摘 要
諸羅樹蛙（Rhacophorus arvalis Lue, Lai, and Chen, 1995），為台灣特有兩棲類，目前僅在台灣南部的雲林北港溪、嘉義朴子溪及台南(麻豆)呈不連續分布，主要棲息環境為竹林、果園、稻田，棲息地易受人為的干擾，由於本身移動能力不佳，加上雄蛙對棲地有相當程度忠誠度（site fidelity）行為，是適合探討受棲地零碎衝擊的物種。現今野外諸羅樹蛙分佈地點，依照水系流域，可分成北港溪北岸、北港溪南岸、朴子溪與曾文溪(麻豆)四個不同群族，利用120隻個體增幅粒線體COX1、cyto b與D-loop部分序列，根據單基因型(haplotype)、核酸歧異度、族群分化、交流指數、AMOVA(analysis of molecular variance)、最小關聯網狀樹狀圖(minimum spanning network)與NCA( Nested clade analysis)方法，進行族群遺傳結構探討，結果顯示諸羅樹蛙最大種內變異程度僅0.004，為國內目前已發表兩棲類中最低者，其中麻豆族群內的遺傳變異為0。另外各地族群皆從很小的有效繁殖族群而來，推測可能和歷史棲地不可測(Stochastic)的環境壓力與人為干擾有關。北港溪南、北流域族群呈現小族群快速成長，而朴子溪與麻豆族群則是受長期瓶頸效應的小族群。從地理親緣關係來看，朴子溪流域的族群，過去可能從北港溪周遭擴散遷移而來。“過去零碎”(past fragmentation) —溪流間陸域的的隔離，導致四條流域諸羅樹蛙族群缺乏基因交流，在瓶頸效應下喪失基因多樣性，存在顯著的族群結構，成為四個不同的族群。目前麻豆族群建議為短期優先保育的地區，北港溪周遭流域族群為長期保育的熱點(hotspot)。

Abstract
Farmland green tree frog, Rhacophorus arvalis is an endemic species of Taiwan. Its main habitats include rice fields, bamboo plantations, and orchards. These environments are frequently disturbed through various types of human activities. Currently it is found only in limited farmlands in Yunlin, Chiayi, and Tainan counties. Apparently this species is greatly threatened by habitat fragmentations and other human activities. Farmland green tree frogs have limited dispersal capabilities due to low mobility and site fidelity. It is one of the best candidate animals to study the impacts of habitat fragmentations on wildlife. Based on their current distribution, farmland green tree frogs can be separate into four populations: north bank of Peikang stream、south bank of Peikang stream、Potz stream and Madou. A total of 120 individuals were used to analyze with partial mitochondrial COX1, cyto b and D-loop gene fragments. Haplotypes, nucleotide diversity, FST, Nm, AMOVA (analysis of molecular variance), minimum spanning network and NCA (Nested clade analysis) methods were applied. The Farmland green tree frog’s maximum intra-specific genetic variance is 0.004, which is the lowest among amphibian being investigated in Taiwan. Even the genetic variance of Madou population is 0. Molecular data showed that four populations all derived from small effective breeding populations. We suspected that the historical stochastic factors and human disturbance might play important roles in this aspect. The north and south bank of Peikang stream populations both were rapid growing. Nevertheless, Potz and Madou population have experienced a period of bottleneck. The phylogeographic evidences indicate that Potz population might come from Peikang population. Data also revealed a consistent phylogeographic pattern, and indicated the results of “past fragmentation”. This leads to low gene flow among populations and bottleneck effect. Genetic variations were lost greatly among population and formed four significantly different populations. We suggest madou population given the higher priority for species conservation in short term. Population of Peikang stream is the genetic hotspot for conservation.

參考文獻
Aizen, M. A., and P. Feinsinger. 1994. Forest fragmentation, pollination, and plant reproduction in a Chaco dry forest, Argentina. Ecology 75: 330–351.
Alford, R. A., and S. J. Richards. 1999. Global amphibian declines: a problem in applies ecology. Annual Review of Ecology and Systematics 30: 133-165.
