OBJECTIVEThe aim of this study was to optimize the testing methods for seed quality, and to provide a basis for establishing seed testing rules and quality grading standard of Glycyrrhiza uralensis.

METHODReferring to the International Seed Testing Rules made by ISTA and Rules for agricultural seed testing (GB/T 3543-1995) issued by China, the seed quality of G. uralensis from different collection areas was measured.

RESULT AND CONCLUSIONThe seed testing methods for quality items of G. uralensis, including sampling, purity analysis, verification of genuineness, weight of 1 000 seeds, percentage germination, moisture content and seed viability of G. uralensis had been initially established.

OBJECTIVETo study the growth of Glycyrrhiza uralensis under different planting density for providing theoretic evidence for reasonable planting density.

METHODThe field experiment was designed by single factor randomized block.

RESULTThe plant height, root diameter, main root length and branches of main stem were all decreased with increasing planting density of G. uralensis. Meanwhile, dry matter accumulation on root, leaf, aerial part and rhizome were all decreased with increasing planting density.

CONCLUSIONTwo hundred and seventy thousand plants/hm2 was feasible planting density.

The present study investigated the chemical constituents from the aerial parts of Glycyrrhiza uralensis. The ethanol extract of the aerial parts of G. uralensis was separated and purified by different column chromatographies such as macroporous resin, silica gel, and Sephadex LH-20, and through preparative HPLC and recrystallization. Thirteen compounds were isolated and identified as(2S)-6-[(Z)-3-hydroxymethyl-2-butenyl]-5,7,3'-trihydroxy-4'-methoxy-dihydroflavanone(1),(2S)-8-[(E)-3-hydroxymethyl-2-butenyl]-5,7,3',5'-tetrahydroxy-dihydroflavanone(2), α,α'-dihydro-5,4'-dihydroxy-3-acetoxy-2-isopentenylstilbene(3), 6-prenylquercetin(4), 6-prenylquercetin-3-methyl ether(5), formononetin(6), 3,3'-dimethylquercetin(7), chrysoeriol(8), diosmetin(9),(10E,12Z,14E)-9,16-dioxooctadec-10,12,14-trienoic acid(10), 5,7,3',4'-tetrahydroxy-6-prenyl-dihydroflavanone(11), naringenin(12), dibutylphthalate(13). Compounds 1-3 are new compounds, and compounds 10 and 13 are isolated from aerial parts of this plant for the first time.
Glycyrrhiza uralensis/chemistry* ; Plant Components, Aerial/chemistry*

OBJECTIVEIn order to understand the glycyrrhizin content range in the wild and cultivated Glycyrrhiza uralensis in China and to find the related influencing factors of glycyrrhizin content.

METHODThe glycyrrhizin content of 165 wild and 1 013 cultivated G. uralensis samples from 37 countries in 9 provinces was determined by HPLC, and the effects of the producing region, medicinal parts, cultivation years, soil type and texture on the glycyrrhizin content were analyzed.

RESULT AND CONCLUSIONThe average glycyrrhizin content was (4.43 +/- 1.32)% in the wild G. uralensis population, and (1.51 +/- 0.49)% in the cultivated and the glycyrrhizin content in the cultivated was less than the minimum sandards in the Chinese Pharmacopoeia. The glycyrrhizin content was significant different in the wild and cultivated G. uralensis in different producing regions, respectively. The glycyrrhizin content in roots and rhizome of the wild G. uralensis had no significant difference, it had no significant difference in the cultivated G. uralensis from 1 to 4 years and it increased rapidly after 5 years, and the effects of the soil types and texture on it were significant.

OBJECTIVETo find out the licorice(Glycyrrhiza uralensis) distribution area, resource complexion and resource reserves of northeast China, to analyze the cause of the swing of the pendulum of resources, to put forward the countermeasure of resource protection and to provide evidence for the establishment of relative statutes.

METHODCombination of visit-inquisition and sample-square investigation involving the resource complexion of the 32 counties and cities in the northeast China wasmade.

RESULT AND CONCLUSIONThe east distribution boundary and the whole distribution current of licorice in the northeast China were determined the northeast licorice distributing region was compartmentalized into three typical sub-regions, and the licorice population character and artificial disturbing status in main counties of every sub-region were described. The licorice reserves were also figured out. At the same time, the nature and the artificial factors that influenced the swing of the pendulum of licorice resource were analyzed, and the correlative safeguard measure was brought forward.

OBJECTIVEWe studied the licorice provenance rule under the manual simulative water stress conditions, discussed geographical variation pattern and the ecology mechanism and laid the theoretical basis for the licorice seed regionalization and the seed allocation.

METHODOn the spot we investigated and collected seed materials. Setting up water potential gradient by PEG, we observed the seed germination characteristic through the experiment of indoor germination.

RESULTThe sensitivity of various provenance seed germinative energy show quite remarkable difference for water potential reducing. Along with geographic longitude increasing, the seed germinates variation sensitivity increasing gradually to the water stress. The results of climatic factors' correlation analysis indicated that provenance seed germinate energy to the water stress is sensitive, in which the annual average temperature is low and annual precipitation is abundant. But it is insensitive in which the annual precipitation is little, the annual average temperature, average temperature in July and the annual average ground temperature is high.

CONCLUSIONUnder the water stress condition the licorice provenance seed germination characteristic exists remarkable geographical variation. Geographical variation is the result of natural selection.

