5-Azacytidine combined with 2,4-D improves somatic embryogenesis of Acca sellowiana (O. Berg) Burret by means of changes in global DNA methylation levels

Fraga, Hugo Pacheco de Freitas; Vieira, Leila N.; Caprestano, Clarissa Alves; Steinmacher, Douglas André; Micke, Gustavo Amadeu; Spudeit, Daniel A.; Pescador, Rosete; Guerra, Miguel Pedro

doi:10.1007/s00299-012-1327-8

Cited by 84 publications

(52 citation statements)

References 44 publications

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“…In line with this postulate, we observed a significantly decreased level of 5 mC in the embryogenic culture of Arabidopsis and the reduction in the DNA methylation level has also been associated with SE induction in other plants (Santos and Fevereiro 2002;Xu et al 2004;Noceda et al 2009;Rodríguez-Sanz et al 2014;Teyssier et al 2014). However, in some plants, an increased (Fraga et al 2012;Rival et al 2013), differentially modulated (Leljak-Levanić et al 2004;Nic-Can et al 2013) and steady (Parra et al 2001) level of DNA methylation during SE has been reported. The diversity in the SE-associated DNA methylation levels suggests that various exo-and endogenous culture factors rather than the induction of embryonic development per se appear to contribute to the global DNA methylation level in cultured tissue (Huang et al 2012;Rival et al 2013;Li et al 2014;Rathore et al 2014).…”

Section: Se Induction Is Associated With the Decreased Level Of Globasupporting

confidence: 84%

“…Consistent with the diverse effects of 5-AzaC on gene expression level, the hypo-and hypermethylation of DNA in response to this chemical were reported in an embryogenic culture of Acca sellowiana, which were dependent on the plant genotype and the supplementation of the culture medium with 2,4-D (Fraga et al 2012).…”

Section: -Azac Treatment and Dna Methylase-related Mutations Affect supporting

confidence: 69%

“…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.…”

Section: Introductionmentioning

confidence: 99%

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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

et al. 2018

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Epigenetic processes including DNA methylation play a pivotal role in regulating the genes that control plant development. In contrast to in planta development, the contribution of DNA methylation to the morphogenic processes that are induced in vitro are much less recognised. Hence, in the present study, we analysed the impact of DNA methylation on somatic embryogenesis (SE) that was induced in Arabidopsis. The results demonstrated a decrease in the global DNA methylation level during SE that contrasted with the up-regulation of MET1 and CMT3 DNA methylases and the down-regulation of DNA demethylases (ROS1, DME and DML2). Hence, the global DNA methylation level appears not to correlate with the transcriptional activity of the genes encoding DNA methylases/demethylases, thereby implying the complexity of the regulatory mechanism that controls the DNA methylation status of the SE-epigenome. Moreover, distinct changes in the expression level of the SE-regulatory genes were indicated in the 5-AzaC-treated and DNA methylase mutant cultures. Accordingly, a significant repression of the LEC2, LEC1 and BBM genes was found in the 5-AzaC-treated culture that was incapable of SE induction. In contrast, the distinct up-regulation of these genes was observed in the drm1drm2 and drm1drm2cmt3 mutant cultures with an improved embryogenic response. The modulated expression of DNA methylase genes and the significantly modified embryogenic response of the met1 and drm mutants imply that both the maintenance and the de novo pathway of DNA methylation are engaged in the regulation of SE in Arabidopsis.

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Section: Se Induction Is Associated With the Decreased Level Of Globasupporting

confidence: 84%

Section: -Azac Treatment and Dna Methylase-related Mutations Affect supporting

confidence: 69%

Section: Introductionmentioning

confidence: 99%

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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

et al. 2018

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“…Nucleic acids digestion procedures were based on the method described by Johnston et al (2005) and Fraga et al (2012). High performance liquid chromatography (HPLC) analysis was performed according to Johnston et al (2005).…”

Section: Plant Materialsmentioning

confidence: 99%

<b>Dynamics in global DNA methylation and endogenous polyamine levels during protocorm-like bodies induction of <i>Cattleya tigrina</i> A. Richard

Almeida

Fraga

Navarro

et al. 2017

Acta Sci. Biol. Sci.

