Abstract
The genus Berchemia comprises important Chinese plants with considerable medicinal value; however, these plants are often misidentified in the herbal medicinal market. To differentiate the various morphotypes of Berchemia species, a proficient method employing the screening of universal DNA barcodes was used in this work. Three candidate barcoding loci, namely, psbA-trnH, rbcL, and the second internal transcribed spacer (ITS2), were used to identify an effective DNA barcode that can differentiate the various Berchemia species. Additionally, PCR amplification, efficient sequencing, intra- and inter-specific divergences, and DNA barcoding gaps were employed to assess the ability of each barcode to identify these diverse Berchemia plants authentically; the species were differentiated using the Kimura two-parameter and maximum composite likelihood methods. Sequence data analysis showed that the ITS2 region was the most suitable candidate barcode and exhibited the highest interspecific divergence among the three DNA-barcoding sequences. A clear differentiation was observed at the species level, in which a maximum distance of 0.264 was exhibited between dissimilar species. Clustal analysis also demonstrated that ITS2 clearly differentiated the test species in a more effective manner than that with the two other barcodes at both the hybrid and variety levels. Results indicate that DNA barcoding is ideal for species-level identification of Berchemia and provides a foundation for further identification at the molecular level of other Rhamnaceae medicinal plants.
Highlights
Berchemia is a genus of plants in the Rhamnaceae family, which comprises 32 deciduous woody plants located in Asia, South America, and Africa (Huxley and Griffiths 1999)
Amplification and sequence analysis Genomic DNA was extracted from 55 samples belonging to seven species of Berchemia
The PCR amplification size for ITS2, psbA-trnH, and rbcL ranged within 491–561, 364–470, and 729–757 bp, respectively
Summary
Berchemia is a genus of plants in the Rhamnaceae family, which comprises 32 deciduous woody plants located in Asia, South America, and Africa (Huxley and Griffiths 1999). In China, Berchemia consists of 19 native species (Chen and Dong 2006), which are primarily distributed in the south, southwestern, and eastern regions (Sinicae 1988). These species include climbing plants or small- to medium-sized trees, several of which are endangered but offer significant medicinal values; these important plants include B. lineatai (Shen et al 2010) and B. berchemiafolia (Kitamura and Murata 1984; Fu and Jin 1992; Ohwi 1984). Leioclada are closely related in terms of microstructure and microscopic characteristics These three species can be distinguished on the basis of the characteristics of their leaf edge; the leaf edge cells of B. lineata are round, and the cell walls are not thickened, or thickening is not obvious. The leaf edge cells of B. polyphylla var. leioclada are round, and the cell walls
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