Belmondo Simone, Calcagno Cristina, Genre Andrea, Puppo Alain, Pauly Nicolas, Lanfranco Luisa
Dipartimento di Scienze della Vita e Biologia dei Sistemi, Università degli Studi di Torino, Via Accademia Albertina 13, 10123, Turin, Italy.
Université Nice Sophia Antipolis, Institut Sophia Agrobiotech, 06900, Sophia Antipolis, France.
Planta. 2016 Jan;243(1):251-62. doi: 10.1007/s00425-015-2407-0. Epub 2015 Sep 24.
Our study demonstrated that the NAPDH oxidase gene MtRbohE is expressed in arbusculated cells and plays a role in arbuscule development. Plant NADPH oxidases, known as respiratory burst oxidase homologs (RBOH), belong to a multigenic family that plays an important role in the regulation of plant development and responses to biotic and abiotic stresses. In this study, we monitored the expression profiles of five Rboh genes (MtRbohA, MtRbohB, MtRbohE, MtRbohG, MtRbohF) in the roots of the model species Medicago truncatula upon colonization by arbuscular mycorrhizal fungi. A complementary cellular and molecular approach was used to monitor changes in mRNA abundance and localize transcripts in different cell types from mycorrhizal roots. Rboh transcript levels did not drastically change in total RNA extractions from whole mycorrhizal and non-mycorrhizal roots. Nevertheless, the analysis of laser microdissected cells and Agrobacterium rhizogenes-transformed roots expressing a GUS transcriptional fusion construct highlighted the MtRbohE expression in arbuscule-containing cells. Furthermore, the down regulation of MtRbohE by an RNAi approach generated an altered colonization pattern in the root cortex, when compared to control roots, with fewer arbuscules and multiple penetration attempts. Altogether our data indicate a transient up-regulation of MtRbohE expression in cortical cells colonized by arbuscules and suggest a role for MtRbohE in arbuscule accommodation within cortical cells.
我们的研究表明,NAPDH氧化酶基因MtRbohE在丛枝细胞中表达,并在丛枝发育中发挥作用。植物NADPH氧化酶,即呼吸爆发氧化酶同源物(RBOH),属于一个多基因家族,在植物发育调控以及对生物和非生物胁迫的响应中发挥重要作用。在本研究中,我们监测了模式植物蒺藜苜蓿根部在丛枝菌根真菌定殖后五个Rboh基因(MtRbohA、MtRbohB、MtRbohE、MtRbohG、MtRbohF)的表达谱。采用了一种互补的细胞和分子方法来监测mRNA丰度的变化,并将转录本定位到菌根根的不同细胞类型中。在从整个菌根和非菌根根中提取的总RNA中,Rboh转录水平没有发生剧烈变化。然而,对激光显微切割细胞和表达GUS转录融合构建体的发根农杆菌转化根的分析突出了MtRbohE在含丛枝细胞中的表达。此外,与对照根相比,通过RNAi方法下调MtRbohE会导致根皮层中定殖模式改变,丛枝数量减少且有多次穿透尝试。我们的数据总体表明,MtRbohE在被丛枝定殖的皮层细胞中表达有短暂上调,并暗示MtRbohE在皮层细胞内丛枝容纳中发挥作用。