ABSTRACT
Objective: To analyze the genome survey of medicinal and edible plant Alpinia katsumadai and complete its genome genetic information. Methods: This study was based on high throughput sequencing platform Illumina, and K-mer analysis was applied to estimate the genome size and heterozygosity rate of A. katsumadai. Meanwhile, simple sequence repeat (SSR) loci that were suitable as markers were identified by MISA software. Results: The estimated genome size of A. katsumadai was 1.60 Gb, with a 0.44% heterozygosity rate and 72.72% repeats; In the genome sequence, 364 395 simple sequence repeats (SSRs) were detected by SSR molecular marker analysis, among which mono-nucleotide, di-nucleotide and tri-nucleotide repetitive motifs ranked the higher percentages of 64.25%, 24.05% and 10.31%, summed up to 98.61%; From the 350 bp library obtained by sequencing, 10 000 single-end reads were randomly selected and blasted with NT bank, the results showed that its genetically close species Alpinia zerumbet and Elettaria cardamomum were blasted with the reads of 12.89% and 12.36% in NT bank. Conclusion: The genome size, heterozygosity rate and SSR molecular marker analysis' genome survey study on A. katsumadai indicated that the genome of A. katsumadai species was a complex, highly repetitive and large genome, which provided genetic information support for the resource protection, genetic diversity analysis and variety breeding of A. katsumadai.
ABSTRACT
OBJECTIVE: To establish the high-performance thin layer chromatographic(HPTLC) fingerprints of volatile oil and flavonoids in Alpinia katsumadai Hayata so as to provide scientific information for its quality control, and determine the fingerprints similarity of with its related species. METHODS: The separation was performed on the pre-coated HPTLC GF254 silica gel plates. The volatile oil was developed with solvent system of toluene-ethyl acetate(9: 1). The relative humidity was 18%. The spots were visualized with 5% vanillin sulfuric acid solution. The flavonoides compounds were developed with solvent system of toluene-ethyl acetate-acetic acid (8: 1.5: 0.5). The spots were visualized with 5% aluminium chloride ethanol solution which needed to be observed at 365 nm. The common patterns of HPTLC fingerprints were obtained by CHROMAP 1.5 solution software, and authentication and quality assessment were performed by similarity and principle component analysis. RESULTS: The common patterns of the volatile oil and flavonoids consisted of 10 characteristic peaks and 7 characteristic peaks, respectively. Eucalyptol and alpinetin were identified by chemical reference substances. CONCLUSION: The qualities of Alpinia katsumadai Hayata collected from different areas are not distinctly different. Obvious difference exists in the chemical compositions of the volatile oil and flavonoids between Alpinia katsumadai Hayata and its counterfeit and other related species. This method is simple and rapid, which can serve as an effective identification and quality assessment method for Alpinia katsumadai Hayata.
ABSTRACT
Objective: The aim of the study was to establish a high speed counter-current chromatography (HSCCC) method for the isolation and purification of alpinetin and cardamomin from Alpinia katsumadai. Methods: Two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (5 : 5 : 7 : 3) was used. The flow rate of the mobile phase was 2.0 mL/min, the revolution speed was 800 r/min, the separation temperature was controlled at 25 °C, the reservation ratio of the stationary phase was 50%, and the detection wavelength was 300 nm. Results: Alpinetin (17.2 mg) and cardamomin (25.1 mg) could be obtained from 100 mg of the crude extract in one-step separation by the method. The purities of them were 98.1% and 99.2%, respectively, as determined by HPLC and their chemical structures were identified by 1H-NMR and 13C-NMR. Conclusion: The traditional method, column elution, could not eliminate irreversible adsorption, while the HSCCC method used for the isolation and purification of alpinetin and cardamomin from A. katsumadai has many advantages, such as facility, high efficiency, and high recovery as well.