Cytogenetic and pollen identification of genus Gagnepainia (Zingiberaceae) in Thailand.

Gagnepainia godefroyi K. Schumann, 1904 and G. harmandii K. Schumann, 1904 belong to the genus Gagnepainia K. Schumann, 1904 of the Ginger family. They have the potential to be developed as medicinal and attractive ornamental plants. To date, the knowledge on the cytological and reproductive aspects of Gagnepainia have not been publicly available. Therefore, the aims of this research are to investigate the cytogenetic and pollen characters of Gagnepainia species using light, fluorescence, and scanning electron microscopes. The regular meiotic figures of 15 bivalents are found in both species and presented for the first time. These evidences indicate that Gagnepainia is diploid and contains 2n = 2x = 30 with basic number of x = 15. The mean nuclear DNA contents range from 1.986 pg in Gagnepainia sp., 2.090 pg in G. godefroyi to 2.195 pg in G. harmandii. Pollens of all species are monad, inaperturate, prolate with bilateral symmetry, and thick wall with fossulate exine sculpturing. The pollen size of G. harmandii (74.506 ± 5.075 µm, 56.082 ± 6.459 µm) is significantly larger than that of G. godefroyi (59.968 ± 3.484 µm, 45.439 ± 2.870 µm). Both 2C DNA content and pollen size are the effective characteristics for species discrimination. The reproductive evidence of high meiotic stability and normal pollen production indicate that both Gagnepainia species have high fertility and seed productivity, which are in accordance with the broad distribution. The present study provides good cytogenetic and pollen characters not only for plant identification, but also plant fertility assessment through plant genetic resource management and improvement of Gagnepainia.

Anti-HIV-1 activities of constituents from the rhizomes of Boesenbergia thorelii.

A new lignan, thoreliin A (1), and a new bisnorlignan, thoreliin B (2), were isolated from a MeOH extract of the rhizomes of Boesenbergia thorelii. In addition, the known bisnorlignans 3 and 4, neolignan 5, phenylpropanoids 6-15, as well as benzenoids 18-21 were also obtained from the same source. The structures were elucidated based on their spectroscopic data. By single crystal X-ray analysis, the relative stereochemistry of 1 was confirmed. All isolated compounds were evaluated for anti-HIV-1 activities. Among them, thoreliin A (1) exhibited anti-HIV-1 activities on both HIV-1 reverse transcriptase (41.43% inhibition at 200 µg/mL) and syncytium reduction assays (EC50 20.6 µM, SI 3.7), while compounds 3-6, 9 and 11-21 showed anti-HIV-1 activity only in the anti-syncytium assay (EC50 6.6-454.1 µM, SI >1.32-7.75).

Medicinal plants used with Thai Traditional Medicine in modern healthcare services: a case study in Kabchoeng Hospital, Surin Province, Thailand.

ETHNOPHARMACOLOGICAL RELEVANCE: Thai Traditional Medicine (TTM) is available in many modern hospitals in Thailand. However, there have been difficulties in integrating TTM, particularly the practices of the use of herbal medicines, into modern healthcare services. Kabchoeng Hospital is one hospital that has been able to overcome these difficulties. Thus, this study aimed to document the successful utilization of herbal medicine at Kabchoeng Hospital. The documentation focused on both the knowledge of medicinal plants and the success factors that facilitated the utilization of herbal medicine in the context of a modern hospital in Thailand. MATERIALS AND METHODS: Kabchoeng Hospital was intentionally selected for this case study. Participatory observation was used for the data collection. There were six groups of key informants: three applied Thai Traditional Medicine practitioners (ATTMPs), a pharmacist, two physicians, two folk healers, the head of an herbal cultivation and collection group, and 190 patients. The plant specimens were collected and identified based on the botanical literature and a comparison with authentic specimens; these identifications were assisted by microscopic and thin layer chromatography (TLC) techniques. RESULTS: Eighty-nine medicinal plants were used for the herbal preparations. The ATTMPs used these plants to prepare 29 standard herbal preparations and occasional extemporaneous preparations. Moreover, in this hospital, seven herbal preparations were purchased from herbal medicine manufacturers. In total, 36 preparations were used for 10 groups of symptoms, such as the treatment of respiratory system disorders, musculo-skeletal system disorders, and digestive system disorders. Four success factors that facilitated the utilization of herbal medicine at Kabchoeng Hospital were determined. These factors included a proper understanding of the uses of herbal medicines, the successful integration of the modern and TTM healthcare teams, the support of an herbal cultivation and collection group, and the acceptance of the local people. CONCLUSIONS: The practices that support the use of herbal medicine at Kabchoeng Hospital illustrated the successful application of TTM and also represented a model for the integration of TTM, and particularly the use of herbal medicine, into modern hospitals. This integration will be beneficial for sustainable healthcare systems in Thailand and in other countries where modern medicine is the mainstream medical system.

Micropropagation of a Thai medicinal plant for women's health, Curcuma comosa Roxb., via shoot and microrhizome inductions.