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ABSTRACT. Cattleya tigrina is endemic to the Atlantic forest biome and classified as vulnerable in the Red Book of Brazilian Flora. In vitro techniques comprise valuable tools for the conservation of endangered plant species. The aim of the present study was to evaluate the morphological features, global DNA methylation levels and free polyamines during protocorm-like bodies (PLBs) induction of C. tigrina. Along with that, an efficient protocol for in vitro propagation of this species is proposed. The first evidences of PLBs induction in C. tigrina occurred at seven days in culture, starting from the basal portion of the leaf abaxial surface. A hypomethylation marked the beginning of cell differentiation, followed by an increased global DNA methylation at 35 days in culture, coinciding with a subtle change in the structures morphogenetic development. During PLBs induction, putrescine exhibited higher levels as compared to spermidine and spermine, and apparently presents a major role during the PLBs induction in C. tigrina. Due to the apparent secondary PLBs formation, this protocol can represent a highly efficient method for in vitro propagation of this species.Keywords: orchids, micropropagation, in vitro culture, epigenetics, PLBs.Dinâmica da metilação do DNA global e níveis endógenos de poliaminas durante a indução de estruturas semelhantes a protocormos de Cattleya tigrina A. Richard RESUMO. Cattleya tigrina é uma espécie endêmica do bioma Mata Atlântica e classificada como vulnerável no Livro Vermelho da Flora Brasileira. As técnicas in vitro compreendem ferramentas valiosas a serem empregadas na conservação de espécies de plantas ameaçadas. O objetivo do presente estudo foi avaliar as características morfológicas, os níveis globais de metilação do DNA e as poliaminas livres durante a indução de estruturas semelhantes a protocormos (ESPs). Paralelamente, um protocolo eficiente para a propagação in vitro desta espécie é apresentado. As primeiras evidências de indução de ESPs em C. tigrina foram observadas aos sete dias de cultivo, a partir da porção basal da superfície abaxial da folha. Uma hipometilação foi observada concomitante ao início da diferenciação celular, e um aumento da metilação global do DNA foi encontrada aos 35 dias de cultivo, coincidindo com uma sutil mudança no desenvolvimento morfogenético das estruturas. Durante a indução de ESPs, a putrescina exibiu níveis aumentados em comparação a espermidina e espermina e, aparentemente, apresenta um papel importante durante a indução dessas estruturas em C. tigrina. Devido à aparente formação secundária de ESPs, este protocolo pode representar um método altamente eficiente para a propagação in vitro desta espécie.Palavras-chave: orquideas, micropropagação, cultivo in vitro, epigenética, ESPs.

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“…Application of 2,4-D in somatic embryogenesis had been reported on guava (Fraga et al 2012), chrysanthemum (Naing et al 2013), tagara herbal plant (Chen et al 2014a), and China rose (Chen et al 2014b); whereas picloram had been applied on pineapple (Roostika 2012), grapes (Dai et al 2015) and Ledebouria (Baskaran et al 2016). Nevertheless, application of 2,4-D to induce mangosteen somatic embryos resulted scarcely regenerated calli (Te-chato 1998; Rohani et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Effect of Different Sources of Plant Growth Regulator on the Induction and Development of Mangosteen Somatic Embryos

Joni¹,

Prihatini²,

Efendi³

et al. 2017

Indones. J. Agric. Sci.

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Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regulators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l -1 ) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l -1 ) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l -1 . The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l -1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l -1 TDZ and 0.7 mg l -1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.

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5-Azacytidine combined with 2,4-D improves somatic embryogenesis of Acca sellowiana (O. Berg) Burret by means of changes in global DNA methylation levels

Abstract: 5-Azacytidine combined with 2,4-D increases the number of Acca sellowiana somatic embryos. Global DNA methylation is directly affected by these compounds.

Cited by 84 publications

References 44 publications

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

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

<b>Dynamics in global DNA methylation and endogenous polyamine levels during protocorm-like bodies induction of <i>Cattleya tigrina</i> A. Richard

Effect of Different Sources of Plant Growth Regulator on the Induction and Development of Mangosteen Somatic Embryos

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