ABSTRACT
Objective:To investigate the differences of chemical constituents of Salviae Yunnanensis Radix et Rhizoma and its closely related species,and to provide reference for the clinical application of Salviae Yunnanensis Radix et Rhizoma. Method:Fourier transform infrared (FTIR) spectroscopy combined with principal component analysis (PCA),cluster analysis (HCA) and partial least squares discriminant analysis (PLS-DA) were used to study the differences of components between Salviae Yunnanensis Radix et Rhizoma and its closely related species,and high performance liquid chromatography (HPLC) was used to compare the differences of water-soluble components between them. Result:There were some differences between Salviae Yunnanensis Radix et Rhizoma and its closely related species acrcording to FTIR spectroscopy and HPLC fingerprint,especially the water-soluble polar components were more abundant in Salviae Yunnanensis Radix et Rhizoma than other species. The chemical components of Salvia trijuga,S. przewalskii and S. bulleyana were more similar in terms of their genetic relationship. The result showed that the Salviae Yunnanensis Radix et Rhizoma and its closely related species can be clearly distinguished by FTIR combined with chemometrics method. Conclusion:Compared with its closely related species,Salviae Yunnanensis Radix et Rhizoma has a unique chemical composition,which has great therapeutic potential for blood stasis. The FTIR combined with chemometrics model can be used for rapid identification of Salviae Yunnanensis Radix et Rhizoma and its closely related species.
ABSTRACT
Objective To identify Cynanchum auriculatum and its closely related species using the ITS2 barcode. Methods A total of 54 samples of C. auriculatum and its related species were collected. Genomic DNAs were extracted from these samples. The ITS2 sequences of these samples were amplified and bidirectional sequenced by PCR. The obtained sequences were submitted to the Gen Bank and the ITS2 sequences of 47 samples belonging to 15 species were downloaded from the GenBank, and ITS2 sequences were annotated by ITS2 database. A total of 101 ITS2 sequences were aligned and the intraspecific and interspecific distances were calculated using the MEGA 5.0. Identification analyses were performed using the similarity search method and nearest distance method, and were presented intuitively by constructing neighbor-joining (NJ) tree. Results The length of all ITS2 sequences of C. auriculatum was 249 bp, which was a haplotype and was close to Cynanchum. There was a significant difference between the interspecific and intraspecific genetic distances of the ITS2 sequences. The NJ tree showed that C. auriculatum obviously differed from its closely related species, which showed high monophyly. According to the secondary structure of ITS2, it was also possible to distinguish between C. auriculatumi and Asclepiadaceae related species. Conclusion As a DNA barcode, ITS2 sequences can stably and accurately distinguish C. auriculatum from its closely related species and also provide a new technique to ensure the clinical safety in utilization of Chinese materia medica.
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Objective: The aim of present study was to identify Atractylodes lancea and its closely related species (Atractylodes chinesis and Atractylodes macrocephala) using the ITS2 barcode. Methods: The total genomic DNAs were extracted from twenty-nine samples of A. lancea and its closely related species from different habitats. The ITS2 sequences of these samples were amplified and bidirectional sequenced by PCR. The obtained sequences were submitted to the GenBank and the ITS2 sequences of 45 samples belonging to ten species were downloaded from the GenBank. Total 73 ITS2 sequences were aligned and the genetic distances were analyzed using the MEGA 5.1. Identification analyses were performed using the BLAST1 and the nearest distance methods, and were presented intuitively by constructing Neighbor-joining (NJ) tree. Results: The lengths of all ITS2 sequences of A. lancea were 229 bp presented as one haplotype pattern. There was significant divergence between the interspecific and intraspecific genetic distances of the ITS2 sequences. The NJ tree showed that A. lancea could differed obviously from its closely related species, which showed high monophyly. The secondary structures of ITS2 in the helical regions displayed clear differences in stem loop number, size, position, and screw angle among the medicinal plants of Compositae. Conclusion: As a DNA barcode, ITS2 sequences can stably and accurately distinguish A. lancea from its closely related species and also provide a new technique to ensure the clinical safety in utilization of Chinese materia medica.
