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Agrobacterium-mediated transformation of embryogenic cell suspension cultures and plant regeneration in Lilium tenuifolium oriental × trumpet ‘Robina’

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Abstract

A method has been developed for embryogenic cell suspension cultures, plant regeneration and transformation of the important ornamental lily genotype (Lilium tenuifolium oriental × trumpet ‘Robina’). Bulb scales, filaments, ovaries and stem axis tissues were used as explants for callus induction in Murashige and Skoog (MS) medium with additions of growth regulators: picloram on its own, or in combination with 1-naphthaleneacetic acid (NAA), and thidiazuron (TDZ). The results show that the optimum medium for callus induction in bulb scale and filament tissue is MS + picloram 1.0 mg L−1, and for the ovary, it is MS + picloram 1.5 mg L−1. The stem axis had the highest rate (89.2 %) of callus induction with MS + NAA 2.2 mg L−1 + TDZ 0.1 mg L−1. The suspension cultures were established with the combination of NAA and TDZ with 2–5 mm cell clusters. These took a long time compared with suspension cultures established by picloram with 1–3 mm cell clusters. In three suspension cultures induced by picloram, the best callus from the point of view of proliferation and regeneration was derived from filaments. For plant regeneration, the growth rate of suspension cultures from the stem axis was higher than from the other three suspension culture induced by picloram. Vector pCAMBIA1301 with the β-glucuronidase (GUS) gene as reporter was transformed by Agrobacterium mediation into suspension cultures initiated from filament and stem axis material. After co-cultivation, the numbers of blue spots in material from the two sources were 26.8 ± 4.3 and 24.0 ± 4.7, respectively (difference not significant). Hygromycin-resistant callus was successfully regenerated into plantlets on plant growth regulator-free MS medium. Transgenic plants were also confirmed by the GUS histochemical assay, polymerase chain reaction.

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Acknowledgments

We would like to thank Dr Alexander (Sandy) Lang in New Zealand for professional editing services of the manuscript. This work was supported by the National High Technology Research and Development Program of China (863 Program) (Grant No. 2007AA10Z182).

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Authors and Affiliations

  1. College of Horticulture, Northwest A & F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Yinyan Qi & Yuejin Wang

  2. College of Forestry, Northwest A & F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Lingjuan Du, Yonghui Quan, Feifei Tian & Yali Liu

  3. Key Laboratory of Biology and Genetic Improvement of Horticultural Crops (Northwest Region), Ministry of Agriculture, Yangling, 712100, Shaanxi, People’s Republic of China

    Yinyan Qi, Lingjuan Du, Yonghui Quan, Feifei Tian, Yali Liu & Yuejin Wang

  4. State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A&F University, Yangling, 712100, Shaanxi, People’s Republic of China

    Yinyan Qi, Lingjuan Du, Yonghui Quan, Feifei Tian, Yali Liu & Yuejin Wang

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  1. Yinyan Qi
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  2. Lingjuan Du
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  3. Yonghui Quan
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  4. Feifei Tian
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  5. Yali Liu
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Correspondence to Yali Liu.

Additional information

Communicated by B. Borkowska.

Y. Qi and L. Du have contributed equally to this work.

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Qi, Y., Du, L., Quan, Y. et al. Agrobacterium-mediated transformation of embryogenic cell suspension cultures and plant regeneration in Lilium tenuifolium oriental × trumpet ‘Robina’. Acta Physiol Plant 36, 2047–2057 (2014). https://doi.org/10.1007/s11738-014-1582-0

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  • DOI: https://doi.org/10.1007/s11738-014-1582-0

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