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SEEDS CONSERVATION
IN GENE BANK
ElSayed Mohamed ElAzazi
Plants Ecology and Range Management Dept. 2015
North Sinai research Station Dec. 2014, by Dr. Elsayed Elazazi
Justifications
Genetic erosion
Habitat loss
Invasive species
Pollution
Climate changes
Photo from SK. Reservation
Conservation: the link between genetic
resources and utilization
Plant Genetic
Resources
Diversity
Conservation
Utilization
(immediate
or potential)
Maxted et al., 1997a).
Utilisation Products
(New varieties, new crops,
pharmaceutical uses, pure and
applied research, on-farm diversity,
aesthetic pleasure, etc
The exploration model
Classification
Evaluation
Utilization
Exploration
Conservation
Marshall and Brown, 1975
Conservation methods
In-situ conservation Ex-situ conservation
Conservation Products
(Seed, live plants, in vitro explants, DNA, pollen, data
In-situ
conservation
 nature reserves
 managed areas
 Biosphere reserves
 on-farm conservation
 Home gardens
 Seed Bank
 Field gene bank
 In vitro slow growth
 Cryobank,
cryopreservation
 Pollen storage
 DNA storage
 Botanical Gardens
Ex-situ
conservation
Seed Bank storage conditions
Long -
term
Base room
(–22 ºC,
≥ 100 years
Short-
term
Active
room (+4ºC,
40% RH)
≥ 25 years
Medium-
term
Freezing
room (–5
ºC, no frost
≥ 50 years
All data in Gene bank by accession number
All material exchange should by under MTA
Seed Bank
In vitro bank, slow
growth
Field Gene Bank
Cryopreservation bank
Seed Bank
Seed Processing
 Sample registration
 Seed cleaning
 Seed drying
 Seed moisture content
 Seed viability
 Seed packaging
 Seed storage
 Monitoring accessions
 Distribution of germplasm
 Regeneration
Photo by: M. Ehsan Dulloo
Registration
 In Field
 In all Seeds processing steps in seed bank
Seed
cleaning
North Sinai research Station 2014, by Dr. Elsayed Elazazi
Seed cleaning
 Collections in cloth and
paper bags held in drying
rooms.
 Collections cleaned to
remove empty or infested
seed and to reduce their bulk
- aim: 95% pure seeds
 All cleaning done by hand,
carefully.
Photo by: Kew 2011
Seed cleaning equipment
 Sieves, bungs & brush
 Dust extraction system (masks)
 Seed aspirator (blower)
 Stereoscope & dissection
equipment
 Washing facility
 Debris disposal
Photo by: ElAzazi
Photo by: KIB, China
Sieves Bag Crushing
Crushing
Photos by: Kew
Clean under running water method
Zig-zag Aspirator Handsorting
Photo by: ElAzazi Photo by: Kew
Seed drying
North Sinai research Station 2014, by Dr. Elsayed Elazazi
Why is it important to understand
seed-air moisture relations?
• Seed storage behaviour – response to water
removal
• Seed ripening – maturation drying
• Seed viability – impact of seed moisture
status on seed longevity
• Seed drying – how seeds dry, monitoring
seed moisture status
Water in seeds
•Determining seed
equilibrium humidity
(eRH) using a digital
humidity sensor
• Air and seed at
equilibrium inside
chamber
Seed drying methods
•Natural drying (sun/shade)
•Dry room
•Drying cabinet
•Incubator drier
•Silica gel /dried rice /
charcoal
Photo by: Kew
Sun/Shade drying
Sun drying in thin layers in
open crates allowing
plenty of air movement.
Protecting seeds from direct
sun to reduce risk of heat
damage
Photo by: Kew
Dry room
•Room Condition at 10-15% RH
and 10 - 25C (FAO / IPGRI Gene
bank Standards)
•Specifically designed for purpose,
taking into account maximum
• seed loading (cleaned & un
cleaned seed) and several other
factors.
10-15% RH and 10 - 25C (FAO / IPGRI Genebank Standards)
Photo from KIB China
Drying cabinets
10-15% RH and 10 - 25C (FAO / IPGRI Genebank Standards)
Photo by: Kew
Cooled incubators
(incubator driers)
•Cost effective way of drying small amounts
(up to 10 kg) of seed
•Usually run at 18ºC, giving an RH of ~15%.
•Dries 10kg of fresh seeds to 7% mc in ~ 3
weeks
•Incubator must run with refrigeration system
on constantly and must have auto defrost.
