KNOX1is expressed and epigenetically regulated duringin vitro conditions inAgave spp

Background:The micropropagation is a powerful tool to scale up plants of economical and agronomical importance, enhancing crop productivity. However, a small but growing body of evidence suggests that epigenetic mechanisms, such as DNA methylation and histone modifications, can be affected under thein vitro conditions characteristic of micropropagation. Here, we tested whether the adaptation to differentin vitro systems (Magenta boxes and Bioreactors) modified epigenetically different clones ofAgave fourcroydesandA. angustifolia. Furthermore, we assessed whether these epigenetic changes affect the regulatory expression ofKNOTTED1-like HOMEOBOX(KNOX) transcription factors. Results:To gain a better understanding of epigenetic changes duringin vitro andex vitroconditions inAgave fourcroydes andA. angustifolia, we analyzed global DNA methylation, as well as different histone modification marks, in two different systems: semisolid in Magenta boxes (M) and temporary immersion in modular Bioreactors (B). No significant difference was found in DNA methylation inA. fourcroydesgrown in either M or B. However, whenA. fourcroydes was compared withA. angustifolia,there was a two-fold difference in DNA methylation between the species, independent of thein vitro system used. Furthermore, we detected an absence or a low amount of the repressive mark H3K9me2 in ex vitroconditions in plants that were cultured earlier either in M or B. Moreover, the expression ofAtqKNOX1andAtqKNOX2,onA. fourcroydesandA. angustifoliaclones, is affected duringin vitro conditions. Therefore, we used Chromatin ImmunoPrecipitation (ChIP) to know whether these genes were epigenetically regulated. In the case ofAtqKNOX1,the H3K4me3 and H3K9me2 were affected during in vitro conditions in comparison withAtqKNOX2. Conclusions:Agave clones plants with higher DNA methylation duringin vitro conditions were better adapted to ex vitroconditions. In addition,A. fourcroydesandA. angustifoliaclones displayed differential expression of the KNOX1gene duringin vitro conditions, which is epigenetically regulated by the H3K4me3 and H3K9me2 marks. The finding of an epigenetic regulation in key developmental genes will make it important in future studies to identify factors that help to find climate-resistant micropropagated plants.

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Bibliographic Details
Main Author: Clelia De-la-Peña
Format: info:eu-repo/semantics/article biblioteca
Subjects:info:eu-repo/classification/cti/6,
Online Access:http://cicy.repositorioinstitucional.mx/jspui/handle/1003/156
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Summary:Background:The micropropagation is a powerful tool to scale up plants of economical and agronomical importance, enhancing crop productivity. However, a small but growing body of evidence suggests that epigenetic mechanisms, such as DNA methylation and histone modifications, can be affected under thein vitro conditions characteristic of micropropagation. Here, we tested whether the adaptation to differentin vitro systems (Magenta boxes and Bioreactors) modified epigenetically different clones ofAgave fourcroydesandA. angustifolia. Furthermore, we assessed whether these epigenetic changes affect the regulatory expression ofKNOTTED1-like HOMEOBOX(KNOX) transcription factors. Results:To gain a better understanding of epigenetic changes duringin vitro andex vitroconditions inAgave fourcroydes andA. angustifolia, we analyzed global DNA methylation, as well as different histone modification marks, in two different systems: semisolid in Magenta boxes (M) and temporary immersion in modular Bioreactors (B). No significant difference was found in DNA methylation inA. fourcroydesgrown in either M or B. However, whenA. fourcroydes was compared withA. angustifolia,there was a two-fold difference in DNA methylation between the species, independent of thein vitro system used. Furthermore, we detected an absence or a low amount of the repressive mark H3K9me2 in ex vitroconditions in plants that were cultured earlier either in M or B. Moreover, the expression ofAtqKNOX1andAtqKNOX2,onA. fourcroydesandA. angustifoliaclones, is affected duringin vitro conditions. Therefore, we used Chromatin ImmunoPrecipitation (ChIP) to know whether these genes were epigenetically regulated. In the case ofAtqKNOX1,the H3K4me3 and H3K9me2 were affected during in vitro conditions in comparison withAtqKNOX2. Conclusions:Agave clones plants with higher DNA methylation duringin vitro conditions were better adapted to ex vitroconditions. In addition,A. fourcroydesandA. angustifoliaclones displayed differential expression of the KNOX1gene duringin vitro conditions, which is epigenetically regulated by the H3K4me3 and H3K9me2 marks. The finding of an epigenetic regulation in key developmental genes will make it important in future studies to identify factors that help to find climate-resistant micropropagated plants.