1.ITS2 Sequences as Barcodes for Identifying and Analyzing Pseudolaricis Cortex
Ting GAO ; Xunzhi ZHU ; Jingyuan SONG
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(3):387-392
Pseudolaricis Cortex and its adulterants were identified using DNA barcoding technique in this study. The sequences of ITS2 of Pseudolaricis Cortex and its five adulterants including 37 samples were analyzed. Se-quences were assembled using CodonCode Aligner 3.5.7. K2P distances were calculated and NJ tree was per-formed applying MEGA5.0. The ITS2 secondary structure was predicted using the ITS2 database and website. The results showed that the haplotypes of ITS2 regions of Pseudolaricis Cortex were the same as that of the original plant leaves. The inter-specific K2P distances of ITS2 were obviously higher than the intra-specific ones. Sam-ples of Pseudolaricis Cortex were clustered into a single clade in the NJ tree. The secondary structure of ITS2 of Pseudolaricis Cortex was significantly different from its adulterants. Therefore, ITS2 could powerfully discriminate Pseudolaricis Cortex and its adulterants.
2.Suspension culture of protocorm in Dendrobium candidum
Jingyuan SONG ; Shunxing GUO ; Peigen XIAO ;
Chinese Traditional and Herbal Drugs 1994;0(09):-
Object To study the feasibility of suspension culture of protocorm in Dendrobium candidum Wall ex Lindl and effect of inoculum and medium volume on the growth of protocorm Methods Effect of four basic media MS, 1/2 MS, 67 V, and B 5, inoculum and medium volume on the growth of protocorm were studied by completely random experimental design and orthogonal test design Results The growth of D candidum protocorms in liquid medium was markedly better than that in solid medium (P0 05), B 5 was much better than 1/2 MS (P
4.Characterization and Primer Design for Simple Sequence Repeat Loci in Salvia miltorrhiza Genome
Haibin XU ; Bo WANG ; Chao SUN ; Lifang XIE ; Jingyuan SONG
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(3):367-370
Salvia miltorrhiza Bge. is a perennial deciduous flowering plant. Its medicinal root and rhizomes part is widely used in the treatment of various diseases. In this study, bioinformatics analysis was performed to identify 4832 genome SSR loci with length longer than or equal to 40 bp from the draft genome assembly of S. miltorrhiza. The re-sults showed that the dinucleotide repeat motifs and trinucleotide repeat motifs constitute the main types of genome SSR loci, accounting for 37.3% and 61.3% respectively. SSR types enriched with A/T bases showed significantly higher abundance than other types, including AT/TA AAT/ATT, ATA/TAT, TAA/TTA, accounting for 30.5%, 21.6%, 17.1%, 20.4% of the total number of SSR loci, respectively. 1079 primer pairs were designed for these genome SSR loci. These primers can be used for genomic diversity analysis, genetic map construction, genetic marker screening. These data could lay the foundation for population genetics and genomics research of S. miltorrhiza.
5.Comparative studies on codon usage bias of Ganoderma lucidum based on analysis of genomic and transcriptomic data.
Xiaoxuan ZHU ; Yingjie ZHU ; Jingyuan SONG ; Chao SUN ; Shilin CHEN
Acta Pharmaceutica Sinica 2014;49(9):1340-5
Codon usage bias is an important characteristic of genetic information transfer in organisms. Analysis of codon usage bias of different species is important for understanding the rules on genetic information transfer. The previous method for analysis of codon usage bias is mainly based on genomic data. However, this method is greatly limited, because the genome sequences of higher organisms are still not available up to now. In this study, we found that we could obtain the same optimal codons of Ganoderma lucidum (Curtis: Fr.) P. Karst based on its whole genomic data or large-scale transcriptomic data from its liquid-cultured hyphae, primordium and fruiting body, separately. This result indicated the feasibility to understand the codon usage bias based on the large-scale transcriptomic data. By calculating the proportion of rare codons of Escherichia coli and Saccharomyces cerevisiae in 26 terpene synthases (TS) of G. lucidum, we found that the rare codons of S. cerevisiae have a higher proportion in TS genes, while the rare codons of E. coli have relatively lower, suggesting that the TS genes of G. lucidum are possibly more difficult to be expressed in S. cerevisiae than in E. coli. Chemical synthesis of TS genes according to the yeast optimal codons will be an effective way to solve the problem on the mismatch of gene codon bias between the foreign genes and the host strain.
6.RNA-Seq and genuine traditional Chinese medicine.
Xiaoyue WANG ; Jingyuan SONG ; Caixiang XIE ; Jianping HAN ; Shilin CHEN
Acta Pharmaceutica Sinica 2014;49(12):1650-7
RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.
7.Construction of the coexpression vector containing key element GLCYP450 involved in Ganoderma triterpene biosynthesis and its reductase gene GLNADPH.