10-15% RH and 10 - 25C (FAO / IPGRI Genebank Standards)
Photo by: Kew
Desiccant
Air
Seed
Drying with desiccants
Photo by: Kew
Drying with desiccants:
silica gel bin drier
•Use a suitable container with a
tightly fitting lid (e.g. a plastic box or
plastic / metal drum).
•Add 20% by (container) volume
dried (10 - 20% RH) silica gel.
•Place bags of seed in container
•Check the RH of the silica gel
weekly; if too high, oven dry a quarter
part at 100ºC and re-mix with the
remaining gel.
•Drying time (~one month) depends on the initial
moisture content of the seeds, the amount of seeds,
the type of seeds and the dryness of the silica gel.
Photo by: Kew
North Sinai research Station 2014, by Dr. Elsayed Elazazi
What is ‘viability’?
• A viable seed is a living seed that has the
capability of germinating
• Empty seeds and seeds without embryos are
non-viable but they should not be called dead
seeds because they never were viable
• Dead seeds are seeds that were viable but
have subsequently died
Rao, N.K.; Hanson, J.; Dulloo, M.E.; Ghosh, K.; Nowell, A.; larinde, M 2006a.
Manual of Seed Handling in Genebanks8.
Why should seed viability be
determined?
1. It is very important that seeds stored in the genebank are
capable of producing plants when sown in the field.
2. They must have high viability at the start of storage and
maintain it during storage.
3. Seeds with a high initial viability will also survive longer in
storage.
4. Seed viability declines slowly at first and then rapidly as seeds
age.
5. It is important to know when this decline occurs in order to
take action to regenerate the accession. Excessive deterioration
will lead to loss of material.
How should viability be determined
Viability
Germination
test
Biochemical
tests
How many seeds should
be tested?
200 Seeds
(Rep. 1) 100Seeds
(Rep. 2) 100Seeds
R1
R2
R3
R4
R1
R2
R3
R4
G% 90%
seed germination test substrate
Top Paper (TP)
North Sinai research Station 2014, by Dr. Elsayed Elazazi
Between Paper (BP)
Agar method
North Sinai research Station 2014, by Dr. Elsayed Elazazi
Sand
North Sinai research Station 2014, by Dr. Elsayed Elazazi
Always dissect a few seeds before you start
North Sinai research Station 2015, by Dr. Elsayed Elazazi
ELAZAZI_GENEBANK@YAHOO.COM
+201002684667

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Genetic Resources, The role of the egyptian deserts gene bank to Conservation plant genetic resources lec 1

  • 1. SEEDS CONSERVATION IN GENE BANK ElSayed Mohamed ElAzazi Plants Ecology and Range Management Dept. 2015 North Sinai research Station Dec. 2014, by Dr. Elsayed Elazazi
  • 3. Genetic erosion Habitat loss Invasive species Pollution Climate changes
  • 4. Photo from SK. Reservation
  • 5. Conservation: the link between genetic resources and utilization Plant Genetic Resources Diversity Conservation Utilization (immediate or potential) Maxted et al., 1997a). Utilisation Products (New varieties, new crops, pharmaceutical uses, pure and applied research, on-farm diversity, aesthetic pleasure, etc
  • 7. Conservation methods In-situ conservation Ex-situ conservation Conservation Products (Seed, live plants, in vitro explants, DNA, pollen, data
  • 8. In-situ conservation  nature reserves  managed areas  Biosphere reserves  on-farm conservation  Home gardens  Seed Bank  Field gene bank  In vitro slow growth  Cryobank, cryopreservation  Pollen storage  DNA storage  Botanical Gardens Ex-situ conservation
  • 9. Seed Bank storage conditions Long - term Base room (–22 ºC, ≥ 100 years Short- term Active room (+4ºC, 40% RH) ≥ 25 years Medium- term Freezing room (–5 ºC, no frost ≥ 50 years All data in Gene bank by accession number All material exchange should by under MTA
  • 10. Seed Bank In vitro bank, slow growth Field Gene Bank
  • 13. Seed Processing  Sample registration  Seed cleaning  Seed drying  Seed moisture content  Seed viability  Seed packaging  Seed storage  Monitoring accessions  Distribution of germplasm  Regeneration Photo by: M. Ehsan Dulloo
  • 14. Registration  In Field  In all Seeds processing steps in seed bank
  • 15. Seed cleaning North Sinai research Station 2014, by Dr. Elsayed Elazazi
