Abstract
Syringin, sinapyl alcohol 4-O-glucoside, is well known as a plant-derived bioactive monolignol glucoside. In Arabidopsis, recombinant chimeric protein UGT72E3/2 has been previously reported to lead to significantly higher syringin production than the parental enzymes UGT72E2 and UGT72E3. To enhance syringin content in Korean soybean (Glycine max L. ‘Kwangan’), we cloned the UGT72E3/2 gene under the control of the β-conglycinin or CaMV-35S promoter to generate β-UGT72E3/2 and 35S-UGT72E3/2 constructs, respectively, and then transformed them into soybean to obtain transgenic plants using the modified half-seed method. Real-time semi-quantitative PCR (RT-PCR) analysis showed that the UGT72E3/2 gene was expressed in the leaves of the β-UGT72E3/2 and 35S-UGT72E3/2 transgenic lines. HPLC analysis of the seeds and mature tissues of the T2 generation plants revealed that the β-UGT72E3/2 transgenic seeds accumulated 0.15 µmol/g DW of total syringin and 0.29 µmol/g DW of total coniferin, whereas coniferin and syringin were not detected in non-transgenic seeds. Moreover, coniferin and syringin also accumulated at high levels in non-seed tissues, particularly the leaves of β-UGT72E3/2 transgenic lines. In contrast, 35S-UGT72E3/2 lines showed no differences in the contents of coniferin and syringin between transgenic and non-transgenic soybean plants. Thus, the seed-specific β-conglycinin promoter might be an effective tool to apply to the nutritional enhancement of soybean crops through increased syringin production.
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Acknowledgements
This work was supported by the Next-Generation BioGreen 21 Program (Code PJ011150 and Code PJ011111022016 granted to Y.S. Chung and J. Nam, respectively), Rural Development Administration, and Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (Code 112124-5 granted to Y.S. Chung), and National Research Foundation (NRF-2017R1C1B5017598 granted to T. Kwon) in Republic of Korea.
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11816_2017_464_MOESM1_ESM.jpg
Verification of the introduced genes and expression levels in 35S-UGT72E3/2 transgenic plants (T0) using PCR and RT-PCR. a Genomic DNAs were extracted from T0 transgenic leaf tissues. Lane 1, UGT72E3/2 gene; Lane 2, Bar gene; Lane 3, DNA region between the 35S promoter and UGT72E3/2 gene; Lane 4, DNA region between the left border and Bar gene; Lane 5, DNA region between UGT72E3/2 gene and right border. b Total RNA was extracted from T0 transgenic leaf tissues. The TUB gene was used as a normalization control for leaf RNA levels. NT, non-transgenic plant; #1-#10, 35S-UGT72E3/2 transgenic lines (T0) (JPG 145 KB)
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Kwon, T., Kim, H.J., Yun, S.Y. et al. Enhancement of syringin contents in soybean seeds with seed-specific expression of a chimeric UGT72E3/E2 gene. Plant Biotechnol Rep 11, 439–447 (2017). https://doi.org/10.1007/s11816-017-0464-5
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DOI: https://doi.org/10.1007/s11816-017-0464-5