Avise, J. C. 1986. A note for molecular genetics in the recognition and conservation of endangered species. Trend in Ecology and Evolution 4: 279-281.
Avise, J. C. 2000. Phylogeography: The history and formation of species. Harvard University Press, Cambridge.p59-60.
Barber, P. H. 1999. Patterns of gene flow and population genetic structure in the canyon treefrog, Hyla arenicolor (Cope). Mol. Ecol. 8: 563-576.
Blaustein, A.R., and J.M. Kiesecker. 2002. Complexity in conservation: lessons from the global decline of amphibian populations. Ecology Letters 5: 597-608。
Bossart, J. L., and D. P. Prowell. 1998. Genetic estimates of population structure and gene flow: Limitations, Lessons and new directions. Trend in Ecology and Evolution 13: 202-206.
Bouzat, J. L., H. A. Lewin, and K. N. Paige. 1998. The ghost of genetic diversity past: historical DNA analysis of the Greater Prairie Chicken. Am. Nat., 152: 1–6.
Buza, L., A. Young, and P. Thrall. 2000. Genetic erosion, inbreeding and reduced fitness in fragmented populations of the endangered tetraploid pea Swainsona recta. Biological Conservation 93: 177–186.
Caizergues, A. 2003. Population genetic structure of male black grouse (Tetrao tetrix L) in fragmented vs. continuous landscapes. Mol. Ecol 12: 2297-2306.
Clarke, G. M. 1999. Golden sun moth Synemon plana draft recovery plan. Sydney: NSW National parks and Wildlife Service.
Clarke, G. M., C. O’Dwyer. 2000. Genetic variability and populations structure of the endangered golden sun moth, Synemon plana. Biological Conservation 92： 371-381.
Crandall, K. A., and A. R. Templeton. 1993. Empirical tests of some predictions from coalescent theory with applications to intraspecific phylogeny reconstructure. Genetics 134: 959-969.
Desplanque, B., F. Viard, J. Bernard, D. Forcioli, P. Saumitou-Laprade, J. Cuguen, and H. van. Dijk. 2000. The linkage disequilibrium between chloroplast DNA and mitochondrial DNA haplotypes in Beta vulgaris ssp. Maritime （L.）: the usefulness of both genomes for population genetic studies. Mol. Ecol. 9: 141-154.
Dodd, C. K. 1994. The effects of drought on population structure, activity, and orientation of toads （Bufo quercicus and B. terrestris） at a temporary pond. Ethology, Ecology and Evolution 6: 331-349.
Driscoll, D. A. 1998. Genetic structure of the frogs Geocrinia Lutea and Geocrinia Rosea reflects extreme population divergence and range changes, not dispersal barriers. Evolution 52（4）: 1147-1157.
Ehrlich, P. R., and D. D. Murphy. 1987. Conservation lessons from long-term studies of checker spot butterflies. Conservation Biology 1: 122-131.
Excoffier, L., P. E. Smouse, and J. M. Quattro. 1992. Analysis of molecular variance inferred from metric distances among DNA haplotypes: Application to human mitochondrial DNA restriction data. Genetics 131: 479-491.
Excoffier, L., P. E. Smouse, and J. M. Quattro. 1993. Analysis of Molecular Variance. Version 1.55. Genetics and Biometry Laboratory. University of Geneva.
Excoffier, L., and P. E. Smouse. 1994. Using allele frequencies and geographic subdivision to reconstruct gene tree within a species: Molecular variance parimony. Genetics 136: 343-359.
Fahrig, L., J. H. Pedlar, S. E. Pope, P. D. Taylor, and J. F. Wegner. 1995. Effect of road traffic on amphibian density. Biological Conservation 73: 177-182.
Felsenstein, J. 1981. Evolutionary trees from DNA sequences: a maximum likelihood approach. Mol. Ecol 17: 368-376.
Futuyma, D. J. 1998. The geography of evolution. In：Evolutionary biology, 3rd ed. Sinauer Associates, Inc., Sunderland.
