Microbial Ecology

, Volume 78, Issue 4, pp 914–926 | Cite as

Functional Fungal Endophytes in Coleus forskohlii Regulate Labdane Diterpene Biosynthesis for Elevated Forskolin Accumulation in Roots

  • Anthati Mastan
  • RKB Bharadwaj
  • Ramesh Kumar Kushwaha
  • Chikkarasanahalli Shivegowda Vivek BabuEmail author
Plant Microbe Interactions


Coleus forskohlii is a perennial medicinal shrub cultivated mainly for its forskolin content. The plant has been used since ancient times in ayurvedic traditional medicines for the treatment of hypertension, glaucoma, asthma, congestive heart failures, obesity, and cancer. Use of endophytic microorganisms presents a special interest for the development of value-added bioactive compounds through agriculture. Limited investigations have been undertaken on in planta enhancement of forskolin content using endophytic fungus in sustainable agriculture. Here we report specific roles of three fungal endophytes, Fusarium redolens (RF1), Phialemoniopsis cornearis (SF1), and Macrophomina pseudophaseolina (SF2), functionally acting as plant probiotic fungus, regulating secondary metabolite (forskolin) biosynthesis in C. forskohlii. The root endophyte, RF1, and shoot endophytes, SF1 and SF2, were found to enhance forskolin content by 52 to 88% in pot and 60 to 84% in field experiments as compared to uninoculated control plants. The three endophytes also enhanced total biomass owing to plant growth promoting properties. The expression of diterpene synthases (CfTPSs) like CfTPS1, CfTPS2, CfTPS3, and CfTPS4 were significantly upregulated in endophyte-treated C. forskohlii plants. Elevated expression of key diterpene synthases (CfTPS2) in the forskolin biosynthesis pathway, exclusively present in the root cork of C. forskohlii, was observed following SF2 endophyte treatment. Furthermore, endophyte treatments conferred a variety of antagonistic activity against nematode galls (80%) and plant pathogens like Fusarium oxysporum, Colletotricum gloeosporioides, and Sclerotium rolfsii. RF1 and SF1 fungal endophytes showed positive for IAA production; however, SF1 also indicated phosphate solubilization activity. Overall, the qualitative and quantitative improvement of in planta forskolin enhancement represents an area of high commercial interest, and hence, our work focused on novel insights for the application of three fungal endophytes for in planta enhancement of forskolin content for C. forskohlii cultivation by a sustainable approach.


Fungal endophyte Forskolin Coleus forskohlii Agriculture Diterpene synthases 



The authors wish to thank the Director, CSIR- Central Institute of Medicinal and Aromatic Plants, Lucknow, India, for providing necessary facilities and encouragement during the course of investigation.

Funding Information

This work was supported by, Council of Scientific and Industrial Research (CSIR), India funded projects BSC 117 and BSC-203. Department of Science and Technology (DST –INSPIRE), Council of Scientific and Industrial Research (CSIR), and Indian Council of Medical Research (ICMR), New Delhi, India, provided financial support in the form of fellowships to A.M., R.K.B.B., and R.K.K., respectively.

Supplementary material

248_2019_1376_MOESM1_ESM.docx (2 mb)
ESM 1. Effect of endophytes RF1, SF1 and SF2 on C. forskohlii plants grown for 180 daysin pot condition and comparison with control plant. (DOCX 2084 kb)
248_2019_1376_MOESM2_ESM.docx (3.2 mb)
ESM 2. Effect of selected endophytes (RF1, SF1 and SF2) on plant growth response of C. forskohlii in field condition. The control plants bed (a), RF1 (b), SF1 (c), and SF2 (d). The photographic images (Beds) represent the field view of endophytes treated C. forskohlii plants (n = 16) at the time of harvesting. (DOCX 3290 kb)
248_2019_1376_MOESM3_ESM.docx (3.2 mb)
ESM 3. The three fungal endophytes isolated from Coleus forskohlii which could regulates biosynthesis of forskolin in roots. (DOCX 3318 kb)
248_2019_1376_MOESM4_ESM.docx (336 kb)
ESM 4. Analysis of forskolin content in fresh roots of endophyte treated and control plants under pot condition. Std: forskolin standard, M: mixture of forskolin types (If: isoforskolin, 7d: 7 deacetyl forskolin, FK: forskolin and 1,9 DF: 1,9 dideoxy forskolin), C : control, 1: RB1, 2: LB1, 3: RF1, 4: RF2, 5: RF3, 6: RF4, 7: SF1, 8: SF2, 9: SF3, 10: SF5 and 11: LF1. (DOCX 335 kb)
248_2019_1376_MOESM5_ESM.docx (637 kb)
ESM 5. Quantification of forskolin by HPLC method. (DOCX 636 kb)
248_2019_1376_MOESM6_ESM.docx (4.3 mb)
ESM 6. Antagonistic activity of endophytes [E1-Fusarium redolens (RF1), E2- Phialemoniopsis cornearis (SF1) and E3-Macrophomina pseudophaseolina (SF2)] against potent plant pathogens (P1- F. oxysporum, P2- S. rolfsiiand P3-C. gloeosporioides). (DOCX 4413 kb)
248_2019_1376_MOESM7_ESM.docx (15 kb)
ESM 7. List of primers used in this study. (DOCX 14 kb)
248_2019_1376_MOESM8_ESM.docx (14 kb)
ESM 8. Colonization of endophytes in C. forskohlii plants (DOCX 14 kb)
248_2019_1376_MOESM9_ESM.docx (14 kb)
ESM 9. NCIM accession numbers of fungal endophytes and their NCBI GenBank accession number(s) for ITS nucleotide sequence(s). (DOCX 14 kb)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Anthati Mastan
    • 1
    • 2
  • RKB Bharadwaj
    • 1
    • 2
  • Ramesh Kumar Kushwaha
    • 1
    • 2
  • Chikkarasanahalli Shivegowda Vivek Babu
    • 1
    • 2
    Email author
  1. 1.Microbial Technology LaboratoryCSIR- Central Institute of Medicinal and Aromatic Plants, Research CenterBangaloreIndia
  2. 2.Academy of Scientific and Innovative Research(AcSIR)GhaziabadIndia

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