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http://dx.doi.org/10.5010/JPB.2016.43.2.231

DNA microarray analysis of RNAi plant regulated expression of NtROS2a gene encoding cytosine DNA demethylation  

Choi, Jang Sun (Vegetable Research Division, National Institute of Horticultural & Herbal Science, RDA)
Lee, In Hye (Department of Horticulture, Hankyong National University)
Jung, Yu Jin (Institute of Genetic Engineering, Hankyong National University)
Kang, Kwon Kyoo (Institute of Genetic Engineering, Hankyong National University)
Publication Information
Journal of Plant Biotechnology / v.43, no.2, 2016 , pp. 231-239 More about this Journal
Abstract
To study the transcript levels of epigenetically regulated genes in tobacco, we have developed a transgenic line OX1 overexpressing NtROS2a gene encoding cytosine DNA demethylation and a RNAi plant line RNAi13. It has been reported that salt- and $H_2O_2$-stress tolerance of these transgenic lines are enhanced with various phenotypic characters (Lee et al. 2015). In this paper, we conducted microarray analysis with Agilent Tobacco 4 x 44K oligo chip by using overexpression line OX1, RNAi plant line RNAi 13, and wild type plant WT. Differentially expressed genes (DEGs) related to metabolism, nutrient supply, and various stressed were up-regulated by approximately 1.5- to 80- fold. DEGs related to co-enzymes, metabolism, and methylation functional genes were down-regulated by approximately 0.03- to 0.7- fold. qRT-PCR analysis showed that the transcript levels of several candidate genes in OX1 and RNAi lines were significantly (p < 0.05) higher than those in WT, such as genes encoding KH domain-containing protein, MADS-box protein, and Zinc phosphodiesterase ELAC protein. On the other hand, several genes such as those encoding pentatricopeptide (PPR) repeat-containing protein, histone deacetylase HDAC3 protein, and protein kinase were decreased by approximately 0.4- to 1.0- fold. This study showed that NtROS2a gene encoding DNA glycosylase related to demethylation could regulate adaptive response of tobacco at transcriptional level.
Keywords
DNA demethylation; methylation; microarray analysis; NtROS2a transgenic tobacco; transcriptional level;
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