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Efficient transient protein expression in tomato cultivars and wild species using agroinfiltration-mediated high expression system

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Abstract

Key message

The new transient protein expression system using the pBYR2HS vector is applicable to several tomato cultivars and wild species with high level of protein expression.

Abstract

Innovation and improvement of effective tools for transient protein expression in plant cells is critical for the development of plant biotechnology. We have created the new transient protein expression system using the pBYR2HS vector that led to about 4 mg/g fresh weight of protein expression in Nicotiana benthamiana. In this study, we validated the adaptability of this transient protein expression system by agroinfiltration to leaves and fruits of several tomato cultivars and wild species. Although the GFP protein was transiently expressed in the leaves and fruits of all tomato cultivars and wild species, we observed species-specific differences in protein expression. In particular, GFP protein expression was higher in the leaves and fruits of Micro-Tom, Solanum pimpinellifolium (0043) and S. pimpinellifolium (0049-w1) than in those of cultivars and wild species. Furthermore, Agrobacterium with GABA transaminase enhanced transient expression in tomato fruits of Micro-Tom. Taken together with these results, our system is applicable to several tomato cultivars and species as well as a model tomato, even though characteristics are often different among tomato cultivars or species. Thus, the system is an effective, simple, and valuable tool to achieve rapid transgene expression to examine gene function in tomato plant cells.

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Abbreviations

GABA:

γ-Aminobutyric acid

gabT:

GABA transaminase

GFP:

Green fluorescence protein

RDR:

RNA-dependent RNA polymerases

TMV:

Tobamovirus

TRV:

Tobacco rattle virus

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Acknowledgements

We thank Ms. Yuri Nemoto at University of Tsukuba for technical support. All cultivars and wild species were provided by Tsukuba-Plant Innovation Research Center (T-PIRC), University of Tsukuba, through the National Bio-Resource Project (NBRP) of the Japan Agency for Research and Development (AMED), Japan. This work was supported by the Cabinet Office, Government of Japan, Cross-ministerial Strategic Innovation Promotion Program, “Technologies for creating next-generation agriculture, forestry and fisheries” (funding agency: Bio-oriented Technology Research Advancement Institution, NARO), by Grants-in-Aid for KAKENHI (JP16K07390) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, and by a Cooperative Research Grant from the Plant Transgenic Design Initiative, Gene Research Center, University of Tsukuba.

Author contribution statement

Conceived and designed the experiments: KH HE KM. Performed the experiments: KH SF NR KE TY KM. Contributed reagents/materials/analysis tools: KH NS HE KM. Wrote the paper: KH HE KM.

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  1. Ken Hoshikawa

    Present address: Japan International Research Center for Agricultural Sciences, 1-1 Ohwashi, Tsukuba, Ibaraki, 305-8686, Japan

Authors and Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8572, Japan

    Ken Hoshikawa, Satoshi Fujita, Na Renhu, Kentaro Ezura, Tsuyoshi Yamamoto, Satoko Nonaka, Hiroshi Ezura & Kenji Miura

  2. Tsukuba-Plant Innovation Research Center, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8572, Japan

    Satoko Nonaka, Hiroshi Ezura & Kenji Miura

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  1. Ken Hoshikawa
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Correspondence to Hiroshi Ezura or Kenji Miura.

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Communicated by Fumihiko Sato.

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Hoshikawa, K., Fujita, S., Renhu, N. et al. Efficient transient protein expression in tomato cultivars and wild species using agroinfiltration-mediated high expression system. Plant Cell Rep 38, 75–84 (2019). https://doi.org/10.1007/s00299-018-2350-1

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