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PREMISE OF THE STUDY: Ephedragerardiana (Ephedraceae), occurring in the Himalayan ranges, is an important plant species used in Tibetan medicine. Due to the lack of molecular markers to characterize genetic diversity, knowledge for conservation and uses of E. gerardiana resources is limited; we therefore developed microsatellite markers for use in this species.METHODS AND RESULTS: Using Illumina MiSeq sequencing technology, we developed 29 polymorphic microsatellite loci suitable for E. gerardiana, of which 15 loci also showed polymorphisms in two related Ephedra species, E. saxatilis and E. monosperma. The average number of effective alleles per locus ranged from two to six. The observed and expected heterozygosity ranged from 0.23 to 0.83 and 0.44 to 0.86, respectively, in E. gerardiana populations.
CONCLUSIONS: The developed 29 microsatellite markers are effective for the study of genetic structure and genetic diversity of E. gerardiana, and 15 of these markers are suitable for related Ephedra species.
Introduction - Radix Gentianae Macrophyllae, a traditional Chinese medicine, has been frequently used to dispel rheumatism and ease pain. There are four species of <i>Gentiana </i>(<i>G</i>. <i>macrophylla</i>, <i>G</i>. <i>straminea</i>, <i>G</i>. <i>dahurica</i> and <i>G</i>. <i>crassicaulis</i>) recorded as herbal drugs in the <i>Chinese Pharmacopoeia </i>and two other <i>Gentiana</i> species (<i>G</i>. <i>officinalis</i> and <i>G</i>. <i>siphonantha</i>) are often used as substitutes. Currently, the LC fingerprint comparison among different species and evidence for the equivalent application of these herbs are lacking.<br>Objective - To develop an HPLC method for the simultaneous determination of four iridoid and secoiridoid glycosides and a comparative study of six species of <i>Gentiana</i>.<br>Methodology - HPLC analysis was performed on a C<sub>18</sub> column (Phenomenex, 150 × 4.6 mm, 5 µm particle size) with gradient elution using 0.4% aqueous phosphoric acid and methanol at 242 nm.<br>Results - The proposed method was precise, accurate and sensitive enough for simultaneous quantitative evaluation of four iridoid and secoiridoid glycosides (loganic acid, swertiamarin, gentiopicroside and sweroside) in the six species of <i>Gentiana</i>. Contents of the four marker compounds varied from each other even among the samples from the same species and the LC chromatograms of the six species of <i>Gentiana</i> showed high similarities.<br>Conslusion - The close similarity of LC chromatograms and chemical composition of the four genuine <i>Gentiana</i> species explain their popular usage as Radix Gentianae Macrophyllae in Chinese medicine. By comparing the four genuine <i>Gentiana</i> species, it is suggested that the two substitutes could be used as Radix Gentianae Macrophyllae to relieve the scarcity of resources. Copyright © 2010 John Wiley & Sons, Ltd.<br>An HPLC method has been developed for simultaneous determination of four iridoid and secoiridoid glycosides and the fingerprints of the six <i>Gentiana</i> species was analyzed and compared. The close similarity of LC chromatograms and chemical composition of the four <i>Gentiana</i> species (<i>G. macrophylla</i>, <i>G. straminea</i>, <i>G. dahurica</i> and <i>G. crassicaulis</i>) explain their popular usage as Radix Gentianae Macrophyllae in Chinese medicine. By comparing the four genuine <i>Gentiana</i> species, it is suggested that <i>G. officinalis</i> and <i>G. siphonantha</i> could be used as Radix Gentianae Macrophyllae to relieve the scarcity of resources.