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Efficient Agrobacterium tumefaciens-mediated transformation using embryogenic suspension cultures in sweetpotato, Ipomoea batatas (L.) Lam.

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Abstract

Efficient Agrobacterium tumefaciens-mediated transformation was achieved using embryogenic suspension cultures of sweetpotato (Ipomoea batatas (L.) Lam.) cv. Lizixiang. Cell aggregates from embryogenic suspension cultures were cocultivated with the A. tumefaciens strain EHA105 harboring a binary vector pCAMBIA1301 with gusA and hygromycin phosphotransferase II gene (hpt II) genes. Selection culture was conducted using 25 mg l−1 hygromycin. A total of 2,218 plants were regenerated from the inoculated 1,776 cell aggregates via somatic embryogenesis. β-glucuronidase (GUS) assay and PCR, dot blot and Southern blot analyses of the regenerated plants randomly sampled showed that 90.37% of the regenerated plants were transgenic plants. The number of integrated T-DNA copies varied from 1 to 4. Transgenic plants, when transferred to soil in a greenhouse and a field, showed 100% survival. No morphological variations were observed in the ex vitro transgenic plants. These results exceed all transformation experiments reported so far in the literature in quantity of independent events per transformation experiment in sweetpotato.

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Abbreviations

ABA:

Abscisic acid

AS:

Acetosyringone

Carb:

Carbenicillin

CTAB:

Cetyltrimethylammonium bromide

2,4-d :

2,4-Dichlorophenoxyacetic acid

GUS:

β-Glucuronidase

hpt II:

Hygromycin phosphotransferase II gene

Hyg:

Hygromycin

LB:

Luria–Bertani

MS:

Murashige and Skoog

npt II:

Neomycin phosphotransferase II gene

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Acknowledgments

This work was supported by Outstanding Younger Science Foundation of China (no. 30225028) and The National Science and Technology Project of China (no. 2006BAD01A06).

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Authors and Affiliations

  1. Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture, China Agricultural University, Beijing, 100094, China

    Bo Yu, Hong Zhai, Yuping Wang, Ning Zang & Qingchang Liu

  2. Beijing Key Laboratory of Crop Genetic Improvement, China Agricultural University, Beijing, 100094, China

    Bo Yu, Hong Zhai, Yuping Wang, Ning Zang & Qingchang Liu

  3. Laboratory of Crop Heterosis and Utilization, Ministry of Education, China Agricultural University, Beijing, 100094, China

    Shaozhen He & Qingchang Liu

Authors
  1. Bo Yu
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  2. Hong Zhai
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  3. Yuping Wang
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  4. Ning Zang
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  5. Shaozhen He
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  6. Qingchang Liu
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Correspondence to Qingchang Liu.

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Yu, B., Zhai, H., Wang, Y. et al. Efficient Agrobacterium tumefaciens-mediated transformation using embryogenic suspension cultures in sweetpotato, Ipomoea batatas (L.) Lam.. Plant Cell Tiss Organ Cult 90, 265–273 (2007). https://doi.org/10.1007/s11240-007-9265-9

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