Preliminary Study On The The Regulation Of Auxin Signaling Pathway By N~6-methyladenosine In Arabidopsis Thaliana

Epigenetics is a discipline that emerged gradually in the 1980 s and developed gradually in the process of studying many life phenomena that do not conform to the classical Mendelss law of inheritances.Epigenetic inheritance exists in every growth and development process, and epigenetic modification regulation is often accompanied by the transformation of cell fate.In recent years,RNA modification has become a research hotspot in the field of epigenetics.RNA modification is abundant in animal,plant,yeast and virus species.A wide range of chemical modifications are present on mostRNAs,including m6A modifications in messengerRNA(mRNA),ribosomalRNA(rRNA)and small nuclearRNA(snRNA).Recently,it has been reported that m6A plays an important biological role in the plant growth and development However,the relationship between phytohormone signaling pathways and m6A modification remains unclear.Therefore,we want to explore the crosstalk between Auxin signaling pathway and m6A modification.Auxin is a key signal controlling plant growth and development.Loss of function of FIP37,a key component of methyltransferase modified by m6A modification,caused auxin related defects.The relationship between auxin signaling pathway and m6A modification was explored through dot blot experiment.In order to study the functional relationship between auxin and m6A modification,we examined the m6A modification level of totalRNA in the wild type(Colombia-0)treated with auxin analogue(NAA).The results showed that m6A was significantly elevated after auxin treatment.Based on the results,we suspected that there was a relationship between auxin pathway and m6A modification.Since Auxin plays an important role in the formation of callus from plant explants,we did tissue culture of wild-type and m6A defective mutants to further explore the relationship between auxin signaling pathway and m6A modification..It was found that most of the roots and leaves could form callus in the wild type.Therefore,we verified the correlation between auxin signaling pathway and m6A modification.In order to better prove the relationship between m6A modification and auxin signaling pathway,we treated the wild type and fip37-4LEC1:FIP37 mutant with exogenous auxin and performedRNA-seq.We found the auxin related genes can be induced by NAA treatment in wild type seedlings but not in the fip37-4LEC1:FIP37 mutant,suggesting m6A modification is essential for auxin signaling pathway.Using the same samples,we performed m6A-seq.It was found that most of m6A targeted transcripts induced by auxin were related to auxin stimulation and auxin activation pathways.This result confirmed the relationship between auxin signaling pathway and m6A modification.In summary,these preliminary results provide clues to explore the relationship between auxin signaling pathway and m6A modification in Arabidopsis and help people to understand function of m6A modification in plant signal transduction.