  • 16. Seed cleaning  Collections in cloth and paper bags held in drying rooms.  Collections cleaned to remove empty or infested seed and to reduce their bulk - aim: 95% pure seeds  All cleaning done by hand, carefully. Photo by: Kew 2011
  • 17. Seed cleaning equipment  Sieves, bungs & brush  Dust extraction system (masks)  Seed aspirator (blower)  Stereoscope & dissection equipment  Washing facility  Debris disposal Photo by: ElAzazi Photo by: KIB, China
  • 19. Clean under running water method
  • 20. Zig-zag Aspirator Handsorting Photo by: ElAzazi Photo by: Kew
  • 21. Seed drying North Sinai research Station 2014, by Dr. Elsayed Elazazi
  • 22. Why is it important to understand seed-air moisture relations? • Seed storage behaviour – response to water removal • Seed ripening – maturation drying • Seed viability – impact of seed moisture status on seed longevity • Seed drying – how seeds dry, monitoring seed moisture status
  • 23. Water in seeds •Determining seed equilibrium humidity (eRH) using a digital humidity sensor • Air and seed at equilibrium inside chamber
  • 24. Seed drying methods •Natural drying (sun/shade) •Dry room •Drying cabinet •Incubator drier •Silica gel /dried rice / charcoal Photo by: Kew
  • 25. Sun/Shade drying Sun drying in thin layers in open crates allowing plenty of air movement. Protecting seeds from direct sun to reduce risk of heat damage Photo by: Kew
  • 26. Dry room •Room Condition at 10-15% RH and 10 - 25C (FAO / IPGRI Gene bank Standards) •Specifically designed for purpose, taking into account maximum • seed loading (cleaned & un cleaned seed) and several other factors. 10-15% RH and 10 - 25C (FAO / IPGRI Genebank Standards) Photo from KIB China
  • 27. Drying cabinets 10-15% RH and 10 - 25C (FAO / IPGRI Genebank Standards) Photo by: Kew
  • 28. Cooled incubators (incubator driers) •Cost effective way of drying small amounts (up to 10 kg) of seed •Usually run at 18ºC, giving an RH of ~15%. •Dries 10kg of fresh seeds to 7% mc in ~ 3 weeks •Incubator must run with refrigeration system on constantly and must have auto defrost. 10-15% RH and 10 - 25C (FAO / IPGRI Genebank Standards) Photo by: Kew
  • 30. Drying with desiccants: silica gel bin drier •Use a suitable container with a tightly fitting lid (e.g. a plastic box or plastic / metal drum). •Add 20% by (container) volume dried (10 - 20% RH) silica gel. •Place bags of seed in container •Check the RH of the silica gel weekly; if too high, oven dry a quarter part at 100ºC and re-mix with the remaining gel. •Drying time (~one month) depends on the initial moisture content of the seeds, the amount of seeds, the type of seeds and the dryness of the silica gel. Photo by: Kew
  • 31. North Sinai research Station 2014, by Dr. Elsayed Elazazi
  • 32. What is ‘viability’? • A viable seed is a living seed that has the capability of germinating • Empty seeds and seeds without embryos are non-viable but they should not be called dead seeds because they never were viable • Dead seeds are seeds that were viable but have subsequently died Rao, N.K.; Hanson, J.; Dulloo, M.E.; Ghosh, K.; Nowell, A.; larinde, M 2006a. Manual of Seed Handling in Genebanks8.
  • 33. Why should seed viability be determined? 1. It is very important that seeds stored in the genebank are capable of producing plants when sown in the field. 2. They must have high viability at the start of storage and maintain it during storage. 3. Seeds with a high initial viability will also survive longer in storage. 4. Seed viability declines slowly at first and then rapidly as seeds age. 5. It is important to know when this decline occurs in order to take action to regenerate the accession. Excessive deterioration will lead to loss of material.
  • 34. How should viability be determined Viability Germination test Biochemical tests
  • 35. How many seeds should be tested? 200 Seeds (Rep. 1) 100Seeds (Rep. 2) 100Seeds R1 R2 R3 R4 R1 R2 R3 R4 G% 90%
  • 36. seed germination test substrate Top Paper (TP) North Sinai research Station 2014, by Dr. Elsayed Elazazi
  • 37. Between Paper (BP) Agar method North Sinai research Station 2014, by Dr. Elsayed Elazazi
  • 38. Sand North Sinai research Station 2014, by Dr. Elsayed Elazazi
  • 39. Always dissect a few seeds before you start North Sinai research Station 2015, by Dr. Elsayed Elazazi