[Comparative analysis on seedling growth dynamics and physiological indices of Bupleurum smithii var. parvifolium and B. scorzonerifolium].

With Bupleurum smithii var. parvifolium and B. scorzonerifolium as test objects, in order to provide a theoretical basis for the introduction and domestication of B. smithii var. parvifolium, the growth and development dynamics of seedlings, biomass accumulation, the content of malonaldehyde(MDA), the activity of antioxidase such as SOD, POD, CAT and APX between them were comparatively analyzed by direct sowing culture in the open field. The results indicated that the morphological index and the biomass accumulation of B. smithii var. parvifolium such as root diameter, root length, plant height and leaf number were inferior to B. scorzonerifolium, the antioxidase SOD and POD activity of B. smithii var. parvifolium was significantly inferior to B. scorzonerifolium (P<0.05), the antioxidase CAT and APX activity of B. smithii var. parvifolium was inferior to B. scorzonerifolium but the difference wasn't significant, while MDA content was superior to B. scorzonerifolium(P<0.05). Thus, compared with cultivated B. scorzoneri folium, the plant growth velocity of wild B. smithii var. parvifolium was relatively slower and its resistance was relatively weaker after introduction and domestication.

[Research on topographic factors of ecology suitability regionalization of Bupleuri Radix in Hebei province].

To study the correlation between the content of saikosaponins ingredient of Bupleuri Radix and topographic factors, we researched the ecology suitability regionalization of topographic of Bupleuri Radix from Hebei province to provide a scientific basis for selecting artificial planting. Based on 43 samples of Bupleuri Radix from Hebei province, the variation of the content of saikosaponins in different conditions of topographic factors and the influence of slope, altitude and aspect were comprehensively analyzed by SPSS 21.0. Then we studied topographic factors of ecology suitability regionalization of Bupleuri Radix on the basis of the relationship between the saikosaponins and topographic factors by ArcGIS. The most suitable conditions of topographic for cultivation of saikosaponins are as follows：altitude 600 m above, slope 4.00-5.50 degrees, aspect to the sun. In Hebei province, it is suitable for growth of Bupleuri Radix in the Taihang Mountains and the Yanshan Mountains where the content of saikosaponins is higher.

Toxic polyacetylenes in the genus Bupleurum (Apiaceae) - Distribution, toxicity, molecular mechanism and analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: The genus Bupleurum includes approximately 200 species that are widely distributed in the Northern Hemisphere, Eurasia and North Africa. Certain species of this genus have long been used as antiphlogistic, antipyretic and analgesic agents in traditional folk medicine. As described in the Chinese Pharmacopoeia, the roots of Bupleurum chinense DC. and B. scorzonerifolium Willd. are the herbal materials that compose Chaihu (Radix Bupleuri), a well-known TCM herb. AIM OF THE REVIEW: This review aims to provide up-to-date and comprehensive information regarding the distribution, toxicity, molecular mechanism and relatively new methods for the qualitative and quantitative determination of polyacetylenes in different Bupleurum species. METHOD: The information needed for this paper were sourced from publishing sites such as Elsevier, science Direct, PubMed; electronic search engines such as Scopus and Web of Science, Google scholar; other scientific database sites for chemicals such as ChemSpider, PubChem, SciFinder, and also from on line books. RESULTS: Polyacetylenes, which are widely distributed in genus Bupleurum of the Apiaceae family, have high toxicity. Among polyacetylenes, bupleurotoxin, acetylbupleurotoxin and oenanthotoxin have strong neurotoxicity. Through previous research, it was found that the toxicity of Bupleurum polyacetylenes manifested as epileptic seizures, with the target of toxicity being the brain. The neurotoxicity of polyacetylenes exhibits a relationship with the γ-aminobutyric acid (GABA) receptor pathway, and polyacetylenes have been shown to inhibit GABA-induced currents (IGABA) in a competitive manner. CONCLUSIONS: The plants of genus Bupleurum have been used in traditional medicine for thousands of years. However, certain species of this genus are poisonous, and it was attributed to the high content of polyacetylenes. The present review indicates that certain polyacetylenes in the genus Bupleurum have highly neurotoxic effects. The major challenge with regard to toxic polyacetylenes is to test their neurotoxic effects in vivo as well as in further preclinical studies, which will require large amounts of purified polyacetylenes. More reference substances should be prepared, and sophisticated analytical technologies should be developed to comprehensively assess the quality of Radix Bupleuri herbs. These investigations will be helpful for further utilization of the plants of genus Bupleurum.

