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首页> 外文期刊>BMC Genomics >Global characterization of Artemisia annua glandular trichome transcriptome using 454 pyrosequencing
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Global characterization of Artemisia annua glandular trichome transcriptome using 454 pyrosequencing

机译：使用454焦磷酸测序技术对青蒿的腺毛状转录组进行全局鉴定

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Background Glandular trichomes produce a wide variety of commercially important secondary metabolites in many plant species. The most prominent anti-malarial drug artemisinin, a sesquiterpene lactone, is produced in glandular trichomes of Artemisia annua. However, only limited genomic information is currently available in this non-model plant species. Results We present a global characterization of A. annua glandular trichome transcriptome using 454 pyrosequencing. Sequencing runs using two normalized cDNA collections from glandular trichomes yielded 406,044 expressed sequence tags (average length = 210 nucleotides), which assembled into 42,678 contigs and 147,699 singletons. Performing a second sequencing run only increased the number of genes identified by ~30%, indicating that massively parallel pyrosequencing provides deep coverage of the A. annua trichome transcriptome. By BLAST search against the NCBI non-redundant protein database, putative functions were assigned to over 28,573 unigenes, including previously undescribed enzymes likely involved in sesquiterpene biosynthesis. Comparison with ESTs derived from trichome collections of other plant species revealed expressed genes in common functional categories across different plant species. RT-PCR analysis confirmed the expression of selected unigenes and novel transcripts in A. annua glandular trichomes. Conclusion The presence of contigs corresponding to enzymes for terpenoids and flavonoids biosynthesis suggests important metabolic activity in A. annua glandular trichomes. Our comprehensive survey of genes expressed in glandular trichome will facilitate new gene discovery and shed light on the regulatory mechanism of artemisinin metabolism and trichome function in A. annua.
机译：背景技术腺毛在许多植物物种中产生了多种具有重要商业意义的次生代谢产物。最著名的抗疟药青蒿素是倍半萜内酯，是在青蒿的腺毛中产生的。但是，目前在这种非模型植物物种中只有有限的基因组信息。结果我们提出了使用454焦磷酸测序对A. annua腺毛状体转录组进行全局表征。使用来自腺毛的两个标准化的cDNA集合进行测序运行，产生了406,044个表达的序列标签（平均长度= 210个核苷酸），这些标签被组装成42,678个重叠群和147,699个单子。进行第二次测序只能将鉴定出的基因数量增加约30％，这表明大规模平行的焦磷酸测序提供了对A.annua trichome转录组的深度覆盖。通过BLAST搜索NCBI非冗余蛋白数据库，推定功能被分配给28,573个unigene，包括以前未描述的可能参与倍半萜生物合成的酶。与从其他植物的毛状体集合中获得的EST进行比较后，发现不同植物物种中共有功能类别的表达基因。逆转录-聚合酶链反应（RT-PCR）分析证实了选定的单基因和新的转录本在青蒿腺毛状体中的表达。结论存在与类萜和类黄酮生物合成酶相对应的重叠群，表明在环孢菌腺毛体中重要的代谢活性。我们对腺毛状体中表达的基因的全面调查将促进新基因的发现，并阐明青蒿素在青蒿中代谢和毛状体功能的调控机制。

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