Flowering induced by 5‐azacytidine, a DNA demethylating reagent in a short‐day plant, Perilla frutescens var. crispa

Abstract

Treatment with 5-azacytidine, a DNA demethylating reagent, induced flowering in Perilla frutescens (L.) Britton var. crispa (Thunb. ex Murray) Decne. ex L. H. Bailey, an absolute short-day plant under long days. The 5-azacytidine treatment induced slight suppression of vegetative growth but had no obvious effect on any other phenotypes. The Southern hybridization analysis of the genomic DNA isolated from the leaves of 5-azacytidine-treated plants and digested with restriction enzyme, methylation-insensitive Msp I or methylation-sensitive Hpa II with P. frutescens 25S-18S rDNA intergenic spacer probe indicated that the 5-azacytidine treatment caused demethylation of the genomic DNA. The 5-azacytidine-induced flowering was delayed as compared with the short day-induced flowering. Flowers were formed even at the lower nodes which had not been directly treated with 5-azacytidine. The results suggest that DNA demethylation induced flowering by inducing the production of a transmissible flowering stimulus in P. frutescens.

Exogenous steroid hormones stimulate full development of autonomous endosperm in Arabidopsis thaliana

“…In animal cultures of established cell lines, 5-azaC is used, inter alia, to investigate the effects of the chemotherapeutic agent in the treatment of tumors [29][30][31]. In plants, 5-azaC is helpful in understanding the mechanisms that control the gene silencing [32]. Active DNA demethylation occurs in plant genome in response to biotic and abiotic stresses suggesting that DNA methylation/demethylation control the mechanisms of plant stress response [33].…”

“…In turn, overexpression of an antisense transcript of DNA methyltransferase [antisense-cDNA methyltransferase (MET1)] induces flowering and reduces FLC gene expression, which is one of the main inhibitors of flowering in Arabidopsis [32]. The use of 5-azaC as DNA demethylating agent in experimental conditions allows the study on the regulation of the activity of the genes responsible for flowering (FLC, VIN, VRN) and seed development (like FIE [36]) through the mechanism of methylation/demethylation [32].…”

Azacitidine (5-AzaC)-treatment and mutations in DNA methylase genes affect embryogenic response and expression of the genes that are involved in somatic embryogenesis in Arabidopsis

“…Thus, it appears that the enhanced activity of the MET1 and CMT3 genes is generally associated with embryonic development and might reflect a requirement for the propagation of the DNA methylation pattern in the intensively dividing pro-embryonic and embryonic cells. In support of this assumption, the period with the highest expression of the MET1 and CMT3 genes coincides with the time of the intensive cell divisions that are associated with the acquisition of embryonic fate by the explant cells (Kurczyńska et al 2007).…”

“…The contrasting embryogenic capacity of the cmt3 and met1 mutants suggests that the contribution of the CMT3 and MET1 methylases to SE appears to be different. The role of CMT3 in the callus-mediated shoot regeneration of Arabidopsis (Shemer et al 2015) might suggest that in the SE system of Arabidopsis in which marginal callus production accompanies the direct development of somatic embryos from explant cells (Gaj 2001;Kurczyńska et al 2007), the function of CMT3 might be not critical for SE induction. In contrast, the importance of MET1 activity for SE induction might be related with the role of the enzyme in the hormone-related processes that play a central role in SE induction (Nic-Can and Loyola-Vargas 2016).…”

“…5-AzaC is randomly incorporated into a newly synthesised DNA strand instead of cytosine and, as a result, a dose-and time-dependent decrease in the DNA methyltransferase activity can be observed that is followed by genome hypomethylation at random sequences (Christman 2002;Issa and Kantarjian 2009). Treatment with 5-AzaC has been shown to induce various plant phenotypes including dwarfism, early flowering and an inhibition of vegetative growth (Kondo et al 2006). Although 5-AzaC treatment has also been indicated to impact the morphogenic processes that are induced in vitro including the capacity of a tissue for SE (Santos and Fevereiro 2002;Yamamoto et al 2005;Tokuji et al 2011;Fraga et al 2012;Teyssier et al 2014;Solís et al 2015), our knowledge about the role of this modification in the epigenetic regulation of the genes that control the developmental plasticity of somatic plant cells is still limited.…”