Identification of genes related to agarwood formation: transcriptome analysis of healthy and wounded tissues of Aquilaria sinensis.
Identification of genes related to agarwood formation: transcriptome analysis of healthy and wounded tissues of Aquilaria sinensis.
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沉香形成相关基因的鉴定：沉香健康和受伤组织的转录组分析。
DOI：
10.1186/1471-2164-14-227
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发表时间：
2013-04-08
期刊：
影响因子：
4.4
通讯作者：
Li W
中科院分区：
文献类型：
--
作者：
Xu Y;Zhang Z;Wang M;Wei J;Chen H;Gao Z;Sui C;Luo H;Zhang X;Yang Y;Meng H;Li W
沉香是一种昂贵的沉香心材，来源于沉香属植物，广泛用于传统医药、熏香和香水。只有受伤的树木才能生产沉香，对沉香产品的巨大需求导致所有沉香属植物。濒危物种，并被列入CITES附录II（http：www.cites.org）。沉香的主要成分是倍半萜和苯乙基色酮。由于缺乏基因组信息，创伤诱导的倍半萜合成和沉香形成的分子基础仍然未知。为了筛选与沉香形成相关的主要基因，我们对健康和受伤的沉香cDNA文库进行了测序。[医]中华吉格共获得89,137个unigene，平均长度为678.65 bp，并在生物信息学水平上进行了详细的注释。在与沉香形成相关的酶中，有30种purone编码的酶参与了倍半萜的生物合成途径，少数转录因子和蛋白激酶与创伤信号转导有关。从大肠杆菌中克隆并表达了三种倍半萜烯脱氢酶（ASS 1 -3）的全长cDNA，经酶活性测定，表明它们是有活性的酶，主要产物为δ-愈创木烯。茉莉酸甲酯（MJ）诱导实验表明，MJ能显著诱导ASS的表达，并相应提高倍半萜类化合物的产量。MJ还诱导了一些转录因子和蛋白激酶的表达，特别是MYB 4、WRKY 4、MPKK 2和MAPK 2，并与ASS的表达有协同作用，提示它们可能是ASS的正调控因子。本研究为沉香属植物的转录组研究提供了丰富的信息。为阐明创伤诱导的沉香倍半萜类化合物生物合成及其调控机制提供了有价值的线索。
Agarwood is an expensive resinous heartwood derived from Aquilaria plants that is widely used in traditional medicines, incense and perfume. Only wounded trees can produce agarwood, and the huge demand for the agarwood products has led all Aquilaria spp. being endangered and listed in the Appendix II of the CITES (http://www.cites.org). The major components of agarwood are sesquiterpenes and phenylethyl chromones. Owing to a lack of genomic information, the molecular basis of wound-induced sesquiterpenes biosynthesis and agarwood formation remains unknown. To identify the primary genes that maybe related to agarwood formation, we sequenced 2 cDNA libraries generated from healthy and wounded A. sinensis (Lour.) Gilg. A total of 89,137 unigenes with an average length of 678.65 bp were obtained, and they were annotated in detail at bioinformatics levels. Of those associated with agarwood formation, 30 putatively encoded enzymes in the sesquiterpene biosynthesis pathway, and a handful of transcription factors and protein kinases were related to wound signal transduction. Three full-length cDNAs of sesquiterpene synthases (ASS1-3) were cloned and expressed in Escherichia coli, and enzyme assays revealed that they are active enzymes, with the major products being δ-guaiene. A methyl jasmonate (MJ) induction experiment revealed that the expression of ASS was significantly induced by MJ, and the production of sesquiterpenes was elevated accordingly. The expression of some transcription factors and protein kinases, especially MYB4, WRKY4, MPKK2 and MAPK2, was also induced by MJ and coordinated with ASS expression, suggesting they maybe positive regulators of ASS. This study provides extensive transcriptome information for Aquilaria spp. and valuable clues for elucidating the mechanism of wound-induced agarwood sesquiterpenes biosynthesis and their regulation.
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