Frond transformation system mediated by Agrobacterium tumefaciens for Lemna minor
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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.
KeywordsDuckweed Lemna minor Frond transformation system Agrobacterium tumefaciens Plant bioreactor Stigmasterol
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).
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.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Bertran K, Bertran K, Thomas C, Guo X, Bublot M, Pritchard N, Regan JT, Cox KM, Gasdaska JR, Dickey LF, Kapczynski DR, Swayne DE (2015) Expression of H5 hemagglutinin vaccine antigen in common duckweed (Lemna minor) protects against H5N1 high pathogenicity avian influenza virus challenge in immunized chickens. Vaccine 33:3456–3462CrossRefGoogle Scholar
- Chang WC, Chiu PL (1976) Induction of callus from fronds of duckweed (Lemna gibba L.). Bot Bull Acad Sin 17:106–109Google Scholar
- Dominguez A, Cervera M, Perez RM, Romero J, Fagoaga C, Cubero J, Lopez MM, Juarez JA, Navarro L (2004) Characterisation of regenerants obtained under selective conditions after Agrobacterium-mediated transformation of citrus explants reveals production of silenced and chimeric plants at unexpected high frequencies. Mol Breeding 14:171–183CrossRefGoogle Scholar
- Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. Calif Agric Exp Stn Circ 347:1–32Google Scholar
- Les DH, Crawford DJ, Landolt E, Gabel JD, Kimball RT (2002) Phylogeny and systematics of Lemnaceae, the duckweed family. Syst Bot 27:221–240Google Scholar
- Moreira AR (2007) The evolution of protein expression and cell culture. Biopharm Int 20:56–68Google Scholar