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Planta

, Volume 248, Issue 5, pp 1277–1287 | Cite as

Biochemical characterization and spatio-temporal analysis of the putative l-DOPA pathway in Mucuna pruriens

  • Susheel Kumar Singh
  • Sunita Singh Dhawan
  • Raj Kishori Lal
  • Karuna Shanker
  • Manju Singh
Original Article

Abstract

Main conclusion

Transcriptome analysis and biochemical characterization of the putative l-3,4-dihydroxyphenylalanine (l-DOPA) pathway in Mucuna pruriens (L.) DC have been performed. Spatio-temporal quantification of the putative l-DOPA biosynthetic pathway genes and its correlation with respective metabolites was established. l-tyrosine, l-DOPA, and dopamine from all plant parts were quantified.

The de novo transcriptome analysis was performed using leaves of the selected M. pruriens mutant T-IV-9 during maturity. The putative l-DOPA pathway and its regulatory genes were retrieved from transcriptome data and the l-DOPA pathway was biochemically characterized. The spatial and temporal gene expression for the l-DOPA pathway was identified with respect to the chemical constituents. l-tyrosine, l-DOPA, and dopamine contents were highest in leaves during maturity (about 170-day-old plants). The polyphenol oxidase (PPO) was highly expressed in tender stems (230-fold higher as compared to seeds) as well as a high l-DOPA content. The PPO gene was highly expressed in leaves (3367.93 in FPKM) with a 79-fold increase compared to control plants during maturity. l-DOPA was found in every part with varied levels. The highest l-DOPA content was found in mature dried seed (3.18–5.8%), whereas the lowest amount was recorded in mature and dried leaves. The reproductive parts of the plant had a higher amount of l-DOPA content (0.9–5.8%) compared to the vegetative parts (0.2–0.91%). Various amino acid transporters and permeases were expressed in M. pruriens. The transcripts of dopa decarboxylase (DDC) were found in almost all parts of the plant, but its higher content was limited to the leaf.

Keywords

Dopamine l-DOPA Mucuna pruriens Transcriptome 

Abbreviations

AD

Arogenate dehydrogenase

DDC

Dopa/tyrosine decarboxylase

L-DOPA

l-3, 4-dihydroxyphenylalanine

PPO

Polyphenol oxidase

Notes

Acknowledgements

We gratefully acknowledge the support and facilities provided by the Director CSIR-CIMAP Lucknow, India and senior research fellowship to the first author from ICMR (Indian Council Medical Research), New Delhi, India. The funding for this work was acknowledged from AGTEC (BSC0110- a CSIR network programme). We are highly thankful to Dr. M.M. Gupta for his kind support.

Compliance with ethical standards

Conflict of interest

Authors declare no conflict of interests.

Supplementary material

425_2018_2978_MOESM1_ESM.docx (3.7 mb)
Supplementary material 1 (DOCX 3764 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Biotechnology DivisionCSIR-Central Institute of Medicinal and Aromatic Plants P.O.-CIMAPLucknowIndia
  2. 2.Genetics and Plant Breeding DivisionCSIR-Central Institute of Medicinal and Aromatic Plants P.O.-CIMAPLucknowIndia
  3. 3.Analytical Chemistry DivisionCSIR-Central Institute of Medicinal and Aromatic Plants P.O.-CIMAPLucknowIndia

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