1. Challenges for Botanic Garden Development in Megadiverse Countries: China Example Hongwen Huang Wuhan Botanical Garden, CAS

2. Introduction  Chinese Botanical Garden network: a brief introduction of main botanical gardens within the Chinese Academy of Sciences as leading role among 140 gardens.  Current efforts in conservation of native plants in China and case studies of rare and endangered plants  Sustainable development of germplasm resources in the Chinese botanical gardens

3. Chinese BG Network 12 CAS botanical gardens in different geographic regions over China Historical and contemporary botanical research institutes in China Leading roles in botanical research, conservation and public education among 140 gardens in China Designated national centers for science, environment and natural heritage education Annual visitation is 3 millions

4. Beijing BG Wuhan BG Huanan BG Xisuanbanla tropical BG CAS-BG Geographically structured Network BG centers Featured BG 东亚植物区 泛北极植物区 古地中海植物区 古热带植物区 青藏高原亚区 中国 — 马来西亚亚区 中 - 日植物亚区

5. Shengyang Arb. Nanjing BG Dinghushan Arb. Huaxi sub Mt BG Guiling BG CAS-BG Geographically structured Network Lushan BG Kunming BG 东亚植物区 泛北极植物区 古地中海植物区 古热带植物区 青藏高原亚区 中国 — 马来西亚亚区 中 - 日植物亚区 BG centers Featured BG Tulufan desert BG

6. Main CAS Botanical Gardens Beijing BG ：  Conservation focus: North China temperate and plain forests; >4000 species conserved  Research focus: Integrative biology and agricultural sustainability  Public education:oriented to raise public understanding of value of plant diversity and preventing desertification in Northern China

7. Main CAS Botanical Gardens Wuhan BG ：  Conservation focus: aquatic and endemic plants in eastern central China; 6000 species conserved  Research focus: Conservation genetics, Aquatic plant biology and Ag-forest ecology  Public education: for public awareness of relationships b/w water quality and aquatic plant integrity

8. Main CAS Botanical Gardens South China BG ：  Conservation focus: Southern subtropical flora; 7000 species conserved  Research focus: Plant systematics, Restoration ecology and Agriculture  Public education: understanding of plant richness and economical and agricultural values

9. Main CAS Botanical Gardens Xisuanbanla tropical BG ：  Conservation focus: Tropical rain forests; 10,000 species conserved  Research focus: Ecology, Conservation biology, economic tree crops  Public education: public understanding critical value of rain forests and global climate changes and sustainability

10. CAS BG Efforts in Conserving Chinese Native Plants About 30,000 vascular plants in China, 1/10 of the world ’ s total. One of the richest country with plant diversity The most conservative estimated 20% in status of rare and endangered The large country with many different ecosystems and complicated threatened factors Progresses have been made since 80 ’ s, but research on basic principles and methods for conservation are urgently needed few successful case studies in restoration and recovery

11. CAS Current Project and Progress A key CAS project: “ Mechanisms and Conservation Principles for Rare and Endangered Plants ” Goals ： 1) To determine factors impacting main rare and endangered plants represented in China 2) To provide scientific guidelines for sampling strategies and conservation practice Research:10 well selected plant species 1 ） Genetic diversity of R & E plants; 2 ） Cytogenetics of R & E plants; 3 ） Ecology of R & E plants; 4 ） Seed physiology of R & E plants; 5 ） Conservation principles and practice

12. Fern —Isoetes sinensis CAS Current Project and Progress — Ten critical endangered species

13. CAS Current Project and Progress — Ten critical endangered species Abies chensiensis Cupressus chengiana Gymnosperm— Abies and Cupressus

14. CAS Current Project and Progress — Ten critical endangered species Herbaceous angiosperm Taihangia rupestris Panax stipuleanatus

15. CAS Current Project and Progress — Ten critical endangered species Woody angiosperm Ammopiptanthus mongolicus /A. nanus Manglietia patungensis

16. CAS Current Project and Progress — Ten critical endangered species Woody angiosperm Berchemiella wilsonii Trigonobalanus doichangensis Parashorea chinensis

17. CAS Current Project and Progress — Geographic regions and habitats Geographical and climatologically regions represented in China: North, Northwest, Central, East and Southwest regions Habitat niche significance: unique ecosystem and special habitats of desert, mountain, tropical rainforest, dry river valleys and southern wetland

18. Case study — Isoetes sinensis （ Natural habitat and ecology ） A relic and perennial wetland fern, rapidly decreasing natural ranges Very few populations remained ， listed as national top priority for protection Natural range: Zhejiang, Anhui, Jiangxi and Jiangsu provices; extinct in many locations Individuals in wild<3000 ； The largest population < 300 m 2

