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Efficient production of Agrobacterium rhizogenes-transformed roots and composite plants in peanut (Arachis hypogaea L.)

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Abstract

Recalcitrance of most large-seeded legumes, such as peanut, to regeneration and genetic transformation has hampered studies on gene function and efforts for genetic improvement. Agrobacterium rhizogenes-mediated transformation provides a system for rapid and efficient transformation of plant tissues. In this study, embryonic axes along with cotyledons of peanut were injected with a suspension culture of A. rhizogenes using microliter syringes. The influence of several factors such as plant genotype, A. rhizogenes culture stage, co-culture period of A. rhizogenes, and acetosyringone concentration in the co-cultivation medium have been evaluated. It is found that A. rhizogenes-mediated transformation of peanut is genotype-independent. Up to 61% transformation was recorded when embryonic axes were co-cultivated with 5 × 107 A. rhizogenes cells from logarithmic phase for 2 days on co-culture medium containing 50 μmol l−1 acetosyringone. Composite plants with transgenic roots were harvested after 45 days of treatment. Furthermore, this method was applied to assess the insecticidal activity of a synthetic cry8Ea1 gene against Holotrichia parallela in transgenic roots of peanut.

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Abbreviations

DIRC:

Days of induced roots emerging after co-cultivation

NC:

Non-transgenic control

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Acknowledgments

This study was sported by 973 Projects of China (2009CB118902) and National Science and Technology Major Project (2009ZX08009-030B). We thank the Australian Research Council for Centre of Excellence funding to PMG. PMG also is a Senior Professorial Fellow of the CAS.

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Authors and Affiliations

  1. State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Lili Geng, Lihong Niu, Changlong Shu, Fuping Song & Jie Zhang

  2. ARC Centre of Excellence for Integrative Legume Research, The University of Queensland, St. Lucia, Brisbane, QLD, 4072, Australia

    Peter M. Gresshoff

  3. Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Dafang Huang

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  1. Lili Geng
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  2. Lihong Niu
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  3. Peter M. Gresshoff
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  4. Changlong Shu
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  5. Fuping Song
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  6. Dafang Huang
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  7. Jie Zhang
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Correspondence to Jie Zhang.

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Geng, L., Niu, L., Gresshoff, P.M. et al. Efficient production of Agrobacterium rhizogenes-transformed roots and composite plants in peanut (Arachis hypogaea L.). Plant Cell Tiss Organ Cult 109, 491–500 (2012). https://doi.org/10.1007/s11240-012-0113-1

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