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CstMYB1R1, a REVEILLE-8-like transcription factor, regulates diurnal clock-specific anthocyanin biosynthesis and response to abiotic stress in Crocus sativus L

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Abstract

Key message

CstMYB1R1 acts as a positive regulator of Crocus anthocyanin biosynthesis and abiotic stress tolerance which was experimentally demonstrated through molecular analysis and over-expression studies in Crocus and Nicotiana.

Abstract

Regulatory mechanics of flavonoid/anthocyanin biosynthesis in Crocus floral tissues along the diurnal clock has not been studied to date. MYB proteins represent the most dominant, functionally diverse and versatile type of plant transcription factors which regulate key metabolic and physiological processes in planta. Transcriptome analysis revealed that MYB family is the most dominant transcription factor family in C. sativus. Considering this, a MYB-related REVEILLE-8 type transcription factor, CstMYB1R1, was explored for its possible role in regulating Crocus flavonoid and anthocyanin biosynthetic pathway. CstMYB1R1 was highly expressed in Crocus floral tissues, particularly tepals and its expression was shown to peak at dawn and dusk time points. Anthocyanin accumulation also peaked at dawn and dusk and was minimum at night. Moreover, the diurnal expression pattern of CstMYB1R1 was shown to highly correlate with Crocus ANS/LDOX gene expression among the late anthocyanin pathway genes. CstMYB1R1 was shown to be nuclear localized and transcriptionally active. CstMYB1R1 over-expression in Crocus tepals enhanced anthocyanin levels and upregulated transcripts of Crocus flavonoid and anthocyanin biosynthetic pathway genes. Yeast one hybrid (Y1H) and GUS reporter assay confirmed that CstMYB1R1 interacts with the promoter of Crocus LDOX gene to directly regulate its transcription. In addition, the expression of CstMYB1R1 in Nicotiana plants significantly enhanced flavonoid and anthocyanin levels and improved their abiotic stress tolerance. The present study, thus, confirmed positive role of CstMYB1R1 in regulating Crocus anthocyanin biosynthetic pathway in a diurnal clock-specific fashion together with its involvement in the regulation of abiotic stress response.

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Data availability

The sequence of CstMYB1R1 gene and CstLDOX promoter sequence used in this study have been submitted to NCBI Gene Bank through BankIt sequence submission tool under the accession numbers OQ436455 and OQ436454, respectively. All other data is given in manuscript and supplementary material.

Abbreviations

RVE8:

REVIELLE-8

Y1H:

Yeast one hybrid

LDOX:

Leucoanthocyanidin oxygenase

qRT-PCR:

Quantitative real-time PCR

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Acknowledgements

Zahid Yaqoob Bhat is highly grateful to University Grants Commission (UGC), New Delhi for providing him financial support as Junior and Senior Research Fellowships (JRF and SRF). The manuscript bears institutional Id: CSIR-IIIM/IPR/00637.

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No funding was received for the work given in this manuscript.

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Authors and Affiliations

  1. Plant Biotechnology Division, CSIR—Indian Institute of Integrative Medicine (IIIM), Sanat Nagar190005, Srinagar, Jammu and Kashmir, India

    Zahid Yaqoob Bhat, Javid Ahmad Mir & Nasheeman Ashraf

  2. Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, UP, India

    Zahid Yaqoob Bhat, Javid Ahmad Mir & Nasheeman Ashraf

  3. Quality Control and Quality Assurance Lab, Quality, Management and Instrumentation Division, CSIR- Indian Institute of Integrative Medicine, Canal Road, Jammu Tawi, 180001, India

    Arvind Kumar Yadav & Deepika Singh

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  1. Zahid Yaqoob Bhat
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  5. Nasheeman Ashraf
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Contributions

The current study was conducted under the supervision of NA. She along with ZYB wrote the manuscript. ZYB and JAM carried out the experimentation and performed their analyses. AKY and DS performed HPLC analysis.

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Correspondence to Nasheeman Ashraf.

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Bhat, Z.Y., Mir, J.A., Yadav, A.K. et al. CstMYB1R1, a REVEILLE-8-like transcription factor, regulates diurnal clock-specific anthocyanin biosynthesis and response to abiotic stress in Crocus sativus L. Plant Cell Rep 43, 20 (2024). https://doi.org/10.1007/s00299-023-03082-x

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