19. Case study — Isoetes sinensis (Genetic diversity — Allozymes ） Sampling in natural range 10 enzymes ， 16 loci in total, 40 alleles, 13 polymorphic Population genetic diversity (species) P=50% (81%), A=1.6 (2.4), He=0.25 (0.30), G ST =0.2953; N m =0.596 Obs. Ho (0.430) > exp. He (0.252), fixed heterozygosity by allotetraploity Few private and rare genes Only 18 multi-locus genotypes identified in total 150 individuals, with 3 dominant genotypes, indicating a high clonal reproduction

20. Case study — Isoetes sinensis (Genetic diversity — AFLPs ） EcoRI/MseI degestion 8 of 48 selective primers resolved high resolution patterns Specie level ： P= 61.6% Population level ： P=35.2% ， He=0.147 ， G ST =0.559, N m =0.1973 AMOVA(nested): 46.9% between groups; 23.2% between Populations Significant correlation found between population area and genetic diversity

21. Case study — Isoetes sinensis (Genetic diversity — AFLPs ） Cluster analysis revealed clear isolation between individuals, populations by geographic isolation Relationship between genetic diversity and fitness of individuals and populations. for example ： growth rate, tillering ability, spores reproduction, seedling survival rate E-ACT/M-CTG

22. Case study — Isoetes sinensis (Cytogenetics ） Allotetraploid (2n=4X) Normal mitosis Meiosis: irregular behaves observed in Microspore and Macrospore mother cells Chromosome bridge, fragments and lagging chromosomes observed Defect cell wall in microspore mother cells, nucleoplasm leakage or cytomisis during meiosis E-ATG/M-CATE-ACT/M-CTG

23. Case study — Isoetes sinensis ( Threatened factors and conservation strategy ) Ex situ normal reproduction by tillering and spores germination Habitat loss by anthropogenic activities and water pollution directly caused reduction of natural populations Remaining populations are in disadvantage to compete with companion species, such as Juncus effuses etc. dominant wetland species. It is highly possible to be replaced by these species An proposed approach for integrated conservation including both in situ and ex situ conservation E-ATG/M-CATE-ACT/M-CTG

24. Case study- Berchemiella wilsonii （ Natural habitat and ecology ） B. wilsonii was first identified and described at western Hubei in 1907 It has not been found in past about 100 years until two trees were rediscovered in 2002 at Houhe Natural Reserve, Hubei Province Its closely related B. wilsonii var. pubescentibus distributed ： Anhui and Zhejing provinces Its natural habitat continues to be deteriorating, the species is in critical endangered

25. Case study- Berchemiella wilsonii （ Genetic diversity — Allozymes, AFLP ） Nine enzymes resolved 20 loci and 42 alleles, 17 polymorphic loci; 8 AFLP primers ： 122 bands, of which 45 polymorphic Several fixed heterozygosity across populations Very a few alleles with low frequency (<.01) ： one in Ruikou ， 3 in Maxiao, Huoshan and Shucheng, respectively Pop.NAP (AFLP)HoHe (AFLP) Ruikou251.8570 (25).580.349 (.099) Maxiao181.8570 (26).592.345 (.105) Shucheng241.8575 (26).563.324 (.159) Huoshan311.8070 (30).545.308 (.177) Mean24.51.8471.3 (26.8).590.348 (.135) Overall2.1085 (36.9).567.378 (.149)

26. Case study- Berchemiella wilsonii （ Difference in genetic differentiation ） FitFstFisNm Overall population -0.4940.130**-0.7171.68 Within population Zhejiang group -0.5530.078*-0.6832.97 Anhui group -0.7280.014 NS -0.75317.28

27. Case study- Berchemiella wilsonii （ Cytogenetics ） Meiosis: obvious mal- behavior, many lagging chromosomes and micronucleolus observed during Anaphase II Cell membrane of microspores depressed, irregularly developed male gametes

28. Case study- Berchemiella wilsonii （ Seed physiology and Conservation ） Seeds collected from all natural ranges were studied, most embryos were not developed and very poor seed germination ， only two seedlings obtained Propagation by hard cuttings unsuccessful Tissue culture successful, complete plantlets obtained

29. A successful case study — Conservation and restoration of the three gorge endemic Myricaria laxiflora Natural range ： along riverside of water fluctuation zone < 130 m of Yangtze River valley in 12 counties from Chongqing – Yichang, Hubei, 31 populations identified, about 90,000 individuals Central range ： 11 populations at Zigui county, Hubei Entire range submerged by three gorge dam raising water level 175 m high

30. The three gorge - Myricaria laxiflora （ Evolutionary history and species traits) 13 species in genus Myricaria grown in northern temperate Euro-Asia, a typical alpine genus distributed in Tibet and neighboring regions as Himalayas center 10 species native to China, mainly distributed plateau areas of southwest and Northeast China, at typical habitats of river/lakeside and low place of desert sand dunes Only Myricaria laxiflora growing at low altitude region of Yangtze river

