SEED VIABILITY EQUATIONS

AND NOMOGRAPHS
 SEED SURVIVAL CURVE
 It is a typical curve produced plotting
germination (%) against time
 VIABILITY EQUATIONS

They are mathematical models developed to predict storage life

Roberts (1973) noted that the relationship between temperature ,

moisture content and period of viability in 5 orthodox species

The seed life span was normally distributed

Upon losing viability at given moisture ,temperature the relation

between germination(%) and storage is sigmoidal
VIABILITY EQUATIONS
 VIABILITY EQUATIONS
 Conversion of germination (%) in Probits
 By this ,the sigmoidal curve is linearised
Now the relation between probit germination and storage is

Basic equation
υ = ki – P/ϭ 1

where υ = germination (%) in probits

p = mean viability period
ϭ = reciprocal of slope
ki = intercept
VIABILITY EQUATIONS
 VIABILITY EQUATIONS
 Linear relation between log of moisture and
time taken to fall by one probit is

Log ϭ = K – C w logm 2

 Similarly relative effect of temperature on

longevity is given as

Log ϭ = K – CH t – CQ t2 3
 VIABILITY EQUATIONS
where
K = intercept on Y axis
C w = moisture constant
m = moisture content
CH , CQ = temperature constants
t = temperature(o C)
 VIABILITY EQUATIONS
 Combining equations 2 & 3
Log ϭ = KE - C w log m - CH t – CQ t2 4

Substitute equation 4 in 1
υ = ki – P/10 KE - C w log m – C t – C t2
H Q

This is called “improved viability equation”

 VIABILITY EQUATIONS
 Where
υ = final viability (%)
K = initial viability
KE ,C w = moisture constant
m = moisture content
CH , CQ = temperature constants
t = temperature
Uses of viability equations

Estimate the
final viability

Estimate the designing and

storage life uses managing of seed
banks

estimate the
Estimate the storage equilibrium
temperature required moisture content
 NOMOGRAPHS
 Nomograph shows relationship between scales for mean
longevity and required viability at the end of storage under
given temperature and moisture
 This makes predicting seed survival easier than using three
equations
 Its value being limited by a number of impacting features :
1) Differences in longevity under same storage conditions .
2) Effect of pre-storage conditions such as high temperature post
harvest drying and mechanical damage .
3) Change in oxygen partial pressure during hermetic storage .
METHOD OF USING NOMOGRAPH
To estimate the time taken for viability at a
given temperature and moisture content
 Put a ruler on required temperature and
moisture
 Note the value indicated on scale c( mean
viability period )
 Using this point as pivot , move the ruler to
indicate required viability (%) on scale e
 The value on scale d is time taken for viability
NOMOGRAPHS
 PREDICTION OF RELATIVE
STORABILITY AND LONGEVITY
OF SEED LOTS
WHY PREDICTION OF STORAGE IS
IMPORTANT ?
 It helps to anticipates loss of seed quality
and replenishes old seed stocks
 Helpful to seed companies and germplasm
repositories
Relation of RH, temperature and
moisture in linseed
LIMITATIONS

 Cultivar and harvest variability

 Pre and post harvest conditions
 Oxygen pressure during storage
 Fluctuating environment conditions
Different changes associated with
loss of vigour and viability during
storage
Viability
It is the capacity of seed to germinate
and produce normal seedling
Vigour
The properties of seed with rapid ,
uniform emergence and development of
normal seedlings under wide range of
field conditions
 • Non viable seeds develop off colour
Physical
changes

Physiologica • Germination
l and • respiration
biochemical
changes

• Disruption of membrane bound cell organelles and

enzymes
Membrane
permeability
Changes associated with vigour loss
Physiological
Physical and Membrane
changes biochemical permeability
changes

Germination Disruption
Non viable of
seeds membrane
develop off respiration bound cell
colour organelles
and enzymes
Changes associated with vigour loss

Micro flora and

Cytogenetic changes
deterioration

Increased mutations Low quality seeds

or chromosomal affected by pests and
abberations diseases
 Physical changes

Non viable seeds

develop off colour
Physiological and biochemical changes

Delay of radicle Slower seedling

emergence growth

GERMINATION

Reduced tolerance
Abnormal seedling
to stress
 MEMBRANE PERMEABILITY

Lipid
peroxidation

Disruption of
membrane
integrity

Free radical Inactivation

of protein
CYTOGENETIC CHANGES
Increased temperature ,
moisture , oxygen

Causes Chromosomal
abberations and mutations

Leads to pollen abortion in A1

Causes phenotypic mutations

in A2 , A3
MICRO FLORA AND DETERIORATION

 Germination depends on interaction

between seed quality and microflora
 Seeds affected by fungi shows reduced
germination or storability
 MICRO FLORA AND DETERIORATION

Lacks
Low quality Release of
membrane
seeds exudates
integrity

Invades seeds
Attracts soil
and
microflora
deteriorates

Poor quality Chemical Optimum

seeds treatment population
GENETIC BASIS OF SEED
LONGEVITY
• Seed testa structure and composition
• Protection against Reactive Oxygen species
(ROS)

• Heat Shock Proteins

• Antioxidants

• Lipid breakdown
• Sugars and Glassy state
SEED TESTA STRUCTURE AND COMPOSITION

 In Arabidopsis mutants , seeds deteriorated faster than

wild types by natural ageing .
 But accumulation of flavonoids enhance seed coat
imposed longevity.
PROTECTION AGAINST REACTIVE
OXYGEN SPECIES (ROS)

 ROS species affects longevity

 But deficiency of ROS scavenging pathway
shown less effect in Arabidopsis mutants
 HEAT SHOCK PROTEINS

 HSP reduces longevity.