[Effects of hormone treatment, light and temperatures on sprouting characteristics of Bupleurum chinense].

Seeds of Bupleurum chinense cultivar, Zhongchai No. 1, were sowed in plastic pots which used the arable layer soil as the nursery bed and putted in the artificial climate incubator at various temperatures (15, 20, 25, 15-25 degrees C) and light (8,12 h) to germinate, respectively. The lower constant temperature (15 degrees ) and the higher constant temperature (25 "C) were not conducive to the sprouting characteristics of B. chinese. While they were able to enhance root activity to some extent; The seeding growth of B. chinese was significantly better in the variable temperature than correspondence in the constant temperature, significantly. The emergence speed, emergence index, vigor index and root activity of Bupleurum were improved under the 12 h of light-time, but the germination rate was not improved. The sprouting of Bupleurum's seeds could be improved to some extent by soaking with hormone, such as gibberellin, cytokinin, salicylic acid. Gibberellin promoted seeds' sprouting and seedings's root activity of Bupleurum, while salicylic acid increased the root activity of seeding. There is a significant influence of light, temperatures and hormone treatment on the germination of Zhongchai No. 1 seeds, and all three are remarkably interacted; It is beneficial to promote seed germination by the temperature (20 + 5) degrees C, lighting (8 h) and gibberellin concentration (10 x 10(-6)).

[Grain filling dynamics and germination characteristics of Bupleurum chinense seeds].

Bupleurum chinense used in the study were cultivated in the experimental fields of Gansu agricultural University for three years. The seeds of B. chinense were collected every 3 days 10 d after the blossom. The result showed that the 1 000-grain fresh weight reached the maximum 43 d after the blossom and then decreased rapidly, at the mature period the fresh weight of seeds were falling to the same level of the dry weight. The dynamic change of the grain dry matter accumulation showed as an S-shape curve, the rapid increase stage was 25-34 d following the flower, and the grain filling was ended 46 d after blossom. Grain filling rate was under the law "fast-slow-fast-slow". And there were two peaks of grain filling rate appeared, after reached the second peak 28 d after the flower the filling rate decreased rapidly and stayed steadily 43 d after flowering. The dehydration rate was also measured at its maximum 43 d following flower. The indexes of seeds all reached the top 52 days following the blossom, when the germination rate reached the peak (34.33%) and water content of seeds was near 10%. The rate of germination and the 1 000-graid weight of seed showed significant positive correlation, while the water content of seeds was found significant negatively correlation with germination percentage. So the best time for harvest should be 52 d after flowering (9 month), the seeds collected at that time showed both high quality and germination rate.

[Induction of hairy roots and plantlet regeneration of Bupleurum chinense DC].

In this study, the induction of hairy roots of Bupleurum chinense DC. was explored and established after experiments at different conditions: A. rhizogenes A4 was used to infect the leaves bases of B. chinense tube seedlings. The explants were co-cultured on Phytagel-solidified media for 3 days and then, were turned into solid media, similar with the co-culture media except that bacteriostat was added. After 10 days, rootlets began to appear and after 4 to 5 weeks, rootlets can be converted into liquid shaking culture stage. Plants regeneration from hairy root was useful for the research of new germplasm production and the variety improvement breeding. In the present study, the regenerated plants were obtained. One approach was to continuously culture under light conditions the seedlings which parting off spontaneously from the hairy roots during liquid shaking culture. The other approach was to culture the callus-like tissues produced by hairy roots with the optimized regeneration media for the induction of regenerated plants. The results of present study provide a technique to induce hairy roots and plantlet regeneration of B. chinense and this technique is helpful for the researches on metabolism, especially on the Agrobacterium-mediated genetic transformation of B. chinense.

[Comprehensive yield and quality comparison of Bupleurum chinense' s seeds of different harvesting time from Qingchuan County with Topsis approach].

In order to investigate the comprehensive quality differences of the seeds harvested in different growing time, and offer theoretical guide for the optimization of seeds' production technology, we analyzed the apparent size, 1 000-grain weight, water absorbency, germination indexes, postharvest embryo rate change, relatively contents of coumarins and the yield of single plant of its seeds of different harvesting time, and compared their comprehensive quality by Topsis analysis. The results showed that there existed obvious differences in yield and quality between seeds of 3 harvesting times. While the harvesting time postponed, the yield of single plant raised, and the shrunken seeds rate decreased, the quality of seedlings increased, while the contents of coumarins showed a steady increase, and the germination rate decreased. The comprehensive quality of the seeds harvested in the black ripe time rank the first place, followed by the brown ripe time and the yellow ripe time. As the harvesting time delays, the seeds' comprehensive quality increases, therefore, we could put off the seeds' harvesting time properly for the high efficient seed production.

