This document summarizes germplasm conservation strategies for oil palm. It discusses the need for conservation given oil palm's narrow genetic base. It describes the genetics and species of oil palm, as well as qualitative traits that have been identified. The document outlines India's field gene bank containing 650 palms of 16 accessions. It discusses various conservation strategies including conservation of seed, cryopreservation of zygotic embryos, somatic embryos, polyembryoids, shoot tip culture, and pollen. Successful protocols have been developed for cryopreserving somatic embryos, polyembryoids, shoot tip culture, and long-term storage of pollen. Challenges remain in assessing survival rates and clonal propagation.

1. Germplasm
conservation in
Oil Palm
P. Murugesan
M. Shareef &
P. Masilamani*
Indian Institute of Oil Palm Research
* AC & RI, Madurai
National seminar on Challenges and innovative
approaches in crop improvement, 16-17 December, 2014

2. CONTENT
• Need for conservation
• Genetics of Oil Palm
• Oil Palm origin and species
• Qualitative traits – selected palms
• Field gene bank
• Conservation strategies
• Conclusion & challenges ahead

3. Need for germplasm conservation
• 5% of oil acreage, palm oil accounts for 33% of
vegetable oil, 45% of edible oil
• 12 million hectares planted , production of 45 MT
• Oil improved from 6.3 to 11.2 t/ha in 4 decades
• Breeding relay on restricted breeding base
• Deli dura - narrow genetic base
• Finding qualitative traits difficult.

4. Genetics of Oil Palm
• Shell thickness - Single gene
• Dominant homozygote (Sh+ Sh+) forms Dura
(D) Recessive homozygote (Sh- Sh-) - shell-
less pisifera (P) and Heterozygote (Sh+ Sh-)
forms tenera (T).

5. Genetics of Oil Palm

6. Oil Palm - two species
Elaeis guineensis
Elaeis oleifera

7. Origin of Oil Palm
1. Elaeis guineensis (African Oil Palm)
2 . E. Oleifera (American Oil Palm)

8. View of Deli Dura planted
During 1848, Indonesia

9. Qualitative traits identified ...
Dwarf... Virescence...fruit size----long
stalk.......small kernel----thorn
less.................

10. Indian Oil Palm Genetic base and selected palms
50 years
Small kernel & thick mesocarp

11. Fruit and Nut Variability in African germplasm – to be introgressed

12. Long bunch stalk- easy harvest

13. Wild oleifera population at Palode

14. Dwarf oleifera & early fruit maturity
Age – 33 years

15. Dwarf, inter specific hybrid
Age – 24 years

16. Dwarf tenera hybrid
Age – 33 years

17. Thorn less Oil Palm

18. Strong Orange colour fruits -
easy assessment of harvest maturity

19. Germplasm conservation in
field- difficulties & alternatives

20. Field gene bank at Palode
Total 650 palms of 16 accessions planted in
Germplasm Block ‘B’ during 1981-1998

21. MULTILOCATION EVALUATION OF
AFRICAN GERMPLASM

22. View of field gene bank at Athirapalli , Kerala

23. Little Andaman –Nigerian material

24. Conservation strategies
• Conservation of seed
• Cryopreservation of Zygotic embryos
• Cryopreservation of somatic embryos
• Vitrification of polyembryoids
• Cryopreservation of SPC
• Cryopreservation of pollen
• Successful protocol of pollen storage

25. Seed technology - relevance to
conservation
• Storage behaviour – intermediate
• Seed dormancy
• Easy methods for breaking dormancy
• How long seed could be stored ?
• Zygotic embryos desiccation tolerance

26. Section of Oil Palm Fruit –
Seed dormancy mechanism

27. Dry heat method – 4 months to get sprouts

28. De-operculuation dormancy breaking method

29. Cryopreservation of seed
• Whole seed - cryopreserved after desiccation to
critical mc (10%)
• If extracted from rehydrated kernels, 65% of
embryos desiccated to around 0.3 g H2O/ g DW
developed into plantlets after cryopreservation.
In contrast, only 25% embryos (at 0.12g H2O/ g
DW) extracted from cryopreserved dry kernels
developed into plantlets. However, this value was
increased to 63% if kernels were partially
rehydrated before freezing until water content of
embryos reached 0.3 g H2O/ g DW

30. Cryopreservation of somatic embryos
• Shiny white, finger-like somatic embryos could
be used for cryopreservation.
• Its efficiency can be markedly improved by
completing the 7-day pre growth period on
0.75 M sucrose by an additional dehydration
period carried out either by placing the
embryos in the air current of the laminar flow
cabinet or in an air tight box containing silica
gel

31. True seeds (Kernel) of Oil Palm

32. Vitrification of polyembryoids
• In vitro grown polyemryoids of oil palm were
successfully embryo preserved by vitrification
with 45% survival.
• Poly embyoids were subjected to plant
vitrification solution-2 (PVS2) 30% (w/v) glycerol
plus 15% (w/v) EG plus 15% (w/v) DMSO plus
0.4M sucrose exposure for 5 min at 26+ 20 C
subsequently plunged into liquid nitrogen.
• Thawed polyembryoids resumed growth within 8
days of culture and shoot development was
recorded 25 days of growth

33. Cryopreservation of SPC
• Cryopreserved stabilized polyembryonic cultures
(SPCs) of six elite oil palm clones was monitored
for nursery and field growth up to 12 years after
transfer in Cote d'Ivoire.
• Six clones tested showed an average recovery of
34% after freezing in liquid nitrogen. Palm trees
originating from control SPCs were found to
flower early than preheated, dehydrated and
cryopreserved SPCs.
• This delay in flowering disappeared progressively
and all palms had flowered 3 years after planting.

34. Cryopreservation of pollen
• Pollen kept in screw-cap polypropylene
cryovials (2ml capacity) was directly immersed
in liquid nitrogen (at -1960 C).
• Pollen viability was retained as high as 54+
1.72% as assessed by FDA with in vitro
germinability of 49+ 1.2%.

35. Successful protocol for pollen
• National Bureau of Plant Genetic Resources
(NBPGR) has already demonstrated the
potential of cryopreservation for successful
storage of oil palm pollen after 8 years of
storage

36. Male inflorescence
at anthesis
At emergence

37. Conclusion & challenges ahead
• Field gene bank important desirable traits
available
• Pollen- successful protocol available
• Variation in survival- palm to palm variation
• In vitro – survival rate of growth – to be
assessed
• Clonal propagation