Download - Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006 Group 1 Presentation 1 Proposed Conservation

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



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?



D. celebica



Taxonomy

Kingdom PlantaeDivision MagnoliophytaClass MagnoliopsidaOrder EricalesFamily EbenaceaeGenus DiospyrosSpecies Celebica

http://en.wikipedia.org/wiki/Plant

http://en.wikipedia.org/wiki/Flowering_plant

http://en.wikipedia.org/wiki/Dicotyledon

http://en.wikipedia.org/wiki/Ericales

http://en.wikipedia.org/wiki/Ebenaceae



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



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.



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



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



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.



Legend: Natural distribution of D. celebica in Indonesia (Overall natural distribution)



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



Threats & Utilization

D. celebica is threatened by heavy exploitation since it is an important source of streaked ebony



D. celebica timber is used for piano keys, carvings, brush backs, inlaying, and parts of stringed instruments.



Ecological Approach In Conserving Ebony

Population Dynamics & Demographic Studies

Phenology Spatial

Distribution

Germination

Study

Population

Survey



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.



Demography

Involves population dynamics, species recruitment and

mortality

Field survey

Set-up plot

Long-term & short-term survey



2. Spatial Distribution

Information

Soil

Topography

Climate

To get information on habitat preference of ebony (ridge, valley, slope) from established plot



•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

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



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



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.



Pollination

BiologyObservation Pollinators Study on

Pollination Biology

Dispersal

strategyObservation

Wind

Animals

Conserve animals that are seed dispersal agents

Conservation strategy



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



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.



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



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



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.



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



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



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



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



3. Population genetic structure

Erikson & Ekberg 2001



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



if high population differentiation

Mean: --low gene flow --high inbreeding --low genetic diversity --high variation among population

more populations need to be conservation



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



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)



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



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



GENETICSECOLOGY

• Distribution & habitat

• Demography

• Germination

• Phenology

CONCLUSION

• Level of genetic diversity

•Spatial genetic structure

• Population differential

• Mating system

•Minimum population size



Dr. Kelvin and Dr. Lee

K LK L



Demography

Download - Training Workshop on Forest Biodiversity Conservation & Management of Forest Genetic Resources; June 5-16 2006 Group 1 Presentation 1 Proposed Conservation

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