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An improved method for transformation of Actinidia arguta utilized to demonstrate a central role for MYB110 in regulating anthocyanin accumulation in kiwiberry

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Abstract

Actinidia arguta and related species produce small edible kiwifruit (kiwiberry) with attractive consumer features and high nutritional value. As a recently developed crop, kiwiberry can be improved further by classical breeding or genetic manipulation. The currently available Agrobacterium-mediated transformation protocol is only applicable to a limited number of kiwiberry genotypes such as the female cultivar ‘Hortgem Tahi’. We developed protocols for regeneration and Agrobacterium-mediated transformation of two A. arguta genotypes, female AA06-01 and male AA05-06, also applicable to ‘Hortgem Tahi’. Altered composition of basal salts in the callus-induction media was sufficient to prevent callus browning in AA06-01 and ‘Hortgem Tahi’, but not in AA05-06. Altering the hormone composition to avoid a prolonged callus stage was successfully used to prevent browning and induce shoot development in all three genotypes. Using our improved protocols, Agrobacterium-mediated transformation of a binary vector carrying the neomycin phosphotransferase II (nptII) expression cassette gave rise to kanamycin-resistant plants of both AA06-01 and AA05-06 and the presence of the nptII transgene was confirmed by genomic PCR. The improved protocols were also used to ectopically overexpress the anthocyanin-related transcription factor AcMYB110 in ‘Hortgem Tahi’ giving rise to purple callus with elevated anthocyanin accumulation. AcMYB110 expression and increased levels of anthocyanins were detected in mature leaves of established transgenic plants compared to controls, clearly demonstrating the important role for MYB110 in regulation of anthocyanin accumulation in kiwiberry. The protocols developed during this study provide tools for further functional analyses and genetic manipulation of kiwiberry genotypes.

Key message

Using a newly developed transformation method we were able to generate transgenic lines of three Actinidia arguta genotypes and demonstrate an important role for MYB110 in anthocyanin accumulation in kiwiberry.

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Acknowledgements

We thank Geeta Chhiba for media preparation, Monica Dragulescu and the glasshouse staff for management of transgenic plants and Cath Kingston for critically reading the manuscript. This work was funded by the New Zealand Ministry of Business, Innovation and Employment Grant C11X1602.

Funding

This work was funded by the NZ Ministry of Business, Innovation and Employment Grant C11X1602.

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Authors and Affiliations

  1. The New Zealand Institute for Plant and Food Research Limited (Plant & Food Research) Mt Albert, Auckland Mail Centre, Private Bag 92169, Auckland, 1142, New Zealand

    Dinum Herath, Tianchi Wang, Yongyan Peng, Andrew C. Allan, Joanna Putterill & Erika Varkonyi-Gasic

  2. School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand

    Dinum Herath, Andrew C. Allan & Joanna Putterill

Authors
  1. Dinum Herath
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  2. Tianchi Wang
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  3. Yongyan Peng
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  4. Andrew C. Allan
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  5. Joanna Putterill
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  6. Erika Varkonyi-Gasic
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Contributions

DH, TW, ACA, JP and EV-G conceived and designed research. DH and YP conducted experiments and analysed data. TW contributed stock plants. DH, YP and EV-G wrote the manuscript. All authors read, edited and approved the manuscript.

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Correspondence to Erika Varkonyi-Gasic.

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The authors declare that they have no conflict of interest.

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Communicated by Henryk Flachowsky.

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Herath, D., Wang, T., Peng, Y. et al. An improved method for transformation of Actinidia arguta utilized to demonstrate a central role for MYB110 in regulating anthocyanin accumulation in kiwiberry. Plant Cell Tiss Organ Cult 143, 291–301 (2020). https://doi.org/10.1007/s11240-020-01915-1

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  • DOI: https://doi.org/10.1007/s11240-020-01915-1

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