星野黑皮海綿(Terpios hoshinota)是一種與行光合作用的藍綠菌共生的平鋪狀海綿(Rutzler and Muzik 1993)。此種海綿會覆蓋石珊瑚，造成珊瑚的死亡。台灣於2006年珊瑚礁總體檢報告中，發現綠島的石珊瑚疑似受到黑皮海綿的感染，而2008-2010在蘭嶼與綠島的野外調查顯示，黑皮海綿在綠島及蘭嶼的淺海域有大量繁生的現象。黑皮海綿群體可能擴散的方式，可分為自行播種(Self-Seeding) 及遠方擴散(Long-range dispersal)兩假說。本研究使用核糖核苷酸 與粒線體核苷酸序列做為分子標記，探討黑皮海綿在台灣周邊海域與日本琉球群島和中國西沙島族群的擴散之方式是屬於自行播種或是遠方擴散。我們於2008年至2009年間採集了台灣墾丁萬里桐、 綠島、蘭嶼 ，日本之宮古島 (Miyako)、今歸仁 (Nakijin)、可米島 (Arahama Kumejima)、沖繩 (Bise Okinawajima)、石垣 (Shiraho Ishigakijima) 、沖永良部島(Yakomo Okinoerabu) 、德之島 (San Tokunoshima)與中國西沙永興島的黑皮海綿，總共110份獨立樣本，使用粒線體核苷酸序列細胞色素C氧化酶（COI）和核糖核苷酸之第二轉錄區間序列（ITS2）變異探討黑皮海綿之族群遺傳結構。

黑皮海綿 COI 序列在台灣綠島和蘭嶼、日本與中國的樣本之中無任何位點的差異，然而台灣萬里桐的黑皮海綿有三個位點上的差異，顯示萬里桐的黑皮海綿可能為不同種。 ITS2分析顯示黑皮海綿之單型多樣性在所有族群之中皆偏高，然而綠島(Green Island)與日本可米島(Arahama Kumejima)內，卻分別僅有單一單型存在。族群分化指數 (Fst) 測試顯示，族群之間有顯著的差異，推測黑皮海綿主要的散播方式以自行播種為主，但伴隨著少數的遠方擴散事件發生於Bise、Shiraho與蘭嶼之間。 分子變方分析顯示黑皮海綿族群之間的差異大於族群內的差異。TCS分析顯示，綠島的單一黑皮海綿單型起源於蘭嶼，而日本Bise地點之黑皮海綿遍佈於日本、台灣等地之族群，綜合核甘酸多樣性、單行多樣性分析與黑皮海綿野外發現之歷史資料，顯示Bise為本研究地點中的黑皮海綿起源。巢式支序分析顯示黑皮海綿族群間之基因交流受到限制，但仍然有某族群呈現鄰近擴張狀態。單型頻率分析則顯示在大部分的黑皮海綿族群之中皆共享一個優勢種類之黑皮海綿單型，該優勢種不僅在當地族群中佔了絕大多數的族群數量，且相對於其他海綿單型來說，該優勢單型可能擁有較佳的散播能力或繁殖能力，使得優勢單型能出現於大多數之黑皮海綿族群之中。

根據結果，星野黑皮海綿並非容易在島嶼之間快速散播的物種，該海綿主要以自行播種的方式在散播，伴隨著偶然發生的遠方擴散事件發生，並減少因距離產生隔離之現象。綠島之黑皮海綿主要是源自於蘭嶼的族群，並且以自行播種的方式在該地繁殖，於數年之內發生族群爆發而大量佔據了綠島，蘭嶼之海綿族群則可能來自於日本地區之Bise、Shiraho與Yakomo，並且也正經歷著族群爆發之現象。日本的黑皮海綿族群之間有明顯的基因差異，並且經歷過奠基者效應與快速的族群增長，而Bise地區則為本研究中黑皮海綿之起源。在黑皮海綿族群之中，並非所有的黑皮海綿都有能力成為該族群的優勢種，在大部分台灣與日本的族群之中，顯示其中某一種單型的黑皮海綿個體有能力成為該族群的優勢種，且該優勢單型不管在族群數量或是族群範圍皆大於其他單型的星野黑皮海綿個體，而該優勢單型也橫跨了台灣、日本的島嶼之間，成為多數地區的優勢單型。