31. The three gorge - Myricaria laxiflora （ Important species traits) Tolerance to flooding: deep rooted, root well developed; survived in completely covered by sands Tolerance to water submerging ： dormancy when flooding for maximum 6 months Rapid vegetative growth after flooding retreated, 2m new grwoth observed Long and continuous flowering High yielding seeds, water float

32. The three gorge - Myricaria laxiflora （ Ex situ Conservation – entire range survey) Simple community ： Single species, or +Salix variegata, +Distyliun chinensis, + Salix variegata + Cynodon dectylon, etc. Habitat environment ： topography, climate, soils, etc. Natural range ： Chongqing - Yichang Population size and structures: 31 population surveyed

33. The three gorge - Myricaria laxiflora （ Breeding system and propagation strategy) Wind/insect pollination, mainly outcrossing species Environment adapted twice flowering per year Unterminate inflorescence and continuous flowering and seed set Wind and water seed dispersal Short life of seed, not well stored Cuttings ： Low part of spring shoot/sand media > 90% rooted, 50 ppm IBA 20 hr. 100% rooted

34. The three gorge - Myricaria laxiflora （ Ex situ Conservation — Genetic diversity) 13 main wild populations, 20-30 trees/population and one ex situ population (WBG ） High genetic diversity: Allozyme: P=87.7%; A=1.8; He=0.317 AFLP: P=41.25%; He=0.115 Also significant difference among populations; excess heterozygotes 60-85% of total diversity resided within population (G ST = 0.1514 – 0.397) Gene flow (Nm=0.7581-1.401)

35. The three gorge - Myricaria laxiflora （ Sampling strategy and increase propagation ） Focused sampling from the populations with higher genetic diversity and large size: Zigui, Xintan, Wuxia and Fuling in order to encompass >95% of the total genetic diversity Sampling rare and local alleles in: Xinling, Niukou and Nanmuyuan Allozymes and AFLPs confirmed all genetic parameters of ex situ populations > individual wild population Increase propagation ： at WBG and Sixi site within three gorges, wild plants transplanting, cuttings and seedling

36. The three gorge - Myricaria laxiflora （ Restoration ） New habitat selection: Water fluctuation and submerge 3-6months Growing season: dormancy during flooding period (May – Oct.); growing during water down (Oct. – April) Climate ： warm winter, moisture summer, weak competitor Suitable locations at up and down stream from the dam, and branch rivers

37. The three gorge - Myricaria laxiflora （ Restoration ）

38. BG Sustainable Development — from germplasm to sustainability Beijing BG: grape cultivar development Wuhan BG: kiwifruit cultivar development Southern China BG: Santalum album Xishuanbanla BG:

39. A case example of sustainable development at Beijing BG: A series commercial wine grape cultivars were developed by hybridization European grapes and Amur (V. amurensis) genotypes based on extensive collections and evaluation of wild grape germplasm, Amur grape native to northern China Grape juice cultivars were developed from hybridization of European grapes and wild germplasm (V. thunbergii ) 北紫北紫 Beifeng BeichunBeimei Beizi

40. A case example of sustainable development at Wuhan BG Actinidia: 66 species and 118 taxa, in the world Sixty-two species in China Most species: edible fruits Two main species and two minor species are in commercial production: A. deliciosa, A. chinensis A. eriantha, A. arguta A. deliciosa A. chinensis A. erianthaA. arguta

41. Other Species with Horticultural and Commercial Significance A. chinensis var. rufopulpa A. arguta var. purpurea A. macrosperma A. valvata A. kolomikta A. rufa A. latifolia

42. Main cultivars developed from the genetic resources in China Kuimi 魁蜜 Jinfeng 金丰 Lushanxiang 庐山香 Zaoxian 早鲜 Miliang-I Jinkui

43. A case example of sustainable development at Wuhan BG Jintao: a novel yellow-fleshed kiwifruit (A. chinensis) developed from the breeding program at the Wuhan BG patented in Europe and South America. The propagation right was licensed for 28 years Royalty JINTAO

44. A case example of sustainable development at Southern China BG Santalum album native to India and Malaysia, but widely used as perfume and traditional Chinese medicine dated back to thousand year ago in China. China has been the largest importing country for S. album materials 12 seeds introduced in 1962, propagation and selection program successfully developed at SCBG and commercial production protocols established for this semiparasite plants

45. A case example of sustainable development at Xishuangbanla BG Dragon blood (resin derived from Dracaena draco, D. cinnabari and D. loureiri) is high value medicine, and widely used as traditional Chinese medicine dated back to 1500 year ago in China In 70s, native Chinese species D. cochinnensis was found in southwest China and used for production of Dragon blood Natural resource has been depleting rapidly. GAP protocol was developed and commercial manufacture of capsules and other products established