Hou Nan, Li Chaoshuo, He Jieqiang, Liu Yu, Yu Sisi, Malnoy Mickael, Mobeen Tahir Muhammad, Xu Lingfei, Ma Fengwang, Guan Qingmei
State Key Laboratory of Crop Stress Biology for Arid Areas/Shaanxi Key Laboratory of Apple, College of Horticulture, Northwest A&F University, No. 3 Taicheng Road, Yangling, Shaanxi, 712100, China.
Research and Innovation Centre, Fondazione Edmund Mach, Via E. Mach, San Michele all'Adige, 38010, Italy.
New Phytol. 2022 May;234(4):1294-1314. doi: 10.1111/nph.18069. Epub 2022 Mar 26.
Although the N -methyladenosine (m A) modification is the most prevalent RNA modification in eukaryotes, the global m A modification landscape and its molecular regulatory mechanism in response to drought stress remain unclear. Transcriptome-wide m A methylome profiling revealed that m A is mainly enriched in the coding sequence and 3' untranslated region in response to drought stress in apple, by recognizing the plant-specific sequence motif UGUAH (H=A, U or C). We identified a catalytically active component of the m A methyltransferase complex, MdMTA. An in vitro methyl transfer assay, dot blot, LC-MS/MS and m A-sequencing (m A-seq) suggested that MdMTA is an m A writer and essential for m A mRNA modification. Further studies revealed that MdMTA is required for apple drought tolerance. m A-seq and RNA-seq analyses under drought conditions showed that MdMTA mediates m A modification and transcripts of mRNAs involved in oxidative stress and lignin deposition. Moreover, m A modification promotes mRNA stability and the translation efficiency of these genes in response to drought stress. Consistently, MdMTA enhances lignin deposition and scavenging of reactive oxygen species under drought conditions. Our results reveal the global involvement of m A modification in the drought response of perennial apple trees and illustrate its molecular mechanisms, thereby providing candidate genes for the breeding of stress-tolerant apple cultivars.
尽管N -甲基腺苷(m⁶A)修饰是真核生物中最普遍的RNA修饰,但干旱胁迫下的全局m⁶A修饰图谱及其分子调控机制仍不清楚。全转录组m⁶A甲基化组分析表明,在苹果中,响应干旱胁迫时,m⁶A主要富集在编码序列和3'非翻译区,通过识别植物特异性序列基序UGUAH(H = A、U或C)。我们鉴定出了m⁶A甲基转移酶复合物的一个具有催化活性的组分MdMTA。体外甲基转移试验、斑点印迹、液相色谱-串联质谱以及m⁶A测序(m⁶A-seq)表明,MdMTA是一个m⁶A写入酶,对m⁶A mRNA修饰至关重要。进一步研究表明,苹果耐旱性需要MdMTA。干旱条件下的m⁶A-seq和RNA-seq分析表明,MdMTA介导了参与氧化应激和木质素沉积的mRNA的m⁶A修饰及转录本。此外,m⁶A修饰可促进这些基因在干旱胁迫下的mRNA稳定性和翻译效率。同样,MdMTA在干旱条件下增强了木质素沉积和活性氧清除。我们的结果揭示了m⁶A修饰在多年生苹果干旱响应中的全局参与情况,并阐明了其分子机制,从而为培育耐胁迫苹果品种提供了候选基因。