OBJECTIVETo study genetic diversity and genetic relationships among Lonicera macranthoides cultivars.

METHODFive cultivars were estimated by ISSR and SRAP. The data of amplified bands were analyzed by Treeconw software. The system diagram of genetic relationship was built by UPGMA.

RESULTTwenty ISSR primers amplified 186 bands with 103 (54.63%) polymorphic bands and 58 SRAP primer combinations amplified 591 bands with 347(55.46%) polymorphic bands. Genetic distance ranges were 0.058 4-0.230 8 (by ISSRs) and 0.1071-0.2611 (by SRAPs). Both ISSR and SRAP analyses revealed a middle level of genetic diversity in L. macranthoides cultivars. The dendrograms based on SRAP and ISSR markers were not all the same.

CONCLUSIONThe genetic diversity of L. macranthoides cultivars is middle. ISSR and SRAP markers can be effectively applied to genetic analysis in L. macranthoides cultivars.

Genetic Variation ; Lonicera ; genetics ; Polymorphism, Genetic ; Software

Twenty-three histone methyltransferase genes were obtained from transcriptome dataset of Lonicera japonica. The nucleotide and proteins characteristics, subcellular localization, senior structural domains and conservative forecasting were analyzed. The result of phylogenetic tree showed that 23 histone methyltransferases were mainly divided into two groups: lysine methyltransferase and arginine methyltransferases. The result of gene expression showed that 23 histone methyltransferases showed preference in terms of interspecies and organs. They were more expressed in buds of L. japonica than in L. japonica var. chinensis and lower in leaves of L. japonica than in L. japonica var. chinensis. Eight genes were specific expressed in flower. These results provided basis for further understanding the function of histone methyltransferase and epigenetic regulation of active ingredients of L. japonica.
Computational Biology ; Gene Expression ; Histone-Lysine N-Methyltransferase ; genetics ; Lonicera ; enzymology ; genetics ; Phylogeny

In order to study the regulation mechanism of secondary metabolites biosynthesis in Lonicera macranthoides, the key genes involved in the regulation of biosynthesis and the mechanism of differential metabolites were explored. In this study, high-throughput sequencing technology was used for transcriptome sequencing of L. macranthoides at different development stages. By using Liquid chromatography-tandem mass spectrometry(LC-MS/MS) technology, the laws of qualitative, quantitative and synthetic accumulation of its metabolites were studied, and the key enzyme genes for the biosynthesis of phenolic acid and flavonoids were screened out according to the differentially expressed genes. A total of 111 differentially accumulate metabolites(DAM) and 6 653 differentially expressed genes(DGE) were obtained by metabonomics and transcriptomics analysis. The metabolites and key enzyme genes in the Erqing(KE) were significantly different from those in the Dabai(KD) and Yinhua(KY) stages. In the phenylalanine biosynthesis pathway, the ion abundance of chlorogenic acid, naringin, quercetin, rutin, coniferol and other metabolites decreased with the development of flowers, while the ion abundance of ferulic acid, coumarin and syringoside increased with the development of flowers. Key enzyme genes such as CHS, HCT, CCR, FLS and COMT positively regulate the downstream metabolites, while PAL, C4H and 4CL negatively regulate the downstream metabolites. This study provides candidate genes and theoretical basis for the further exploration of key enzymes in the biosynthesis of secondary metabolites and for the regulation of the accumulation of secondary metabolites in L. macranthoides by molecular biological methods.
Chromatography, Liquid ; Flowers/genetics* ; Lonicera/genetics* ; Metabolomics ; Proteomics ; Tandem Mass Spectrometry

OBJECTIVEThis study aimed to analyze the genetic diversity and genetic relationship of germplasm resources of Lonicera japonica in main producing areas of China and provide reference for developing new varieties of L. japonica.

METHODUsing 6 primer combinations, 13 germplasm of L. japonica were analyzed by AFLP marker. The genetic distance was worked out by using DPS V3.01 software, and the cluster was conducted based on UPGMA.

RESULTA total of 435 bands were obtained including 191 polymorphic ones. The average polymorphic frequency was 43.9%. Cluster analysis showed that the relationship of cultivated variety from the same genuine area was near, and the classification result based on AFLP marker of germplasm of L. japonica from Shandong province was basically consistent with those on their morphological character.

CONCLUSIONAFLP marker can indicate the abundant genetic diversity of L. japonica and provide theoretical evidence for reasonable utilization and breeding new cultivar of L. japonica in molecular level.

Amplified Fragment Length Polymorphism Analysis ; China ; Genetic Variation ; Lonicera ; classification ; genetics ; Phylogeny ; Polymorphism, Restriction Fragment Length

Sixty-three morphological traits from 743 specimens of the 41 taxa within the cultivated Lonicera japonica were observed and measured, such as the height of plants, the length of leaf, the width of leaf, the length of anther, the alabastrum's number of one branch, the color of alabastrum and so on. A numerical taxonomy is presented by using the cluster analysis, principal components analysis (PCA) and factor analysis. Sixteen of 63 characters were screened by means of PCA and used for cluster analysis of 41 taxa with the method of Ward linkage and average euclidean distance. The cluster analysis showed that the 41 taxa could be divided into 5 groups when the Euclidean distance coefficient was 11.84. The factor analysis indicated that the shape of leaf, color of alabastrum, the pilosity and color of twiggery were of significance for the cultivated L. japonica classification. The results of this study will be a base for the core collection and breeding of L. japonica.
Breeding ; China ; Flowers ; chemistry ; classification ; genetics ; Lonicera ; chemistry ; classification ; genetics ; growth & development ; Plant Leaves ; chemistry ; classification ; genetics ; Quantitative Trait Loci