[Studies on high efficient plant regeneration system from anther callus of Bupleurum chinense].

The callus of Bupleurum chinense with anthers at the stage of uninucleate was induced. After several subcultures, anther calli of B. chinense were cultured at 20 MS culture mediums with different plant hormones to differentiate into plantlets. Differentiation of callus was detected after 21 and 49 days to select the most effective medium. There were 19 culture mediums in which anther callus could differentiate into plantlets with differentiation rate range from 3% to 60% , and most less than 20%. MS + KT 0.5 mg x L(-1) + sucrose 30 g c L(-1) + phytagel 5 g x L(-1) was the best differentiation medium with the differentiation rate of 60%, followed by MS + ZT 1.0 mg x L(-1) + sucrose 30 g x L(-1) + phytagel 5 g x L(-1) with the differentiation rate of 58%. Then plantlets were transferred to rooting medium to obtain whole plant. All plantlets could root in the rooting medium of MS + sucrose 30 g x L(-1) + phytagel 5 g x L(-1) and 1/2 MS + NAA 0.5 mg x L(-1) + sucrose 30 g x L(-1) + phytagel of 5 g L(-1) with the rooting rate of 100%. As a result, the high efficient and stable plant regeneration system was established from anther callus of B. chinense.

[Investigation on bupleurum resources in northwest area of Hubei Province].

OBJECTIVE: To investigate the species and the distribution of Bupleurum genus in northwest of Hubei province and provide basis for the standardization planting of Bupleurum in the region. METHODS: Investigated and collected specimens on the spot, com- pared them with literatures and specimen collected before. RESULTS: The main species of Bupleurum plants was B. marginatum, only very few of them was B. scorzonerifolium, no B. chinense was found in northwest of Hubei province. CONCLUSION: B. marginatum is the mainstream species of Bupleurum plants and the roots of B. marginatum are the actual source of commodity with the name of "Beichaihu (the roots of B. chinense)" in northwest of Hubei province.

Large-scale production of saikosaponins through root culturing of Bupleurum falcatum L. using modified airlift reactors.

Modification of internal configuration of a bubble column, airlift and stirred tank reactor (10-200 L) was made for root cultures of Bupleurum falcatum L. Agitation with an impeller covered with partition mesh was ineffective for a 10-L modified reactor, because it caused intensive foaming and subsequent overflow of the culture medium even at a low rotation speed of 50 rpm and a low aeration rate of 0.1 vvm (volume per volume of medium). In contrast, efficient aeration through a ceramic sparger placed at the bottom of a 20-L bubble column reactor yielded approximately 25 g/L of dry roots and 500 mg/L of saikosaponin-a and saikosaponin-d over 42 days. On a 200-L scale, however, the roots became flocculated under the upper perforated plate initially positioned near the middle of the reactor, forming a firm disk of roots and a large empty space between the disk and the medium. Thus, the roots had poor contact with the medium, which severely suppressed their growth. To avoid this flocculation, a bottom perforated plate and draft tube were installed as a partitioning device separating the culturing area (outside the draft tube) from the aeration area (inside the draft tube). The draft tube was made of a stainless steel mesh rather than a solid material, and the tube greatly increased the root yield in the 20-L reactor. This configuration was successfully applied at the 200-L scale, yielding 500-600 mg/L of saikosaponin-a and saikosaponin-d over 56 days.

[Advances of studies on seed biology of Bupleurum chinense].

Bupleurum chinense is a commonly used Chinese medicinal material, which has been used medicinally in China for over 2000 years, the development of it is of great value. There have been great advances of studies on its embryology and seed biology in recent years. In this paper, we make a review of the growth of its embryo and fruit, the characteristics of dormancy and storage and the germination of its seed. Besides, we summarize the reasons of its low germination rate and the achievements in improving the situation, for the purpose of providing reference in research on seed physiology of B. chinense.

[Cropping patterns of Bupleurum chinense in semi-arid region of Loess Plateau].

