Comparison of metabolites and variety authentication of Amomum tsao-ko and Amomum paratsao-ko using GC-MS and NIR spectroscopy.

Amomum tsao-ko, as an edible and medicinal variety, has been cultivated for more than 600 years in China. Recently, two cultivars, A. tsao-ko and Amomum paratsao-ko, were found in A. tsao-ko planting area. The two cultivars are often confused because of the similar phenotype and difficult to distinguish through sensory judgment. In this study, the non-targeted gas chromatography-mass spectrometry (GC-MS) metabolomics combined with near-infrared spectroscopy (NIRS) were used for dissecting the two cultivars with phenotypic differences. According to principal component analysis (PCA) loading diagram and orthogonal partial least squares discriminant analysis (OPLS-DA) S-plot of the metabolites, the accumulation of major components including 1,8-cineole, α-phellandrene, (E)-2-decenal, (-)-ß-pinene, (E)-2-octenal, 1-octanal, D-limonene, and decanal, were present differences between the two cultivars. Seven metabolites potential differentiated biomarkers as ß-selinene, decamethylcyclopentasiloxane, (E,Z)-2,6-dodecadienal, (E)-2-hexenal, (E)-2-decenal, isogeranial, 1,8-cineole and ß-cubebene were determined. Although A. tsao-ko and A. paratsao-ko belong to the same genera and are similar in plant and fruit morphology, the composition and content of the main components were exposed significant discrepancy, so it is necessary to distinguish them. In this study, the discriminant model established by GC-MS or NIRS combined with multivariate analysis has achieved a good classification effect. NIRS has the advantages of simple, fast and nondestructive and can be used for rapid identification of varieties and fruit tissues.

Chemotaxonomic Monitoring of Genetically Authenticated Amomi Fructus Using High-Performance Liquid Chromatography-Diode Array Detector with Chemometric Analysis.

Amomi Fructus is widely used to treat digestive disorders, and Amomum villosum, A. villosum var. xanthioides, and A. longiligulare are permitted medicinally in national pharmacopeias. However, there are a variety of adulterants present in herbal markets owing to their morphological similarities to the genuine Amomum species. Forty-two Amomi Fructus samples from various origins were identified using internal transcribed spacer and chloroplast barcoding analyses, and then their chromatographic profiles were compared using chemometric analysis for chemotaxonomic monitoring. Among the Amomi Fructus samples, A. villosum, A. longiligulare, A. ghaticum, and A. microcarpum were confirmed as single Amomum species, whereas a mixture of either these Amomum species or with another Amomum species was observed in 15 samples. Chemotaxonomic monitoring results demonstrated that two medicinal Amomum samples, A. villosum and A. longiligulare, were not clearly distinguished from each other, but were apparently separated from other non-medicinal Amomum adulterants. A. ghaticum and A. microcarpum samples were also chemically different from other samples and formed their own species groups. Amomum species mixtures showed diverse variations of chemical correlations according to constituent Amomum species. Genetic authentication-based chemotaxonomic monitoring methods are helpful in classifying Amomi Fructus samples by their original species and to distinguish genuine Amomum species from the adulterants.

Comparison and Phylogenetic Analysis of Chloroplast Genomes of Three Medicinal and Edible Amomum Species.

Amomum villosum is an important medicinal and edible plant with several pharmacologically active volatile oils. However, identifying A. villosum from A. villosum var. xanthioides and A. longiligulare which exhibit similar morphological characteristics to A. villosum, is difficult. The main goal of this study, therefore, is to mine genetic resources and improve molecular methods that could be used to distinguish these species. A total of eight complete chloroplasts (cp) genomes of these Amomum species which were collected from the main producing areas in China were determined to be 163,608-164,069 bp in size. All genomes displayed a typical quadripartite structure with a pair of inverted repeat (IR) regions (29,820-29,959 bp) that separated a large single copy (LSC) region (88,680-88,857 bp) from a small single copy (SSC) region (15,288-15,369 bp). Each genome encodes 113 different genes with 79 protein-coding genes, 30 tRNA genes, and four rRNA genes. More than 150 SSRs were identified in the entire cp genomes of these three species. The Sanger sequencing results based on 32 Amomum samples indicated that five highly divergent regions screened from cp genomes could not be used to distinguish Amomum species. Phylogenetic analysis showed that the cp genomes could not only accurately identify Amomum species, but also provide a solid foundation for the establishment of phylogenetic relationships of Amomum species. The availability of cp genome resources and the comparative analysis is beneficial for species authentication and phylogenetic analysis in Amomum.

[Macroscopic and microscopic identification of Amomum tsao-ko and Amomum paratsao-ko fruits].

OBJECTIVE: To compare macroscopic and microscopic characteristics of the fruits of Amomum tsao-ko and Amomum paratsao-ko. METHODS: Transverse section and powder of pericarp and seeds of these two species were identified. RESULTS: The fruits of Amomum tsao-ko and Amomum paratsao-ko had similar microscopic characteristics. They could be distinguished from each other by the peri-carp color, furrows and ribs, and the shape of seeds. CONCLUSION: Above characteristics can be used as basis of pharmacognostical study of Amomum tsao-ko and Amomum paratsao-ko.

[Studied on the identification of Fructus amomi from its confusable varieties by Fourier transform infrared spectroscopy].

This paper uses Fourier transform infrared spectrometer with OMNI sampler to distinguish Fructus amomi from their confusable varieties, i. e. Amomum aurantiacum H. T. Tsai et S. W. Zhao, Amomum chinense Chun ex T. L. Wu, Alpinia chinensis (Ketz.) Rosc and Alpinia japonica (Thunb.) Miq. IRs of Amomum villosum Lour., Amomum longiligulare T. L. Wu and Amomum villosum Lour. Var xanthioides T. L. Wu et Senjen are resemble, and they are markedly different from the FTIR of the confusable varieties. Repeat experiments were processed with different samples of the same set, and the probability is 1.000. The result shows that FTIR can be directly used to distinguish Fructus amomi from their confusable varieties.

[Analysis of Amomun villosum species and some adulterants of zingiberaceae by RAPD].

Different Amomum villosum Lour. species and some their adulterants of Zingiberaceae were researched using RAPD. The PCR indicates favourable differentiation of the reaction. The Amomum villosum Lour. species have the similar DNA fingerprints while it is obviously differ from adulterants. There are 12.17 percent of primers which appear polymorphism. We can identify different species according to these dissimilarity. We also construct the tree of the molecular evolution through NJ software, the hereditary distance of the graph shows different relationships between Amomum villosum Lour. species and their adulterants. The result created by software is analogous the traditional methods.

[Studies on quality standard of Fructus Amomi].

OBJECTIVE: To study the quality standard of different parts and species of Fructus Amomi from different producing. METHOD: GC method, ash determination and water determination. RESULT: The contents of bornyl acetate in different parts and species of Fructus Amomi from different producing areas were obviously different, but the traditional curative effects remained unchanged. The standards for the total ash content, acid-insoluble ash content and water content in Fructus Amomi have been set up. CONCLUSION: Bornyl acetate with a definite content limit should be chosen as the index in studying the quality standard of Fructus Amomi.