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Cloning and expression analysis of chalcone synthase gene from Coleus forskohlii

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Abstract

Flavonoids are an important class of secondary metabolites that play various roles in plants such as mediating defense, floral pigmentation and plant–microbe interaction. Flavonoids are also known to possess antioxidant and antimicrobial activities. Coleus forskohlii (Willd.) Briq. (Lamiaceae) is an important medicinal herb with a diverse metabolic profile, including production of a flavonoid, genkwanin. However, components of the flavonoid pathway have not yet been studied in this plant. Chalcone synthase (CHS) catalyses the first committed step of flavonoid biosynthetic pathway. Full-length cDNA, showing homology with plant CHS gene was isolated from leaves of C. forskohlii and named CfCHS (GenBank accession no. KF643243). Theoretical translation of CfCHS nucleotide sequence shows that it encodes a protein of 391 amino acids with a molecular weight of 42.75 kDa and pI 6.57. Expression analysis of CfCHS in different tissues and elicitor treatments showed that methyl jasmonate (MeJA) strongly induced its expression. Total flavonoids content and antioxidant activity of C. forskohlii also got enhanced in response to MeJA, which correlated with increased CfCHS expression. Induction of CfCHS by MeJA suggest its involvement in production of flavonoids, providing protection from microbes during herbivory or mechanical wounding. Further, our in silico predictions and experimental data suggested that CfCHS may be posttranscriptionally regulated by miR34.

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Acknowledgements

PA and VM are supported by CSIR-Senior Research Fellowship. SGG acknowledges the financial support for this work from CSIR 12th FYP project ‘PMSI’ (BSC0117) of Council of Scientific and Industrial Research (CSIR) and Fast-Track ‘OYS’ project of SERB (DST, New Delhi).

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    1. Indian Institute of Integrative Medicine (CSIR-IIIM), Council of Scientific and Industrial Research, Canal Road, Jammu, 180 001, India

      PRAVEEN AWASTHI, VIDUSHI MAHAJAN, VIJAY LAKSHMI JAMWAL, NITIKA KAPOOR, YASHBIR S. BEDI & SUMIT G. GANDHI

    2. Academy of Scientific and Innovative Research, Anusandhan Bhawan, 2 Rafi Marg, New Delhi, 110 001, India

      VIDUSHI MAHAJAN, YASHBIR S. BEDI & SUMIT G. GANDHI

    3. Shri Mata Vaishno Devi University, Katra, 182 301, India

      PRAVEEN AWASTHI & SHAFAQ RASOOL

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    Awasthi P., Mahajan V., Jamwal V. L., Kapoor N., Rasool S., Bedi Y. S. and Gandhi S. G. 2016 Cloning and expression analysis of chalcone synthase gene from Coleus forskohlii. J. Genet. 95, xx–xx

    Praveen Awasthi and Vidushi Mahajan contributed equally to this work. PA did gene expression profiling using real time PCR, quantification of flavonoid content and measurement of antioxidant activity. VM carried out cloning work and wrote the manuscript. VLJ and NK did the analysis of gene structure and splice variants. YSB and SR provided critical inputs for the study as well as during preparation of manuscript. SGG designed the study, analysed the results and edited the manuscript and figures.

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    AWASTHI, P., MAHAJAN, V., JAMWAL, V.L. et al. Cloning and expression analysis of chalcone synthase gene from Coleus forskohlii . J Genet 95, 647–657 (2016). https://doi.org/10.1007/s12041-016-0680-8

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