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

시토신 탈메틸화 관련 NtROS2a 유전자 발현을 제어한 RNAi 식물의 DNA microarray 분석

Choi, Jang Sun;Lee, In Hye;Jung, Yu Jin;Kang, Kwon Kyoo
최장선;이인혜;정유진;강권규

  • Received : 2016.06.24
  • Accepted : 2016.06.26
  • Published : 2016.06.30

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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Cited by

  1. Overexpression ofNtROS2agene encoding cytosine DNA demethylation enhances drought tolerance in transgenic rice vol.43, pp.3, 2016, https://doi.org/10.5010/JPB.2016.43.3.376

Acknowledgement

Grant : Gene editing기술을 이용한 농업형질개량 유용작물개발

Supported by : 농촌진흥청