Graur, D., and W. H. Li. 2000. Fundamentals of Molecular Evolution, 2nd ed. Sinauer Associates, Inc., Sunderland.
Grativol, A. D., J. D. Ballou, and R. C. Fleischer. 2001. Microsatellite variation within and among recently fragmented populations of the golden lion tamarin（Leontopithecus rosalia）.Conservation Genetic 2: 1-9.
Gross, C. L. 2001. The effect of introduces honeybees on native bee visitation and fruit-set in Dillwynia Juniperina （Fabaceae） in a fragmented ecosystem. Biol. Conservation 102: 89-95.
Grant, W. A. S., and B. W. Bowen. 1998. Shallow population histories in deep evolutionary lineages of marine fishes: insights from sardines and anchovies and lessons for conservation. Heredity 89: 415-426.
Golding, G. B. 1987. The detection of deleterious selection using ancestors inferred from a phylogenetic history. Genet. Res. 49: 71-82.
Hale, M. L., P. W. Lurz, M. D. Shirley, S. Rushton, R. M. Fuller, and K. Wolff. 2001. Impact of landscape management on the genetic structure of red squirrel population. Science 293: 2246-2248.
Harrison, S., and A. Hastings. 1996. Genetic and evolutionary consequences of metapopulation structure. Trend in Ecology and Evolution Ⅱ: 180-183.
Hans, V. D., and E. Matthysen. 1999. Habitat fragmentation and insect flight: a changing “design” in a changing landscape? Trend in Ecology and Evolution 14: 172-174.
Haila, Y. 2002. A conceptual genealogy of fragmentation research: from island biogeography to landscape ecology. Ecological Application 12(2): 321-334.
Hudson, R. R. 1990. Gene genealogies and the coalescent process. Oxford University Press, New York.
James, F. C. 1991. Signs of trouble in the largest remaining population of red-cockaded woodpeckers. Auk 108:419-423.
Joanna, S., J. Tim, P. David, and M. Craig. 2004. Limited effect of anthropogenic habitat fragmentation on molecular diversity in a rain forest skink, Gnypetoscincus queenslandiae. Mol. Ecol. 13: 259-269.
Jules, E. S. 1998. Habitat fragmentation and demographic change for a common plant: Trillium in old-growth forest. Ecology 75(5): 1645-1656.
Kaufmann, J. H. 1992. Habitat use by wood turtle in central Pennsylvania. Journal of Herpetology 26: 315-321.
Kimura, M. 1980. A simple method for estimating evolutionary rates of base substitutions trough comparative studies of nucleotide sequence. Mol. Evol 16: 111-120.
Kimura, M. 1983. Rare variant alleles in the light of the neutral theory. Molecular biology evolution 1: 84-93.
Kumar, S., K. Tamura, I. B. Jakobsen, and M. Nei. 2001. MEGA2: Molecular Evolutionary Genetics Analysis. Arizona State University, Tempe, Arizona, USA.
Land, D. E., and R. C. Lacy. 2000. Introgression level achieved through Florida panther genetic restoration. Endangered Species update 17: 99-103.
Laurance, W. F. 1998. Rain forest fragmentation and the dynamics of Amazonian tree communities. Ecology 79: 2032–2040.
Laurie, W. C., and L. Fahrig. 2001 Effect of road traffic on two amphibian species of differing vagility. Conservation Biology 15(4): 1071–1078.
Lamont, B. B. 1993. Population fragmentation may reduce fertility to zero in Banksia goodii a demonstration of the allee effect. Oecologia 94: 46-450.
Lidicker, W. Z. 2002. From dispersal to landscapes: progress in our understanding of population dynamics. Acta Theriologica 46: in press.
Lue, K. Y., J. S. Lai, and S. L. Chen. 1994. A new Rhacophorus（Anura: Rhacophoridae from Taiwan. Herpetology 50(3): 303-308.
Lynch, M., and B. Walsh. 1998. Genetics and analysis of Quantitative Traits. Sinauer Associates, Inc., Sunderland.
Nei, M., and F. Tajima. 1983. Maximum likelihood estimation of the number of nucleotide substitutions from restriction sites data. Genetics 105: 205-217.