 AB13 protein activates HsfA9 promoter which
in turn activates Hsf genes.
 ANTIOXIDANTS

 Tocopherols more in seeds

 They lipophilic antioxidants
 Deficiency in vitamin E reduces longevity
 This is due to mutation at loci (VTE1 and VTE 2)
 LIPID BREAKDOWN

 Phospholipase D (PLDα1) affects ageing in

Arabidopsis

SUGARS AND GLASSY STATE

 Sucrose non-fermenting related kinase (SnRK 1)

complex showed reduced longevity
 It impairs accumulation of sucrose content
APPLICATION OF PHYSIOLOGICAL
AND BIOCHEMICAL TECHNIQUES
FOR EVALUATION OF SEED AGEING
 SEED AGEING

 It is a natural process which involves cytological,

physiological, biochemical and physical changes in
seeds
 These changes reduce viability and ultimately
cause death of the seed.
 SEED AGEING

Seed viability

Seed ageing in
gene banks is
assesed through

vigour germination
TOOLS FOR ASSESSING SEED AGEING

• Cold tests
Physiological • Accelerated ageing
tests •
•
Hiltner test
Cool germination test

• TZ test
Biochemical • Conductivity test
• Vital colouring test
tests
PHYSIOLOGICAL TESTS
 METHODS FOR TESTING SEED
DETERIORATION
ACCELERATED AGEING TEST
 This test is used to determine germination capacity
 It determines the period of safe seed storage.
 The seed is exposed for a short period to double
stress condition like high temperature, and high
relative humidity
 High vigour seed will withstand these extreme
conditions
 Compare the AA germination with standard
germination
 ACCELERATED AGEING TEST

 AA germination similar to standard

germination – high vigour
 AA germination less than standard
germination – low vigour
 Hiltner test
 It used to determine the seed-borne
infection by Fusarium spp.
 This test is based on the fact that damaged
seeds is often weak and unable to
withstand adverse conditions during
germination
1) a test for seed health testing
2) A test to determine other types of
damages
 COLD TEST
 It is widely used for maize in North
America
 seeds are sown when temperature are
low (100 C – 25 0 )
 seed germination obtained by standard
laboratory test is not in positive
correlation with field emergence
 germination results obtained with applied
cold test represent the most precise
indicators of maize germination in the field
COLD TEST
 COOL TEMPERATURE TEST
 It is widely used measure of seed vigour to
evaluate cotton
 It determines if seed lots are suitable for
planting in cool soils or less than ideal
situations
 Cotton can germinate about 120C – 400C
 The test is conducted at 18 °C and incubate
for 7 days
COOL TEMPERATURE TEST
Normal seedlings are defined as
combined radicle + hypocotyl length of
4cm or longer
 Seed lots with greater than 60% are
considered good to excellent.
BIOCHEMICAL TESTS
 BIOCHEMICAL TESTS
(A)TETRAZOLIUM TEST
 This method was developed by Professor
Georg Lakon.
Objective : rapid assessment of seed viability
 It distinguishes between viable and dead tissues
of the embryo based on relative respiration
rate
 Only living cells contain dehydrogenase enzyme
 TETRAZOLIUM TEST

Bisect the seed sample

Treat the seeds with 1% of 2,3,5 – triphenyl tetrazolium chloride

Non viable seeds

Viable seeds
(pink colour) (colour less)
TETRAZOLIUM TEST
 Electrical Conductivity Test
 This test measures leakage of solutes mainly K+
 As the seed get aged , cell membrane become
less rigid
 This allows loss of solutes which increases
electric conductivity
 It provides a rapid indication of seed viability
for seed lots
 VITAL COLOURING TEST
 The principle is differential colouration of
live and dead tissues when dyes are applied
 Dyes like sulphuric acid , indigo caramine ,
aniline are used
 Stained tissue (blue)- dead tissue
 Unstained tissue (colourless) – live tissue
Possible questions
1)Write about viability equations?
2)Write about nomographs?
3)What are the changes associated with loss
of vigour and viability during storage ?
4)Write about physiological and biochemical
techniques to asses seed ageing
 REFERENCES
 SEED SCIENCE AND TECHNOLOGY – N.C.SINGHAL
 SEED QUALITY - BASRA
 SEED SCIENCE AND TECHNOLOGY – LAWRENCE O.
COPELAND AND MILLER B. MC DONALD