Callus of East Indian sandalwood co-cultured with fungus Colletotrichum gloeosporioides accumulates santalenes and bisabolene
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The inducible accumulation of desired products via in vitro cultures provides an experimental system for researching secondary metabolism in woody plants. This system is convenient, because environmental conditions can be strictly controlled. This is particularly important for East Indian sandalwood (Santalum album L.), a tree with desired sandal oil products that are restricted to the heartwood. In this study, we established a method to induce and proliferate callus from sandalwood shoot explants. Thidiazuron (TDZ) (0.2–1.5 mg/l) could induce the formation of callus, which proliferated rapidly within a month following three successive subcultures in liquid shake culture on Murashige and Skoog (MS) basal medium supplemented with 0.8–1.0-mg/l TDZ. Callus cultured in this liquid medium for 7 days was co-cultured with fungi, either Colletotrichum gloeosporioides or Penidiella kurandae. Gas chromatography–mass spectrometry (GC–MS) analysis of the solvent extract by chemical anhydrous diethyl ether of callus co-cultured with C. gloeosporioides showed the presence of santalenes and bisabolene, which are the precursors of santalol. However, another fungus, P. kurandae, could not induce santalenes or bisabolene. This study provides an opportunity to further studies on the santalene and bisabolene biosynthetic signaling pathway and the fungal endophyte–plant interaction in sandalwood.
KeywordsCallus induction Fungus Liquid shake culture Sandalwood Santalol Santalum album Secondary metabolites Thidiazuron
This work was financially supported by the National Natural Science Foundation of China (Grant numbers 31470685, 31270720, and 31100498), the Natural Science Foundation of Guangdong Province (S2012010009025), and a Guangdong Science and Technology project (2015B020231008).
Compliance with ethical standards
Conflict of interest
The authors declare no conflicts of interest.
- Celedon JM, Chiang A, Yuen M, Diaz-Chavez ML, Madilao LL, Finnegan PM, Barbour EL, Bohlmann J (2016) Heartwood-specific transcriptome and metabolite signatures of tropical sandalwood (Santalum album) reveal the final step of (Z)-santalol fragrance biosynthesis. Plant J 86:289–299CrossRefGoogle Scholar
- Choi D, Bostock RM, Avdiushko S, Hildebrand DF (1994) Lipid-derived signals that discriminate wound-and pathogen-responsive isoprenoid pathways in plants: methyl jasmonate and the fungal elicitor arachidonic acid induce different 3-hydroxy-3-methylglutaryl-coenzyme A reductase genes and antimicrobial isoprenoids in Solanum tuberosum L. Proc Nat Acad Sci USA 91:2329–2333CrossRefGoogle Scholar
- Jones CG, Moniodis J, Zulak KG, Scaffidi A, Plummer JA, Ghisalberti EL, Barbour E, Bohlmann J (2011) Sandalwood fragrance biosynthesis involves sesquiterpene synthases of both the terpene synthase TPS-a and TPS-b subfamilies, including santalene synthases. J Biol Chem 286:17445–17454CrossRefGoogle Scholar
- Kuramae-Izioka EE (1997) A rapid, easy and high yield protocol for total genomic DNA isolation from Colletotrichum gloeosporioides and Fusarium oxysporum for RAPD. Rev Unimar 19:683–689Google Scholar
- Misra BB, Dey S (2015) Biological activities of East Indian sandalwood tree, Santalum album. PeerJ 1. https://doi.org/10.7287/peerj.preprints.96v1
- Misra BB, Dey S (2016) Culture of East Indian sandalwood tree somatic embryos in air-lift bioreactors for production of santalols, phenolics and arabinogalactan proteins. AoB Plants S5:plt025Google Scholar
- Peeris M, Senarath W (2015) In vitro propagation of Santalum album L.. Eur J Biochem 213:743–748Google Scholar
- Phillips MA, Walter MH, Ralph SG, Dabrowska P, Luck K, Urós EM, Boland W, Strack D, Rodríguez-Concepción M, Bohlmann J, Gershenzon J (2007) Functional identification and differential expression of 1-deoxy-d-xylulose 5-phosphate synthase in induced terpenoid resin formation of Norway spruce (Picea abies). Plant Mol Biol 65:243–257CrossRefGoogle Scholar
- Valluri J (2009) Bioreactor production of secondary metabolites from cell cultures of periwinkle and sandalwood. In: Jain SM, Saxena PK (eds) Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants. Methods in molecular biology (methods and protocols), vol 547. Humana Press, Totowa, pp 325–335Google Scholar
- White TJ, Bruns T, Lee S, Taylor JW (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, Inc., New York, pp 315–322Google Scholar
- Zhang YY, Yan HF, Niu MY, Cheng QW, Zhang XH, Teixeira da Silva JA, Ma GH (2018) Multiple strategies for increasing yields of essential oil and obtaining sandalwood terpenoids by biotechnological methods in sandalwood. Trees 32:21–29Google Scholar