Mining genes associated with furanocoumarin biosynthesis in an endangered medicinal plant, Glehnia littoralis

The endangered medicinal plant Glehnia littoralis is one of the important natural source of furanocoumarin, which has been used as mucolytic, antitussive, antitumour and antibacterial. However, the genetic information of furanocoumarin biosynthesis in G. littoralis is scarce at present. The objective of this study was to mine the putative candidate genes involved in the biosynthesis pathway of furanocoumarin and provide references for gene identification, and functional genomics of G. littoralis. We carried out the transcriptome analysis of leaves and roots in G. littoralis, which provided a dataset for gene mining. Psoralen, imperatorin and isoimperatorin were detected in G. littoralis by high performance liquid chromatography analysis. Candidate key genes were mined based on the annotations and local BLAST with homologous sequences using BioEdit software. The relative expression of genes was analysed using quantitative real-time polymerase chain reaction. Further, the CYP450 genes were mined using phylogenetic analyses using MEGA 6.0 software. A total of 156,949 unigenes were generated, of which 9021 were differentially-expressed between leaves and roots. A total of 82 unigenes encoding eight enzymes in furanocoumarin biosynthetic pathway were first obtained. Seven genes that encoded key enzymes in the downstream furanocoumarin biosynthetic pathway and expressed more in roots than leaves were screened. Twenty-six candidate CYP450 unigenes expressed abundantly in roots and were chiefly concentrated in CYP71, CYP85 and CYP72 clans. Finally, we filtered 102 differentially expressed transcription factors (TFs) unigenes. The transcriptome of G. littoralis was characterized which would help to elucidate the furanocoumarin biosynthetic pathway in G. littoralis and provide an invaluable resource for further study of furanocoumarin.

Blast injuries to carotid artery: development of a finite element model and biomechanical analysis / 中华创伤杂志

Objective To construct a three-dimensional finite element model to investigate the biomechanical mechanism of carotid blast injuries.Methods Based on the head and neck CT angiography data of a healthy male volunteer,the 3D geometric model was extracted by Mimics software.The 3D solid model was obtained by fitting the geometric model to the non-uniform rational B-splines (NURBS) by Geomagic Studio software.The mesh of blood vessels,blood and soft tissue was divided by HyperMesh software to obtain the three-dimensional finite element model of the carotid artery.The material parameters and boundary conditions were set,and the vessel wall rupture damage threshold was 1 MPa.The dynamic process of carotid injury caused by MK3A2 grenade explosion shock wave at the distance of 60,70 and 80 cm to the neck was simulated using the LS-DYNA,generating the shock waveform and peak overpressure.The stress cloud map was used to analyze the stress distribution and damage morphology,and the stress curve was used to analyze the mechanical changes.Results The peak values of shock wave overpressure were 0.45,0.63 and 0.96 MPa at the distance of 80,70 and 60 cm away from the explosion center,respectively.At 80 cm,the peak stress of vessel wall was 0.43 MPa,and the vessel wall was not ruptured;at 70 cm,the peak stress of anonyma was greater than 1 MPa,which resulted in small rupture;at 60 cm,the peak stress of both anonyma the ascending aorta were greater than 1 MPa,leading to obvious rupture.The root part of the common carotid artery,anonyma and the arch of the aorta were high stress concentration areas,manifested as high-prevalence areas of damage and rupture.Conclusions The finite element model of explosive carotid artery injury is successfully constructed,which can be used to analyze the mechanical response and damage mechanism of carotid blast injuries.The main cause of injury and rupture is that the sudden change of stress in the process of explosion shock reaches or exceeds the threshold of vascular wall injury.Carotid artery rupture will occur when the vessel wall stress peak is greater than 1 MPa at 60 and 70 cm away from the explosion center,providing references for the clinical treatment and injury prevention.

Cloning and Bioinformatics Analysis of GI4CL Gene in Glehnia littoralis / 世界科学技术-中医药现代化

This study was aimed to clone and analyze the open reading frame (ORF) of 4-coumarate:coenzyme A ligase (Gl4CL) gene in Glehnia littoralis.Based on the high-throughput sequencing of G.littoralis,the full-length cDNA of Gl4CL gene was cloned by the rapid amplification of cDNA ends (RACE) method.Physical and chemical properties,secondary structure and three-dimensional structure of Gl4CL protein were predicted.Real-time PCR was used to detect the expression of Gl4CL gene in roots and leaves of G.littoralis.A total of 1951 bp full-length cDNA of Gl4CL gene was obtained,which encoded a protein of 544 amino acids with a predicted molecular weight of 59.481 kDa and the isoelectric point of 8.20.The cDNA of Gl4CL gene included 1 635 bp of ORF,153 bp of 5'untranslated regions (5'UTR) and 163 bp of 3'UTR.The result of real-time PCR showed that Gl4CL gene was both expressed in roots and leaves of G.littoralis,while the expression of gene in roots was significantly higher than that in leaves.It was concluded that the study will lay the foundation for further study of Gl4CL gene in function and gene regulation.Through in-depth study of the relationship between the expression of Gl4CL gene and lignin,as well as the plant growth phenotypes,it is expected to obtain high yield and quality lines of Glehniae Radix with strong resistance to diseases and insect pests.

ITS2 Sequences as Barcodes for Identifying and Analyzing Pseudolaricis Cortex / 世界科学技术-中医药现代化

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.