Mertens Jan, Van Moerkercke Alex, Vanden Bossche Robin, Pollier Jacob, Goossens Alain
Department of Plant Systems Biology, VIB, B-9052 Gent, Belgium.
Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Gent, Belgium.
Plant Cell Physiol. 2016 Dec;57(12):2564-2575. doi: 10.1093/pcp/pcw168. Epub 2016 Oct 1.
Plants produce many bioactive, specialized metabolites to defend themselves when facing various stress situations. Their biosynthesis is directed by a tightly controlled regulatory circuit that is elicited by phytohormones such as jasmonate (JA). The basic helix-loop-helix (bHLH) transcription factors (TFs) bHLH iridoid synthesis 1 (BIS1) and Triterpene Saponin Activating Regulator (TSAR) 1 and 2, from Catharanthus roseus and Medicago truncatula, respectively, all belong to clade IVa of the bHLH protein family and activate distinct terpenoid pathways, thereby mediating monoterpenoid indole alkaloid (MIA) and triterpene saponin (TS) accumulation, respectively, in these two species. In this study, we report that promoters of the genes encoding the enzymes involved in the specific terpenoid pathway of one of these species can be transactivated by the orthologous bHLH factor from the other species through recognition of the same cis-regulatory elements. Accordingly, ectopic expression of CrBIS1 in M. truncatula hairy roots up-regulated the expression of all genes required for soyasaponin production, resulting in strongly increased levels of soyasaponins in the transformed roots. Likewise, transient expression of MtTSAR1 and MtTSAR2 in C. roseus petals led to up-regulation of the genes involved in the iridoid branch of the MIA pathway. Together, our data illustrate the functional similarity of these JA-inducible TFs and indicate that recruitment of defined cis-regulatory elements constitutes an important aspect of the evolution of conserved regulatory modules for the activation of species-specific terpenoid biosynthesis pathways by common signals such as the JA phytohormones.
植物在面对各种胁迫情况时会产生许多具有生物活性的特殊代谢产物来保护自身。它们的生物合成由一个严格控制的调控回路引导,该回路由茉莉酸(JA)等植物激素引发。来自长春花和蒺藜苜蓿的基本螺旋-环-螺旋(bHLH)转录因子(TFs),即bHLH环烯醚萜合成1(BIS1)以及三萜皂苷激活调节因子(TSAR)1和2,均属于bHLH蛋白家族的IVa进化枝,并分别激活不同的萜类途径,从而在这两个物种中分别介导单萜吲哚生物碱(MIA)和三萜皂苷(TS)的积累量。在本研究中,我们报告称,其中一个物种特定萜类途径中参与酶编码的基因启动子,可被来自另一物种的直系同源bHLH因子通过识别相同的顺式调控元件而反式激活。相应地,长春花的CrBIS1在蒺藜苜蓿毛状根中的异位表达上调了大豆皂苷产生所需的所有基因的表达,导致转化根中大豆皂苷水平大幅增加。同样,蒺藜苜蓿的MtTSAR1和MtTSAR2在长春花花瓣中的瞬时表达导致MIA途径中环烯醚萜分支相关基因的上调。总之,我们的数据说明了这些JA诱导型TFs的功能相似性,并表明特定顺式调控元件的募集构成了由JA植物激素等共同信号激活物种特异性萜类生物合成途径的保守调控模块进化的一个重要方面。