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Agrobacterium-mediated cassava transformation for the Asian elite variety KU50

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Abstract

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Cassava genetic transformation has mostly been reported for African cassava varieties, but not for Asian varieties. This is the first report of cassava transformation in Asian elite varieties using friable embryogenic calli.

Abstract

Agrobacterium-mediated cassava transformation via friable embryogenic calli (FEC) has enabled the robust production of transgenic cassava. So far, mostly the model cassava variety 60444 and African varieties have been transformed because of their good production and regeneration from embryogenic tissues. It is important to develop transformation methods for elite Asian cassava varieties to meet the changing needs in one of the world’s major cassava production areas. However, a suitable transformation method for the Asian elite variety Kasetsart 50 (KU50) has not been developed. Here, we report a transformation method for KU50, the cultivar with the highest planting area in Thailand and Vietnam. In cassava transformation, the preparation of FEC as the target tissue for transgene integration is a key step. FEC induction from KU50 was improved by using media with reduced nutrients and excess vitamin B1, and somatic embryo and plant regeneration optimized by manipulation of naphthalene acetic acid (NAA), and benzylamino purine (BAP). The transformation efficiency for KU50 was 22%, approximately half that of 60444 at 45%. Transcriptome analysis indicated that the expression of genes related to cell-wall loosening was upregulated in FEC from KU50 compared with 60444, indicating that cell-wall production and assembly were disproportionate in the Asian variety. The transformation system for KU50 reported here will contribute to the molecular breeding of cassava plants for Asian farmers using transgenic and genome-editing technologies.

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Data Availability

The novel data generated in this study, including microarray data, has been deposited in the National Center for Biotechnology Information under the accession number GSE169685.

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Acknowledgements

This work was supported by the following funding: Strategic Funds for the Promotion of Science and Technology, Japan Society for the Promotion of Science (JSPS) Program for Advancing Strategic International Networks to Accelerate the Circulation of Talented Researchers, EIG CONCERT-Japan 4th Call under the Strategic International Research Cooperative Program of the Japan Science and Technology Agency (JST, JPMJSC16C4), the Science and Technology Research Partnership for Sustainable Development (SATREPS) in collaboration with the Japan Science and Technology Agency (JST, JPMJSA1508) and the Japan International Cooperation Agency (JICA), and the grant from RIKEN Center for Sustainable Resource Science (CSRS). We would like to thank Editage (www.editage.com) for English language editing.

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Authors and Affiliations

  1. RIKEN Center for Sustainable Resource Science, Plant Genomic Network Research Team, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan

    Yoshinori Utsumi, Chikako Utsumi, Maho Tanaka, Yoshie Okamoto, Satoshi Takahashi, Hiroki Tokunaga & Motoaki Seki

  2. RIKEN Cluster for Pioneering Research, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan

    Maho Tanaka & Motoaki Seki

  3. Agricultural Genetic Institute, Km 2, Pham Van Dong Road, Tuliem, Hanoi, Vietnam

    Tong Thi Huong, Anh Vu Nguyen & Nguyen Van Dong

  4. Donald Danforth Plant Science Center, St. Louis, MO, 63132, USA

    Nigel Taylor

  5. Kihara Institute for Biological Research, Yokohama City University, 641-12 Maioka-cho, Totsuka-ku, Yokohama, Kanagawa, 244-0813, Japan

    Motoaki Seki

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  1. Yoshinori Utsumi
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  2. Chikako Utsumi
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  3. Maho Tanaka
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  4. Yoshie Okamoto
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  5. Satoshi Takahashi
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  6. Tong Thi Huong
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  7. Anh Vu Nguyen
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  8. Nguyen Van Dong
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  9. Hiroki Tokunaga
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  10. Nigel Taylor
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Contributions

YU and MS supervised the experiments, and YU, MS, and NJT wrote the manuscript. YU, CU, and MT performed the experiments and analyzed the data. ANV, NVD, and HT provided constructive comments. YU, MT, and ST performed transcriptome analysis. CU, YO, MT, and TTH managed the plants. The NJT provided critical suggestions on methodologies.

Corresponding authors

Correspondence to Yoshinori Utsumi or Motoaki Seki.

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Utsumi, Y., Utsumi, C., Tanaka, M. et al. Agrobacterium-mediated cassava transformation for the Asian elite variety KU50. Plant Mol Biol 109, 271–282 (2022). https://doi.org/10.1007/s11103-021-01212-1

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