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Comparative transcriptome mining for terpenoid biosynthetic pathway genes in wild and cultivated species of Plantago

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Abstract

Plantagos are important economical and medicinal plants that possess several bioactive secondary metabolites, such as phenolics, iridoids, triterpenes, and alkaloids. Triterpenoids are the ubiquitous and dynamic secondary metabolites that are deployed by plants for chemical interactions and protection under biotic/abiotic stress. Plantago ovata, a cultivated species, is the source of psyllium, while Plantago major, a wild species, has significant therapeutic potential. Wild species are considered more tolerant to stressful conditions in comparison to their cultivated allies. In view of this, the present study aimed to decipher the terpenoid biosynthetic pathway operative in P. ovata and P. major using a comparative transcriptomics approach. Majority of terpenoid biosynthetic genes were observed as upregulated in P. major including rate limiting genes of MVA (HMGR) and MEP (DXR) pathways and genes (α-AS, BAS, SM, and CYP716) involved in ursolic acid biosynthesis, an important triterpenoid prevalent in Plantago species. The HPLC output further confirmed the higher concentration of ursolic acid in P. major as compared to P. ovata leaf samples, respectively. In addition to terpenoid biosynthesis, KEGG annotation revealed the involvement of differentially expressed unigenes in several metabolic pathways, aminoacyl-tRNA biosynthesis, biosynthesis of antibiotics, and biosynthesis of secondary metabolites. MYB was found as the most abundant transcription factor family in Plantago transcriptome. We have been able to generate valuable information which can help in improving terpenoid production in Plantago. Additionally, the present study has laid a strong foundation for deciphering other important metabolic pathways in Plantago.

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Data availability

The Illumina sequence data have been submitted to NCBI sequence read archive (SRA) under accession number PRJNA687882.

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Acknowledgements

SG acknowledge “National Postdoctoral Fellowship” scheme of the Science & Engineering Research Board (SERB) for their financial support. The authors would like to acknowledge the Director CSIR-Indian Institute of Integrative Medicine for providing the research facilities and infrastructure. We thank Negenome Biosolutions Pvt. Ltd. for their help. The authors are thankful to Amit Kumar, CSIR IIIM, Jammu for facilitating HPLC analyses.

Funding

The work is supported by DST SERB National Postdoctoral Fellowship (NPDF) award number PDF/2017/001741.

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  1. Ravail Singh, Arti Sharma and Gulzar A. Rather contributed equally to this work.

Authors and Affiliations

  1. Plant Biotechnology Division, CSIR-Indian Institute of Integrative Medicine, Canal Road, Jammu Tawi, Jammu, 180001, India

    Suruchi Gupta, Ravail Singh, Arti Sharma, Gulzar A. Rather & Surrinder K. Lattoo

  2. Genome Research Laboratory, School of Biotechnology, University of Jammu, Jammu, 180006, India

    Manoj K. Dhar

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  1. Suruchi Gupta
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Supplementary Information

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709_2021_1663_MOESM1_ESM.docx

Supplementary file1: Sequence of primers used to amplify the genes of terpenoid and iridoid pathways using quantitative real-time PCR (qRT-PCR). (DOCX 13 KB)

709_2021_1663_MOESM2_ESM.xlsx

Supplementary file 2: Annotation of assembled transcripts satisfying the criteria of E-value (10-5) and minimum query coverage (40%) against Uniprot plant database (XLSX 11075 KB)

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Supplementary file 3: BLASTX hit of unigenes of P.major and P. ovata showing sequence homology with NCBI genomes (XLSX 20 KB)

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Supplementary file 4: Gene ontology annotation information of differentially expressed unigenes (biological process; sheet 1, cellular components; sheet 2, molecular functions; sheet 3) (XLSX 82 KB)

709_2021_1663_MOESM5_ESM.xlsx

Supplementary file 5: KEGG pathway analysis of 526 differentially expressed unigenes. Pathways are arranged in descending order of unigenes number. (XLSX 10 KB)

709_2021_1663_MOESM6_ESM.xlsx

Supplementary file 6: List of upregulated genes in P.major in comparison to P. ovata with their fold change value and general gene information retrieved from Uniprot database. (XLSX 641 KB)

709_2021_1663_MOESM7_ESM.xlsx

Supplementary file 7: List of downregulated genes in P.major in comparison to P. ovata with their fold change value and general gene information retrieved from Uniprot database. (XLSX 605 KB)

709_2021_1663_MOESM8_ESM.jpg

Supplementary Fig. 1: Heat map showing comparative expression analysis of genes related to terpenoid (20 genes) and iridoid (10 genes) pathways in two species of Plantago (P. ovata and P. major) based on FPKM values. Colors correspond with expression values; Dark pink indicates low expression, red indicates average expression, and light pink color indicates high expression levels. (JPG 44 KB)

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Gupta, S., Singh, R., Sharma, A. et al. Comparative transcriptome mining for terpenoid biosynthetic pathway genes in wild and cultivated species of Plantago. Protoplasma 259, 439–452 (2022). https://doi.org/10.1007/s00709-021-01663-9

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