PURPOSE: The Glycyrrhiza uralensis species (Leguminosae) as a medicinal biocompound, and one of its root components, isoliquritigenin (ISL), which is a flavonoid, has been reported to have anti-tumor activity in vitro and in vivo. However, its function in bone formation has not been studied yet. In this study, we tested the effect of Glycyrrhiza uralensis (ErLR) and baked Glycyrrhiza uralensis (EdLR) extracts on osteoblast proliferation, alkaline phosphatase (ALP) activity, and bone-related gene expression in osteoblastic MC3T3-E1 cells. METHODS: MC3T3-E1 cells were cultured in various levels of ErLR (0, 5, 10, 15, 20 µg/mL), EdLR (0, 5, 10, 15, 20 µg/mL), or ISL (0, 5, 10, 15, 20 µM) in time sequences (1, 5, and 20 days). Also, isoliquritigenin (ISL) was tested for comparison to those two biocompound extracts. RESULTS: MTT assay results showed that all three compounds (ErLR, EdLR, and ISL) increased osteoblastic-cell proliferation in a concentration-dependent manner for one day. In addition, both ErLR and EdLR compounds elevated the osteoblast proliferation for 5 or 20 days. Extracellular ALP activity was also increased as ErLR, EdLR, and ISL concentration increased at 20 days, which implies the positive effect of Glycyrrhiza species on osteoblast mineralization. The bone-related marker mRNAs were upregulated in the ErLR-treated osteoblastic MC3T3-E1 cells for 20 days. Bone-specific transcription factor Runx2 gene expression was also elevated in the ErLR- and EdLR-treated osteoblastic MC3T3-E1 cells for 20 days. CONCLUSION: These results demonstrated that Glycyrrhiza uralensis extracts may be useful for preventing osteoporosis by increasing cell proliferation, ALP activity, and bone-marker gene expression in osteoblastic cells.
Alkaline Phosphatase* ; Cell Proliferation* ; Gene Expression ; Glycyrrhiza uralensis* ; Glycyrrhiza* ; In Vitro Techniques ; Miners ; Osteoblasts* ; Osteogenesis ; Osteoporosis ; RNA, Messenger ; Transcription Factors

The transplants of one-year-old Glycyrrhiza uralensis Fisch. were subjected to five concentrations of MnSO4-H2O (0, 1.81, 18.1, 36.2 and 54.3 mg·L(-1)) culturing in vermiculite. qRT-PCR and HPLC were respectively used to measure the relative expression of SQS1 gene and the content of glycyrrhizic acid of G. uralensis in different concentrations of MnSO4·H2O. This is to explore discuss the effects of the expression of SQS1 gene and the accumulation of glycyrrhizic acid by Mn treatment. The results showed both the expression of SQS1 gene and the content of glycyrrhzic acid of G. uralensis tended to rise after the fall of the first with the increase of concentration of Mn treatment. And they were of very significant positive correlation (P<0.01, r=0.737). Relative expression of SQS1 gene reached the highest 7.90 under 18.1 mg·L(-1) MnSO4·H2O treatment. It was very significantly different between 18.1 mg·L(-1) concentration of MnSO4·H2O treatment and CK (0 mg·L(-1)), 1.81, 36.2 and 54.3 mg·L(-1) (P<0.01), and 1.75, 1.37, 1.37, 2.33 times respectively. The content of glycyrrhizic acid reached the highest under 1.81 and 18.1 mg·L(-1) MnSO4·H2O treatment, and there were not significant difference (P>0.05). It was very significantly different between them and other concentrations of MnSO4·H2O treatment (P<0.01). This study suggests the appropriate concentration of Mn treatment could certain promote the expression of SQS1 gene and the accumulation of glycyrrhizic acid of G. uralensis.
Chromatography, High Pressure Liquid ; Genes, Plant ; Glycyrrhiza uralensis ; chemistry ; genetics ; Glycyrrhizic Acid ; analysis ; Manganese

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

OBJECTIVETo estimate the broad-sense heritability of the production of Glycyrrhiza uralensis and the content of glycyrrhizin as well as the genetic relationship of various growth indexes and biomass indexes, and provide the scientific basis for establishment of high quality licorice cultivate technology system.

METHODThe randomized method was used to assign the provenance trial, the content of glycyrrhizin was determined by HPLC, and the method of classic genetics was applied to estimate the broad-sense heritability and genetic correlation coefficient.

RESULT AND CONCLUSIONThe content of glycyrrhizin is influenced by the growth environment and gene, but the growth environment is the dominant factor. The estimated result of single sites about broad-sense heritability (h2) showed that the production of G. uralensis (W(u)) and the content of glycyrrhizin was controlled by gene which the broad-sense heritability was 0.663 2, 0.751 1 respectively, they had some potential on genetic modification. The results of genetic analysis correlation showed that the plant height and the stem diameter was positive (P < 0.01) correlated significantly with the production (W(u)) either on phenotype or on genetic, it suggests that the plant height and the stem diameter could be the index above ground to assessment the production of the G. uralensis. The content of glycyrrhizin had a positive correlation with the number of lateral root (P < 0.05), but it had a negative correlation with the plant height, stem diameter, diameter of root top (D(r)), the total biomass (W(t)) and the biomass underground (W(u)) on inheritance. It is suggested that it was difficult to achieve both high content and high yield simultaneously in the genetic improvement, so we should have a deeply thought about the specific improvement target when making the reformed scheme.