Xu GUO ; Chao SUN ; Jingyuan SONG ; Hongmei LUO ; Shilin CHEN
Acta Pharmaceutica Sinica 2013;48(2):206-10
Cytochrome P450 (CYP450) is a key element in the Ganoderma triterpenoid biosynthetic pathway. The catalytic reaction process for CYP450 requires NADPH / NADH for electron transfer. After searching the genome dataset of Ganoderma lucidum, the unique sequence encoding CYP450 and NADPH were discovered, separately. The open reading frames of GLCYP450 and GLNADPH were cloned separately using RT-PCR strategy from G lucidum. The appropriate restriction enzyme cutting sites were introduced at the 5' and 3' ends of gene sequence. The genes of GLCYP450 and GLNADPH were recombined into the yeast expression vector pESC-URA, leading to the formation of the yeast expression plasmid pESC-GLNADPH-GLCYP450. This study provides a foundation for researching Ganoderma triterpene biosynthesis using the approach of synthetic biology.
8.Evaluation of polypoidal choroidal vasculopathy with optical coherence tomography angiography
Song XIA ; Jingyuan YANG ; Xinyu ZHAO ; Youxin CHEN
Chinese Journal of Experimental Ophthalmology 2021;39(1):54-58
Objective:To investigate the characteristics of polypoidal lesions and abnormal branching vascular network (BVN) in patients with polypoidal choroidal vasculopathy (PCV) by optical coherence tomography angiography (OCTA).Methods:A case series study was conducted.Untreated 45 eyes of 42 patients with initial diagnosis of PCV from May 2016 to April 2017 in Peking Union Medical College Hospital were enrolled.There were 26 males (27 eyes) and 16 females (18 eyes) with the average age of (68.59±8.52) years.All patients underwent fundus fluorescein angiography (FFA), indocyanine green angiography (ICGA) and OCTA.The OCTA image features and detection rate of polypoidal lesion and BVN were analyzed.The study protocol was approved by the Ethics Committee of Peking Union Medical College Hospital (No.S-K631). Written informed consent was obtained from each patient prior to any examination.Results:ICGA images showed obvious polyps with higper fluorescence in all PCV patients, and BVN structure was observed in the ICGA images of 38 eyes.Ring, circle, cluster or node like hyper-reflective structures were detected in polypoidal lesions in the en face OCTA images of 35 eyes, and the detection rate was 77.8% (35/45). The BVN structure was detected in the OCTA images of all the 38 eyes, with the 100% (38/38) detection rate.Conclusion:OCTA can detect polypoidal lesions and BVN of PCV non-invasively, which can locate the lesion in combination with en face image and B-scan image.
9.Contrucion of Salvia miltorrhiza Protein/Peptide Database for Mass Spectrometry Analysis
Haibin XU ; Jingyuan SONG ; Jun QIAN ; Bo WANG ; Lifang XIE ; Hongmei LUO
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(3):339-341
Salvia miltorrhiza Bge is a medicinal herb in traditional Chinese medicines. Until recently, dozens of compound ingredients with medicinal properties have been identified from S. miltorrhiza. Proteome analysis on medicinal ingredients synthesis mechanisms could provide a theoretical basis for S. miltorrhiza genetic improve-ment. In this study, a S. miltorrhiza specific protein/peptide sequence database was constructed using Illumina high-throughput transcriptome sequencing data of multiple types of S. miltorrhiza tissues. The database could act as a key component to carry out the proteome analysis in S. miltorrhiza.
10.Universality Assessment of matK Primer Pairs in Seed Plants
Linchun SHI ; Jinxin LIU ; Hui YAO ; Xiaolan XU ; Tianyi XIN ; Caixiang XIE ; Jingyuan SONG
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(3):381-386
matK is one of the core DNA barcode markers for plant DNA barcode identification and its universality using single makers has been in controversy. However, the universalities of different matK primer pairs in same seed plant group (order) and same matK primer pairs in different seed plant groups (order) are lack of systematic research. In this study, we collected 14563 full-length matK sequences of 11429 species of 3292 genera in 239 families belonging to 36 orders in seed plants. The universalities of 13 matK primer pairs and its 78 primer com-binations have been assessed using bioinformatics methods. The results indicated that xf/5r, 1F/8R, 390F/1326R and 3F_KIM/1R_KIM were the four most universality primer pairs. The four markers' universalities were 91.18%, 84.65%, 79.81% and 80.94% respectively in all 11429 seed plants. The most universality primer pairs in different orders were different. For each order, the primer pair with maximum universality was different. the xf/5r was the basal primer pair for primer combination and 1F/8R, 1F/1R, M3/M4 and 3F_KIM/1R_KIM could be the complementary primer pairs. This study could be a valuable resource for the primer selection of the research DNA barcoding identification in seed plants.