Field experiments were conducted to investigate the suitable cropping patterns of grain crops and medical plant Bupleurum chinense in the semi-arid region of the Loess Plateau. Three cropping patterns were designed, including corn-Bupleurum chinense intercropping (corn/Bupleurum chinense), wheat followed by soybean-Bupleurum chinense intercropping (wheat-soybean/Bupleurum chinense), and sequential planting of Bupleurum chinense (wheat-Bupleurum chinense). Of the three patterns, wheat-soybean/Bupleurum chinense was the best in economic yields and ecological benefits. Among the cultivars (cv. Lingchuan, Zuoquan and Wanrong) of Bupleurum chinense tested, the cv. Wanrong originated from relatively high temperature region exhibited the highest yield, while the cv. Zuoquan originated from relatively low temperature region had the lowest one, indicating that introducing Bupleurum chinense cultivars from the areas with higher temperature to lower temperature areas could increase their yield. The cropping pattern of wheat-soybean/Bupleurum chinense was the most effective one, with the yield being 11.6% and 16.8% higher than that of corn/Bupleurum chinense and wheat-Bupleurum chinense, respectively.

[Study on main inheritable parameters of test-tube plants population of Bupleurum chinense in different growing periods].

OBJECTIVE: To determining the growth status and medicine quality of test-tube plants of Bupleurum chinense. METHODS: Measuring the field characters of the labeled plants at fixed period after turning green,such as plant height, number of tiller, number of leaves. Using statistical methods to analyse the measured data RESULTS: Each genetic characters standard deviation and relative standard deviation are all smaller at vegetative growth phase, bud forming phase and seed ripening phase. The test-tube plants population of Bupleurum chinense grows consistently. Its uniformity is higher, variability is low. Bupleurum chinense length and weight of root relative standard deviation are all low. The average root length and weight are more predominant than ordinary seed plants. The roots have high content of effective components. Bupleurum chinense population pharmaceutical character is excellent. The medicinal quality is consistent. The number of branch stem and effective fruit bunch is bigger, The seed production is higher. The aerial part yield is large, The economic value of test-tube plants of Bupleurum chinense is higher. The correlation between each characters of test-tube plants of Bupleurum chinense is different from the ordinary seed plants in different growing periods. CONCLUSION: The botanical and pharmacognostic characters of test-tube plants of Bupleurum chinense are excellent.

[Breeding of new varieties "zhongchai no. 2" and "zhongchai no. 3" of Bupleurum chinense].

OBJECTIVE: To breed new varieties with better uniformity and commercial quality as well as higher saikosaponin contents. METHOD: The excellent germplasm resources were selected from "zhongchai no. 1" population. Single plant method was applied to get better varieties. All the breeding material was investigated according to morphological characters, agronomic characters and the contents of saikosaponin a and saikosaponin d. The experiments of comparative test and varieties regional test were carried out. RESULT: The bred new varieties of "zhongchai No. 2" and "zhongchai No. 3" had better uniformity. The dark brown roots ratios of the two varieties were 83.2%, 89.9%, respectively. The contents of saikosaponins (a + d) of the two varieties reached 1.31%, 1.02%, respectively. CONCLUSION: "zhongchai No. 2" and " zhongchai No. 3" both had the advantages of better uniformity, darker brown roots and higher saikosaponin contents.

[Study on dynamic accumulation of index components from Bupleurum chinense in various collecting periods].

OBJECTIVE: To study the dynamic change law of volatile oil, saikosaponin a, d and alcohol-extract from Bupleurum chinense at Songxian region in Henan province, and to explore the optimal harvest period of Bupleurum chinense. METHODS: With the contents of saikosaponin a and d, absorbance of volatile oil and percentage of alcohol-extract as indexes, HPLC-ELSD and ultraviolet spectrophotometry were successively used to analyze them. RESULTS: There are obvious differences among the contents of volatile oil, saikosaponin a, d and alcohol-extract in various collecting periods sample, the absorption of volatile oil in distillation was the highest in October, the content of saikosaponin a was the highest in September, the saikosaponin d in December and the percentage of alcohol-extract in October. CONCLUSION: The optimal harvest period of Bupleurum chinense at Songxian region in Henan is identified, which can provide scientific basis for crude drug production and processing.

[Studies on fingerprints of Bupleurum chinense and vinegar-baked Bupleurum chinense from Hubei province].

OBJECTIVE: To establish the HPLC Fingerprint of Bupleurum chinense from GAP provenance of Hubei province Baokang, and discuss the changes before and after processing. METHODS: Using HPLC detection method equipped with Hypersil ODS column (5 microm, 4.6 mm x 250 mm), using acetonitrile-water as mobile phase gradient elution, flow as 1.0 ml/min, UV detector set at 203 nm, column temperature was at 25 degrees C. RESULTS: Similarity of processed Bupleurum chinense and Bupleurum chinense exceeded 0.95, and there were 13 characteristic fingerprints of processed Bupleurum chinense, 15 characteristic fingerprints of Bupleurum chinense. CONCLUSION: The HPLC Fingerprint of Bupleurum chinense from GAP provenance of Hubei province Baokang are established, and the difference between Bupleurum chinense before and after processing is studied initially.

