Phylogenetic analysis and development of molecular markers for five medicinal Alpinia species based on complete plastome sequences.

BACKGROUND: Alpinia species are widely used as medicinal herbs. To understand the taxonomic classification and plastome evolution of the medicinal Alpinia species and correctly identify medicinal products derived from Alpinia species, we systematically analyzed the plastome sequences from five Alpinia species. Four of the Alpinia species: Alpinia galanga (L.) Willd., Alpinia hainanensis K.Schum., Alpinia officinarum Hance, and Alpinia oxyphylla Miq., are listed in the Chinese pharmacopeia. The other one, Alpinia nigra (Gaertn.) Burtt, is well known for its medicinal values. RESULTS: The four Alpinia species: A. galanga, A. nigra, A. officinarum, and A. oxyphylla, were sequenced using the Next-generation sequencing technology. The plastomes were assembled using Novoplasty and annotated using CPGAVAS2. The sizes of the four plastomes range from 160,590 bp for A. galanga to 164,294 bp for A. nigra, and display a conserved quadripartite structure. Each of the plastomes encodes a total of 111 unique genes, including 79 protein-coding, 28 tRNA, and four rRNA genes. In addition, 293-296 SSRs were detected in the four plastomes, of which the majority are mononucleotides Adenine/Thymine and are found in the noncoding regions. The long repeat analysis shows all types of repeats are contained in the plastomes, of which palindromic repeats occur most frequently. The comparative genomic analyses revealed that the pair of the inverted repeats were less divergent than the single-copy region. Analysis of sequence divergence on protein-coding genes showed that two genes (accD and ycf1) had undergone positive selection. Phylogenetic analysis based on coding sequence of 77 shared plastome genes resolves the molecular phylogeny of 20 species from Zingiberaceae. In particular, molecular phylogeny of four sequenced Alpinia species (A. galanga, A. nigra, A. officinarum, and A. oxyphylla) based on the plastome and nuclear sequences showed congruency. Furthermore, a comparison of the four newly sequenced Alpinia plastomes and one previously reported Alpinia plastomes (accession number: NC_048461) reveals 59 highly divergent intergenic spacer regions. We developed and validated two molecular markers Alpp and Alpr, based on two regions: petN-psbM and psaJ-rpl33, respectively. The discrimination success rate was 100 % in validation experiments. CONCLUSIONS: The results from this study will be invaluable for ensuring the effective and safe uses of Alpinia medicinal products and for the exploration of novel Alpinia species to improve human health.

Screening of analgesic and anti-inflammatory active component in Fructus Alpiniae zerumbet based on spectrum-effect relationship and GC-MS.

Fructus Alpiniae zerumbet is widely used in Guizhou province as a miao folk herb with anti-inflammatory, analgesic, protection against cardiovascular diseases, antihypertension and antioxidant activities. To further investigate the chemical material basis, the spectrum-effect relationship was established using gray relational analysis between the chromatographic fingerprint and its bioactivities. Herein, the fingerprints of essential oils from Fructus Alpiniae zerumbet (EOFAZ) from various sources were determined by gas chromatography mass spectrometry, and the analgesic and anti-inflammatory bioactivities were investigated using the mouse model of acetic acid-induced writhing test and dimethylbenzene-induced mouse ear edema test. Finally, 17 common peaks were identified from nine batches of A. zerumbet, by comparison with the standard mass spectra in Nist2005, Wiley275 library. Meanwhile, the results showed significant analgesic and anti-inflammatory effects in all of the different sources of EOFAZ. In particularly, peak 1 (α-pipene), peak 3 (ß-pinene), peak 9 (camphor) and peak 16 (α-cadinol) might be the main bioactive ingredients for analgesic and anti-inflammatory activities. The model of the spectrum-effect relationships of EOFAZ was successfully discovered, which provided a novel platform for finding the bioactive components, a theoretical foundation for its further study and helping to establish quality control of Fructus A. zerumbet.

Anti-Oxidant, Anti-Aging, and Anti-Melanogenic Properties of the Essential Oils from Two Varieties of Alpinia zerumbet.

