Identification of Bupleurum Seeds by ITS2 Genetic Barcoding and Seed Morphology / 中国实验方剂学杂志

Objective:To accurately identify Bupleurum seeds by traditional morphological identification method combined with DNA barcoding technique. Method:A total of 41 seed samples on the market were collected and 75 ribosomal DNA internal transcribed spacer 2 (ITS2) sequences of 15 varieties were downloaded from the GenBank database as experimental materials. The seeds were measured and observed by stereomicroscope and vernier caliper, and their 1 000-grain weights were calculated. Genomic DNA was extracted from the seeds and used as a template, and ITS2 sequences were amplified using polymerase chain reaction (PCR) and bidirectional sequencing. Species identification was conducted based on BLAST method, neighbor-joining (NJ) phylogenetic tree method, Kimura two-parameter model (K2P) genetic distance method, and secondary structure of ITS2 sequence. Result:There were slight differences in the length, width, cross-section, and 1 000-grain weight among Bupleurum seeds from different origins. The ITS2 sequences of B. chinense seeds had 2 intraspecific variable sites and 3 haplotypes, the maximum intraspecific genetic distance (0.009) was far smaller than the minimum interspecific genetic distance (0.032). B. chinense and B. scorzonerifolium in the NJ phylogenetic tree were clustered into independent branches with good monophyletic property. The secondary structure of ITS2 sequences could make up for the shortcomings of NJ tree in identifying variants. The collected 41 seeds included 30 B. chinense seeds, 3 B. scorzonerifolium seeds, 5 B. falcatum seeds, 2 B. marginatum var. stenophyllum seeds, and 1 B. smithii var. parvifolium seeds. Conclusion:The B. chinense seeds on the market have problems of diverse sources and chaotic origins. Based on the combination of ITS2 gentic barcoding and seed morphological identification, the Bupleurum seeds can be accurately identified, which provides scientific bases for establishing the quality standard of Bupleurum seeds, standardizing the cultivation of B. chinense, and solving the quality problems of B. chinense from the source, and provides a reference for the accurate identification of other medicinal plant seeds or seed medicinal materials.

Advances in Bioculture Technology of Bupleuri Radix and Biosynthesis Pathways of Saikosaponins / 中国实验方剂学杂志

Synthetic biology is an emerging discipline that analyzes the biosynthesis pathways of active constituents in traditional Chinese medicine and explores genes involved in biosynthesis. Bupleuri Radix is one of the most commonly used Chinese medicinal materials with remarkable medicinal value, its index component is saikosaponins, which has significant anti-inflammatory, anti-viral and anti-tumor activities. However, the current wild resources of Bupleuri Radix have been destroyed, and there were some problems in the process of artificial cultivation. The application of biological culture technology and synthetic biology can expand the sources of saikosaponins and protect resources of Bupleuri Radix. The culture conditions of different plants can be followed without a fixed pattern, and the biosynthetic pathways of different medicinal active ingredients are also inconsistent. At present, there is no review report on the culture technology of Bupleuri Radix and the research on the biosynthesis pathway of saikosaponins. This paper introduces the research progress of biological culture techniques, such as callus culture, adventitious root culture, hairy root culture and suspension cell culture used in synthetic biology, and the biosynthesis pathway of saikosaponins and its key enzyme functional genes. It is suggested to optimize the biological culture technology of Bupleuri Radix by referring to the tissue culture technology of other traditional root medicinal materials, so as to provide a reference for the in-depth study on the biosynthesis pathway and metabolic regulation of saikosaponins.

Study on correlation between trace elements and active ingredient in glycyrrhizae radix et rhizome / 中国中药杂志

