is acceptable to import and sell seed lots

which are physiologically deteriorated, and
therefore highly likely to have trouble
performing once sown”
 Should seed quality legislation exist at all?

• New Zealand – no seed laws apart from

biosecurity for imports
– The onus is on the purchaser to check the
quality first
– Do this by asking to see the Seed Analysis
Certificate

• Most other countries have historically

considered seed laws important and still have
them
Do seed germination standards serve any
purpose?

Yes, if exclude very poor germinating lots

Yes, if standard set at a biologically acceptable

level (e.g. 90%)

Probably not if standard set too low (e.g. 60%,

70%, 80%)
Bio-Protection Research Centre
PO Box 85084
Lincoln University
Lincoln 7647, New Zealand
P + 64 3 423 0932
F + 64 3 325 3864
www.bioprotection.org.nz