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中国南部沿海条纹斑竹鲨和尖头斜齿鲨种群遗传学研究 【作者】陈骁 【导师】杨圣云

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中国南部沿海条纹斑竹鲨和尖头斜齿鲨种群遗传学研究
~$sr+l~Lgq X0Population Genetics Research of Chiloscyllium Plagiosum and Scoliodon Laticaudus in China's Southern Coast

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F5E,I _kN8xPYL0作者陈骁导师杨圣云
&C#Y:rM7wldP&d0【作者基本信息】厦门大学,海洋生物学,2008年,博士水利论文R;jh? ^g)r8o i

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adb ?!QTj0【中文摘要】 以鲨、鳐类为代表的软骨鱼类主要生活在海洋中,大约有1100种。作为食物链的顶级捕食者和海洋生态系统的重要组成部分,软骨鱼类会对许多其他海洋生物的种群结构产生影响。因此,当软骨鱼类遭到过渡捕捞后将严重影响海洋生态系统中生物群落组成及生态环境。研究发现,相对于硬骨鱼类,大部分鲨、鳐类具有生长缓慢、成熟期长、生殖力低的特点。这些特点导致其具有较低的内禀增长率以及在捕捞压力下较低的种群恢复能力。因此,大部分软骨鱼类种群只能承受中等捕捞强度的渔业开发,否则将导致种群数量急剧减少甚至种群崩溃,并且在种群数量急剧减少后无法象许多硬骨鱼类那样快速恢复原有种群数量和规模。当前鲨鱼渔业在世界范围内快速发展,全球鲨鱼资源已经遭到严重破坏,近十年来鲨鱼种群数量急剧下降,渔业捕捞有可能在自然选择之前造成鲨鱼的灭绝。在这种情况下,鲨鱼的种群遗传学研究显得十分重要。但是由于鲨鱼外部形态特征较少,无法反映其种群个体间的遗传差异,所以难以运用传统的标志放流观察和测量等方法进行种群遗传学研究。从1960年代同工酶电泳技术的发展开始,分子标记技术越来越多地应用在鲨鱼种群遗传学的研究中。分子标记直接针对基因进行分析,不受环境...更多因素、生物个体发育阶段及器官组织变化的影响,能直接揭示鲨鱼个体间和群体间的遗传差异,成为鲨鱼种群遗传学研究的有力工具。目前国外已经有大量关于软骨鱼类种群遗传学的研究报道,但是国内在这方面的研究还未见报道。因此作者利用分子标记技术对我国南部沿海两种有代表性的鲨鱼种类(条纹斑竹鲨和尖头斜齿鲨)的种群遗传学特征进行研究,充实我国软骨鱼类种群遗传学的基础理论,并为我国软骨鱼类保护和资源合理开发提供科学依据。1采用随机引物扩增多态性DNA(RAPD)技术分析了中国南部沿海闽东(MD)、闽南(MN)、粤西(YX)3个条纹斑竹鲨地理群体遗传多样性和遗传结构.结果表明,34个随机引物在这3个群体中共检测出316个位点,各群体检测出的位点数分别为308、308、303,其中多态位点数分别为49、52、42,多态位点比例分别为15.90%、16.88%、13.86%.3个群体的Shannon多样性指数分别为0.0998、0.1055、0.0936:表明条纹斑竹鲨群体的遗传多样性水平较低.遗传距离和UPGMA聚类分析结果显示条纹斑竹鲨的基因交流模式属于沿海岸线的距离隔离模式,遗传差异大小与地理距离远近相关。分子方差分析(AMOVA)结果显示群体问存在显著的遗传结构差异(Fst=0.04404,P<0.05),变异主要发生在群体内部(95.6%)。