[Effects of nitrogenous phosphorus and potassium fertilizer on yield of Bupleurum scorzonerifolium].

OBJECTIVE: To study the effect of different fertilizer assortment (nitrogenous, phosphorus and potassium) on Bupleuram scorzonerifolium yield. METHODS: Three factors were designed by quadratic regression and D-optimum designation. RESULTS: As far as the biomass concerned, the best design of fertilizer was N 120. 3 - 333. 3 kg/hm(2) P2O5 0-939. 45 kg/hm(2), K2O 0 - 227. 25 kg/hm(2). As to R/T, the best design was N 62. 66 -333. 3 kg/hm(2), P2O5 0 -922. 54 kg/hm(2) , K2O 0 -227. 25 kg/hm(2). And for the root weight, the best design was N 34. 5 -333. 3 kg/hm(2), P2O5, 0 -939. 45 kg/hm2 , K2O 0-227. 25 kg/hm(2). CONCLUSION: Application quality of nitrogenous, phosphorus and potassium fertilizer are closely related with yield of B. Scorzonerifolium.

[Preliminary study on the characteristics of Bupleurum chinense in nutrition uptake].

It was studied that the characteristics of Bupleurum chinense DC. in nutrition uptake based on potted and field experiment to provide a properly fertilizer application measure. The result showed that the absorption amounts of N, P and K were in the order of K > N > P under field condition. For one-year-old B. chinesnse DC., the average N, P2O5 and K2O absorption per 100 kg dry matter of root were 11.77 kg, 2.43 kg, 14.07 kg respectively; for two-year-old, the average N, P2O5 and K2O absorption amount per 100 kg dry matter of root were 12.11 kg, 2.74 kg, 18.39 kg respectively. The total absorption amount of N, P2O5 and K2O were lower in the first year, while the percentage of content was higher, that was to say, the demand for N, P and K was urgent in the first year. The accumulation peak of N, P2O5 and K2O occurred regreening to later elongation stage in the second year. The ratio of N, P and K were differential in different growth stages.

[Preliminary study on N, P, K fertilizer to control of root rot of Bupleurum chinense].

The application of N and K fertillizer could improve the sensibility of Bupleurvum chinense DC. to Root Rot, while large application of P fertilizer could decrase the sensibility. The fertilizer measure which could obtain highest yield but could not increase its disease resistense. To protect Bupleurum chinense against root rot, more phosphorous fertilizer, certain nitrogen and potassium ferilizer should be applied in early elongation stage in the second growing year.

Seasonal branch nutrient dynamics in two Mediterranean woody shrubs with contrasted phenology.

BACKGROUND AND AIMS: Mediterranean woody plants have a wide variety of phenological strategies. Some authors have classified the Mediterranean phanaerophytes into two broad phenological categories: phenophase-overlappers (that overlap resource-demanding activities in a short period of the year) and phenophase-sequencers (that protract resource-demanding activities throughout the year). In this work the impact of both phenological strategies on leaf nutrient accumulation and retranslocation dynamics at the level of leaves and branches was evaluated. Phenophase-overlappers were expected to accumulate nutrients in leaves throughout most of the year and withdraw them efficiently in a short period. Phenophase-sequencers were expected to withdraw nutrients progressively throughout the year, without long accumulation periods. METHODS: To test this hypothesis, variations in phenology and leaf NPK in the crown of a phenophase-overlapper Cistus laurifolius and a phenophase-sequencer Bupleurum fruticosum were monitored monthly during 2 years. KEY RESULTS: Changes in nutrient concentration at the leaf level were not clearly related with the different phenologies. Nitrogen and phosphorous resorption efficiencies were lower in the phenophase-overlapper, and accumulation-retranslocation seasonality was similar in both species. Changes in the branch nutrient pool agreed with the hypothesis that the phenophase-overlapper accumulated nutrients from summer until the bud burst of the following spring, recovering a large nutrient pool during massive leaf shedding. The phenophase-sequencer did not accumulate nutrients from autumn until early spring, achieving lower nutrient recovery during spring leaf shedding. CONCLUSIONS: It is concluded that phenological demands influence branch nutrient cycling. This effect is easier to detect by assessing changes in the branch nutrient pool rather than changes in the leaf nutrient concentration.