The encrusting sponge Terpios hoshinota is a cyanobacteriosponge with symbiotic photosynthetic cyanobacteria. It covers live corals causing their death. Corals at Green Island were suspected to be infected by Terpios hoshinota in 2006, and field investigations indicated there was massive propagation of the species in both Green Island (Lyudao) and Orchid Island (Lanyu) in 2008 to 2010. We propose two hypotheses, either by Self-Seeding or by Long-Range Dispersal, that explain the fast propagation of Terpios hoshinota in the islands offshore of southeastern of Taiwan. We use ribosomal DNA and mitochondria DNA as molecular markers to investigate how the sponge disperses locally and in a greater geographic scale. A total of 110 samples, from Taiwan: Green Island, Orchid Island, and Kenting (Wanlitong). Japan: Okinawa, Nakijin, Miyako, Bise, Shiraho, Arahama Kumeshima, Yakomo (Okinoerabu), San (Takunoshima), and Xisha Island of China, were collected. Internal Transcribed Spacer 2 (ITS2) from ribosomal DNA and cytochrome oxidase I (COI) from mitochondria DNA are used as markers to infer population structure of Terpios hoshinota.

No genetic variation within COI sequence over all sponges from Taiwan to Japan and China was found, although the only sponge sample from Wanlitong in Kenting had three variable sites, which suggest different species of Terpios hoshinota. Based on ITS2 analysis, haplotype diversity (h) is commonly high among most populations, but with different single haplotype found at Green Island and Arahama (Japan). Pairwise population differentiations (FST) are usually high and significant among populations supporting self-seeding, although Bise, Shiraho and Lanyu populations showed no significant differentiation that supports long-range dispersal. Analysis of Molecular Variance (AMOVA) shows no population subdivision; however, genetic differentiations among populations are significantly greater than within populations. TCS analysis indicates that single haplotype in Green Island is originated from Lanyu, and populations in Bise are widely dispersed over other sponge populations in Taiwan and Japan regions. By evidence of TCS analysis with nucleotide diversity, haplotype diversity and field investigation, Bise is the origin of Terpios hoshinota among populations within this study. Frequency of sequence haplotypes indicates one dominant haplotype is shared among most of the sponge populations, and the dominated sponge haplotype takes highest proportions of local populations. The existence of dominant haplotype may result from better dispersal or reproduction ability than other haplotype in populations. Nested clade analysis shows that populations mainly have restricted gene flow with some clade have contiguous range expansion.

We suggest that populations of Terpios hoshinota propagate mainly by self-seeding method with occasional long-range dispersal event that leading to genetic connection among populations and obscuring evidence of isolation by distance in these populations. In Green Island, we consider local populations as undergoing explosion within past several years and propagate by self-seeding method coming from single lineage of Lanyu. Populations in Lanyu may come from Bise, Shiraho, and Yakomo, yet may still in status of population explosion. Populations in Japan may underwent founder effect with rapid population growth, while most populations are rarely interact with each other showing deep genetic differentiation among islands, and Bise is the origin of Terpios hoshinota in this study. Not all of the sponge individuals have ability to dominate local populations, expect for one special haplotype of Terpios hoshinota is capable of dominating local population in both range and quantity, which also has capability of spreading across islands as larger distances than its habitats range in Taiwan and Japan.

1.Introduction:01-06

2.Materials and Methods:07-15

3.Results:16-21

4.Discussion:22-3

5.Appendix1:31-38

6.Appendix2:39-40

7.References:41-52

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