2对中国南部沿海5个海域的条纹斑竹鲨群体线粒体控制区序列多态性进行研究,获得长度为1094~1096bp的线粒体控制区完整序列,经过序列比对,发现了6个多态性核苷酸位点,定义了8个单倍型。5个群体的单倍型多样度(h=0.5425~0.7448)、平均核苷酸差异(K=0.625287~1.073563)和核苷酸多样度(π=0.000571~0.000980)均处于较低水平。分子方差分析(AMOVA)结果显示群体间存在显著的遗传结构差异(Fst=0.21626,P<0.0001),变异主要发生在群体内部(78.4%)。遗传距离分析显示各群体间的遗传差异大小与地理距离远近相关。系统地理学分析显示闽南群体最古老,其余群体是由闽南群体扩张发育而来。3采用随机引物扩增多态性DNA(RAPD)技术对中国南部沿海闽东霞浦-浙南瑞安海域(MD)、闽南-台湾浅滩渔场(MN)、粤西湛江-阳江海域(YX)、北部湾海域(BBW)4个尖头斜齿鲨地理群体遗传多样性进行分析,29个随机引物在4个群体中共检测出282个位点。4个群体各自检测出的位点数分别为273、274、272、276,其中多态位点数分别为54、57、52、45,多态位点比例分别为19.78%、20.80%、19.12%、16.30%。Shannon多样性指数分别为0.104、0.107、0.103、0.090,表明尖头斜齿鲨群体的遗传多样性水平较低。遗传距离和UPGMA聚类分析结果显示尖头斜齿鲨的基因交流模式属于距离隔离模式,遗传差异大小与地理距离远近相关。分子方差分析(AMOVA)结果显示群体间存在微弱但显著的遗传结构差异(Fst=0.019,p<0.05),变异主要发生在群体内部(98.06%)。4对中国南部沿海4个海域的尖头斜齿鲨群体线粒体控制区序列多态性进行研究,获得长度为1064bp的线粒体控制区完整序列,经过序列比对,发现了12个多态性核苷酸位点,定义了19个单倍型。4个群体的单倍型多样度(h=0.7829~0.8301)、平均核苷酸差异(K=1.314943~1.492473)和核苷酸多样度(π=0.001236~0.001403)均处于较低水平。分子方差分析(AMOVA)结果显示群体间存在微弱但显著的遗传结构差异(Fst=0.02078,P<0.05),变异主要发生在群体内部(97.92%)。遗传距离分析显示各群体间的遗传差异大小与地理距离远近相关。系统地理学分析显示祖先单倍型在各群体中的出现频率相近,各海域群体都拥有各自独特的单倍型分枝。北部湾群体的单倍型进化分枝特别明显,可能与北部湾海域较封闭的地理特征以及尖头斜齿鲨的恋出生地行为有关。5对北部湾海域尖头斜齿鲨4个季节群体线粒体控制区序列进行比较,发现了9个多态性核苷酸位点,定义了15个单倍型。4个季节群体的单倍型多样度(h=0.7540~0.8309)、平均核苷酸差异(K=1.128736~1.503171)和核苷酸多样度(π=0.001061~0.001413)均处于较低水平,其中夏季群体的多样性最低,秋季群体最高。分子方差分析(AMOVA)结果显示群体间不存在显著的遗传结构差异(Fst=-0.00752,P>0.05)。6对条纹斑竹鲨和尖头斜齿鲨的研究结果表明这两种鲨鱼的种群遗传多样性均较低,这主要是由于软骨鱼类线粒体DNA进化速率较低的普遍规律造成的,但是高强度的捕捞作业导致种群数量减少对种群遗传多样性的降低也有影响。7条纹斑竹鲨和尖头斜齿鲨的的采样海域基本相同,但是表现出来的群体间遗传结构变化趋势却不同,分别与它们的行为特点保持一致。条纹斑竹鲨营底栖生活,行动缓慢游泳能力差,因此地理距离对其群体间基因交流的限制作用比较强,导致闽东、闽南、粤西群体间表现出显著的遗传结构差异。但是在同样的地理距离尺度下,活动能力较强的尖头斜齿鲨闽东、闽南、粤西群体间未表现出显著的遗传结构差异。  还原

