Conservation genetics in Amentotaxus formosana Yu-Chung Chiang 1 1 Department of Biological Science, National Sun Yat-sen University, Kaohsiung 804 Ya-Zhu Ko 1, Mei-Hui Chen 2, Tzen-Yuh Chiang 3 2 Conservation Division, Forestry Bureau, Council of Agriculture, Executive Yuan, Taipei 100, Taiwan. 3 Department of Life Sciences, National Cheng Kung University, Tainan 701, Taiwan 2015/09/28
OUTLINE Dilemma on endangered conifer species in Taiwan Past problem Recent problem Ornamental values Commercial values Habitat destruction Conservation strategies based on population genetics
Gymnosperms in Taiwan listed on The IUCN Red List of Threatened Species 28 species in total, 12 species listed on CR, EN, or VU. 2 species listed on NT. Amentotaxus formosana -- Critically Endangered, CR Cycas taitungensis -- Endangered, EN Keteleeria davidiana var. formosana -- Endangered, EN Podocarpus nakaii -- Endangered, EN Calocedrus formosana -- Endangered, EN Podocarpus fasciculus -- Vulnerable Pseudotsuga sinensis var. sinensis -- Vulnerable
Amentotaxus formosana Southern Taiwan Central Mountain less than 20 km Dawu Taiwan Amentotaxus Nature Reserve Pingtung 6 39 28 29 30 Taitung 5 3 4 2 Chachayalaishan Major Wildlife Habitat 1 5 watershed
A I B I D I E I C II C III C IV C I chlorotype Cycas taitungensis After Wurm Ice Ages (1-11Mya) restricted on southeastern Taiwan 3.70 % 2.47 % 2.47% 2.47% 2.47% 3.70% 3.70% 0 6KM Coastal Area 9.52% 90.48% C RL 79.02% Taitung Cycad Nature Reverse Pacific Ocean Two extant populations: 30 km apart
Keteleeria davidiana var. formosana Taitung New Taipei City Taitung county Two separate populations apart Northern and Southern Taiwan Less than one thousand individuals
Migrant-pool migration model Ice Age refuge
OUTLINE Dilemma on endangered species in Taiwan Past problem Recent problem Ornamental values Commercial values Habitat destruction Conservation strategies based on population genetics
Ornamental values and Commercial values 50 individuals of Taiwan Cow-tail Fir cost 10 million NT dollars One individual with 1 to 2 meter high of Juniperus chinensis var. tsukusiensis costs 2800 NT dollars http://www.herbs-garden.idv.tw/userfiles/image/e477%20(6).jpg http://twimg.edgesuite.net/images/renews/20141203/640_1ede1cb93a5babe2bd998c01b90d5438.jpg
Habitat destruction Original broad-leaf or Conifer forests Wulong tea Cabbage Betel nut
OUTLINE Dilemma on endangered conifer species in Taiwan Past problem Recent problem Ornamental values Commercial values Habitat destruction Conservation strategies based on population genetics
Conservation strategy Wild population management Phylogenetic relationship Inventory Study Evaluation for genetic variation Past Population dynamic Conservation genetics Distinct genetics units ex situ Conservation Seed collection Cutting propagation
Evaluate the Evolutionarily Significant Unit (ESU) 1. Current geographic separation 2. Genetic differentiation at neutral among related ESUs caused by past restriction of gene flow 3. Locally adapted phenotypic traits caused by differences in selection.
Amentotaxus distribution Species Red List status A. formosana CR Assam Yunnan Szechwan Hubei Kwangtung A. yunnanensis EN Vietnam Hong Kong Taiwan A. argotaenia VU A. poilanei VU Distribution of extant Amentotaxus in the Far East (Ferguson et al. 1978) 14
Distributions and frequencies of clades of organelle DNA in the populations of Amentotauxs species
The two, three, and six clusters (K = 2, 3, and 6) were detected from STRUCTURE analyses The effective population size fluctuation through time atpi/h rps16/trnk nad intron Microsatellites Sum Var (%) Sum Var (%) Sum Var (%) Sum Var (%) Among species 109.08 44.11 147.24 17.77 254.26 28.30 525.93 34.19 Among populations 112.20 41.06 345.08 68.46 458.17 63.49 316.11 14.68 Within populations 47.51 14.83 82.66 13.78 70.96 8.22 1211.13 51.13 K=2 K=3 K=6 Evolutionarily Significant Unit
Wild population management Inventory Study Amentotaxus formosana
In situ conservation Nature Reserve National Park Wildlife Refuge Seabird Refuge Forest Reserve Fish Refuge Protected Areas Wildlife Habitat
Conservation strategy Wild population management Phylogenetic relationship Inventory Study Evaluation for genetic variation Past Population dynamic Conservation genetics Distinct genetics units ex situ Conservation Seed collection Cutting propagation
