Plant Biotechnology, Faculty of Biology, University of Freiburg, Schaenzlestr. 1, Freiburg, 79104, Germany.
Cell Biology, Faculty of Biology, University of Freiburg, Schaenzlestr. 1, Freiburg, 79104, Germany.
New Phytol. 2017 Oct;216(2):455-468. doi: 10.1111/nph.14506. Epub 2017 Mar 6.
In seed plants, strigolactones (SLs) regulate architecture and induce mycorrhizal symbiosis in response to environmental cues. SLs are formed by combined activity of the carotenoid cleavage dioxygenases (CCDs) 7 and 8 from 9-cis-β-carotene, leading to carlactone that is converted by cytochromes P450 (clade 711; MAX1 in Arabidopsis) into various SLs. As Physcomitrella patens possesses CCD7 and CCD8 homologs but lacks MAX1, we investigated if PpCCD7 together with PpCCD8 form carlactone and how deletion of these enzymes influences growth and interactions with the environment. We investigated the enzymatic activity of PpCCD7 and PpCCD8 in vitro, identified the formed products by high performance liquid chromatography (HPLC) and LC-MS, and generated and analysed ΔCCD7 and ΔCCD8 mutants. We defined enzymatic activity of PpCCD7 as a stereospecific 9-cis-CCD and PpCCD8 as a carlactone synthase. ΔCCD7 and ΔCCD8 lines showed enhanced caulonema growth, which was revertible by adding the SL analogue GR24 or carlactone. Wild-type (WT) exudates induced seed germination in Orobanche ramosa. This activity was increased upon phosphate starvation and abolished in exudates of both mutants. Furthermore, both mutants showed increased susceptibility to phytopathogenic fungi. Our study reveals the deep evolutionary conservation of SL biosynthesis, SL function, and its regulation by biotic and abiotic cues.
在种子植物中,独脚金内酯(SLs)通过对环境信号的响应来调节结构并诱导共生真菌。SLs 的形成是由 9-顺式-β-胡萝卜素的类胡萝卜素双加氧酶(CCDs)7 和 8 的联合活性产生的,导致卡尔拉酮,然后由细胞色素 P450(711 类;拟南芥中的 MAX1)转化为各种 SLs。由于Physcomitrella patens 具有 CCD7 和 CCD8 同源物,但缺乏 MAX1,我们研究了 PpCCD7 是否与 PpCCD8 一起形成卡尔拉酮,以及这些酶的缺失如何影响生长和与环境的相互作用。我们在体外研究了 PpCCD7 和 PpCCD8 的酶活性,通过高效液相色谱(HPLC)和 LC-MS 鉴定了形成的产物,并生成和分析了ΔCCD7 和ΔCCD8 突变体。我们将 PpCCD7 的酶活性定义为立体特异性的 9-顺式-CCD,将 PpCCD8 定义为卡尔拉酮合酶。ΔCCD7 和ΔCCD8 系表现出增强的原丝体生长,添加 SL 类似物 GR24 或卡尔拉酮可使其恢复。野生型(WT)分泌物可诱导 Orobanche ramosa 的种子发芽。这种活性在磷酸盐饥饿时增加,并在两个突变体的分泌物中被消除。此外,两个突变体对植物病原菌真菌的敏感性增加。我们的研究揭示了 SL 生物合成、SL 功能及其对生物和非生物线索的调节的深度进化保守性。