The DNA demethylase genes are widespread in plants. Four DNA demethylase genes (LJDME1, LJDME2, LJDME3 and LJDME4) were obtained from transcriptome dataset of Lonicera japonica Thunb by using bioinformatics methods and the proteins' physicochemical properties they encoded were predicted. The phylogenetic tree showed that the four DNA demethylase genes and Arabidopsis thaliana DME had a close relationship. The result of gene expression model showed that four DNA demethylase genes were different between species. The expression levels of LJDME1 and LJDME2 were even more higher in Lonicera japonica var. chinensis than those in L. japonica. LJDME] and LJDME2 maybe regulate the active compounds of L. japonica. This study aims to lay a foundation for further understanding of the function of DNA demethylase genes in L. japonica.
Computational Biology ; DNA, Plant ; chemistry ; Genes, Plant ; Lonicera ; enzymology ; genetics ; Oxidoreductases, O-Demethylating ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Transcriptome

This study analysed the tissue specific expression of critical genes involved in chlorogenic acid and luteolin biosynthesis, for exploiting the molecular mechanism of components biosynthesis in Lonicera confusa. Expression of PAL, 4CL, C4H, CHS, CHI, FNS and HQT gene families of chlorogenic acid and luteolin biosynthesis-related genes in buds and leaves of L. confusa were analyed by Real-time PCR. Expressions of PAL1, C4H1, 4CL1, CHS1, CHI3 and HQT2 in buds were lower than that in leaves, and expressions of PAL3, 4CL2, CHI2 and FNS2 in buds were higher than that in leaves. The results indicated that that PAL3 and 4CL2 may be associated with accumulation of chlorogenic acid, and the expression patterns of PAL1, CHS1, CHI3 and HQT2 in buds and leaves of L. confusa were different with L. japonica. This study provided some theoretical basis for the further research on genetic mechanism of active components differences in L. confusa and L. japonica.
Biosynthetic Pathways ; Chlorogenic Acid ; metabolism ; Gene Expression Regulation, Plant ; Lonicera ; genetics ; metabolism ; Luteolin ; biosynthesis ; Multigene Family ; Plant Proteins ; genetics ; metabolism

To simply and rapid authenticate Lonicera japanica. Rapid allele-specific PCR primer was designed base on trnL-trnF 625 G/T Single nucleotide polymorphism and the PCR reaction systems including annealing temperature was optimized; optimized results were performed to authenticate L. japanica and its 9 adulterants. When 100 x SYBR Green I was added in the PCR product of 87 degrees C initial denatured 1 min; 87 degrees C denatured 5 s, 68 degrees C annealing 5 s, 30 cycle; L. japanica visualize strong green fluorescence under 365 nm UV lamp whereas adulterants without. The results indicate rapid allele-specific PCR could authenticate L. japanica and its adulterants rapidly and simply.
Alleles ; DNA Primers ; genetics ; Drug Contamination ; prevention & control ; Lonicera ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Quality Control

OBJECTIVETo provide theoretical evidence for dividing and breeding cultivars of Lonicera japonica, the botanical character of laminas leaves observed and compared.

METHODUsing the morphological method, the main character of leaves of 11 farm cultivars of L. japonica were systematically observed and the data were comparatively studied with statistical means. It included the length, width and the ratio of the length to the width of blade, density, length and thickness of the nonglandular hair in the epidermis.

RESULTThe results showed that there were obvious differences in length, width and the ratio of the length to the width of blade, density and length of the nonglandular hair in the epidermis.

CONCLUSIONOn the basis of the differences of morphological character of leaves, the various farm cultivars could be identified.

Breeding ; Drugs, Chinese Herbal ; Lonicera ; anatomy & histology ; classification ; genetics ; growth & development ; Plant Leaves ; anatomy & histology ; genetics ; growth & development

OBJECTIVETo explore the new method of discriminating Gelsemium elegans from Lonicera japonica and its close species by using specific PCR amplification.

METHODThirteen samples of the different G. elegans materials and 58 samples of L. japonica, L. macranthoides and L. dasystyla were collected. The total DNA of the samples were extracted, and the DNA of G. elegans, L. japonica and L. macranthoides water extracts were extracted. PsbA-rnnH sequence from G. elegans was amplified by PCR and sequenced unidirectionally, ClustulW was used to align psbA-trnH sequences of the G. elegans and L. japonica and its close species from GenBank database.

RESULTAll samples were amplified by PCR with specific primer, DNA from G. elegans would be amplified 97 bp whereas PCR products from all of Lonicera samples had not bands.

CONCLUSIONSpecific PCR amplification can be used to identify G. elegans from L. japonica and its close species successfully and is an efficient molecular marker for authentication of G. elegans and L. japonica and its close species.

DNA, Plant ; genetics ; isolation & purification ; Drugs, Chinese Herbal ; analysis ; Gelsemium ; chemistry ; genetics ; Lonicera ; chemistry ; genetics ; Phylogeny ; Plant Extracts ; chemistry ; genetics ; Polymerase Chain Reaction ; Water ; chemistry