Xu Xuan, Guerriero Gea, Domergue Frederic, Beine-Golovchuk Olga, Cocco Emmanuelle, Berni Roberto, Sergeant Kjell, Hausman Jean-Francois, Legay Sylvain
Environmental Research and Innovation (ERIN) Department, Luxembourg Institute of Science and Technology, Esch-Sur-Alzette, Luxembourg.
Université de Bordeaux, Centre National de la Recherche Scientifique (CNRS) - Unité Mixte de Recherche (UMR) 5200, Laboratoire de biogenèse Membranaire, Bâtiment A3 - Institut Natitonal de la Recherche Agronomique (INRA) Bordeaux Aquitaine, Villenave d'Ornon, France.
Front Plant Sci. 2023 Mar 20;14:1143961. doi: 10.3389/fpls.2023.1143961. eCollection 2023.
Apple russeting is mainly due to the accumulation of suberin in the cell wall in response to defects and damages in the cuticle layer. Over the last decades, massive efforts have been done to better understand the complex interplay between pathways involved in the suberization process in model plants. However, the regulation mechanisms which orchestrate this complex process are still under investigation. Our previous studies highlighted a number of transcription factor candidates from the Myeloblastosis (MYB) transcription factor family which might regulate suberization in russeted or suberized apple fruit skin. Among these, we identified , which was co-expressed with number of well-known key suberin biosynthesis genes.
To validate the MdMYB68 function, we conducted an heterologous transient expression in Nicotiana benthamiana combined with whole gene expression profiling analysis (RNA-Seq), quantification of lipids and cell wall monosaccharides, and microscopy.
MdMYB68 overexpression is able to trigger the expression of the whole suberin biosynthesis pathway. The lipid content analysis confirmed that MdMYB68 regulates the deposition of suberin in cell walls. Furthermore, we also investigated the alteration of the non-lipid cell wall components and showed that MdMYB68 triggers a massive modification of hemicelluloses and pectins. These results were finally supported by the microscopy.
Once again, we demonstrated that the heterologous transient expression in coupled with RNA-seq is a powerful and efficient tool to investigate the function of suberin related transcription factors. Here, we suggest MdMYB68 as a new regulator of the aliphatic and aromatic suberin deposition in apple fruit, and further describe, for the first time, rearrangements occurring in the carbohydrate cell wall matrix, preparing this suberin deposition.
苹果锈果病主要是由于角质层出现缺陷和损伤后,细胞壁中木栓质积累所致。在过去几十年里,人们付出了巨大努力来更好地理解模式植物中木栓质化过程所涉及的途径之间的复杂相互作用。然而,协调这一复杂过程的调控机制仍在研究中。我们之前的研究突出了髓细胞组织增生(MYB)转录因子家族中的一些转录因子候选物,它们可能调控锈果或木栓质化苹果果皮中的木栓质化过程。其中,我们鉴定出了 ,它与许多著名的关键木栓质生物合成基因共表达。
为了验证MdMYB68的功能,我们在本氏烟草中进行了异源瞬时表达,并结合全基因表达谱分析(RNA测序)、脂质和细胞壁单糖定量以及显微镜观察。
MdMYB68的过表达能够触发整个木栓质生物合成途径的表达。脂质含量分析证实,MdMYB68调控木栓质在细胞壁中的沉积。此外,我们还研究了非脂质细胞壁成分的变化,结果表明MdMYB68引发了半纤维素和果胶的大量修饰。这些结果最终得到了显微镜观察的支持。
我们再次证明,在 中进行异源瞬时表达并结合RNA测序是研究木栓质相关转录因子功能的强大而有效的工具。在此,我们提出MdMYB68是苹果果实中脂肪族和芳香族木栓质沉积的新调控因子,并首次进一步描述了碳水化合物细胞壁基质中为木栓质沉积所发生的重排。