Molecular dissection of mercury-responsive transcriptome and sense/antisense genes in Medicago truncatula.
Molecular dissection of mercury-responsive transcriptome and sense/antisense genes in Medicago truncatula.
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DOI：
10.1016/j.jhazmat.2013.02.011
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发表时间：
2013-05
影响因子：
13.6
通讯作者：
Z. Zhou;Shengzhou Yang;Hua Li;C. Zhu;Zhaolei Liu;Z. Yang
中科院分区：
文献类型：
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作者：
Z. Zhou;Shengzhou Yang;Hua Li;C. Zhu;Zhaolei Liu;Z. Yang
我们描述了一种新开发的方法，即下一代标签测序，以确定全球基因转录本和重金属调控的复杂性在紫花苜蓿。以有毒重金属汞(II)处理的元宝菊幼苗为材料，构建了两个cDNA文库。随着大量读图基因的产生，我们观察到大多数基因在两个文库之间存在差异表达。此外，还检测到了几类新的转录本，包括转录因子、反义转录本和逆境反应基因。表达水平变化最大的40个基因与对环境压力的耐受性和次生代谢有关。通过定量RT-PCR对基因进行验证，证实了深度测序的结果。大多数编码金属转运蛋白、硫酸盐代谢和细胞壁固化的基因在汞暴露下发生了显著变化。我们还检测了两个文库之间正义和反义(S-AS)转录本表达比率的变化。通过分析阅读序列的链特异性信息，发现经金属处理后的S-AS转录本被富集化。基因本体论(GO)和京都基因和基因组百科全书(KEGG)进一步分析了转录组序列，揭示了金属胁迫下不同的生物学功能和代谢途径。
We described a newly developed approach, namely next-generation tag sequencing, to identify global gene transcripts and complexity regulated by heavy metals in Medicago truncatula. Two cDNA libraries were generated from M. truncatula seedlings: treated and non-treated with the toxic heavy metal mercury Hg(II). With the large number of read-mapped genes generated, we observed that most of the genes were differentially expressed between the two libraries. In addition, several classes of new transcripts including transcription factors, antisense transcripts, and stress responsive genes were detected. The forty genes most altered in expression levels were associated with tolerance to environmental stress and secondary metabolism. Validation of genes by quantitative RT-PCR confirmed the results from deep-sequencing. Most of genes coding for metal transporters, sulfate metabolism, and cell wall solidification were significantly altered by Hg exposure. We also examined altered expression ratios of sense and antisense (S–AS) transcripts between the two libraries. By analyzing strand-specific information of read sequences, S–AS transcripts were found to be enriched with metal treatment. The transcriptome sequences were analyzed further with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) and showed diverse biological functions and metabolic pathways under the metal stress.