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In vitro anther culture and Agrobacterium-mediated transformation of the AP1 gene from Salix integra Linn. in haploid poplar (Populus simonii × P. nigra)

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Abstract

A reliable, efficient anther culture system, the dominant technique for generating haploid plants in breeding programs, that can be used for generating transgenic poplar plants has been needed. In the present study, therefore, an anther culture system was developed using isolated mid- and late-uninucleate anthers of poplar (Populus simonii × P. nigra). From a combination of SSR and ploidy analyses, six double haploid and two haploid lines were characterized from 86 plants grown from 16 regenerated anther cultured lines. After 48 months of development, two plant lines from the regenerated plants maintained their haploid level in vitro for over 2 years. A number of haploid plants from the different lines were transferred to soil. The leaves of these transplants were then used as explants for transformation with the APETALA1 (AP1) gene using Agrobacterium tumefaciens. Overexpression of AP1 in haploid poplar induced early flowering with obvious petals when ectopically expressed. To our knowledge, this is the first report on changes in flowering time in AP1-trangenic poplar, which is important for elucidating the regulatory mechanism of tree flower development.

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Acknowledgements

The Fundamental Research Funds for the Central Universities (2572015EA01), the 111 Project (B16010), the Innovation Project of State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University; Grant Number 2013A04) and Natural Science Fund of Heilongjiang Province (No. QC2015035) supported this study.

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Authors and Affiliations

  1. State Key Laboratory of Tree Genetics and Breeding, Northeast Forestry University, 26 Hexing Road, Harbin, 150040, People’s Republic of China

    Jingli Yang, Kun Li, Chunyan Li, Junxiu Li & Chenghao Li

  2. Department of Pharmacy, Heilongjiang University of Chinese Medicine, Harbin, 150040, People’s Republic of China

    Bo Zhao

  3. Research Center on Life Sciences and Environmental Sciences, Harbin University of Commerce, Harbin, 150076, People’s Republic of China

    Wei Zheng

  4. Annoroad Gene Technology Co., Ltd, Beijing, 100176, People’s Republic of China

    Yuchi Gao

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  1. Jingli Yang
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  2. Kun Li
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  3. Chunyan Li
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  4. Junxiu Li
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  5. Bo Zhao
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  6. Wei Zheng
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  7. Yuchi Gao
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  8. Chenghao Li
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Corresponding author

Correspondence to Chenghao Li.

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Project funding: This work was supported by The Fundamental Research Funds for the Central Universities (2572015EA01), the Innovation Project of State Key Laboratory of Tree Genetics and Breeding (Northeast Forestry University; grant number 2013A04) and Natural Science Fund of Heilongjiang Province (No. QC2015035).

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Corresponding editor: Chai Ruihai.

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Yang, J., Li, K., Li, C. et al. In vitro anther culture and Agrobacterium-mediated transformation of the AP1 gene from Salix integra Linn. in haploid poplar (Populus simonii × P. nigra). J. For. Res. 29, 321–330 (2018). https://doi.org/10.1007/s11676-017-0453-0

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  • DOI: https://doi.org/10.1007/s11676-017-0453-0

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