We studied the effects of explant types, plant growth regulators, and sucrose concentrations on shoot and microrhizome inductions of Curcuma comosa Roxb., an important Thai medicinal plant for women's health. Explant types significantly affected shoot induction of the plant. The maximum shoot multiplication rate of 11.82 ± 1.03 shoots/responding explant was obtained when culturing terminal bud explants on semi-solid Murashige and Skoog (MS) medium supplemented with 18.16 µM thidiazuron for 8 weeks. Subsequently, they were transferred to a semi-solid MS medium without plant growth regulators for 4 weeks. The regenerated shoots produced roots spontaneously. Rooted plantlets were successfully transferred to the soil. Microrhizome induction was significantly influenced by sucrose concentrations, but not by 6-benzyladenine (BA). Liquid MS medium with a combination of 17.76 µM BA and 50 g L(-1) sucrose was optimal for microrhizome induction of C. comosa. After 12 weeks of culture, the microrhizome induction rate was 3.36 ± 0.44 microrhizomes/responding explant. Starch accumulation in microrhizomes increased with higher sugar concentration and with longer duration of culture. The microrhizomes were allowed to germinate under greenhouse conditions and further developed into normal plants. The protocols established will be used for the production of uniform plantlets suitable for field plantation for the herbal industry.

In vitro propagation of Stemona hutanguriana W.Chuakul, an endangered medicinal plant.

In vitro propagation of Stemona hutanguriana W.Chuakul, an endangered species of Stemonaceae, was established. Nodal and internodal explants were cultured on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) alone or in combination with 1-naphthaleneacetic acid (NAA). When cultured on MS medium supplemented with 18.16 µM TDZ with 0.54 µM NAA for 8 weeks and then on MS medium without plant growth regulator (PGR) for 8 weeks, nodal explants demonstrated a responding frequency of 91.67 % and a shoot regeneration rate of 5.46 shoots/responding explant. Furthermore, internodal explants demonstrated a responding frequency of 17.17 % with 11.17 shoots/responding explant. The regenerated shoots were rooted using a two-step protocol by culturing for 4 weeks on MS medium supplemented with various concentrations of NAA and Indole-3-butyric acid (IBA) and 8 weeks on MS medium without PGR. When 5.40 µM NAA was used as a PGR, the maximum root induction rate was 69.45 %. Regenerated plantlets were successfully transferred to soil.

Chromosome number variation of phytoestrogen-producing Curcuma (Zingiberaceae) from Thailand.

Wan-chak-motluk is a Thai local name for native species of Curcuma (Zingiberaceae) that produce phytoestrogen in their rhizomes. Wan-chak-motluk has long been used in Thai traditional medicine for treatment of illness in the uterus and ovarian hormone deficit, and therefore it is cultivated for economic purposes throughout Thailand. The rhizome morphology of wan-chak-motluk is highly variable and this may be related to its medicinal properties. Thus, this study aimed to identify and classify wan-chak-motluk collected from cultivation sites in 16 provinces of Thailand. Chromosome numbers, together with inflorescent, floral, and leaf morphology, were used to separate wan-chak-motluk into five cultivars belonging to three species: C. comosa, C. elata, and C. latifolia. Curcuma comosa has short (2-5 cm) peduncle, glabrous lower leaf, and consists of two cultivars. One cultivar has a cylindrical spike 13-17 cm long and 5-8 cm wide, whereas the cylindrical spike of the other cultivar is shorter (10-15 cm) but larger (8-12 cm). The former cultivar has 2n = 42 and the latter has 2n = 63, seldom 2n = 62 or 64. Both C. elata and C. latifolia have long (10-25 cm) peduncle and pubescent lower leaf. They differ in that C. latifolia has 2n = 63 and 84 with a red path along the midrib, whereas C. elata has 2n = 63 without a red path along the midrib. This study shows that chromosome numbers can be used to accurately verify the taxonomic identification of wan-chak-motluk.

Genetic variation and species identification of Thai Boesenbergia (Zingiberaceae) analyzed by chloroplast DNA polymorphism.

Genetic variation and molecular phylogeny of 22 taxa representing 14 extant species and 3 unidentified taxa of Boesenbergia in Thailand and four outgroup species (Cornukaempferia aurantiflora, Hedychium biflorum, Kaempferia parviflora, and Scaphochlamys rubescens) were examined by sequencing of 3 chloroplast (cp) DNA regions (matK, psbA-trnH and petA-psbJ). Low interspecific genetic divergence (0.25-1.74%) were observed in these investigated taxa. The 50% majority-rule consensus tree constructed from combined chloroplast DNA sequences allocated Boesenbergia in this study into 3 different groups. Using psbA-1F/psbA-3R primers, an insertion of 491 bp was observed in B. petiolata. Restriction analysis of the amplicon (380-410 bp) from the remaining species with Rsa I further differentiated Boesenbergia to 2 groupings; I (B. basispicata, B. longiflora, B. longipes, B. plicata, B.pulcherrima, B. tenuispicata, B. thorelii, B. xiphostachya, Boesenbergia sp.1 and Boesenbergia sp.3; phylogenetic clade A) that possesses a Rsa I restriction site and II (B.curtisii, B. regalis, B. rotunda and Boesenbergia sp.2; phylogenetic clade B and B. siamensis; phylogenetic clade C) that lacks a restriction site of Rsa I. Single nucleotide polymorphism (SNP) and indels found can be unambiguously applied to authenticate specie-origin of all investigated samples and revealed that Boesenbergia sp.1, Boesenbergia sp.2 and B. pulcherrima (Mahidol University, Kanchanaburi), B. cf. pulcherrima1 (Prachuap Khiri Khan) and B. cf. pulcherrima2 (Thong Pha Phum, Kanchanaburi) are B. plicata, B. rotunda and B. pulcherrima, respectively. In addition, molecular data also suggested that Boesenbergia sp.3 should be further differentiated from B. longiflora and regarded as a newly unidentified Boesenbergia species.