Yamamoto Naoki, Tsugane Taneaki, Watanabe Manabu, Yano Kentaro, Maeda Fumi, Kuwata Chikara, Torki Moez, Ban Yusuke, Nishimura Shigeo, Shibata Daisuke
The Graduate School of Life Sciences, Tohoku University, Kazusa-Kamatari 2-6-7, Kisarazu, Chiba 292-0818, Japan.
Gene. 2005 Aug 15;356:127-34. doi: 10.1016/j.gene.2005.04.026.
Laboratory-grown miniature tomato (Lycopersicon esculentum) cultivar Micro-Tom has attracted attention as a host for functional genomics research. In this study, we generated 35,824 expressed sequence tags (ESTs) from leaves and fruits of Micro-Tom. The ESTs comprised 10,287 unigenes (5007 contigs and 5280 singletons), including 1858 novel tomato unigenes. Of the 18 unigenes that shared strong homology with tobacco chloroplast genome sequences, one unigene was likely derived from polyadenylated transcripts of the atpH gene. Interestingly, ESTs for vacuolar invertase, pectate lyase and alcohol acyl transferase were underrepresented in the Micro-Tom data set. From all of the ESTs, we mined 2039 candidate single nucleotide polymorphisms (SNPs) and 121 candidate insertions and deletions (indels) based on homology with four tomato inbred lines, E6203, R11-13, Rio Grande PtoR and R11-12, and a wild relative, L. pennellii TA56, for which sequence data was publicly available with more than 5000 entries. Direct genome sequencing of several SNP or indel sites in Micro-Tom and L. esculentum E6203 suggested that more than 69% of the candidate sites were truly polymorphic, making them useful for the preparation of DNA markers.
实验室培育的微型番茄(Lycopersicon esculentum)品种Micro-Tom作为功能基因组学研究的宿主已受到关注。在本研究中,我们从Micro-Tom的叶片和果实中生成了35,824个表达序列标签(EST)。这些EST包含10,287个单基因(5007个重叠群和5280个单拷贝序列),其中包括1858个新的番茄单基因。在与烟草叶绿体基因组序列具有高度同源性的18个单基因中,有一个单基因可能源自atpH基因的多聚腺苷酸化转录本。有趣的是,液泡转化酶、果胶酸裂解酶和乙醇酰基转移酶的EST在Micro-Tom数据集中的代表性不足。基于与四个番茄自交系E6203、R11-13、Rio Grande PtoR和R11-12以及一个野生近缘种L. pennellii TA56(其序列数据已公开,条目超过5000条)的同源性,我们从所有EST中挖掘出2039个候选单核苷酸多态性(SNP)和121个候选插入缺失(indel)。对Micro-Tom和L. esculentum E6203中几个SNP或indel位点的直接基因组测序表明,超过69%的候选位点是真正多态的,这使得它们可用于制备DNA标记。