Amentotaxus formosana Population differentiation analysis based on AMOVA Source of variation Sum of squares Variance % of Variation Fixation indexes components Among population 279.07 0.44 8.95 * F ST =0.09* Among individuals within populations 2362.44 2.18 44.35 * F IS =0.49* Within individuals 806.00 2.29 46.70 * F IT =0.53* Significant (P<0.05) values are indicated with asterisk (*) 8.95 % variation accumulated between Populations Signification differentiation Positive fixation indexes (F IS & F IT > 0) inbreeding within population the distance of pollen dispersal http://www.epochtimes.com/i6/709212313421119.jpg restricted distance http://www.epochtimes.com/i6/709212313411119.jpg
Amentotaxus formosana Detection effective population size using Isolation with Migration Model Small effective population size and large ancestral population size population concentration effective population size effective population size of ancestral migration rate divergence time(t) Chachayalaishan/ Jinshueiying N 1 N 2 N A μ=3.0 10-5 1830.67 506.67 248424.17
Amentotaxus formosana Detection effective population size using Linkage Disequilibrium Model in Chachayalaishan population (383 samples) Lowest Alleles Frequency used = 0.05 Independent Comparisons Overall r 2 Expected r 2 Ne (95% CIs for Ne) All 1388 0.007 0.003 94.4(81.9-109.1) 1 (74.4-141.0) 2 Cha-1 1096 0.02 0.014 55.9(42.2-77.7) 1 (37.4-92.9) 2 Cha-2 835 0.033 0.021 25.2(19.1-34.1) 1 (15.9-42.9) 2 Cha-3 948 0.02 0.013 45.6(35.2-60.9) 1 (30.2-74.9) 2 Cha-4 959 0.024 0.013 30.0(24.2-37.8) 1 (18.0-53.9) 2 Cha-5 937 0.028 0.021 45.2(31.9-69.4) 1 (28.5-84.1) 2 Lowest Alleles Frequency used = 0.02 Independent Comparisons Overall r 2 Expected r 2 Ne (95% CIs for Ne) All 2109 0.006 0.003 102.1(90.7-115.2) 1 (78.7-134.2) 2 Cha-1 1631 0.02 0.014 55.7(44.1-72.7) 1 (39.1-85.9) 2 Cha-2 912 0.031 0.021 29.9(22.5-41.3) 1 (19.4-50.2) 2 Cha-3 1660 0.019 0.013 57.9(46.3-74.6) 1 (41.1-87.8) 2 Cha-4 1248 0.021 0.013 42.0(33.7-53.5) 1 (25.6-77.6) 2 Cha-5 1038 0.028 0.021 49.7(35.0-77.3) 1 (32.4-88.0) 2 Lowest Alleles Frequency used = 0.01 Independent Comparisons Overall r 2 Expected r 2 Ne (95% CIs for Ne) All 2258 0.006 0.003 105.1(93.6-118.3) 1 (81.8-136.9) 2 Cha-1 1807 0.02 0.014 62.6(49.3-82.3) 1 (42.1-103.8) 2 Cha-2 918 0.031 0.021 30.3(22.8-41.9) 1 (20.0-49.8) 2 Cha-3 1832 0.017 0.013 73.1(57.2-97.4) 1 (51.9-112.1) 2 Cha-4 1275 0.02 0.013 45.7(36.4-58.8) 1 (26.7-91.5) 2 Cha-5 1112 0.026 0.021 66.7(44.5-116.7) 1 (41.6-134.1) 2 Small population with narrow habitat Small effective population size Inbreeding with related individuals Wahlund effect
Amentotaxus formosana Structure within Chachayalaishan population using PCoA Differentiation within population causing by inbreeding with related individuals Spatial structure effects pollen and seed dispersal subdivision within population Z Principal Coordinate Analysis (PCOA)
Amentotaxus formosana MCMC Assignment test + Geographic data Distinct genetic units Conservation Management units Select health and reproductive individuals for in situ or ex situ conservation management CG-8-1 CG-8-2 CG-8-3 CG-8-4 CG-8-5 CG-8-6 CG-8-7 CG-8-8 GENELAND analysis results K =8
Conservation strategy Wild population management Phylogenetic relationship Inventory Study Evaluation for genetic variation Past Population dynamic Conservation genetics Distinct genetics units ex situ Conservation Seed collection Cutting propagation
Amentotaxus formosana in situ or ex situ conservation management Selected individuals Phenological investigation Seed collection Cutting propagation for ex situ conservation
Amentotaxus formosana Cutting propagation for ex situ conservation 19 months old cutting seedling Test the combination and concentration of plant hormone IAA, IBA, and NAA
Summary Phylogenetic and phylogeographic analyses identify Evolutionarily Significant Unit (ESU) Inventory Study discovery the distribution of populations In situ conservation protect endangered species in wild habitats Management units for conservation Distinct genetic units by population genetics Ex situ conservation To avoid extinction
Acknowledgements Funding Forestry Bureau, COA National Science Council Supporting Taitung Forest District Office, Forestry Bureau, COA NSYSU
建 議 與 後 續 研 究 Thank you for your attention