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【英文摘要】 Sharks and their relatives,the rays(subclass Elasmobranchii) are a group of about 1100 species of mostly marine fishes.They can affect population structure of many other marine lifes because of their top predators in marine food chain and an important component of marine ecosystems.Therefore,the composition of biological communities and the ecological environment in marine ecosystems will be seriously affected if they are over-fished.Most sharks and rays that have been studied have slow growth,late maturity and low fecundity compared to bony fishes.These attributes result in low intrinsic rates of increase and low resilience to fishing mortality.Thus,most shark and ray populations can withstand only modest levels of fishing without depletion and stock collapse,and decline more rapidly and are not able rebound as quickly as other bony fishes to population reductions.At present shark fisheries have developed around the world,the global shark resources have been seriously damaged,rapid st...更多ock decline and collapse with decades,so fisheries will become commercially extinct before the targets of those fisheries become biologically extirpated.In these circumstances,studies of population genetics are important to protection of shark resources.But it is difficult to study population genetics of sharks using traditional tagging observation and morphological measurement methods because they exhibits relatively low levels of morphological features.Since the development of isozyme electrophoresis in 1960s, molecular markers have been increasingly used to study population genetics of sharks.It could directly reveal the genetic differences among individuals and populations without being affected by environment、developmental phase、apparatus and tissue differences.Currently,there are a large number of research reports about elasmobranchs population genetics in abroad,but domestic research in this field has not been reported.Thereby we studied population genetics of two representative sharks(Chiloscyllium plagiosum and Scoliodon laticaudus) by molecular marker technique in China's southern coast,enrich the basic theory of elasmobranchs population genetics and provide a scientific information for our protection and rational exploitation of elasmobranchs resources.1 Random amplified polymorphic DNA(RAPD) technique was used to assess genetic diversity and population structure within and among three whitespotted bamboosharks (C.plagiosum) stocks from Mindong(MN),Minnan(MN) and Yuexi(YX) of China's southern coast.Thirty-four primers produced 316 loci in all samples.There were 308,308 and 303 loci in three stocks respectively,in which 49(15.90%),52(16.88%) and 42(13.86%) were polymorphic.Shannon genetic diversity indices of three stocks(0.0998,0.1055,0.0936) indicates that genetic diversity of C.plagiosum is low.In addition,the results of UPGMA cluster analysis and Nei's genetic distances showed that gene exchange between C.plagiosum stocks fits Isolation-By-Distance Model and genetic differences among stocks correlates to their geographical distance.Using Analasis of Molecular Variance(AMOVA),weak but significant population differentiation was detected with a globle fixation index(Fst= 0.04404,P<0.05).It meaning that the greater variance occurred among populations(95.60%) and lesser variance within populations(4.402%).2 The entire mitochondrial control region sequences(1094~1096 nucleotides) were amplified from five whitespotted bamboosharks(C.plagiosum) stocks in coastal waters of southern China then 8 haplotypes were defined by 6 polymorphic sites after sequence alignment.Gene diversity(h=0.5425~0.7448)、mean number of pairwise differences(K= 0.625287~1.073563) and nucleotide diversity(π=0.000571~0.000980) indicates that genetic diversity of C.plagiosum is low.Using Analasis of Molecular Variance(AMOVA), significant population differentiation was detected with a globle fixation index(Fst= 0.21626,P<0.0001).It meaning that the greater variance occurred among populations(78.4%) and lesser variance within populations(21.6%).The results of genetic distances analysis showed that genetic differences among stocks correlates to their geographical distance. Statistical phylogeography analysis indicates that MN stock is more ancient than other stocks.3 Random amplified polymorphic DNA(RAPD) technique was used to assess genetic diversity and population structure within and among four spadenose shark(S.laticaudus) stocks from MD,MN,YX and BBW of China's southern coast.Twenty-nine primers produced 282 loci in all samples.There were 273、274、272 and 276 loci in four stocks respectively,in which 54(19.78%),57(28.80%),52(19.12%)and 45(16.30%)were polymorphic.Shannon genetic diversity indices of four stocks(0.104、0.107、0.103、0.090) indicates that genetic diversity of S.laticaudus is low.In addition,the results of UPGMA cluster analysis and Nei's genetic distances showed that gene exchange between S.laticaudus stocks fits Isolation-By-Distance Model and genetic differences among stocks correlates to their geographical distance.Using Analasis of Molecular Variance(AMOVA),weak but significant population differentiation was detected with a globle fixation index(Fst=0.019,P<0.05).It meaning that the greater variance occurred among populations(98.06%) and lesser variance within populations(1.94%). 4 The entire mitochondrial control region sequences(1064 nueleotides) were amplified from four spadenose shark(S.laticaudus) stocks in coastal waters of southern China then 19 haplotypes were defined by 12 polymorphic sites after sequence alignment.Gene diversity(h =0.7829~0.8301)、mean number of pairwise differences(K=1.314943~1.492473) and nucleotide diversity(π=0.001236~0.001403) indicates that genetic diversity of S.laticaudus is low.Using Analasis of Molecular Variance(AMOVA),significant population differentiation was detected with a globle fixation index(Fst=0.02078,P<0.05).It meaning that the greater variance occurred among populations(97.92%) and lesser variance within populations(2.08%).The results of genetic distances analysis showed that genetic differences among stocks correlates to their geographical distance.Statistical phylogeography analysis indicates that frequency of ancient haplotype in all stocks is similar,but all stocks possess their distinct haplotyes clade.The haplotyes clade of BBW stock is most distinct,it may related to more closed geographical characteristics of Beibu Bay and philopatry in S. laticaudus.5 There 19 haplotypes were defined by 12 polymorphic sites after mitochondrial control region sequences alignment of 4 seasonal stocks in Beibu Bay.Gene diversity(h=0.7540~0.8309)、mean number of pairwise differences(K=1.128736~1.503171) and nucleotide diversity(π=0.001061~0.001413) of 4 seasonal S.laticaudus stocks is low,in which summer stock is lowest and autumn stock is highest.AMOVA result indicates no significant population differentiation was detected with a globle fixation index(Fst=-0.00752,P>0.05).6 The results of low population genetic diversity in C.plagiosum and S.laticaudus mainly due to slow rates of mitochondrial DNA evolution in elasmobranchs,but stock depletion by high intensity fisheries may also lower population genetic diversity.7 Genetic structure changes trends between stocks is different in C.plagiosum and S. laticaudus despite their sampling locations is similarity,it consistent to their behavior characters.C.plagiosum move and swimming slowly because of their benthic life,thus geographical distance strongly restricted their gene flow between stocks and resulted in significant genetic structure between MD,MN and YX stocks.But in similar geographical distance scales,S.laticaudus with stronger movement ability didn't present significant genetic structure between these three stocks.  还原水利论文[hP0Q(Z+E

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【中文关键词】 种群遗传学; 条纹斑竹鲨; 尖头斜齿鲨
#N\;A t-qQ0【英文关键词】 population genetics; Chiloscyllium plagiosum; Scoliodon laticaudus水利论文E!Vr+q1q}_

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