Here, we investigated the anti-oxidant and anti-aging effects of essential oils (EOs) from the leaves of Alpinia zerumbet (tairin and shima) in vitro and anti-melanogenic effects in B16F10 melanoma cells. The anti-oxidant activities were performed with 2,2-diphenyl-1-picrylhydrazyl (DPPH); 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS); nitric oxide; singlet oxygen; hydroxyl radical scavenging; and xanthine oxidase. The inhibitory activities against collagenase, elastase, hyaluronidase, and tyrosinase were employed for anti-aging. The anti-melanogenic was assessed in B16F10 melanoma cells by melanin synthesis and intracellular tyrosinase inhibitory activity. The volatile chemical composition of the essential oil was analyzed with gas chromatography-mass spectrometry (GC/MS). The EO was a complex mixture mainly consisting of monoterpenes and sesquiterpenes. The results revealed that tairin and shima EOs showed strong anti-oxidant activities against DPPH and nitric oxide, hydroxyl radical scavenging activity, and xanthine oxidase inhibition. Compared to shima EO; tairin EO exhibited strong anti-aging activity by inhibiting collagenase, tyrosinase, hyaluronidase, and elastase (IC50 = 11 ± 0.1; 25 ± 1.2; 83 ± 1.6; and 213 ± 2 µg/mL, respectively). Both EOs inhibited intracellular tyrosinase activity; thus, reducing melanin synthesis. These results suggest that tairin EO has better anti-oxidant/anti-aging activity than shima EO, but both are equally anti-melanogenic.

[Microscopic identification of dai medicine alpiniae kwangsiensis rhizoma and its confused species].

OBJECTIVE: To observe microscopic characteristics of rhizome of Alpinia kwangsiensis, Alpinia platychilus, Alpinia blepharocalyx, and to provide basis for their identification. METHODS: Microscopic identification of root transverse section by paraffin tissue section and free-hand section and powder were carried out to distinguish them. RESULTS: The microscopic identification can be made by cell structure of epidermis and endodermis, the existence of nonglandular hair, the lignification degree of vascular bundle fiber, the number and existence style of vascular bundle fiber, the number and size of tube, and so on. The powder identification can be made mainly according to the existence of nonglandular hair and spiral vessel, the shape of starch grain, and so on. CONCLUSION: The microscopic characteristics of rhizome can provide basis for the identification of three Alpinia species.

[AFLP analysis of genetic diversity of Alpinia officinarum].

OBJECTIVE: To explore the genetic diversity and relationship of different Alpinia officinarum germplasm. METHOD: Amplified fragment length polymorphism (AFLP) markers were developed to analyze genetic polymorphism in A. officinarun from eight resources. The amplified fragments were used as primary matrix with NTSYSpc-2.11F software to analyze the similarity between the A. officinarum germplasm and to construct the genetic phylogenetic tree. RESULT: A total of 1,120 fragments were genotyped using AFLP with eight prime combinations. Analysis identified 1,044 polymorphic fragments, accounting for 92.57% of the total detected variation. Genetic phylogenetic tree analysis indicates that three categories can be divided among the eight resources of A. officinanrum. CONCLUSION: Significant polymorphism and genetic diversity can be observed among A. officinarum germplasm resources.

Chemical diversity in the genus Alpinia (Zingiberaceae): comparative composition of four Alpinia species grown in Northern India.

The essential-oil compositions of leaves, flowers, and rhizomes of Alpinia galanga (L.) Willd., Alpinia calcarata Rosc., Alpinia speciosa K. Schum., and Alpinia allughas Rosc. were examined and compared by capillary GC and GC/MS. Monoterpenoids were the major oil constituents identified. 1,8-Cineole, alpha-terpineol, (E)-methyl cinnamate, camphor, terpinen-4-ol, and alpha- and beta-pinenes were the major constituents commonly distributed in leaf and flower essential oils. The presence of endo-fenchyl acetate, exo-fenchyl acetate, and endo-fenchol was the unique feature of rhizome essential oils of A. galanga, A. calcarata, and A. speciosa. On contrary, the rhizome oil of A. allughas was dominated by beta-pinene. Significant qualitative and quantitative variations were observed in essential-oil compositions of different parts of Alpinia species growing in subtemperate and subtropical regions of Northern India. Cluster analysis was performed to find similarities and differences in essential-oil compositions based on representative molecular skeletons. Monoterpenoids, viz., 1,8-cineole, terpinen-4-ol, camphor, pinenes, (E)-methyl cinnamate, and fenchyl derivatives, were used as chemotaxonomic markers.

[Vectorial angle method for studying on GC fingerprint of naphtha in Alpinia officinarum Hance].

OBJECTIVE: To establish GC quality analysis and GC fingerprint spectrum of naphtha in Alpinia officinarum Hance. METHODS: The naphtha in Alpinia officinarum Hance was extracted and analyzed by GC to establish the fingerprint spectrum. The results were analyzed by similarity grade calculate method to compare the fingerprint difference of Alpinia officinarum Hance. RESULTS: The GC fingerprint spectrum of Alpinia officinarum Hance were established. It consisted of 11 peaks. The GC spectrum results were analyzed by similarity grade calculate method which can divide Alpinia officinarum Hance into various habitats. CONCLUSION: The fingerprint spectrum can be used to distinguish Alpinia officinarum Hance and to evaluate its quality.

[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.