, Volume 220, Issue 3, pp 376–383

Expression of terpenoid indole alkaloid biosynthetic pathway genes corresponds to accumulation of related alkaloids in Catharanthus roseus (L.) G. Don

  • Ajaswrata Dutta
  • Jyoti Batra
  • Sashi Pandey-Rai
  • Digvijay Singh
  • Sushil Kumar
  • Jayanti Sen
Original Article


Madagascar periwinkle, Catharanthus roseus (L.) G. Don, a medicinally important plant, produces anticancer dimeric alkaloids, vinblastine and vincristine, in the leaves and accumulates antihypertensive alkaloids, ajmalicine and serpentine, in the roots. This plant grows wild in distant tropical and sub-tropical geographical locations with different agro-climates and shows wide variations in morphological and alkaloid yield-related traits. In order to understand the correlation between the expression of terpenoid indole alkaloid (TIA) pathway genes and accumulation of related alkaloids, six different genetic resources of C. roseus, including the medicinal cultivars Nirmal, Prabal, Dhawal, the mutants gsr-3 and gsr-6, and one horticultural variety, Pacifica blush, were studied. The expression profiles of one early and two late TIA biosynthetic pathway genes, namely, strictosidine synthase, desacetoxyvindoline 4-hydroxylase and deacetyl vindoline 4-O-acetyl transferase were analyzed in these plants. A positive correlation between transcript abundance and accumulation of related alkaloids was observed in the different genetic resources. The potential of these TIA biosynthetic pathway genes for use in screening of high-yielding C. roseus germplasm has been discussed.


Catharanthus Gene expression Genetic resource Morphological and biochemical variations Terpenoid indole alkaloid pathway Alkaloid contents 



Terpenoid indole alkaloid


Strictosidine synthase


Desacetoxyvindoline 4-hydroxylase


Deacetyl vindoline 4-O-acetyl transferase


Glycophytic salinity response


  1. Altschul SF, Madden TL, Schaffe AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedGoogle Scholar
  2. Arnon DJ (1949) Copper enzymes in isolated chloroplasts: polyphenol oxidase in Beta vulgaris. Plant Physiol 24:1–15Google Scholar
  3. Bates LS, Woldren RP, Teare ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–208Google Scholar
  4. Bradford MM (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  5. De Luca V, Laflamme P (2001) The expanding universe of alkaloid biosynthesis. Curr Opin Plant Biol 4:225–233CrossRefPubMedGoogle Scholar
  6. Dwivedi S, Singh M, Singh AP, Singh V, Uniyal GC, Khanuja SPS, Kumar S (2001) Registration of a new variety Prabal of Catharanthus roseus. JMAPS 23:104–106Google Scholar
  7. Endt DV, Kijne JW, Memelink J (2002) Transcription factors controlling plant secondary metabolism: what regulates the regulators? Phytochemistry 61:107–114CrossRefPubMedGoogle Scholar
  8. Fits LD, Memelink J (2000) ORCA3, a jasmonate-responsive transcriptional regulator of plant primary and secondary metabolism. Science 289:295–297CrossRefPubMedGoogle Scholar
  9. Gantet P, Memelink J (2002) Transcription factors: tools to engineer the production of pharmacologically active plant metabolites. Trends Pharm Sci 23:563–569CrossRefPubMedGoogle Scholar
  10. Grieve CM, Gratton SR (1983) Rapid assay for determination of water soluble quaternary ammonium compounds. Plant Soil 70:303–307Google Scholar
  11. Jordan MA, Thrower D, Wilsen L (1991) Mechanism of inhibition of cell proliferation by Vinca alkaloids. Cancer Res 51:2212–2222PubMedGoogle Scholar
  12. Kulkarni RN, Baskaran K, Chandrashekara RS, Kumar S (1999) Inheritance of morphological traits of periwinkle mutants with modified contents and yields of leaf and root alkaloids. Plant Breed 118:71–74CrossRefGoogle Scholar
  13. Kutchan TM (1995) Alkaloid biosynthesis—the basis for metabolic engineering of medicinal plants. Plant Cell 7:1059–1070CrossRefPubMedGoogle Scholar
  14. Levy A (1982) Natural and induced genetic variation in the biosynthesis of alkaloids and other secondary metabolites. In: Improvement of oil seed and industrial crops by induced mutations. IAEA, Vienna, pp 213–222Google Scholar
  15. Levy A, Milo J, Ashri A, Palevitch D (1983) Heterosis and correlation analysis of the vegetative components and ajmalicine content in the roots of the medicinal plant—Catharanthus roseus (L.) G. Don. Euphytica 32:557–564Google Scholar
  16. Memelink J, Verpoorte R, Kijne JW (2001) ORCAnization of jasmonate-responsive gene expression in alkaloid metabolism. Trends Plant Sci 6:212–219CrossRefPubMedGoogle Scholar
  17. Menke FLH, Parchmann S, Mueller MJ, Kijne JW, Memelink J (1999) Involvement of the octadecanoid pathway and protein phosphorylation in fungal elicitor-induced expression of terpenoid indole alkaloid biosynthetic genes in Catharanthus roseus. Plant Physiol 119:1289–1296CrossRefPubMedGoogle Scholar
  18. Mishra P, Uniyal GC, Sharma S, Kumar S (2001) Pattern of diversity for morphological and alkaloid yield related traits among the periwinkle Catharanthus roseus accessions collected from in and around Indian subcontinent. Genet Resour Crop Evol 48:273–286CrossRefGoogle Scholar
  19. Pandey-Rai S, Luthra R, Kumar S (2003) Salt-tolerant mutants in glycophytic salinity response (GSR) genes in Catharanthus roseus. Theor Appl Genet 106:221–230PubMedGoogle Scholar
  20. Pasquali G, Goddijn OJM, de Wall A, Verpoorte R, Schilperoort RA, Hoge JHC, Memelink J (1992) Coordinated regulation of two indole alkaloid biosynthetic genes from Catharanthus roseus by auxin and elicitors. Plant Mol Biol 18:1121–1131PubMedGoogle Scholar
  21. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467PubMedGoogle Scholar
  22. Singh DV, Maithy A, Verma RK, Gupta MM, Kumar S (2000) Simultaneous determination of Catharanthus alkaloids using reversed phase high performance liquid chromatography. J Liq Chromatogr 23:601–607CrossRefGoogle Scholar
  23. St-Pierre B, Laflamme P, Alarco AM, De Luca V (1998) The terminal O-acetyl transferase involved in vindoline biosynthesis defines a new class of proteins responsible for coenzyme A-dependent acyl transfer. Plant J 14:703–713CrossRefPubMedGoogle Scholar
  24. Tikhomiroff C, Jolicoeur M (2002) Screening of Catharanthus roseus secondary metabolites by high-performance liquid chromatography. J Chromatogr 955:87–93CrossRefGoogle Scholar
  25. Vazquez-Flota F, De Carolis E, Alarco A-M, De Luca V (1997) Molecular cloning and characterization of desacetoxyvindoline 4-hydroxylase, a 2-oxoglutarate dependent dioxygenase involved in the biosynthesis of vindoline in Catharanthus roseus (L.) G. Don. Plant Mol Biol 34:935–948CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Ajaswrata Dutta
    • 1
  • Jyoti Batra
    • 1
  • Sashi Pandey-Rai
    • 1
  • Digvijay Singh
    • 1
  • Sushil Kumar
    • 1
  • Jayanti Sen
    • 1
  1. 1.National Centre for Plant Genome ResearchNew DelhiIndia

Personalised recommendations