Study On Anatomy And Histochemistry Of Coptis Chinensis

Coptis chinensis is a kind of perennial herbs and belong to ranunculaceae, whose rhizomes were listed as the top grade medicine of Chinese herbal. Coptis chinensis growth environment is more strict than other plants, as it need multiple soil coverage, and prefer the darkness environment, avoid high temperature and strong light, drought, etc, so that limited its production, and in China only lichuan of hubei provience and chongqing have Coptis chinensis cultivate.The apoplastic barrier structure is a interface protection organization between plants and environment, its including diffusion of suberin outside the epidermis of root, internal and external cortex with casparin bands and suberin,and the secodary wall lignification, or subcutaneous layer thickening being’φ’shaped, the surface of cuticle of hypocotyls, shoots and stalks, endodermis, exodermis, casparin bands and suberin in vascular bundle and the secodary wall lignification, which could prevent water, mineral and oxygen transportion between plant and environment, thereby protect the plant avoid harmage of plant diseases and insect pects, and promoted the protection of plant physiological and defense function. Coptis chinensis accommodate the darkness environment, however, there were little research report on the apoplastic barrier structure and its protect functions.The quantity of Coptis chinensis in highly relatived with its active constituent, especially the concentration of berberine. As the diversity of growth environment, resulting in the effective constituent of berberine concentration has significant difference. At present, the research on the quantity of Coptis chinensis were focused on alkaloids extraction, and there are skimp report on the distribution of berberine in different tissue and organ of plant.Over the view, the present work combibed with the plant cell wall histochemical localization, and through bare-handed slice under the light and fluorescent microscopy observe anatomical structure and barrier characteristics of Coptis chinensis, and the structure of the organ which berberine deposition, to provide the future new theoretical foundation for the improvement of yield and quality at future cultivation.The main results of the study were as follows:(1)Anatomical structure of underground and the barrier development process of the the of Coptis chinensis during the seedling stage. Combined with the research about histochemical method of cell wall, using the roots and hypocotyl of Coptis chinensis at seedlings period to make unarmed slice observe after different staining. Stained by the thiocyanate berberine-aniline blue, casprian bands present vivid yellow colour, lignification cell wall present dull yellow colour. Fat Red 7B staining suberin lamellae present red colour, hydrochloric aid-phloroglucinol staining lignin present cherry-red colour; the primary wall Casparian bands appear wavy by sulfuric acid digestion. The results showed that the Coptis chinensis seedlings inocluding two improtant structure, the primary structure and the secondary structure. The primary structure were located at cotyledon stage, the first true leaf, and the second true leaf stage, the root is included epidermis, exodermis, cortex, endodermis and vascular column. The secondary structure were located at the third, forth and fifth true leaf stage, and the mainly strcuture was the secondary xylem. Hypocotyl anatomic structures including epidermis, cortex, the cortex and xylem. The roots barrier structure of Coptis chinensis in the seedling stage is casparian bands in endodermis, slight lignification and strong suberin, as well as casparian bands in exodermis, lignification and suberin. The barrier structure of hypocotyl is cuticle, casparian bands in endodermis, slight lignification and strong suberin. The barrier structure of Coptis chinensis adventitious develop early, and limited the water, mineral ions and the oxygen absorption, thereby result of Coptis chinensis slow growth during the seedling period.(2)The adventitious root anatomy and barrier structure research of Coptis chinensis in the field cultivated. Using the above methods to observe Coptis chinensis of field cultivated after staining, the results show that, primary structure of adventitious roots composed of vascular cylinder, endodermis, cortex, exodermis and epidermis, secondary structure composed of secondary xylem, secondary phloem and phellem of Coptis Chinensis. The apoplastic barriers in adventitious roots of Coptis chinensis is casparian bands, lignification and suberin in endodermis and exodermis at primary primary structure. The apoplastic barriers in adventitious roots is lignification and suberin in phellem at secondary structure.(3)The tissue or organ research about berberine sedimentary in Coptis chinensis. After the lignified cell wall combined with berberine, the sections stained berberine release yellow fluorescence under bule exciting of fluorescence microscope. According to the fluorescence intensity berberine accumulation and distribution in different organizational structures of Coptis chinensis was determined according to the fluorescence intensity. Primary structure of rhizomes composed of cuticle, cortex and vascular cylinder, secondary structure composed of phellem, and mainly with cortex and vascular cylinder. Petioles composed of pith, vascular bundles, sclerenchyma rings, cortex and cuticle. According to the fluorescence intensity and area of Coptis chinensis adventitious roots, the result showed that berberine mainly accumulate in the xylem, and rest of parenchyma distribute small except secondary phloem and cambium almost have no fluorescence. At the xylem of rhizomes and through the cortex section of adventitious roots, secondary phloem and cambium maybe have no berberine, the other all have a little berberine. And the vascular bunds of petiole and sclerenchyma all distribute, the other structure have little berberine distribute. Therefore, berberine accumulated focused on the xylem of Coptis chinensis roots and rhizomes, and vascular bundles and sclerenchyma of petioles, these structures were important targets for evaluation quality.In short, this paper argues that primary structure of adventitious roots composed of epidermis, exodermis, cortex, endedermis and vascular cylinder. secondary structure composed of vascular cylinder and phellem. The anatomical structure of hypocotyl is epidermis, cortex, exodermis and xylem. Primary structure of rhizomes composed of cuticle, cortex and vascular cylinder; secondary structure composed of phellem, cortex and vascular cylinder,and mainly with cortex and vascular cylinder. The apoplastic barriers in adventitious roots of Coptis chinensis at primary structure is casparian bands in endodermis, slight lignification and strong suberin, casparian bands in exodermis, lignification and suberin. The apoplastic barriers in adventitious roots at secondary structure is lignification and suberin in phellem. The barrier structure of hypocotyl is cuticle, casparian bands in endodermis, slight lignification and suberin.Combined with the reported of Lu Gaolian, the ionic state of berberine in parenchyma cell and Hg-KI form crystals. Parenchyma is the synthesis of berberine. Berberine mainly accumulated in the xylem. So the parenchyma and xylem of Coptis chinensis in various parts all is the structure of berberine synthesis and accumulation. Rapid detection about the distribution of berberine under the bule light, it is worth to determine the quality of Coptis chinensis for future application.The earlier development of barrier structure of Coptis chinensis adventitious root tips, limited the water and ions absorption. Berberine deposed in the primary xylem, berberine may jamed the primary xylem, and prevented the upward transportation of water and ion to the result in lack of available water and ions of plant. Combined with the reported of Yuan Wangjun, the leaf of Coptis chinensis is isolateral, there were no histological differentiation in the internal structure of leaves. These characteristics may be the important reason for Coptis chinensis is not resistant to high temperature, strong light and adapted to darkness environment. The result of the present work have a great significance on how to improve the soil mois ture in promoting the Coptis chinensis production and as well as the quality of Coptis chinensis determination via a fast and easy method to its popularization and application.