Studies On The Anatomy Structure Of Vegetative Organs In Polygonum Cuspidatun And Its Callus And Histochemical Localization Of Resveratrol

Polygonum cuspidatum Sie b.et Zucc, a traditional Chinese medicinal plant, is a kind of perennial herb or inferior arbuscular. It widely distributes in the district of Liaoning, Shanxi, Gansu, Sichuan, Guizhou, Yunnan, the east China and mid-south China, and deserves research and exploitation. Recent researches of polygonum cuspidatum Sie b.et Zucc on secondary metabolites and bioactive compounds have suggested that the content of resveratrol in polygonum cuspidatum Sie b.et Zucc. is the highest. Based on the systematic study on resveratrol,and by means of plant anatomy and phytochemistry, some rules have been explored by the combination of some basic studies as anatomy structure of vegetative organs, redifferentiation of explants, structure of callus, redifferentiation of organs and histochemical localization of resveratrol, in order to provide basis for the introduction and acclimatization of Polygonum cuspidatun Sieb.et Zucc, cultivate new varieties, and exploitation and industrialization of medicinal ingredients.The main results are as follows:1. The Polygonaceae plant Polygonum cuspidatun Sieb.et Zucc, the structures of the rhizomes in terrestrial stem and in rhizome are different. The rhizomes in terrestrial stem is not surrounded by lepises, the rhizomes in rhizome is surrounded by lepises,there are much starch and cluster in the cells. Adventitious roots , terrestrial stem, irhizome have primary growth and secondary growth.Their stucteres are similar to the root and stalk of dualcotyledon herb and have their characteristics . The xylem of the root is tetrarch, lateral root initiate the in thexylem radiating angle opposite to pericycle cell. Adventitious roots and rhizome have much periderm, the suppository level is high; Fibers in the vegetative organs root and stalk are abundant. There are rich pith cavity in the terrestrial stem and parenchyma cells constitute pith in rhizome. Leaves in the growth period is of diversity,and there are not only nail non-glandular hairs on it but also capitate glandular hairs that was new discovered. It was first discovered and reported that there exist extrafloral nectary in petiole basal portion. 2. Through observation of cytohistology in vitro culture of Polygonum cuspidatum Sie b.et Zucc.explants,the callus in the stem part is from two ways:the first is by cell dedifferentiation of parenchyma cells in epidermal and cortex,and the second is by cell dedifferentiation of basic tissue of vascular tissue around incisions. Young callus contains mainly embryogenic cell , embryogenic cell-aggregates and meristem nodules, mature callus, subculture callus and multiple culture callus contains mainly tracheid, vascular tissue nodules and parenchyma cells,and were surrounded by callus periderm with plenty of suppositories yellow substances.By slice comparison of the leaf by in vitro culture and the stem in vitro culture,it was found that the differences are great because of the difference of their morphology.The bud of the callus of Polygonum cuspidatum Sie b.et Zucc, differentiated mainly by the external reasons.3. Based on overall and systematical studies on the anatomy structure of vegetative organs and the callus in Polygonum cuspidatun Sie b.et Zucc.,the histochemical localization of resveratrol was researched. Taking account of the intensity of color reaction,growth development and enviromental conditions,a convenient and reliable histochemical localization method of resveratrol was established. Treated by AlCl₃ and H₂O, observed with fluorescent microscope,it was found that resveratrol in Polygonum cuspidatun Sie b.et Zucc.distribute mainly in cell wall, phloem fiber, wood-fiber, catheter and corkification phellem cells that are highly lignified and fibrosis.Resveratrol histochemical localization in mature callus make it possible to culture and produce resveratrol in batches from Polygonum cuspidatun Sie b.et Zucc, callus.4. The results by HPLC quantitative detection validation are basically in consistence with that by the fluorescent microscope observation. The content of resveratrol in Polygonum cuspidatun Sie b.et Zucc.was xylem > phloem> periderm.There was obvious difference in the content of resveratrol in mature callus from different explants, the content of resveratrol in callus from rhizome bud is the highest (34.60μg·g^-1), the lower orderly is from leaf(7.08μg·g^-1), from stem phloem(3.60μg·g^-1) and from root(3.01μg·g^-1).This result has set a base for the establishment of the system for screening and culture of high-yield cells of Polygonum cuspidatun Sie b.et Zucc.5. In the paper,a new ideas was proposed on morphological features (abundant of rhizome,stong, short internode and dense eustipes) that are helpful to exploit and utilize Polygonum cuspidatun Sie b.et Zucc.and to culture new germplasm(both high-yield of Polygonum cuspidatun Sie b.et Zucc, and high-content of resveratrol).