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

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.

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.

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.

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.

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.

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.

Objective To make the identification of medicinal herbs in Salvia L. quickly and accurately. Methods In this work,DNA barcoding and chemical fingerprint were compared for the identification of herbs in Salvia L. First, the nucleotide sequences of the internal transcribed spacer region two amplified from 48 medicinal plants in Salvia L., and three other groups of medicinal plants in Lamiaceae were sequenced. A molecular phylogeny was constructed using the minimum evolution and maximum parsimony methods according to their sequence diversity. Second, the water-solution bioactive components and lipid soluble components were tested by HPLC. Then a chemical phylogeny was built using HPLC fingerprint data. Comparing the molecular and chemical phylogenetic trees revealed many similarities. Results DNA barcoding was sequencing based and could therefore provide more accurate results within a shorter time especially in large-scale studies. Conclusion The results show that ITS2 region is a novel DNA barcode for the authentication of the species in Salvia L. This is the first work to show the relationship between DNA barcoding and chemical components.

DNA barcoding is a technique in which species identification and discovery are performed by using short and standard fragments of DNA sequences. In this study, eleven species of Paris, including seven varieties, were sampled. Five chloroplast sequences, psbA-trnH, rpoB, rpoC1, rbcL, matK, and one nuclear marker, the second internal transcribed spacer (ITS2) of ribosomal DNA, were amplified and sequenced. The PCR amplification and sequencing efficiency, intra- and inter-specific divergence and barcoding gap were used to evaluate different loci, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The ITS2 sequences in the studied samples of Paris were amplified and sequenced successfully using primers designed by our group, while matK showed low level in the amplification and psbA-trnH was difficult for sequencing because of over 800 bp and poly (A) structure. Analysis of the intra- and inter-specific divergence and barcoding gap showed ITS2 was superior to other loci. The ITS2 showed a much higher percentage of success (100%) in identification than other five loci, none of which indicated more than 50% except matK (52.9%). The 2-locus combination of rbcL+matK didn't improve ability of authentication. In addition, the rate of successful identification with ITS2 kept 100% when the samples were expanded to 67 samples of 29 species. In conclusion, ITS2 can be used to correctly identify medicinal plants of Paris, and it will be a potential DNA barcode for identifying medicinal plants of other taxa.