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Genetic transformation of Gentiana dahurica Fisch by Agrobacterium tumefaciens using zygotic embryo-derived callus

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Abstract

Gentiana dahurica Fisch is one of four important commercial Radix Gentianae stipulated by the Chinese Pharmacopoeia. We have established a rapid and effective regeneration system using zygotic embryo-derived callus of G.dahurica Fisch. Using this regeneration system, Agrobacteriumtumefaciens-mediated transformation of G. dahurica Fisch has been developed. Zygotic embryo-derived callus was infected with an A.tumefaciens strain (GV3130) harboring a pBI121 vector that contained an nptII selective marker gene. In a total of 60 zygotic embryo-derived calli assayed, frequency of calli with nptII gene PCR-positive transgenic plantlets is 5%. Transient glucuronidase (GUS) expression was observed from these transgenic plantlets. Frequency of GUS-positive transgenic plantlets (4/78) was approximately consistent with that of PCR assay (3/60). Our data indicate that we have successfully established a stable G. dahurica Fisch transformation system by A.tumefaciens. In general, this transformation system we have established might be useful for modifying physiological, medicinal and horticultural traits.

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Abbreviations

IM:

Induction medium for callus forming

RM:

Regeneration medium for obtaining regenerants from calli

ROM:

Root medium for induction of regenerants rooting

SM:

Selection medium with 30 or 50 mg l−1 kanamycin sulphate for screening putative transgenic plantlets

6-BA:

6-Benzyladenine

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Authors and Affiliations

  1. GanSu College of Traditional Chinese Medicine, 730000, Lanzhou, China

    Shao-Bo Sun

  2. School of Life Sciences, Tongji University, 200092, Shanghai, China

    Lai-Sheng Meng

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  1. Shao-Bo Sun
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  2. Lai-Sheng Meng
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Correspondence to Shao-Bo Sun or Lai-Sheng Meng.

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Communicated by E. Lojkowska.

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Sun, SB., Meng, LS. Genetic transformation of Gentiana dahurica Fisch by Agrobacterium tumefaciens using zygotic embryo-derived callus. Acta Physiol Plant 32, 629–634 (2010). https://doi.org/10.1007/s11738-009-0439-4

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  • DOI: https://doi.org/10.1007/s11738-009-0439-4

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