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Frond transformation system mediated by Agrobacterium tumefaciens for Lemna minor

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Abstract

The Lemnaceae, known as duckweed, the smallest flowering aquatic plant, shows promise as a plant bioreactor. For applying this potential plant bioreactor, establishing a stable and efficient genetic transformation system is necessary. The currently favored callus-based method for duckweed transformation is time consuming and genotype limited, as it requires callus culture and regeneration, which is inapplicable to many elite duckweed strains suitable for bioreactor exploitation. In this study, we attempted to establish a simple frond transformation system mediated by Agrobacterium tumefaciens for Lemna minor, one of the most widespread duckweed species in the world. To evaluate the feasibility of the new transformation system, the gene CYP710A11 was overexpressed to improve the yield of stigmasterol, which has multiple medicinal purposes. Three L. minor strains, ZH0055, D0158 and M0165, were transformed by both a conventional callus transformation system (CTS) and the simple frond transformation system (FTS). GUS staining, PCR, quantitative PCR and stigmasterol content detection showed that FTS can produce stable transgenic lines as well as CTS. Moreover, compared to CTS, FTS can avoid the genotype constraints of callus induction, thus saving at least half of the required processing time (CTS took 8–9 months while FTS took approximately 3 months in this study). Therefore, this transformation system is feasible in producing stable transgenic lines for a wide range of L. minor genotypes.

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Acknowledgements

This study was supported by the National Key Technology R&D Program of China (2015BAD15B01), the National Natural Science for General Foundation of China (31770395), Key deployment projects of Chinese Academy of Sciences (ZDRW-ZS-2017-2-1), Science and Technology Service Network Initiative of Chinese Academy of Sciences (KFJ-STS-ZDTP-008); Science & Technology Program of Sichuan Province (2017NZ0018 and 2017HH0077), and the Key and Open Fund of Key Laboratory of Environmental and Applied Microbiology, Chinese Academy of Sciences (KLEAMCAS201501, KLCAS-2014-05 and KLCAS-2016-02).

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  1. Gui-Li Yang and Yang Fang have contributed equally to this work and are co-first authors.

Authors and Affiliations

  1. Key Laboratory of Environmental and Applied Microbiology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041, China

    Gui-Li Yang, Yang Fang, Ya-Liang Xu, Li Tan, Qi Li, Yang Liu, Fan Lai, Yan-Ling Jin, An-Ping Du, Kai-Ze He, Xin-Rong Ma & Hai Zhao

  2. Environmental Microbiology Key Laboratory of Sichuan Province, Chengdu, 610041, China

    Gui-Li Yang, Yang Fang, Ya-Liang Xu, Li Tan, Qi Li, Yang Liu, Fan Lai, Yan-Ling Jin, An-Ping Du, Kai-Ze He, Xin-Rong Ma & Hai Zhao

  3. University of Chinese Academy of Sciences, Beijing, 100049, China

    Gui-Li Yang, Ya-Liang Xu, Qi Li, Yang Liu & Fan Lai

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  1. Gui-Li Yang
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  2. Yang Fang
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Contributions

HZ, XRM, GLY and YF conceived the research. GLY and XRM performed experiments. GLY and XRM analyzed the data and wrote the manuscript. YLX, LT, QL, YL, FL, YLJ, APD, and KZH contributed partial experiments.

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Correspondence to Xin-Rong Ma or Hai Zhao.

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Yang, GL., Fang, Y., Xu, YL. et al. Frond transformation system mediated by Agrobacterium tumefaciens for Lemna minor. Plant Mol Biol 98, 319–331 (2018). https://doi.org/10.1007/s11103-018-0778-x

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