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Studies on Relationship between Structures, Development and Medicinal Components Accumulation of Bupleurum Chinense DC. and Bupleurum Scorzonerifolium Willd

Author: TanLingLing
Tutor: HuZhengHai
School: Northwestern University
Course: Botany
Keywords: Bupleurum chinense Angustifolia Bupleurum Bupleurum total saponins Saikosaponin a Flavonoids Histochemical Phytochemicals
CLC: S567.79
Type: PhD thesis
Year: 2008
Downloads: 497
Quote: 4
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Abstract


Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd which belong to Bupleurum L. in Umbelliferae are perennial herbaceous plants, and both of them are original plants of Radix Bupleuri which were collected in "Chinese Pharmacopoeia".The main medical parts are their dry roots, and the main medicinal ingredients are saikosaponin and flavonoid. Anatomical, histochemical and phytochemical methods were used to systematically investigate the structures of organs in adult plants of Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd, the development of vegetative organs of Bupleurum chinense DC, and the localization and content changes of saikosaponin and flavonoid in vegetative organs of Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd, in order to discuss the relationship of structures, development and major medicinal components accumulation in Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. The results are as followes:The development of Bupleurum chinense DC. root conformed to the general developmental law of perennial herbaceous dicotyledons. This progress could be divided into four stages: promeristem, primary meristem, the primary and secondary stages of growth. The promeristem consisted of three groups of primitive cells which had the cytological characteristics of typical meristem cells and the primary meristem was composed of calyptrogens, dermatogen, periblems and pleromes. The primary structure consisted of epidermis, cortex and stele. The primary xylem was diarch. The secondary growth mainly relied on activities of vascular cambium and cork cambium. The cork cambium originated from pericyclic cells that resumed their dividing capabilities. However, before forming the cork cambium, pericyclic cells produced many layers pericyclic parenchyma cells by periclinal division. Therefore, the secondary structure included periderm, pericyclic parenchyma tissue and secondary vascular tissue from the outside to the inside of taproots of Bupleurum chinense DC. In addition, there were many developed wood fibers which distributed in a clumped manner in secondary xylem. The root of Bupleurum scorzonerifolium Willd was similar to that of Bupleurum chinense DC. in basic secondary structure.The development of stem of Bupleurum chinense DC. also included four stages: promeristem, primary meristem, primary and secondary stages of growth. Promeristem was composed of two layers of tunica cells and a group of corpus cells. With the development of promeristem, the primary meristem, including protoderm, ground meristem and procambium, was formed at the apex of the shoot. The primary meristem differentiated into primary structure consisting of epidermis, cortex, vascular system, pith and pith ray. With the progressive development of stem, the activity of vascular cambium began. The fascicular cambium differentiated into secondary xylem and secondary phloem, while the interfascicular cambium only differentiated into wide ray of parenchyma tissue. Later, these parenchyma cells whose cell walls were thickened and lignified became sclerenchyma. Consequently, vascular bundles in stem of Bupleurum chinense DC. always kept discrete, and there were developed sclerenchyma between them. Meanwhile, the cork was not found to form in the stem from beginning to end in its whole life. The stem of Bupleurum scorzonerifolium Willd was also similar to that of Bupleurum chinense DC. in basic secondary structure.The development of Bupleurum chinense DC. leaves could be divided into three stages: promeristem, primary meristem and mature structure. The leaf of Bupleurum chinense DC. was the typical bifacial leaf which composed of blade and stipe. The blade consisted of epidermis, mesophyll and vein. And the mesophyll was developed, with distinct differentiation of palisade tissue and spongy tissue. Palisade tissue was one cell wide and there were three to five layers cells in spongy tissue. Meanwhile, both inhabited separately half of the whole cross section of leaf. However, the leaf of Bupleurum scorzonerifolium Willd was isobilateral leaf and the mesophyll was developed. There were palisade tissues inside both supine surface and superficies inferia of the leaf, and the spongy tissue was distributed between palisade tissues.According to the fact that saikosaponin can react with 5% vanillin-glacial acetic acid-perchloric acid solution to produce characteristic colors, which is from light red to dark red, we use it as a test reagent to test saikosaponin. The results showed that the saikosapinon was distributed in pericycle, vascular cambium, secondary phloem and partial pericyclic parenchyma cells in secondary xylem of the root. In the stem, they were mainly distributed in epidermis and cortex, and there were small amounts in vascular cambium and pericyclic parenchyma cells in secondary phloem. In the leaf, they were mainly located in the epidermis and mesophyll, and were not found in the vein.Flavonoid was another main medicinal