Abstract
Main Conclusion
The novel C-methyltransferase, MaMT1, could catalyze the conversion of piperidine to 2-methylpiperidine, which may be involved in the methylation step of DNJ biosynthesis in mulberry leaves.
Abstract
Mulberry (Morus alba L.) is a worldwide crop with medicinal, feeding and nutritional value, and 1-deoxynojirimycin ((2R, 3R, 4R, 5S)-2-hydroxymethyl-3, 4, 5-trihydroxypiperidine, DNJ) alkaloid, a potent α-glucosidase inhibitor, is its main active ingredient. Our previous researches clarified the biosynthetic pathway of DNJ from lysine to Δ1-piperideine, but its downstream pathway is unclear. Herein, eight differential methyltransferases (MTs) genes were screened from transcriptome profiles of mulberry leaves with significant differences in DNJ content (P < 0.01). Subsequently, MaMT1 (OM140666) and MaMT2 (OM140667) were hypothesized as candidate genes related to DNJ biosynthesis by correlation analysis of genes expression levels and DNJ content of mulberry leaves at different dates. Functional characterization of MaMT1 and MaMT2 were performed by cloning, prokaryotic expression and enzymatic reaction in vitro, and it showed that MaMT1 protein could catalyze the conversion of piperidine to 2-methylpiperidine. Moreover, molecular docking confirmed the interaction of MaMT1 protein with piperidine and S-adenosyl-l-methionine (SAM), indicating that MaMT1 had C-methyltransferase activity, while MaMT2 did not. The above results suggested that MaMT1 may be involved in the methylation step of DNJ alkaloid biosynthesis in mulberry leaves, which is a breakthrough in the analysis of DNJ alkaloid biosynthetic pathway. It is worth mentioning that the novel MaMT1, annotated as serine hydroxymethyltransferase, could rely on SAM to perform C-methyltransferase function. Therefore, our findings contribute new insights into the research of DNJ alkaloid biosynthesis and C-methyltransferase family.
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Data availability
The cDNA sequences of MaMT1 (GenBank accession no. OM140666) and MaMT2 (GenBank accession no. OM140667) are available in the NCBI. Transcriptome sequencing data of mulberry leaves samples (Ma7 and Ma11) are available in the Sequence Read Archive database (SRA accession no. SRP127713). All data generated during this study are included in this article and its supplementary files.
Abbreviations
- DNJ:
-
1-Deoxynojirimycin
- MTs:
-
Methyltransferases
- SAM:
-
S-Adenosyl-l-methionine
- SHMT:
-
Hydroxymethyltransferase
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Funding
This work was supported by the National Natural Science Foundation of China (Grant numbers 81872961, 81573529); the National Key Research and Development Program of China (Grant number 2018YFC1706105); and the Key Project at Central Government Level (Grant number 2060302).
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Wan, J., Liao, Y., Liu, J. et al. Screening, cloning and functional characterization of key methyltransferase genes involved in the methylation step of 1-deoxynojirimycin alkaloids biosynthesis in mulberry leaves. Planta 255, 121 (2022). https://doi.org/10.1007/s00425-022-03901-7
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DOI: https://doi.org/10.1007/s00425-022-03901-7