ALKBH10B mediated epitranscriptomic regulation enhances drought tolerance of Arabidopsis thaliana via hormonal cross-talk and sustained photosynthesis.

The m⁶A demethylase ALKBH10B emerges as a crucial post-transcriptional regulator of hormonal crosstalk, photosynthetic maintenance, and mitochondrial energy metabolism to fine-tune plant stress adaptation to drought. Epitranscriptomic N⁶-methyladenosine (m⁶A) mRNA modification has emerged as an important regulatory layer of gene expression and protein synthesis in plant stress adaptation. In this study, expression profiling revealed that drought stress induces a global reprogramming of the m6A machinery in Arabidopsis thaliana, with the m6A demethylase ALKBH10B exhibiting a more pronounced time-dependent response. ALKBH10B transcription is induced by ABA, most likely via ABRE and MYB cis-elements, whereas jasmonate alleviates this activation. Moreover, ALKBH10B overexpressing plants (OX-1) accumulated wild type (WT) levels of ABA under drought stress but displayed a markedly reduced content in jasmonic acid (JA) and jasmonoyl-isoleucine (JA-Ile). They furthermore exhibit enhanced stomatal closure, improved water-use efficiency, and thus an enhanced drought tolerance. Transcriptome analysis revealed that drought stress induces fewer transcriptional changes in OX-1 plants compared to WT and the drought-sensitive alkbh10b-1 mutant. Differential expression and GO enrichment analyses highlight that photosynthesis-related processes were most affected by ALKBH10B activity, correlating with a preserved chlorophyll content and PSII efficiency in OX-1. Analysis by m⁶A-IP-qPCR showed that ALKBH10B directly demethylates specific photosynthesis-related and drought-responsive genes, while other transcripts are indirectly regulated by ALKBH10B-dependent processes. Moreover, transcripts associated with respiration were enriched in OX-1, indicating that ALKBH10B also supports mitochondrial energy metabolism during drought. Together, our results suggest that ALKBH10B enhances drought tolerance by coordinating m⁶A-dependent transcriptional and post-transcriptional regulation to maintain photosynthetic capacity and mitochondrial energy metabolism, as well as fine-tuning ABA-jasmonate crosstalk.