ingredient in Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. According to the fact that flavonoid can react with 1% magnesium acetate-methanol, NA and 5% sodium hydroxide to produce characteristic colours or fluorescence, we use them as test reagents to test flavonoid. The results showed that flavonoid was mainly distributed in stem and leaf, there were small amounts in roots of Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. The results showed that flavonoid was distributed in epidermis, collenchyma which lied in angularity, partial cortex cells and myelin sheath cells of the stem; In the leaf, they were mainly distributed in epidermis, collenchyma which lay inside the epidermis. However, when slices of the root reacted with color development reagents, it would be difficult to accurately confirm the exact location of flavonoid in the root.According to researches through phytochemical methods, we could conclude that the content of total saikosaponin in the root was the highest in organs of Bupleurum chinense DC. And in Bupleurum scorzonerifolium Willd, the content of total saikosaponin in the mature fruit was the highest and the content of that in the root was also high. As a result, it had scientific gists that the main medical parts were their dry roots in Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd.Through the study on the content of total saikosaponin and saikosaponin-a in different parts of Bupleurum chinense DC, we found the content of saikosaponin in "skin areas" was higher than that in xylem, and the content was higher in lateral roots than that in taproots. Based on these results, the ratio of "skin areas" to xylem can be used as an anatomical criterion to estimate the saikosaponin content. Meanwhile, judging from the external characteristics of the drug, the one with an acerose taproot and more lateral roots was of high quality.Through the study on the content of total saikosaponin and saikosaponin-a in different years roots of Bupleurum chinense DC, we concluded that the content of saikosaponin in one-year-old mature root was higher than that in two-year-old mature root, indicating we should collect one-year-old roots of Bupleurum chinense DC. to get drug which had a higher saikosaponin content and a better quality,. However, with a view of practical yield, the little biologic yield of one-year-old roots could directly bring on a poor economic benefit. Consequently, we can make a correct choice about the appropriate collection of drug according to our practical need.The content of saikosaponin in different seasons in a growth period varied in the root of Bupleurum chinense DC. According to the dynamic change of the total saikosaponin in the whole growth period, we concluded that the content of total saikosaponin was high in both pre-blossom of spring and fruit mature period of autumn. However, we used the yield of activated component as the criterion to determine the best collecting season. As a result, we considered that the best collecting season of Bupleurum chinense DC. was fruit mature period of autumn, when both the dry weight of the root and the content of saikosaponin were high.The content of saikosaponin changed in Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd drugs from different areas. By comparison, in the same drug, the content of its total saikosaponin was high, but the content of saikosaponin-a was not high, and vice verse. Therefore, it is not appropriate to evaluate the quality of the drug according to the content of total saikosaponin or that of saikosaponin-a. The good drug should have high contents of both total saikosaponin and saikosaponin-a. At the same time, the different contents of saikosaponin in the drug from different areas were concerned with the local climate, planting environment, picking period, cultivation methods and other factors. Therefore, in order to ensure the quantity and the quality of the drug, we should carry out further studies to control and improve the drug’s quality and explore the basic factors which affected the quality of the drug.Through phytochemical methods, flavonoid in Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd was found to be abundant in their leaves and reproductive organs and it was less in roots. These results were consistent with findings through histochemical method. And these results may be related with the flavonoid’s character which was the most important UV absorption and also be related to the other physiological functions of the flavonoid.The content of flavonoid in different seasons in a growth period varied in stems and leaves of Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. In the whole growth period, the content of flavonoid was the highest in blossom period. Then through a dynamic variance, it returned to another high level again in pre-withering period in Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. Flavonoid was also an important medicinal ingredient in Bupleurum chinense DC. and Bupleurum scorzonerifolium Willd. Therefore, it was proposed that when roots were collected in pre-withering period, we also collected their aboveground parts, extracted the flavonoid in order to develop and utilize these two medicinal plants comprehensively.From the results above, it can be determined that there is an apparent correlation between structures, development and medicinal components accumulation in Bupleurum chinense DC and Bupleurum scorzonerifolium Willd.

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