To clear the kinds of trace elements which is closely related with the active ingredient, proclaim the effects of trace elements on the quality of the Glycyrrhizae Radix et Rhizoma, provide the theoretical foundation to the further quality control of cultivation, take the advantage of the HPLC to determine the contents of glycyrrhizic acid and the liquiritin according to Chinese pharmacopoeia, use the inductively coupled plasma mass spectrometry to test the contents of Cu, Zn, Mn, Pb, Se, Cd, Ni, La, Na, Cr, M, Fe, Ca, Al, K, Sr, then, use SPSS statistical software for active ingredient and trace elements Correlation Analysis. The result of correlation analysis showed that Liquiritin contents of Glycyrrhizae Radix et Rhizoma have strong a positive correlation with the Mn, Pb contents, well, have a negative correlation with the Cu, Na contents; Glycyrrhizic acid contents showed a positive correlation with Mg, Cd, La contents, however, it showed a negative correlation with K, Fe contents. Comprehensive analysis of the results of the study, a preliminary thought that the active ingredient of Glycyrrhizae Radix et Rhizoma closely related with the trace elements, but the exact conclusion still need further study concentration-response relationship analysis.

Study on geographical variation of morphologic and germination characteristic of different Glycyrrhiza uralensis provenance seeds / 中国中药杂志

<p><b>OBJECTIVE</b>To study the geographical variation of morphologic and germination characteristic of different Glycyrrhiza uralensis provenance seeds, approach the geographical variation mode and ecology mechanism, and laid theoretical foundation for districting and allocating of G. uralensis seeds.</p><p><b>METHOD</b>Field investigation and laboratory analysis were applied. Seed shape and kilosseed weight were sampled randomly, germination rate germination force by general methods.</p><p><b>RESULT</b>The morphologic characteristic of G. uralensis seeds showed roughly longitude variation tendency that the seeds increased gradually from west to east. While the germination characteristic showed roughly altitude variation tendency that the seeds germination rate and germination force increased with the increase of the altitude, and the average germination rate was the same with the seeds morphologic characteristic. The results of analysis correlated with the climatic factors show that the morphologic characteristic of G. uralensis was positive correlated with annual rain-fall of the habitat, and the germination rate was quickened by drought, high temperature and strong sunshine.</p><p><b>CONCLUSION</b>The morphologic and germination characteristic and of G. uralensis seeds present distinguished geographical variation, and the formation of the variation was related to the ecological environment in which the seed provenance adapted.</p>

Effect of drought stress on growth of Glycyrrhiza uralensis / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of drought stress on the growth of Glycyrrhiza uralensis and its drought resistance characteristic.</p><p><b>METHOD</b>The growth in dicators of G. uralensis including leaf, stem, root and biomass were measured when two-years-old G. uralensis had lived in drought stress soil for 60 days.</p><p><b>RESULT</b>The growth of all organs of G. uralensis was restrained because of drought stress, especially to up ground organs.</p><p><b>CONCLUSION</b>It is obvious that the restraining effect of drought stress on G. uralensis possesses organ speciality. Both yield and quality of G. uralensis will be satisfied when it grows in the soil with relative water content of 50%.</p>

The underground part growth distribution pattern of Glycyrrhiza uralensis and its effects on glycyrrhizinic acid content / 中国中药杂志

<p><b>OBJECTIVE</b>To ascertain the relationship between glycyrrhizinic acid content and the underground part growth character of Glycyrrhiza uralensis and provide the theoretical evidence for wild resources protection and artificial cultivation method of G. uralensis.</p><p><b>METHOD</b>Through the analytical investigation on the underground part of G. uralensis and analysis of glycyrrhizinic acid content in different organs, parts, ages, and diameter medicinal materials, the systematic study on the relationship between glycyrrhizinic acid content and the underground part growth character of glycyrrhiza uralensis was carried out.</p><p><b>RESULT</b>The underground part of a G. uralensis seedling consisted of seed root, random root, horizontal underground stem, vertical underground stem and assimilating root. The glycyrrhizinic acid content in horizontal underground stem with the age below two years old or in random root with the diameter below 0.5 cm was low. The difference of glycyrrhizinic acid content among horizontal underground stem, random root and vertical underground stem was obvious, but the difference between horizontal underground stem and random root was not obvious.</p><p><b>CONCLUSION</b>The horizontal underground stem was of G. uralnesis acts as a link that can connect random root, vertical underground and stem assimilating root, so that the whole underground part constructs one huge underground net system. The glycyrrhizinic acid accumulation is a effected by organ type, growth age, root diameter and grow position, and the distribution pattern of random root and vertical underground stem has influence on glycyrrhizinic acid distribution in horizontal underground stem.</p>