Nei, M. 1987. Molecular Evolutionary Genetics. Columbia Univ. Press. New York.
Martof, B. S. 1959. Territoriality in the green frog, Rana Clamitans. Ecology 34(1): 165-174.
Mork, J., N. Ryman, G. Staahl, F. Utter, and G. Sundnes. 1985. Genetic variation in Atlantic cod （Gadus morhua） throughout its range. Can. J. Fish. Aquat. Sci 42: 1580-1586.
Moritz, C. 1999. Conservation units and translocations: strategies foe conserving evolutionary process. Heredity 130: 217-228.
Mossman, C. A., and P. M. Waser. 2001. Effects of habitat fragmentation on population genetic structure in the white-footed mouse （Peromyscus leucopus）. Canadian Journal of Zoology 79: 285-296.
Page, R. D. M., and E. C. Holmes. 1998. Molecular evolution: A phylogenetic approach. Blackwell Science Inc. London.
Paulo, O. C., W. C. Jordan, and M. W. Bruford. 2002. Using nested clade analysis to assess the history of colonization and the persistence of population of an Iberian Lizard. Mol. Ecol. 11: 809-819.
Posada, D., K. A. Crandall, and A. R. Templeton. 2000. GeoDis: a program for the cladistic analysis of the geographical distribution of genetic haplotypes. Mol. Ecol. 9: 487-488.
Reh, W., and A .Seitz. 1990. The influence of land use on the genetic structure of populations of the common frog Rana temporaria. Biological Conservation 54: 239-249
Rozas, J., and R. Rozas. 1997. DnaSP version 2.0: a novel software packed for extensive molecular population genetic s analysis. Comut. Applic. Biosci 13: 307-331.
Robert, A. N., and S. Tina. 2001. Microsatellite variation and fine-scale population structure in the wood frog (Rana sylvatica). Mol. Ecol. 10: 1087-1100.
Santos, T., and J. L. Telleria. 1997. Vertebrate predation on Holm Oak, Quercusilex, acorns in a fragmented habitat: effect on seedling recruitment. Forest Ecology and Management 98(2): 181-187.
Schneider, S., Roessli, D, and Excoffier, L. 2000. Arlequin: A software for population genetics data analysis. Ver. 2.0. Genetics and Biometry Lab, Dept. of Anthropology, University of Geneva.
Schmiegelon, F. K. A., and M. Monkkonen. 2002. Habitat loss and fragmentation in dynamic landscapes: avian perspectives from the boreal forest. Ecological Applications 12(2): 375-389.
Srikwan, S. 1998. Genetic erosion in small mammal populations following rain forest fragmentation in Thailand. PhD thesis, University of California, San Diego.
Slatkin, M. 1985. Gene flow in natural population. Annual Review of Ecology and Systematics 16:393-430.
Slatkin, M. 1993. Isolation by distance in equilibrium and non-equilibrium populations. Evolution 47: 264-279.
Stow, A. D., P. Sunnucks, D. A. Briscoe, and M. G. Gardner. 2001. The impact of habitat fragmentation on dispersal of Cunningham’s skink (Egernia cunninghami): evidence from allelic and genotypic analysis of microsatellites. Mol. Ecol. 10: 867-878.
Tajima, F. 1989. Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics 123: 585-595.
Templeton, A. R., E. Boerwinkle, and C. F. Sing. 1987. A cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping.Ⅰ. Basic theory and an analysis of alcohol dehydrogenase activity in Drosophila. Genetics 117:343-351.
Templeton, A. R., and C. F. Sing. 1993. A cladistic analysis of phenotypic associations with haplotypes inferred from restriction endonuclease mapping. Ⅳ. Nested analyses with cladogram uncertainty and recombination. Genetics 134: 659-669.
Templeton, A. R., E. Routman, and C. A. Phillips. 1995. Separating population structure from population history: a cladistic analysis of the geographical distribution of mitochondrial DNA haplotypes in the tiger salamander, Ambystoma tigrinum. Genetics 140: 767-782.
