ABSTRACT
ObjectiveThe internal transcribed spacer （ITS） 2 region of ribosomal gene， a DNA barcode， was employed to identify 12 medicinal Aconitum species and the genetic relationship among the species was analyzed. MethodA total of 30 samples of the 12 species were collected. The DNA was extracted with spin column plant genomic DNA kit and the universal primers of ITS2 sequence were used for polymerase chain reaction （PCR） amplification， followed by electrophoresis detection and bi-directional sequencing. The yielded sequences were aligned and spliced by CodonCode Aligner 17.0 and sequence variation was analyzed by MEGA 7.0. The secondary structure was predicted by ITS2 Database and the neighbor-joining （NJ） method was applied to generate the phylogenetic tree. ResultThe ITS2 sequences of the 12 species were 220-221 bp， with the average guanine and cytosine （GC） content of 64.09%， 140 variable sites， 137 informative sites， and 81 conservative sites. The intraspecific genetic distance （K2P） was smaller than the interspecific genetic distance. According to the secondary structures of ITS2 sequences and NJ cluster analysis， A. scaposum， A. sinomontanum， and A. barbatum had close genetic relationship， while the rest nine showed close kinship， particularly A. soongaricum and A. yinschanicum. ConclusionITS2 sequence is of great value for the molecular identification and genetic relationship determination of Aconitum， which provides a new method for the study of ethnomedicine.
ABSTRACT
The identification of species primordium has been one of the hot issues in the identification of traditional Chinese medicine. Sea snake is one of the most valuable Chinese medicinal materials in China. In order to understand the origin and varieties of sea snake in the market, we studied the molecular identification of 46 sea snakes by cytochrome B(Cytb). After comparison and manual correction, the sequence length was 582 bp, and the content of A+T(58.9%) was higher than that of G+C(41.1%). There exist 197 variable sites and 179 parsimony-informative sites of the sequence. There are 44 kinds of sequence alignment with consistency equal to 100%, and 2 kinds equal to 96%. A total of 408 Cytb effective sequences were downloaded from GenBank database, with a total of 68 species. Phylogenetic tree of a total of 454 sea snake sequences with the samples in this study were constructed by neighbor-joining trees and Bayesian inference method, respectively, which can identify 42 samples of medicinal materials, while 4 samples can not be identified because of their low node support. The results showed that the species of the sea snake medicine were at least from 2 genera and 5 species, namely, Aipysurus eydouxii, Hydrophis curtus, H. caerulescen, H. curtus, H. ornatus and H. spiralis. This study suggested that the original species of commercial sea snake are very complex and can provide insight into the identification of sea snakes.
Subject(s)
Animals , Bayes Theorem , China , Cytochromes b/genetics , Elapidae , Medicine, Chinese Traditional , PhylogenyABSTRACT
Objective: To investigate the genetic polymorphisms of 24 autosomal short tandem repeats (STR) loci in Gelao and Miao populations dwelled in Guizhou province, and explore the population genetic relationships and evaluate their application value on forensic medicine. Methods: The DNA samples of 732 unrelated individuals (399 Guizhou Gelao population and 333 Guizhou Miao population) were amplified using SureID® PanGlobal kit, and the PCR products were analyzed by electrophoresis through 3500XL genetic analyzer. The fragment sizes of alleles were subsequently analyzed by GeneMapper ID-X v1.5. Allele frequencies and forensic genetic parameters of 24 STR loci were statistically analyzed and compared with the available data of other populations from different races and regions. Results: For Guizhou Gelao and Miao populations, the individual discrimination power (DP) ranged from 0.7833 to 0.9909 and 0.8010 to 0.9909, respectively; the polymorphic information content (PIC) ranged from 0.5608 to 0.9385 and 0.5677 to 0.9414, respectively; the total discrimination power (TDP) were 1-7.6036 × 10-30 and 1-6.8630 × 10-30, respectively, and the cumulative probability of exclusion (CPE) were 1-1.9608 × 10-11 and 1-1.9738 × 10-14, respectively. Analysis with the matrix of Nei's DA genetic distance indicated that the genetic distance was the smallest (0.0205) between Guizhou Gelao and Hubei Han populations, while was the largest (0.0449) between Guizhou Gelao and Yunnan Miao populations; the genetic distance was minimum (0.0033) between Guizhou Miao and Hunan Han populations, while was maximal (0.0363) between Guizhou Miao and Yunnan Miao. Conclusions: The 24 STR loci are abundant in genetic polymorphism in Guizhou Gelao and Miao populations. It is of great significance to study the genetic diversity of different ethnic groups in order to understand their origin, migration and interrelationship.
ABSTRACT
We identified Lacrymaria velutina of the Coprinaceae in Korea. The unusually large and sturdy fruiting body, fibrillose to fibrillose-scaly cap and stalk without a volva with an obscure superior hairy ring zone or hairy annulus, and blackish brown, warted spores distinguished this species from closely related Psathyrella species. An illustrated account of the microscopic traits is presented. Fruiting bodies with obtusely hemispherical caps, 2.5~6 cm, becoming convex with age; surface dry, densely fibrillose-scaly with split margin; stipe, 4.5~6 cm, equal, hollow, fibrillose, dry, whitish above the superior ring zone, light brown below; crowded gills, adnexed, dark black at maturity. Pileipellis typically cellular with the gill edge appearing white and beaded. Blackish brown basidiospores that discolor in concentrated sulfuric acid. Spores elliptical, warted, 9~11 x 6~8 microm, with prominent snout-like germpores. Cheilocystidia abundant, 57~68 x 19~25 microm, and narrowly elongated clavate, often clustered in threes or fours. Pleurocystidia rarely present, 45~47.5 x 12~13 microm, and clavate to utriform. This trait distinguishes our sample as L. velutina from other Psathyrella spp. of the Coprinaceae, which have smooth spores. This taxon was clarified by the observation that Psathyrella spores fade in concentrated sulfuric acid. A molecular phylogenetic study revealed that our specimen was Lacrymria velutipes, which is closely related to Lacrymaria lacrymabunda. Moreover, those two species are clearly distinguishable from other Psathyrella species, which agreed with the morphologically distinctive traits described above. We believe that this is the first report of this taxon, which has not been described in Korea.