Abstract
Interest in the medicinal properties of secondary metabolites of Boesenbergia rotunda (fingerroot ginger) has led to investigations into tissue culture of this plant. In this study, we profiled its primary and secondary metabolites, as well as hormones of embryogenic and non-embryogenic (dry and watery) callus and shoot base, Ultra Performance Liquid Chromatography-Mass Spectrometry together with histological characterization. Metabolite profiling showed relatively higher levels of glutamine, arginine and lysine in embryogenic callus than in dry and watery calli, while shoot base tissue showed an intermediate level of primary metabolites. For the five secondary metabolites analyzed (ie. panduratin, pinocembrin, pinostrobin, cardamonin and alpinetin), shoot base had the highest concentrations, followed by watery, dry and embryogenic calli. Furthermore, intracellular auxin levels were found to decrease from dry to watery calli, followed by shoot base and finally embryogenic calli. Our morphological observations showed the presence of fibrils on the cell surface of embryogenic callus while diphenylboric acid 2-aminoethylester staining indicated the presence of flavonoids in both dry and embryogenic calli. Periodic acid-Schiff staining showed that shoot base and dry and embryogenic calli contained starch reserves while none were found in watery callus. This study identified several primary metabolites that could be used as markers of embryogenic cells in B. rotunda, while secondary metabolite analysis indicated that biosynthesis pathways of these important metabolites may not be active in callus and embryogenic tissue.
Highlights
Boesenbergia rotunda is a member of the Zingiberaceae family
We have determined the relative abundance of fifty-one targeted primary metabolites in shoot base and three callus types of B. rotunda using Ultra Performance Liquid Chromatography– Mass Spectrometry (UPLC-MS) (Table 1)
The high organic acid levels in shoot base (SB) tissue may be closely related to its role in energy production in the Glycine (Gly) Homoserine Glutamine (Gln) Histidine (His) S-adenosyl methionine Spermine Arginine (Arg) Alanine (Ala) Asparagine (Asn) Aspartic acid (Asp) Glutamic acid (Glu) Serine Proline (Pro) Phenylalanine (Phe) Valine (Val) Tyrosine (Tyr) Trptophan (Trp) Hydroxyproline Lysine (Lys) Methionine (Met) Antranilate Adenine Creatine Glycerol-3-phosphate Fructose-6-phosphate Fructose-1,6-phosphate Gluconic acid Erythrose-4-phosphate Xylulose-5-phosphate Ribulose-5-phosphate 6-phosphogluconic acid Putresine GABA Citrulline Ornithine (Orn) Guanine Uracil Thymine Hypoxanthine Ribose-5-phosphate Shikimic acid Shikimate-3-phosphate Malic acid 2-Oxoisovaleric acid cis-Aconitic acid Citric acid
Summary
Boesenbergia rotunda is a member of the Zingiberaceae family. This monocotyledon plant is commonly called fingerroot, Chinese keys or Chinese ginger and is used in food, flavourings and traditional medicines [1]. Several flavonoids and chalcone derivatives have been isolated from extracts of B. rotunda, including pinocembrin, pinostrobin, alpinetin, panduratin, cardamonin, quercetin and kaempferol [2]. These compounds are reported to have various biological effects. Jing et al compared its anti-proliferative effect against five cancer cell lines using extracts from rhizomes, leaves and stems from different Boesenbergia species [8]. They found that extracts from the rhizome of B. rotunda gave the most promising results in cytotoxic activity for all five cancer cell lines
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