IRD, UMR DIADE, BP 64501, Montpellier Cedex 5, France.
Mol Genet Genomics. 2011 Jun;285(6):447-60. doi: 10.1007/s00438-011-0617-0. Epub 2011 Apr 20.
In the study, we developed new markers for phylogenetic relationships and intraspecies differentiation in Coffea. Nana and Divo, two novel Ty1-copia LTR-retrotransposon families, were isolated through C. canephora BAC clone sequencing. Nana- and Divo-based markers were used to test their: (1) ability to resolve recent phylogenetic relationships; (2) efficiency in detecting intra-species differentiation. Sequence-specific amplification polymorphism (SSAP), retrotransposon-microsatellite amplified polymorphism (REMAP) and retrotransposon-based insertion polymorphism (RBIP) approaches were applied to 182 accessions (31 Coffea species and one Psilanthus accession). Nana- and Divo-based markers revealed contrasted transpositional histories. At the BAC clone locus, RBIP results on C. canephora demonstrated that Nana insertion took place prior to C. canephora differentiation, while Divo insertion occurred after differentiation. Combined SSAP and REMAP data showed that Nana could resolve Coffea lineages, while Divo was efficient at a lower taxonomic level. The combined results indicated that the retrotransposon-based markers were useful in highlighting Coffea genetic diversity and the chronological pattern of speciation/differentiation events. Ongoing complete sequencing of the C. canephora genome will soon enable exhaustive identification of LTR-RTN families, as well as more precise in-depth analyses on contributions to genome size variation and Coffea evolution.
在这项研究中,我们开发了用于咖啡属种间系统发育关系和种内分化的新标记。通过对咖啡种质的 BAC 克隆测序,我们分离出了两种新型的 Ty1-copia LTR-逆转录转座子家族:Nana 和 Divo。利用 Nana 和 Divo 标记来检验它们:(1)解析近期系统发育关系的能力;(2)检测种内分化的效率。我们应用序列特异性扩增多态性(SSAP)、逆转座子-微卫星扩增多态性(REMAP)和基于逆转座子的插入多态性(RBIP)方法对 182 个样本(31 个咖啡种和一个 Psilanthus 样本)进行了检测。Nana 和 Divo 标记揭示了不同的转座历史。在 BAC 克隆位点上,对咖啡种的 RBIP 结果表明,Nana 插入发生在咖啡种分化之前,而 Divo 插入则发生在分化之后。综合 SSAP 和 REMAP 数据表明,Nana 可以解析咖啡谱系,而 Divo 在较低的分类水平上则具有较高的效率。综合结果表明,基于逆转座子的标记在突出咖啡遗传多样性和种化/分化事件的时间模式方面具有重要作用。对咖啡种质基因组的全序列测序将很快能够详尽地鉴定 LTR-RTN 家族,以及更精确地深入分析它们对基因组大小变化和咖啡进化的贡献。