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This study aimed to identify Inulae Herba, Inulae Flos and their closely related species using the ITS2 bar-code, and secure the quality and clinical curative effect of these medicinal materials. DNA was extracted from Inula linariifolia, I. japonica, I. britanica, which are the original species of Inulae Herba and Inulae Flos, together with their closely related species. The ITS2 sequence was amplified by PCR and sequenced bidirectionally. Sequence assembly and generation of consensus sequence were conducted by the CodonCode Aligner. The genetic distances were comput-ed using MEGA 5.0 in accordance with the Kimura-2-parameter (K2P) model, and the phylogenetic tree constructed by the neighbor-joining (NJ) method. The results showed that the ITS2 sequences of the various species have stable variable sites. The intraspecific genetic distances among Inulae Herba and Inulae Flos were obviously lower than the interspecific genetic distance among the above two medicinal materials and its adulterants. The NJ tree based on ITS2 sequences can clearly differ from Inulae Herba, Inulae Flos and their closely related species. It is concluded that ITS2 sequence can be used as DNA barcode to identify Inulae Herba, Inulae Flos and their closely related species.
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In this study, the psbA-trnH sequence was used as DNA barcoding to evaluate the accuracy and stability for identification of Bamboo shavings, Concretio Silicea Bambusae and their closely related species. 56 samples were collected and conducted the DNA extraction. Obtained sequences were assembled using the CodonCode Aligner V4.2, the genetic distances and NJ tree were computed and constructed using MEGA 5.0. The results shows that the maximum intra-specific K2P distances were less than the minimum inter-specific K2P distances. The NJ tree indicated that Bamboo shavings, Concretio Silicea Bambusae and their closely related species can be distinguished from each other clearly, except for Bambusa tuldoides Munro and Bambusa textilis, which perhaps attributed to their closely genetic relationship.
ABSTRACT
The spatial and temporal distribution of Sphoeroides greeleyi and Sphoeroides testudineus were established from collections (biological material and environmental data) conducted on a monthly basis from May 2000 to April 2001 in intertidal areas along the north-south axis of the estuarine complex of Paranaguá, Paraná State. In addition to characterizing a north-south spatial gradient, which fluctuates seasonally, the variation in the abiotic factors made possible the division of the estuary into three regions: north, central and south. Spatially, it was found that the number of individuals declines significantly for both species in the north-south direction of the estuary. Moreover, significant differences were found in the size of individuals across the estuarine regions. The largest S. greeleyi individuals were caught in the north, as well as the smallest S. testudineus individuals. The catches with the highest numbers of puffer fish occurred from late spring to early autumn, coinciding with the occurrence of specimens of smaller size and lower mean body mass. The results indicate that spatial and temporal variations in the environment impact the distribution patterns of both puffer fish species, suggesting that the co-occurrence of closely related species functions as a modulating factor in that distribution.
A distribuição espacial e temporal de Sphoeroides greeleyi e Sphoeroides testudineus foram estabelecidas a partir de coletas (material biológico e dados ambientais) realizadas mensalmente de maio/2000 a abril/2001 em áreas intertidais, no eixo norte-sul, do complexo estuarino de Paranaguá, Estado do Paraná. A variação dos fatores abióticos coletados, além de caracterizar um gradiente espacial, no sentido norte-sul, que varia sazonalmente, possibilitou a divisão do estuário em três regiões: norte, central e sul. Espacialmente, verificou-se para ambas as espécies que o número de indivíduos decresce, significativamente, no sentido norte-sul estuarino. Ainda, foram encontradas diferenças significativas no porte dos indivíduos entre as regiões do estuário, no norte ocorreram os maiores indivíduos de S. greeleyi e os menores de S. testudineus. As maiores capturas dos baiacus ocorreram do final da primavera ao início do outono, coincidindo com a ocorrência de exemplares de menor tamanho e de menor massa corporal média. Os resultados indicam que as variações espaciais e temporais do ambiente afetam os padrões de distribuição de ambas as espécies de baiacus, sugerindo que a co-ocorrência de espécies aparentadas age como um fator modulador nesta distribuição.