Key Laboratory of Eco-environments of Three Gorges Reservoir Region, Ministry of Education, Institute of Resources Botany, School of Life Sciences, Southwest University, Chongqing, 400715, China.
Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, 810008, Xining, China.
New Phytol. 2017 Jul;215(1):351-367. doi: 10.1111/nph.14569. Epub 2017 Apr 26.
Proanthocyanidins (PAs) are major defense phenolic compounds in the leaves of poplar (Populus spp.) in response to abiotic and biotic stresses. Transcriptional regulation of PA biosynthetic genes by the MYB-basic helix-loop-helix (bHLH)-WD40 complexes in poplar is not still fully understood. Here, an Arabidopsis TT2-like gene MYB115 was isolated from Populus tomentosa and characterized by various molecular, genetic and biochemical approaches. MYB115 restored PA productions in the seed coat of the Arabidopsis tt2 mutant. Overexpression of MYB115 in poplar activated expression of PA biosynthetic genes, resulting in a significant increase in PA concentrations. By contrast, the CRISPR/Cas9-generated myb115 mutant exhibited reduced PA content and decreased expression of PA biosynthetic genes. MYB115 directly activated the promoters of PA-specific structural genes. MYB115 interacted with poplar TT8. Coexpression of MYB115, TT8 and poplar TTG1 significantly enhanced the expression of ANR1 and LAR3. Additionally, transgenic plants overexpressing MYB115 had increased resistance to the fungal pathogen Dothiorella gregaria, whereas myb115 mutant exhibited greater sensitivity compared with wild-type plants. Our data provide insight into the regulatory mechanisms controlling PA biosynthesis by MYB115 in poplar, which could be effectively employed for metabolic engineering of PAs to improve resistance to fungal pathogens.
原花青素(PAs)是杨树(Populus spp.)叶片中对非生物和生物胁迫的主要防御性酚类化合物。杨树中 MYB-碱性螺旋-环-螺旋(bHLH)-WD40 复合物对 PA 生物合成基因的转录调控尚不完全清楚。本研究从毛白杨中分离出一个拟南芥 TT2 样基因 MYB115,并通过多种分子、遗传和生化方法进行了鉴定。MYB115 恢复了拟南芥 tt2 突变体种皮中的 PA 合成。在杨树中过表达 MYB115 可激活 PA 生物合成基因的表达,导致 PA 浓度显著增加。相反,CRISPR/Cas9 产生的 myb115 突变体表现出降低的 PA 含量和降低的 PA 生物合成基因的表达。MYB115 直接激活 PA 特异性结构基因的启动子。MYB115 与杨树 TT8 相互作用。MYB115、TT8 和杨树 TTG1 的共表达显著增强了 ANR1 和 LAR3 的表达。此外,过表达 MYB115 的转基因植物对真菌病原体 Dothiorella gregaria 的抗性增加,而与野生型植物相比,myb115 突变体表现出更高的敏感性。我们的数据提供了对 MYB115 控制杨树 PA 生物合成的调控机制的深入了解,这可以有效地用于 PA 的代谢工程以提高对真菌病原体的抗性。