Genetic transformation of Populus alba × P. glandulosa ‘84K’ with AtDME1 and its chemical-inducible expression analysis

Wu Xiaojuan; Lu Junqian; Chang Yingying; Zhong Shanchen; Su Xiaohua; Zhang Bingyu

doi:10.12171/j.1000-1522.20190068

Journal of Beijing Forestry University > 2020 > 42(6): 26-32. > DOI: 10.12171/j.1000-1522.20190068

Wu Xiaojuan, Lu Junqian, Chang Yingying, Zhong Shanchen, Su Xiaohua, Zhang Bingyu. Genetic transformation of Populus alba × P. glandulosa ‘84K’ with AtDME1 and its chemical-inducible expression analysis[J]. Journal of Beijing Forestry University, 2020, 42(6): 26-32. DOI: 10.12171/j.1000-1522.20190068

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Genetic transformation of Populus alba × P. glandulosa ‘84K’ with AtDME1 and its chemical-inducible expression analysis

State Key Laboratory of Tree Genetics and Breeding, Key Laboratory of Tree Breeding and Cultivation of National Forestry and Grassland Administration, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, China

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Received Date: January 24, 2019
Revised Date: March 21, 2019
Available Online: May 29, 2020
Published Date: June 30, 2020

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Abstract

Abstract

ObjectiveDNA methylation is an important epigenetic modification that plays an essential role in plant growth and development. In this study, the chemically-induced promoter and the Arabidopsis thaliana demethylation transferase gene AtDME1 were introduced into the genome of Populus alba × P. glandulosa ‘84K’, and the expression of AtDME1 was effectively induced by 17-β-estradiol treatment. The expression characteristics of AtDME1 in transgenic poplar plants were investigated. This study laid a foundation for the establishment of poplar methylation-induced variation system and genetic improvement of poplars.
MethodThe chemically-induced promoter and AtDME1 were transformed into genome of P. alba × P. glandulosa ‘84K’ using Agrobacterium-mediated transformation. Traditional PCR and DNA sequencing were used to identify the transgenic plants in hygromycin resistant plants. The chemical inducer 17-β-estradiol was used to induce expression of AtDME1 in in vitro leaves of a transgenic clone for 0, 3, 6, 12, 24, 48, 96 and 144 hours, and the expression of AtDME1 gene was detected by quantitative real-time PCR (qRT-PCR).
ResultA total of 224 hygromycin resistant buds were obtained, among them six hygromycin resistant plants were screened. Finally, six transgenic plants were identified by molecular methods, which were named as AD-1−6. Results of qRT-PCR showed that the expression of AtDME1 reached its highest level after 3 hours treatment of 17-β-estradiol, and its expression gradually decreased after12 hour’s treatment.
ConclusionThe chemical inducer 17-β-estradiol can rapidly and efficiently induce the expression of AtDME1 gene in transgenic poplar, which lays a solid foundation for further study on the mechanism of DME1 gene in the regulation of poplar genome methylation. It lays a solid foundation for the study of the chemical expression characteristics of poplar.
- Populus alba × Populus glandulosa ‘84K’,
- transgenic plant,
- AtDME1,
- chemical-inducible expression

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Genetic transformation of Populus alba × P. glandulosa ‘84K’ with AtDME1 and its chemical-inducible expression analysis