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In vitro conservation of groundnut
germplasm
by
M.M. Abdulmalik & I.S. Usman
Department of Plant Science, Institute for Ag...
Introduction
• Groundnut (Arachis hypogaea L.) is an important
source of protein and edible oil in the world.
• Nigeria ra...
• Groundnut germplasm are conventionally stored in
gene banks and seeds are the most preferred
propagule used.
• Due to th...
• The IAR maintains its varieties and cultivars
by planting every season.
• Not only is this laborious, time consuming
and...
• Cryopreservation or storage in liquid nitrogen at temperature
of –1960C, at which all the cells are in a state of suspen...
Material and Methods
• Plant material- Seeds of four
groundnut (Arachis hypogaea
L.) varieties.
• Surface sterilized of se...
Vitrification technique
• Preculturing of embryonic axes
on solidified MS medium
supplemented with 0.3M
sucrose for 24hr.
...
Desiccation
• Embryonic axes were excised
and subjected to desiccation
under the air current of a
laminar flow cabinet for...
• Cryovials were
directly immersed into
liquid nitrogen (-
196°C) and held for
1hr.
• Thawing took place in
a water bath at 40°C
for 2min.
• Embryonic axes were
cultured individually in
test tubes containing
10ml of MS medium
(Murashige and
Skoog, 1962)
supplem...
• Cultures were
maintained in a
growth chamber at
26±2°C under 16hr
light/8hr dark photo
period provided by
white inflores...
• Regenerated
microshoot of
groundnut after
4weeks
• Microshoots were
subcultured in MS
media supplemented
with 1mg/L NAA for
rooting
• Hardening-off &
acclimatization
– Conditioning plantlets
to the external env’t•
i.e. before transfer to
field conditions
• Groundnut plantlet in
the screen house
Expected Impact
• Genebank managers have
a complimentary
biotechnology conservation
technique
• Breeders have wide array
o...
THANK
YOU
FOR
LISTENING
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B4FA 2012 Nigeria: Cryopreservation of Groundnut Germplasm in Nigeria - Maimuna Abdulmalik

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Presentation by Maimuna Abdulmalik, Ahmadu Bello University, Zaria, Nigeria
Delivered at the B4FA Media Dialogue Workshop, Ibadan, Nigeria - September 2012
www.b4fa.org

Published in: Science, Business, Technology
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B4FA 2012 Nigeria: Cryopreservation of Groundnut Germplasm in Nigeria - Maimuna Abdulmalik

  1. 1. In vitro conservation of groundnut germplasm by M.M. Abdulmalik & I.S. Usman Department of Plant Science, Institute for Agricultural Research Ahmadu Bello University Zaria
  2. 2. Introduction • Groundnut (Arachis hypogaea L.) is an important source of protein and edible oil in the world. • Nigeria ranks third after India and China in terms of production (FAOSTAT 2010). • Production is constraint by low quality of seeds as these deteriorate rapidly in storage particularly in the tropics (Delouche et al., 1973). • Conventional methods used by our subsistence farmers to store seeds (RMRDC, 2004) are inadequate for long term storage. • Newly improved varieties are fast replacing our traditional varieties.
  3. 3. • Groundnut germplasm are conventionally stored in gene banks and seeds are the most preferred propagule used. • Due to their high lipid content and thin seed coat they cannot tolerate the gene bank conditions for longer periods like true orthodox seeds. • This has led to the suggestion that the groundnut should be considered as suborthodox (Vasquez- Yanes and Arechiga 1996, Gagliardi, et al., 2000). • Studies on groundnut seeds viability in storage in IAR revealed that all the entries evaluated had viability of 50% or less by 1 year and 25% by 2 years of storage.
  4. 4. • The IAR maintains its varieties and cultivars by planting every season. • Not only is this laborious, time consuming and expensive but also plants are exposed to the possible risk of pest, disease and environmental stresses. • Thus, cryopreservation is considered as important complimentary strategies for ex situ conservation.
  5. 5. • Cryopreservation or storage in liquid nitrogen at temperature of –1960C, at which all the cells are in a state of suspended animation, is the most promising method of invitro germplasm storage • advantages – enables long term storage of the plant material – require less space – less labor – it’s cheap at the long run – genetic stability – elimination of viral disease. • Cryopreservation techniques – Desiccation – Vitrification – Encapsulation vitrification – Encapsulation dehydration – Droplet freezing
  6. 6. Material and Methods • Plant material- Seeds of four groundnut (Arachis hypogaea L.) varieties. • Surface sterilized of seeds -5min in70% alcohol, -20min in 10% NaOCl+ 2-3 drops of tween 20 -rinsed thrice with sterile distilled water - 10min in 5% NaOCl + 2-3 drops of tween 20 -washed three times with sterile distilled water. • Seeds were soaked in sterile distilled water for 3hr. • Embryonic axes were excised
  7. 7. Vitrification technique • Preculturing of embryonic axes on solidified MS medium supplemented with 0.3M sucrose for 24hr. • treatment with a loading solution (2M glycerol plus 0.4M sucrose dissolved in MS medium) for 15min at 25°C. • Treated embryonic axes were transferred to 2ml cryovials and 1ml PVS2 was added dehydrated for 2hr. – PVS2 (30% (w/v) glycerol, 15% (w/v) ethylene glycol and 15% (w/v) dimethylsulfoxide (DMSO) in MS medium with 0.4M sucrose)
  8. 8. Desiccation • Embryonic axes were excised and subjected to desiccation under the air current of a laminar flow cabinet for 4hr. • Moisture content was determined on fresh weight basis after drying in a 100°C oven for 24hr (3 replicates of 10 embryonic axes per duration). • Desiccated and non desiccated (control) embryonic axes were placed in 2ml sterile cryovials.
  9. 9. • Cryovials were directly immersed into liquid nitrogen (- 196°C) and held for 1hr.
  10. 10. • Thawing took place in a water bath at 40°C for 2min.
  11. 11. • Embryonic axes were cultured individually in test tubes containing 10ml of MS medium (Murashige and Skoog, 1962) supplemented with 15mg/L 6- benzylaminopurine (BAP) and solidified with 8g/L agar.
  12. 12. • Cultures were maintained in a growth chamber at 26±2°C under 16hr light/8hr dark photo period provided by white inflorescence.
  13. 13. • Regenerated microshoot of groundnut after 4weeks
  14. 14. • Microshoots were subcultured in MS media supplemented with 1mg/L NAA for rooting
  15. 15. • Hardening-off & acclimatization – Conditioning plantlets to the external env’t• i.e. before transfer to field conditions
  16. 16. • Groundnut plantlet in the screen house
  17. 17. Expected Impact • Genebank managers have a complimentary biotechnology conservation technique • Breeders have wide array of groundnut germplasm • Improved groundnut seeds that are high yielding and resistant to pest and diseases and better adapted to changing climates made available to farmers • Groundnut pyramids of Kano flourishing
  18. 18. THANK YOU FOR LISTENING

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