Guyot Romain, de la Mare Marion, Viader Véronique, Hamon Perla, Coriton Olivier, Bustamante-Porras José, Poncet Valérie, Campa Claudine, Hamon Serge, de Kochko Alexandre
UMR GDP, IRD BP 64501, Centre IRD de Montpellier, Montpellier Cedex 5, France.
BMC Plant Biol. 2009 Feb 25;9:22. doi: 10.1186/1471-2229-9-22.
Coffea canephora, also called Robusta, belongs to the Rubiaceae, the fourth largest angiosperm family. This diploid species (2x = 2n = 22) has a fairly small genome size of approximately 690 Mb and despite its extreme economic importance, particularly for developing countries, knowledge on the genome composition, structure and evolution remain very limited. Here, we report the 160 kb of the first C. canephora Bacterial Artificial Chromosome (BAC) clone ever sequenced and its fine analysis.
This clone contains the CcEIN4 gene, encoding an ethylene receptor, and twenty other predicted genes showing a high gene density of one gene per 7.8 kb. Most of them display perfect matches with C. canephora expressed sequence tags or show transcriptional activities through PCR amplifications on cDNA libraries. Twenty-three transposable elements, mainly Class II transposon derivatives, were identified at this locus. Most of these Class II elements are Miniature Inverted-repeat Transposable Elements (MITE) known to be closely associated with plant genes. This BAC composition gives a pattern similar to those found in gene rich regions of Solanum lycopersicum and Medicago truncatula genomes indicating that the CcEIN4 regions may belong to a gene rich region in the C. canephora genome. Comparative sequence analysis indicated an extensive conservation between C. canephora and most of the reference dicotyledonous genomes studied in this work, such as tomato (S. lycopersicum), grapevine (V. vinifera), barrel medic M. truncatula, black cottonwood (Populus trichocarpa) and Arabidopsis thaliana. The higher degree of microcollinearity was found between C. canephora and V. vinifera, which belong respectively to the Asterids and Rosids, two clades that diverged more than 114 million years ago.
This study provides a first glimpse of C. canephora genome composition and evolution. Our data revealed a remarkable conservation of the microcollinearity between C. canephora and V. vinifera and a high conservation with other distant dicotyledonous reference genomes. Altogether, these results provide valuable information to identify candidate genes in C. canephora genome and serve as a foundation to establish strategies for whole genome sequencing. Future large-scale sequence comparison between C. canephora and reference sequenced genomes will help in understanding the evolutionary history of dicotyledonous plants.
卡内弗拉咖啡(Coffea canephora),也被称为罗布斯塔咖啡,属于茜草科,是被子植物的第四大科。这种二倍体物种(2x = 2n = 22)的基因组大小相当小,约为690 Mb,尽管它具有极其重要的经济价值,特别是对发展中国家而言,但关于其基因组组成、结构和进化的知识仍然非常有限。在此,我们报告了首个测序的卡内弗拉咖啡细菌人工染色体(BAC)克隆的160 kb及其精细分析。
该克隆包含编码乙烯受体的CcEIN4基因以及其他20个预测基因,基因密度高达每7.8 kb一个基因。其中大多数与卡内弗拉咖啡表达序列标签完全匹配,或通过对cDNA文库进行PCR扩增显示出转录活性。在该位点鉴定出23个转座元件,主要是II类转座子衍生物。这些II类元件中的大多数是微型反向重复转座元件(MITE),已知它们与植物基因密切相关。该BAC的组成与番茄(Solanum lycopersicum)和蒺藜苜蓿(Medicago truncatula)基因组中富含基因的区域相似,表明CcEIN4区域可能属于卡内弗拉咖啡基因组中富含基因的区域。比较序列分析表明,卡内弗拉咖啡与本研究中所研究的大多数参考双子叶植物基因组之间存在广泛的保守性,如番茄(S. lycopersicum)、葡萄(V. vinifera)、蒺藜苜蓿(M. truncatula)、黑杨(Populus trichocarpa)和拟南芥(Arabidopsis thaliana)。在卡内弗拉咖啡和葡萄之间发现了更高程度的微观共线性,它们分别属于菊类植物和蔷薇类植物,这两个分支在1.14亿多年前就已分化。
本研究首次揭示了卡内弗拉咖啡基因组的组成和进化。我们的数据揭示了卡内弗拉咖啡和葡萄之间微观共线性的显著保守性以及与其他远缘双子叶植物参考基因组的高度保守性。总之,这些结果为鉴定卡内弗拉咖啡基因组中的候选基因提供了有价值的信息,并为建立全基因组测序策略奠定了基础。未来卡内弗拉咖啡与参考测序基因组之间的大规模序列比较将有助于理解双子叶植物的进化历史。