饰纹姬蛙种群分化与谱系地理研究

Population differentiation and some new species has been discovered in Microhyla fissipes, indicating complicated ralatonships embeded in this species;It distributed mainly in southern part of eastern Asia,peninsula north to China,and dwell mainly in plain, hills and mountains below 1400 asl.; its range covered complicated physiognomy and various natural barriers, e.g., sea strait,high mountains,gorges,and large rivers etc.,so this area has been hotspot where research systematics and biogeography. Based on the already work, this project paln to survey this speceis in its range and collect specimens or samples of about 164 populations, record their calls, sequence five mitochondrial and nuclear genes of about 2000 individuals and examine 15 loci of microsatellite in nuclear genome of about 3000 individuals; carry out systematics research,toexplain population differntiations, identify possible new form, determin populaton composition of each major lineage recogized in this work and its geographical distribution patterns; carry out phylogeographical and ecological niche modelling analyses and researches, to estimate divergence dates between different lineages and their relationships, to discover population genetic structure and reconsructed demography history, moddelling their ecological distribution patterns. In the light of the work above, differentation history, geographical distribution patterns and their forming process of lineages recognized in this species will be elucidated, and then promoting systeamtics, zoogeography and adaptive evolution researches in this area.

饰纹姬蛙Microhyla fissipes广泛分布于东亚南部和中南半岛海拔1400m以下的平原、丘陵和山地，种群分化明显，系统关系复杂，可能存在多个隐存种。该区域地貌复杂、天然屏障多样，包括高山、峡谷、海峡和大江大河，是系统学和生物地理学研究的热点地区。本项目拟在已有工作基础上，深入调查饰纹姬蛙分布区各地理单元的代表种群约164个，采集标本和环境生态信息，录取繁殖鸣叫信息，测定约2000个体的5个线粒体和核基因的DNA序列，检测约3000个体的核基因组15个微卫星位点的多态信息。开展系统学研究，阐释其种群分化，识别隐存种，揭示不同支系的种群构成与分布格局。通过谱系地理和生态位模型分析研究，建立种群的网络支系关系、计算主要支系分化时间、分析种群的遗传结构及种口历史、模拟生态分布格局。籍此阐述饰文姬蛙谱系分化历史、地理分布格局及演变过程，进而推动该地区的系统分类、动物地理与适应进化研究。

采集饰纹姬蛙及近缘和同域物种标本285种群3107只标本，筛选饰纹姬蛙4碱基重复微卫星位点15个，检测其53个种群858号标本的微卫星位点的多样性，测定其Cytb基因588bp计706条、COI基因618bp计1002条、16S rRNA基因 869bp计415条，测得其海南岛19种群372只成体7项形态学指标数据，录取其海南岛9种群35只雄体共1077个鸣叫。系统地解析了饰纹姬蛙种群分化与谱系地理问题：①微卫星检测结果识别4个地理种群组，基因流大致从四川盆地，经由云贵高原，向华南地区扩散，同时华东华北地区也向华南地区扩散。②分子系统发育树识别7支系，它们最近共同祖先分化时间是0.25Ma左右，近期有过快速分化，同时种群可能存在近期扩张。③微卫星和线粒体基因检测分析结果均表明四川盆地和海南岛各自构成独特单元。④四川盆地饰纹姬蛙可分为两组，但内部各种群间基因交流频繁，而长江在三峡地区显示出了明显的隔离效应。从末次冰期至今，其适宜分布区范围扩大，提示它们在近期发生了种群扩张，可能自西向东的长江水系发展促进了其种群扩散。⑤形态学、声学和分子系统学分析结果均表明海南岛饰纹姬蛙可分为东北组和西南组；生态位模型分析结果表明这2组的生态位异质性显著，其中最干月降水量是影响海南岛饰纹姬蛙分布的最主要因素。这可能促进了海南岛饰纹姬蛙的种群分化。此外，在饰纹姬蛙基础生物学其它方面取得较多成果，包括构建胚胎发育图谱，揭示甲状腺激素及其受体调控变态发育的基本机制、尾凋亡的转录调控机制，基于饰纹姬蛙蝌蚪的响应过程和机制评价草甘膦的急性毒性及机制，不同光照和背景条件下蝌蚪肤色模式的形成与变化过程及其调节机制。发现姬蛙属2新种、发表相关论文25篇（SCIE论文16篇）、引进青年3名、培养4名博士和2名硕士。这些成果能为饰纹姬蛙的保护、模式化研发与应用提供支撑，促进有关学科的发展。