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1Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Proposed Conservation Strategies Proposed Conservation Strategies for for Diospyros celebicaDiospyros celebica
Prasit Amy Aileen Zue Rao Salwana Tedi Tri Zhuo
Group 1
2Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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Presentation Outline
Background/Rationale What are the critical biological information
(BI) needed? How can we generate the BI? How can we translate these BI to
environmental conservation strategies?
3Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
D. celebica
4Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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Taxonomy
Kingdom PlantaeDivision MagnoliophytaClass MagnoliopsidaOrder EricalesFamily EbenaceaeGenus DiospyrosSpecies Celebica
5Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Population Status and Trends
Status of origin: Endemic to Sulawesi (Minahasa and Bolaang Mongondow/North Sulawesi; Parigi, Poso, Donggala, Toli-toli, Kolonodale and Luwuk/Central Sulawesi; Maros, Barru, Luwu and Mamuju/South Sulawesi)
Floristic element: Eastern provinces of Malesian element
Major ecological region: Sulawesi
Once a widespread species in Sulawesi, it is now comparatively rare, especially in the South
Exported since 18th century When in forests, D. celebica tends to scatter irregularly
6Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Rationale
Rare plant conservation programs must be guided by the species biological attributes.
Ecological and genetic processes often interact synergistically to influence the population viability and to determine the persistence of populations in the long run.
Conservation has a cost and the resources available for conservation programs are always limited. Thus, CS must not only be scientifically justified but also practical in terms of resource availability.
7Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
What Are The Critical BI?
GENETICSECOLOGY
• Distribution & habitat
• Demography
• Germination
• Phenology
• Level of genetic diversity
•Spatial genetic structure
• Population differential
• Mating system
•Minimum population size
8Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Ecological Distribution & Habitat
Ecological interaction between plants and their environment can influence population growth rates via their effects on fecundity, growth, or survivorship of individuals (Blundell and Peart, 2001; Peters, 2003)
The studies on population dynamics and demography patterns will lead to a better understanding of the natural processes that operate within the population
9Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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Distribution and Habitat
Common name
Macassar Ebony Black Ebony Diospyros macassar (synonymous)
Distribution Endemic to Sulawesi Can be found in rain and monsoon forests Can also grow in both humid conditions and
in seasonal climate
Habitat Can survive on a variety of soils Occurs in undulating areas up to 600m above sea
level.
10Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Legend: Natural distribution of D. celebica in Indonesia (Overall natural distribution)
11Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Conservation status & measures
IUCN status available of the Vulnerable (VU) (International Union for Conservation Red Data Book, 1978)
In Sulawesi, D. celebica is protected and there is a quota system in place. The Indonesian government has already started a planting program. It has not, however, been planted on a large commercial scale
12Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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Threats & Utilization
D. celebica is threatened by heavy exploitation since it is an important source of streaked ebony
13Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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D. celebica timber is used for piano keys, carvings, brush backs, inlaying, and parts of stringed instruments.
14Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Ecological Approach In Conserving Ebony
Population Dynamics & Demographic Studies
Phenology Spatial
Distribution
Germination
Study
Population
Survey
15Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
1. Population Survey
Study Plot
200 m
200m • Tagging
• Diameter measurement (DBH> 5cm)
• Mapping & coordinate using GPS & GIS
To know the population status and relative density of ebony in the area.
16Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
17Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Demography
Involves population dynamics, species recruitment and
mortality
Field survey
Set-up plot
Long-term & short-term survey
18Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
2. Spatial Distribution
Information
Soil
Topography
Climate
To get information on habitat preference of ebony (ridge, valley, slope) from established plot
19Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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20Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
•65 km SE Manila
•shifting cultivation, burned and selectively logged
•Mature secondary forest with natural mixed stand
•Dominated by Celtis luzonica and Diplodiscus paniculatus
•Tropical monsoon
Mt. Makiling Forest Reserve
21Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
22
•113 tree species and 6 palms, >5 cm dbh
•Highest density - Celtis
•Highest basal area -Diplodiscus
•Max. dbh by Ficus (balete, strangling fig)
•Max. mean height of 21m
Structural characteristics of main canopy species Table 1. Structural characteristics of main canopy species of the secondary forest of Mt. Makiling Forest Reserve, Philippines.Species Density Min Max Mean Basal area
(90mx140m DBH DBH Height (m2ha-1)
(12,600m2) (cm) (cm) (m)Diplodiscus paniculatus 123 5.0 136.6 13.2 5.5Celtis luzonica 265 5.0 121.3 11.9 4.9Parashorea malaanonan 22 5.0 79.6 13.6 3.2Litsea garciae 5 13.8 77.2 15.4 3.1Casearia fuliginosa 6 5.1 77.0 14.0 3.1Trichadenia philippinensis 3 5.1 71.6 no data 2.9Pisonia umbellifera 16 5.3 69.9 11.6 2.8Drypetes maquilingensis 10 7.6 66.2 21.0 2.6Macaranga bicolor 3 10.7 59.5 19.6 2.4Syzygium nitidum 10 5.4 56.2 11.0 2.2Sapindus saponaria 1 7.6 54.1 19.7 2.2Ahernia glandulosa 10 5.7 53.0 8.0 2.1Ficus congesta 24 5.0 53.2 13.8 2.1Panguim edule 2 13.8 51.9 16.3 2.1Diospyros philippensis 8 5.0 77.3 9.5 2.1Chisocheton cumingianus 55 5.0 47.7 13.9 1.9Planchonia spectabilis 7 8.6 43.4 14.8 1.7Nephelium mutabile 76 5.0 41.7 11.6 1.7Other species 478 5.0 92.0 no data 72.0Total 1187 5.0 140.0 no data 141.5
23Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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3. Pop’n Dynamics & Demographic Studies
10m
10 m
Regeneration Quadrat Plot WHY???
1. Monitor growth
2. Seedling recruitment and mortality of the seedling (3 years; short-term study
3 . First year seedlings will be identified, marked & appearance will be recorded
to know the changes taking place in the life cycle of ebony
24Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
4. Phenology of Ebony
Reproductive biology will determine gene flow (mating system, pollination, fruit dispersal, etc.) & recruitment rate of the species (Lee, 2006 personal comm’n)
STEPS:
Identify the ebony tree from the established plots/
Check and measure every month
Do some ranking such as budding stage, peak bloom and mature seed of
ebony.
25Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Pollination
BiologyObservation Pollinators Study on
Pollination Biology
Dispersal
strategyObservation
Wind
Animals
Conserve animals that are seed dispersal agents
Conservation strategy
26Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Regeneration & species role
Flowering and fruiting occurs at the age of 5-7 years
Seeds remain viable for only a short time
Seeds vectors: Bats Birds Monkeys Found with Homalium
celebicum
27Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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Germination study
From established plot, set up seed trap for seedfall study of ebony
Monthly seedfall collection will be done + seeds will be checked
Correlation of seed weight to germination capacity of ebony
For mature & sound seeds, weighing & germination test will be done tocompare growth & survival from natural forest condition
To know the germination trait of ebony for ex-situ conservation.
28Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
In summary…Topic Possible Outcomes Conservation strategies
Distribution Limited distribution Conserve all remaining populations
Widespread Designated conservation area based on ecological units (different in climate, soil and topography)
Demography 1. Low recruitment rate (normal J distrb’n)
2. High mortality rate (inverse J distrb’n)
Vegetative propagation (Enrichment planting)
Assisted Natural Regeneration
Silvicultural treatment
Germination 1. Recalcitrant
2. Orthodox
1. Immediate planting and establishment of seedling nursery
2. Seed storage
Phenology 1. Regular
2. Irregular
1. -
2. Vegetative propagation and seed storage
29Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Rationale for Genetic Info
Allelic richness could contribute to population growth thru its effect on evolutionary potential, or the ability of a species to respond to changes in its selective environment.
Reduced heterozygosity can result in decrease of population growth due to inbreeding depression.
Therefore, we need to know the genetic diversity partition within and among populations
Levels of Genetic
Diversity
Genetics Approach Genetics Approach Conserving EbonyConserving Ebony
Population Differentiation
Mating System
Spatial Genetic
Structure
Minimum Population
Size
31Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
1. Levels of Genetic DiversityTo estimate level of genetic diversity that we need togenerate information
aCriterion AFLP RAPD SSR Allozymes
Quantity of information High High High Low
Replicability High Variable High High
Resolution of genetic differences
High Moderate High Moderate
Ease of use and development
Moderate Easy Difficult Easy
Development time Short Short Long Short
aThe scoring scheme follows closely those in Hillis et al.2 and Karp and Edwards49.
32Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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Why Choose SSR Marker ? SSR- (microsatellite) marker was developed by (Weber & May 1989
in human and found to be abundant in plant by Morgante & Olivieri 1993)
Very high degree of polymorphism & codominance make them extremely informative
Practical number of loci is 10
Few as 5 or 6 microsatellite loci can often answer many conservation genetic questions (e.g. paternity, pollen flow) that cannot be answered with 30 or more isozyme loci
High reliability (reproducibility)
If the resources is limited, we can use allozyme
33Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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How to Get The Parameters ?
DNA extraction
Microsatellite analysis
Analysis data
Sample Collection•D.celebica leaf samples will collected from 40 populations base on 40 area divide based on soil, climate and slope
•From each population, about 30 adults will be sampled
•Develop primer•PCR amplification•For genotyping, PCR product will be electrophoresed along with GeneScan ROX 400 internal size standard using DNA automated sequencer
•DNA genomic will be extracted using Murray and Thompson (1980) and purified using High Pure PCR Template Preparation Kit
Genescan analysis software and
GENOTYPER software v3.7
•D.celebica leaf samples will collected from 40 populations base on 40 area divide based on soil, climate and slope
•From each population, about 30 adults will be sampled
•Develop primer•PCR amplification•For genotyping, PCR product will be electrophoresed along with GeneScan ROX 400 internal size standard using DNA automated sequencer
•DNA genomic will be extracted using Murray and Thompson (1980) and purified using High Pure PCR Template Preparation Kit
Genescan analysis software and
GENOTYPER software v3.7
34Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Low genetic diversity means not enough sufficient gene pool for short term adaptation and long term evolutionary
Result of Genetic Diversity
Need to enhance by the introductionNeed to enhance by the introduction of new alleles throughof new alleles through
introgression following hybridizationintrogression following hybridization for long term conservationfor long term conservation
35Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
2. Spatial Genetic Structure
To determine genetic structure within a population
Moran I Coefficient analysis statistical analysis
Significantly structured
Need sampling strategy for ex-situ conservation
random Need
capture all Choose and
select
36Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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3. Population genetic structure
Erikson & Ekberg 2001
37Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
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Analysis Data from SSR Analysis
To determine coefficient of population differentiation.
Gst<0.050 low genetic differentiation
0.050<Gst<0.15 moderate genetic differentiation
0.151<Gst<0.250 large gene differentiation
0.250<Gst very large gene differentiation
38Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
if high population differentiation
Mean: --low gene flow --high inbreeding --low genetic diversity --high variation among population
more populations need to be conservation
39Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
If low population differentiation Mean: --high genetic flow --high genetic diversity --low variation among population
--Low population genetic differentiation among population implies no preference in identification of population for in-
situ conservation or germplasma collection for ex situ conservation
need more bigger area, and few population
40Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
4. Mating System and Gene Flow
OUTCROSSING RATEOUTCROSSING RATE
General description (Gregorius 1989)
“Random mating, the environmental influence on mating events, selfing and the consequences of
selfing and other forms of inbreeding, and incompatibility systems”
Data analysis by using multilocus mating system program (MLTR) Ritland (1996)
41Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Example: Predominantly outcrossing Selfing
Ex-situ conservation (field genebank) Outcrossing : collect many seeds from a few
mother trees Selfing : collect seeds from many mother
trees)
Mating System
42
How many individuals in a conserved population are needed to maintain evolutionary potential
of population and to resist to inbreeding depression from
generation to next generation
5. Minimum Population Size
Ht Ht+1
H selection
H mutation
N=Number of individuals in population t =Number of generation Htt=Heterozygosity at time t
Most breeders agree that :A small loss of heterozygosity by 1% from one generation to next generation will not be significant for breeding , and does not result in inbreeding depression.
In other word, we accept Ht+1=99%*Ht and then calculate N=50.
So 50 individuals is often referred to as the basic rule of conservation genetics under the conditions of absence of selection, randomness of mating, each individuals in reproductive phase.
Theoretically
1 t 1 t HH00·· 11-- = H= Htt
2 N2 N
44Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
In fact, a population is in selection and non-randomness of mating, which are very difficult to be quantified, so a population size of 500
reproducible individuals is hoped to represent a safety device and to provide a better chance for the restoration of genetic variation by mutation
against the loss (Hattemer 2005).
If a population has less than 500 individuals, maybe all of them should be conserved.
Minimum Population Size
45Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
GENETICSECOLOGY
• Distribution & habitat
• Demography
• Germination
• Phenology
CONCLUSION
• Level of genetic diversity
•Spatial genetic structure
• Population differential
• Mating system
•Minimum population size
46Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Dr. Kelvin and Dr. Lee
K LK L
47Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006
Group 1 Presentation
Demography