Templeton, A. R. 1998. Nested clade analyses of phylogeographic data: testing hypotheses about gene flow and population history. Mol. Ecol. 7: 381-397.
Thomas, S. 2002. Influence of habitat fragmentation on the genetic structure of polyommatus coridon (Lepidoptera: Lycaenidae）: implications for conservation. Biological Conservation 107: 291-297.
Tomas, R. 2002. So near and yet so far-habitat fragmentation and bird movement. Trend in Ecology and Evolution 17: 61.
Vos, C. C., A. G. Antonisse-De Jong, P. W. Goedhart, and M. J. Smulders. 1998.
Genetic similarity as a measure for connectivity between fragmented populations of the moor frog (Rana arvalis). Journal of Applied Ecology 35: 44-56.
Vrijenhoek, R. C. 1994. Unisexual fish: models for studying ecology and evolution. Annu. Rev. Ecol. Syst. 25: 71–96.
Whitlock, M. C., and N. H. Barton. 1997. The effective size of a subdivided population. Genetics 146: 427-441.
White, G. M., D. H. Boshier, and W. Powell. 2002. Increased pollen flow counteracts fragmentation in a tropical dry forest: an example from Swietenia humilis Zuccarini. PANS 99: 2038~2042.
Wright, S. 1978. Evolution and genetics of populations. Chicago University Press, Chicago.
Young, A. G., A. H. D. Brown, and F. C. Zich. 1999. Genetic structure of fragmented populations of the endangered grassland daisy, Rutidosis Leptorrhynchoides. Conservation biology 13: 256-265.
Young, A. C., and G. M. Clarke. 2000. Genetic, Demography and Viability of Fragmented Population. Cambridge University Press, Cambridge.
江友中，2002。八丈芒分子地理親緣之研究。國立台灣師範大學生物研究所博士論文。
江樹生，1985。梅花鹿與台灣早期歷史關係之研究。內政部營建署墾丁國家公園管理處。
呂光洋，2003。諸羅樹蛙～我終於想通了。台灣濕地 45：18-20。
沈聖峰、李培芬、袁孝維，2003。零碎化棲地的穩定冠羽畫眉族群：食物、巢位與棲地選擇。棲地零碎化、生態廊道及棲息地網研討會論文集。P185-p198。
林思民，2003。台灣及鄰近地區草蜥屬的親緣關係與親緣地理學研究（有鱗目：蜥蜴科）。國立台灣師範大學生物研究所博士論文。
徐堉峰，2003。夸父璀灰蝶的族群棲地分布特性初探。棲地零碎化、生態廊道及棲息地網研討會論文集。P64-p68。
郭瓊華，2002。台灣蜓蜥族群遺傳結構之研究。國立台灣師範大學生物研究所碩士論文。
莊孟憲，2005。麻豆地區諸羅樹蛙生態及其棲息地調查。地方生態資源共管座談會（消失中的諸羅樹蛙），台南。
莊鎮碩，1999。諸羅樹蛙生殖行為之研究。國立台灣師範大學生物研究所碩士論文。
黃鈞漢，2002。以mtDNA D-loop片段探討艾氏樹蛙（溪頭地區）之族群遺傳結構。國立彰化師範大學生物研究所碩士論文。
曾晴賢，2003。台灣櫻花鉤吻鮭族群監測與生態調查(六)。內政部營建署雪霸
 國家公園管理處委託研究報告。
葉文珊，1997。莫氏樹蛙族群地理親源關係之研究。國立台灣大學動物學研究所碩士論文。
楊懿如，1994。台北樹蛙生殖生態與族群基因組成變異。國立台灣大學動物學研究所博士論文。
劉怡里，2000。以RAPD方法分析台灣產樹蛙屬樹蛙之族群遺傳結構。國立台灣大學動物學研究所碩士論文。
蔡長益，1999。以粒線體12S核糖體核酸與細胞色素b序列分析守宮族群親緣關係。國立中山大學生命科學系碩士論文。
謝煒智，2004。諸羅樹蛙群集展示行為研究。國立台灣師範大學生物研究所碩士論文。