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Molecular cloning, characterization, and expression analysis of LeMYB1 from Lithospermum erythrorhizon

  • Original Papers
  • Published:
Biologia Plantarum

Abstract

MYB transcription factors (TFs) are known to have important functions in regulating the biosynthesis of secondary metabolites in plants. In this study, LeMYB1, a member of the MYB gene family of Lithospermum erythrorhizon, was cloned via the rapid amplification of cDNA ends. The alignment of the predicted translations of LeMYB1 with other MYB proteins revealed that LeMYB1 contained an N-terminal R2R3 repeat and a high degree of amino acid identity to NtMYBJS1 which is involved in jasmonic acid signalling and phenylpropanoid biosynthetic pathway regulation. To determine the expression pattern of LeMYB1, its promoter was cloned and the sequence analysis was performed. The results revealed a number of potential regulatory motifs related to tissue-specific gene expression and abiotic and biotic stress responses. Real-time PCR results suggest that LeMYB1 was induced transiently during the early stage when L. erythrorhizon cells were transferred from a B5 growth medium to a M9 production medium for shikonin formation. Exogenous methyl jasmonate (MeJA), an effective inducer of shikonin biosynthesis, induced the rapid LeMYB1 expression. In contrast, a treatment with ibuprofen (IBU), an inhibitor of jasmonate biosynthesis, significantly inhibited the LeMYB1 expression. Another inhibitor of shikonin formation, 2,4-dichlorophenoxyacetic acid (2,4-D), also markedly repressed the expression of LeMYB1. Tissue-specific expression analysis showed that LeMYB1 mRNA was predominantly accumulated in roots where shikonin was synthesized. Thus, the LeMYB1 gene may be a valuable member of the R2R3-MYB family in L. erythrorhizon and is possibly involved in the regulation of shikonin biosynthesis.

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Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

4CL:

4-coumarate:CoA-ligase

GAPDH:

glyceraldehydephosphate dehydrogenase

GPP:

geranylpyrophosphate

HMGR:

3-hydroxy-3-methylglutaryl-CoA reductase

HTH:

helix-turn-helix

IBU:

ibuprofen

MeJA:

methyl jasmonate

NJ:

neighbor-joining

NCBI:

National Center for Biotechnology Information

ORF:

open reading frame

PAL:

phenylalanine ammonia-lyase

PHB:

p-hydroxybenzoic acid

PGT:

p-hydroxybenzoate-3-geranyltransferase

PLACE:

plant cis-acting regulatory DNA elements

RACE:

rapid amplification of cDNA ends

RT-PCR:

reverse transcription polymerase chain reaction

TD-PCR:

touch-down PCR

TFs:

transcription factors

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Authors and Affiliations

  1. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, 210093, P.R. China

    H. Zhao, S. K. Baloch, L. R. Kong, W. J. Zhang, A. L. Zou, X. M. Wang, J. L. Qi & Y. H. Yang

  2. Engineering Technology Research Center of Anti-Aging Chinese Herb, School of Life Sciences, Fuyang Normal College, Fuyang, 236032, P.R. China

    H. Zhao

  3. Department of Biotechnology FCPD, Sindh Agriculture University, Tandojam, 71000, Pakistan

    S. K. Baloch

  4. Huaian High-Tech Research Institute, Nanjing University, Huaian, 223005, P.R. China

    J. L. Qi

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  1. H. Zhao
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  2. S. K. Baloch
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Correspondence to J. L. Qi or Y. H. Yang.

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Acknowledgements: This research was supported by the grants from the Natural Science Foundation of China (NSFC) (31071082, 31170275, 31171161), the Natural Science Foundations of the Jiangsu Bureau of Science and Technology (BK2010053, BK2011414), the Program for Changjiang Scholars and Innovative Research Team in University (IRT1020), the Project of New Century Excellent Talents in University (NCET-11-0234), and the Foundations of Huaian High-Tech Research Institute of Nanjing University (2011Z2).

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Zhao, H., Baloch, S.K., Kong, L.R. et al. Molecular cloning, characterization, and expression analysis of LeMYB1 from Lithospermum erythrorhizon . Biol Plant 58, 436–444 (2014). https://doi.org/10.1007/s10535-014-0411-z

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