Russian Characteristic Medicinal Plant Resources Based on DNA Barcode Technology / 中国实验方剂学杂志

Objective::To investigate the distribution status of medicinal plants in the wild areas of Russian Caucasus and Altai, and clarify the types and efficacy information of medicinal plants in the area, so as to dig deep into new resources and new functions of medicinal plants in the countries along the Belt and Road. Method::Medicinal plants in the wild were searched and collected to make waxy specimens, and sent back to the country to extract the total DNA of the leaves of the leaves. Internal Transcribed Spacer(ITS)sequence universal primers were used for Polymerase Chain Reaction (PCR)amplification. The PCR products were sent for the two-way sequencing, and the sequencing results are spliced by software according to National Center for Biotechnology Information(NCBI). The same ITS sequence of the highest similarity species obtained by database BLAST was analyzed by DNAman software to identify the ITS sequence of the species and the ITS sequence of the same species. The MEGA 7 software was used as the phylogenetic tree, and the Kimura-2 parameter genetic distance was used to construct the neighbor joining(NJ) phylogenetic tree by the neighbor-joining method. The confidence of each branch of the development tree was tested by the bootstrap test method. A total of 2 000 cycles were performed, and the results were identified based on the clustering results. On this basis, the key medicinal plants in the Russian Caucasus and Altay wild areas were summarized and analyzed. Result::After BLAST alignment in NCBI database, the ITS sequences of each specimen were clustered with the login sequences on the NCBI database, which were separated from the outer group. The species classification of the specimens to be identified was determined by combining the characteristics of the specimens. A total of 51 plants were identified from the specimens collected in the field, covering 44 genera of 17 families, and 29 plants had clear efficacy records. The National Drug List of the Russian Federation and the Chinese Pharmacopoeia were retrieved to summarize commonly used medicinal plants in China and conclude that 20 kinds of Chinese and Russian common medicinal materials have different medicinal effects in local areas. This study has a reference significance for expanding the scope and clinical experience of traditional Chinese medicines, and provides a basis for strengthened local species conservation, development and utilization of wild medicinal plant resources.

Identification of pathogenic nonsporulating molds in eyes by internal transcribed spacer sequences alignment / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To identify the species and genus of pathogenic nonsporulating molds(NSM)by internal transcribed spacer(ITS)sequences alignment, and reveal the biodiversity of NSM in Hainan Island with tropical climate. <p>METHODS: Nine teen fungal strains, identified as NSM by conventional method in the laboratory department of Hainan Province Eye Hospital, were involved in this study. All of the strains were isolated from the infectious eye tissues from patients, and cultured with potato-dextrose agar and Sabourand's agar in 37℃ incubator for 7-21d. Any reproductive structure was detected by microscopy for up to 21d. After growth for 3-7d, the genomic DNA of specimens were extracted by grinding method combined with chemical method. Then, ITS sequences in the ribosome were amplified by PCR and analyzed using the National Center for Biological Information(NCBI)GenBank database. Finally, the species and genus were confirmed by sequence alignment. <p>RESULTS: The corresponding target bars could be observed in all 19 specimens after PCR. The results from genetic alignment classified the 19 specimens into 12 species, including Lasiodiplodia theobromae(6 isolates), Curvalaria lunata(1 isolate), Arthrinium sp.(2 isolates), neodeightonia subglabosa(2 isolates), Earliella scabrosa(1 isolate), Hypocreales sp.(1 isolate), phoma multirostrata(1 isolate), Trichophyton rubrum(1 isolate), Aspergillus westerdijkiae(1 isolate), roussoella siamensis(1 isolate), Ceriporia lacerata(1 isolate), Fusarium solqni(1 isolate).<p>CONCLUSION: ITS sequence alignment can identify NSM to genus and species level. The NSM in Hainan Island contains varies species, and is associated with multiple infectious diseases of the eye.

ITS sequence analysis and gene pattern recognition of Cichorium intybus from different habitats / 中草药

Objective: To analyze the ribosomal DNA internal transcribed spacer (rDNA-ITS) sequence of Cichorium intybus and C. glandulosum from different habitats, and to provide DNA molecular marker for the identification of chicory. Methods: The total DNA was extracted from the samples of C. intybus and C. glandulosum from 14 habitats by rapid broad spectrum of plant genomic DNA extraction kit. The ITS sequence was amplified by PCR with universal primer of ITS and then sequenced. The two kinds of ITS sequences were compared by DNAMAN V6 software. The cluster analysis was adopted by SPSS 17.0 after the different ITS bases from all the samples were mathematically treated. Results: The intraspecies identity of ITS sequence was above 99.2% in C. intybus, and that in C. glandulosum was above 99.8%, while the interspecies identity of ITS sequence was below 99.2%. There were various specific information sites in the ITS sequences of the two kinds of chicory. Conclusion: The ITS sequence is an available molecular marker for the identification of C. glandulosum and C. intybus.

Isolation and Identification of Mushroom Pathogens from Agrocybe aegerita

Agrocybe aegerita is an important mushroom cultivated in Korea, with good feel and a peculiar fragrance. A. aegerita can be cultivated throughout the year using culture bottles but is more susceptible to contamination than other mushrooms. Twenty-two pathogens were isolated from the fruiting bodies and compost of A. aegerita, and seven isolates were isolated from Pleurotus ostreatus to compare with the A. aegerita isolates, collected from Gimje, Iksan, Gunsan of Chonbuk, and Chilgok of Gyeongbuk Province in 2009. These isolates were identified based on morphological and molecular characteristics. Of the 29 isolates, 26 were identified as Trichoderma spp. and the remaining three were Aspergillus spp., Mucor spp., and Penicillium spp. A phylogenetic analysis revealed that the 26 isolates of Trichoderma were divided into four taxa, namely T. harzianum, T. pleuroticola, T. longibrachiatum, and T. atroviride. Among the Trichoderma spp., 16 isolates (55.2%) were identified as T. harzianum, six as T. pleuroticola (20.7%), two as T. longibrachiatum, and the remaining two were T. atroviride.