School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London, E1 4NS, UK.
Mol Ecol. 2013 Jun;22(11):3098-111. doi: 10.1111/mec.12131. Epub 2012 Nov 21.
New sequencing technologies allow development of genome-wide markers for any genus of ecological interest, including plant genera such as Betula (birch) that have previously proved difficult to study due to widespread polyploidy and hybridization. We present a de novo reference genome sequence assembly, from 66× short read coverage, of Betula nana (dwarf birch) - a diploid that is the keystone woody species of subarctic scrub communities but of conservation concern in Britain. We also present 100 bp PstI RAD markers for B. nana and closely related Betula tree species. Assembly of RAD markers in 15 individuals by alignment to the reference B. nana genome yielded 44-86k RAD loci per individual, whereas de novo RAD assembly yielded 64-121k loci per individual. Of the loci assembled by the de novo method, 3k homologous loci were found in all 15 individuals studied, and 35k in 10 or more individuals. Matching of RAD loci to RAD locus catalogues from the B. nana individual used for the reference genome showed similar numbers of matches from both methods of RAD locus assembly but indicated that the de novo RAD assembly method may overassemble some paralogous loci. In 12 individuals hetero-specific to B. nana 37-47k RAD loci matched a catalogue of RAD loci from the B. nana individual used for the reference genome, whereas 44-60k RAD loci aligned to the B. nana reference genome itself. We present a preliminary study of allele sharing among species, demonstrating the utility of the data for introgression studies and for the identification of species-specific alleles.
新的测序技术允许为任何具有生态意义的属开发全基因组标记,包括先前由于广泛的多倍体和杂交而难以研究的植物属,如桦木属(桦木)。我们提供了一份贝达那纳(矮桦)的从头参考基因组序列组装,其覆盖度为 66×短读序列,这是一种二倍体,是亚北极灌丛群落的关键木本物种,但在英国却受到保护关注。我们还提供了 100bp PstI RAD 标记用于贝达那纳和相关的桦树物种。通过将 RAD 标记与参考贝达那纳基因组对齐,在 15 个个体中组装 RAD 标记,每个个体产生 44-86k RAD 基因座,而从头组装 RAD 标记则产生每个个体 64-121k 基因座。在从头组装的基因座中,在所有 15 个个体中发现了 3k 个同源基因座,在 10 个或更多个体中发现了 35k 个同源基因座。通过将 RAD 基因座与用于参考基因组的贝达那纳个体的 RAD 基因座目录进行匹配,两种 RAD 基因座组装方法的匹配数量相似,但表明从头组装 RAD 基因座方法可能会过度组装一些同源基因座。在 12 个异源于贝达那纳的个体中,37-47k RAD 基因座与用于参考基因组的贝达那纳个体的 RAD 基因座目录匹配,而 44-60k RAD 基因座与贝达那纳参考基因组本身对齐。我们初步研究了物种间的等位基因共享,证明了这些数据在基因渗入研究和鉴定物种特异性等位基因方面的有用性。