Allanblackia seed

germination protocols

Prepared by Lucy Mwaura

For
Allanblackia domestication
workshop
23rd to 27th Oct 2006
Outline
 AB seed physiology
 Conditions required for germination.
 Possible Allanblackia seed dormancy
 Seed germination treatments
applied by partcipating countries
 Treatment methods that have been
use on a large scale
 Lesson learnt
Allanblackia seed physiology

 Fruit/seed weight

 Seed moisture

 Storage

 Dormancy

 Germination
Fruit/seed weight

-A single seed weight

ranged between 5-
10grams when fresh
-A kilo of fresh seeds
had between 90-100
seeds
-viable seeds per kilo?
( not confirmed)
-fruit weight ranged
between 2.5-5.5kg
Moisture content
 Seed moisture
-AB seed had moisture
content of up to 40% when
fresh.
-Loss of seed moisture was
found to be proportional to
loss in viability (desiccation
tolerance trial/biochemical
test 2004 )
-The rate of moisture loss is
high soon after extraction,
and decreased with time of
exposure to air.
Storage trials
 Storage
- Extracted seeds can be
stored at -20oC in
airtight aluminum bags
for at least one year
without loss of moisture
and viability. (desiccation
tolerance test of 2004) .
-Seeds remained viable
with high moisture while
still enclosed in the fruit
buried, in moist soil/sand
for three months.
Conditions required for
seed germination
 In Tanzania and Kenya use of moist sand
media has been used to pre-germinate seeds
 The germination beds should remain moist
during this period, however minimum watering
is recommended to avoid water logging( seed
would rot)
 Seeds in nursery beds should be put under
shade. (In Tanzania seeds are covered with
dried banana leaves).
 Temperatures should be between 25-30
degrees Celsius.
AB possible seed
dormancy
 Dormancy-is the physiological state that prevents normal growth
of most seeds when all the conditions are favorable.
 Seed coat dormancy: gelatinous sticky substances found
on seeds could contain chemical inhibitory compunds which make the
seed coat impermeable to water and gases, this growth inhibitors must be
leached out to allow germination (Forest Biology journal 2000).
 Embryo dormancy: have growth-inhibitors present and
lack growth promoters.
-The embryo in enclosed in endocarp that contains high oil content, this
could have chemical inhibitory compound that prevent germination
-Abscisic acid contained in the embryo makes the seed dormant until
certain native enzymes are triggered to permit breakdown of the
hormones ( intensive work required here)
Seed germination treatments
applied and results obtained by
specific countries
1. Cameroon.

 Results from an experiment investigating the germination potential of Allanblackia

seeds in a mixture of forest soil and sand (2:1) indicated that it took 18 months for
A floribunda and 19 months for A gabonensis to reach 50% germination level.

 24 months after the trial was set an overall 86% of seeds germinated.
Pretreatments experiments were developed to reduce the germination period

 Preliminary results from pretreated seeds of AB (immersion in 90% gibberellic acid

(GA3) for 72 hours indicate that a germination rate of 48.33 % can be obtained
after 7 months (A. floribunda).

 New experiment investigating the combined effects of GA3 concentration and

duration of immersion on 4800 A. floribunda seeds subjected to 3 immersion
durations and 4 GA3 concentrations in a factorial design; (experimental unit of 100
seeds) is being done in Cameroon.
Seed germination trial in
specific countries
2. Ghana

 Out of 13 accessions sown at the nursery the

overall germination was 0.02% as at June
2005.
 Another 21 accessions were sown in the green
house with three treatments, (with testa,
without testa and soaking in water for 24 hrs) in
early 2005, overall germination was 0.05% by
February 2006.
 ITSC also put 600,000 seeds on nursery beds
of (A parviflora), only 0.1% of seedlings was
transplanted into poly tubes.
Treatments that have been used on a large
scale ( best germination results so far )
3 Tanzania
 Extracted seeds are sown on seed-beds
 Burying the whole fruit in a hole and cover with
forest debris and then transfer germinated
seeds to the polyethylene tubes.
 Collecting newly emerged seeds and raise
in the polyethylene tubes( rat caches)
 Extracted seeds are soaked in water while
floater are discarded, the sinkers are removed
and nicked on the distal end and later soaked
in GA31000ppm( refer to note for GA3
preparation)
Seed germination treatments
from specific countries con’t
4. Kenya

 Germination capacity
-Seed germination capacity was low
and slow
-tap root emerged first within 3-
5months
-a fully developed seedling with two
alternate leaves occurred after 7
months of sowing.
-seeds that did not germinate after 5
months had an enlarged embryo
and remained dormant
-seed stained brilliant red indicating
viable tissues with TTZ test.
Pre-sowing methods

 sowing without treatment

- No seed treatment has been
found effective to improve
germination percentages.
-First trial 2004 done without
treatment (germination was
4%)
 Sowing with treatment
-seeds were stored for a
month, soaked in water 24hrs,
and dried for 2 days then
cracked to open the seed
coat( germination was 1%)
Pre-sowing methods cont’

 Preparation and soaking in GA-3

and KNO3
-Gibberellic acid (GA-3) was dissolved
in 1 ml ethyl-alcohol/water
250ppm, 500ppm,1000ppm( refer to
notes)
-Potassium nitrate(KNO3 )
0.2%,0.5%,and 1%(ref: ISTA news
bulletin No 130 Oct 2005)
-Combined treatment (KNO3 0.2% and
250ppm)
-one seedlot was not treated
Treatments cont’
 Experimental set up
-seeds were put in
various treatment and
soaked for 48hrs at room
temperature.
-Deep flower pots were
filled with sterile sand at
¾ level, sown seeds
were covered with a thin
layer of sand.
-similar set-up was put at
the nursery same time.
Germination progress.
 Sowing
-Dec 2005, seeds were sown
2 cm apart in containers, and
incubated at 25ºC night and
30ºC day temperature, the
sand was kept moist without
logging.
 Monitoring
-Minimum watering with
distilled water was done.
-Daily monitoring and
recording was continuously
done.
Germination chart
A stuhlmannii germination report

30

25

20

%germinated seeds 15

10

0
250ppm GA3 500ppm GA3 1000ppm GA3 KNO3 0.2% KNO3 0.5% KNO3 1.0 % Cold w ater 250ppm GA3 Control
soak 48hrs soak 48hrs soak 48hrs 48hrs 48hrs 48hrs soak 48hrs +KNO3 0.2%
48hrs
Treatments
Lesson learnt
 We still have a big challenge ahead of us to find ways to improve
seed germination.
 Use of local knowledge practice in Tanzania is so far the best
practice that improves seed germination.
 More research needs to be done using KNO3 and GA-3
treatments.
 Bioassay study will be important to find out whether hormones
ratios found in seeds have any influence on physiological seed
activities (germination? dormancy?).

 We need to set-up seed propagators in central nurseries where

environmental conditions can be controlled and temperatures
monitored.