Identification of sex-specific molecular markers using restriction site-associated DNA sequencing
Identification of sex-specific molecular markers using restriction site-associated DNA sequencing
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DOI：
10.1111/1755-0998.12237
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发表时间：
2014-09-01
影响因子：
7.7
通讯作者：
Zarkower, David
中科院分区：
文献类型：
--
作者：
Gamble, Tony;Zarkower, David
性别决定和性染色体进化研究的一个主要障碍是缺乏关于不同物种之间发生的性别决定机制类型的信息。这在大多数物种缺乏视觉上异形的性染色体的群体中尤其成问题，例如鱼类、两栖动物和爬行动物，因为细胞遗传学分析将无法识别这些物种的性染色体。我们描述了使用限制位点相关DNA(RAD)测序，或RAD-SEQ，来识别性别特异的分子标记，并随后确定一个物种是否有雄性或雌性异性恋。为了测试这项技术的准确性，我们检查了蜥蜴Anolis Carolinensis。我们对7个雄株和10个雌株进行了RAD-SEQ分析，发现了一个雄性特异的分子标记。Carolis carolinensis此前已被证明具有雄性异配性，最近公布的A.carolinens基因组有助于性别特异性RAD-seq标记的特征描述。我们通过对其他个体的PCR验证了新标记的雄性特异性，并发现它在其他一些Anolis物种中是保守的。我们讨论了使用RAD-SEQ来鉴定具有隐形或同态性染色体的其他物种的性别决定机制的效用，以及对Anolis中雄性异性配偶制进化的影响。
A major barrier to evolutionary studies of sex determination and sex chromosomes has been a lack of information on the types of sex-determining mechanisms that occur among different species. This is particularly problematic in groups where most species lack visually heteromorphic sex chromosomes, such as fish, amphibians and reptiles, because cytogenetic analyses will fail to identify the sex chromosomes in these species. We describe the use of restriction site-associated DNA (RAD) sequencing, or RAD-seq, to identify sex-specific molecular markers and subsequently determine whether a species has male or female heterogamety. To test the accuracy of this technique, we examined the lizard Anolis carolinensis. We performed RAD-seq on seven male and ten female A. carolinensis and found one male-specific molecular marker. Anolis carolinensis has previously been shown to possess male heterogamety and the recently published A. carolinensis genome facilitated the characterization of the sex-specific RAD-seq marker. We validated the male specificity of the new marker using PCR on additional individuals and also found that it is conserved in some other Anolis species. We discuss the utility of using RAD-seq to identify sex-determining mechanisms in other species with cryptic or homomorphic sex chromosomes and the implications for the evolution of male heterogamety in Anolis.