| Rhizoma alismatis is firstly recorded in "Shen Nong’s Herbal Classic" and is listed in the top grade. According to Pharmacopoeia of China (2010), Rhizoma alismatis is the dried tuber of Alisma orientale (Sam.) Juzep which belongs to alismataceae flora. It has the functions of clearing damp and promoting diuresis, purging heat, melting turbidity and reducing lipid. Rhizoma alismatis is one Chinese herbal medicine, widely used in clinical treatment and Chinese herbal drug production. Shown by the modern pharmacology that rhizoma alismatis has function of diuresis, anti-atherosclerosis, hypolipidemic effect, hypoglycemic effect etc. In order to ensure the high quality of the medical material, this thesis focuses on the kinship and genetic diversity of medical materials in different producing areas, making basis for the breeding of improved varieties; it also makes research on the selection of excellent rhizoma alismatis variety, sets standards for the seeds and germchit of rhizoma alismatis. The results are shown as follows:1. Kinship and Genetic Diversity of Rhizoma Alismatis in Different Producing Areas(1) The results of research on the Chinese materia medica have shown that there are six alismataceae flora recorded in ancient Chinese materia medica, among which four belong to waterplantain, one does not blong to this genus, one can not be classified because of a rough recordation. Two of the four categories which belong to waterplantain show no difference to Alisma orientale (Sam.) Juzep. Recorded in Chinese pharmacopeia (2010). The other two are Alisma nanum D. F. Cui and Alisma canaliculatum A. Braun et Bouche. This indicates that in the ancient times there existed confusion among the varieties of rhizoma alismatis, and one herbal medicine had more than one origin. Even in the ancient times it was thought that rhizoma alismatis were mainly produced in Sichuan and Fujian with a better quality.(2) According to the study of Fertility characteristics and plant morphology, rhizoma alismatis in Sichuan is the first to sprout, the growth and development period of blooming and bearing is shorter than that in other producing areas. Rhizoma alismatis in different producing areas has no visible difference in plant morphology, so it is very difficult to tell which producing area the rhizoma alismatis come from by its appearance. Leaves of rhizoma alismatis in Sichuan mostly arrange in three beams neatly, while rhizoma alismatis in other producing areas has no such distinguished characteristic or only has a low proportion.(3) A study of rhizoma alismatis’HPLC fingerprint makes clear that rhizoma alismatis in four producing areas has eleven common peaks. After the identifying of characteristic peaks with alismoxide, alisol A24-acetate and alisol B23-acetate, it turns out that the peaks are the second, the fifth and the eighth. Provenance sample of rhizoma alismatis in the same producing area has a high degree of similarity, there is only a slight differene among the average value of similarity of provenance sample in different producing areas, but on significant difference in terms of statistics. So it is obvious that HPLC fingerprints of rhizoma alismatis in different producing areas is similar to each other and the structures of chemical elements are almost the same. The difference in the average value of similarity of rhizoma alismatis in different producing materials is mainy caused by the difference of each composition. The selection of improved variety has little influence on the chemical elements composition of rhizoma alismatis.(4) Study of ISSR molecular marker shows that with the selected thirty-five ISSR primers to amplify twenty-three rhizoma alismatis samples, the total percentage of polymorphic loci is70.5%, Nei’s genetic diversity (He) is0.4196, Shannon’s information index is0.6081, which indicate that the genetic diversity of rhizoma alismatis in different producing areas is good. The23samples can be divided into two categories if we have a marking line at similarity degree of0.5349. There are21samples in the first group including all the main origins of Alisma Orientalis. The second category is composed of2collected samples, referring to the one sample in Shuiji town, Jjianyang city, Fujian province and the other in Tong’an town, yongtai county, Fuzhou city, Fujian province, which indicates that the genetic distance of these two samples are relatively closer. Furthermore, the first group can further considered as two large groups, among which Guangxi, Sichuan, Fujian, Jiangxi province belong to the first group while Sichuan and Fujian provinces pertain to the second group. Therefore, there is neither obvious genetic differentiation between different areas nor apparent regional difference in the Alisma Orientalis. And the mutual circulation between the germplasm resources may probably lead to the phenomenon of less obvious genetic regional difference among various areas. The combined features of Alisma Orientalis’s high genetic variety and low genetic distinction provides favorable conditions for the rearing of superior Alisma Orientalis.2. The Study on the Breeding of the Superior Alisma Orientalis(1) The Alisma Orientalis yield determination shows that the plant growing is obviously relevant to the amount of yielding. The rate of three-prismatic plants is closely related with the final production (that is, P<0.01). Those plants with three-prismatic state are more likely to have bigger stems. Hence the conclusion can be made that three-prismatic is of great value to the superior Alisma Orientalis screening and breeding. In comparison of the collected samples, the yielding of the plants from Sichuan province is relatively high whereas those in Fujian, Guangxi and Jiangxi province are varied in their growing state with lower and distinct production. If we change into the superior seeds and cultivate them in the second year, each sample grows better than the previous year. According to the results of variance analysis, the F1generation seed is obviously marked from those of the first year (P<0.05). Among them, the most striking is the turnout is1.5times more than the previous year and the seeds from Sichuan province is nearly1.1-1.2times more. Accordingly, the usage of superior seeds is beneficial for the enhancement of production.(2) The chemical elements determinations indicate that all the samples of Alisma Orientalis contain alismoxide,24-acetyl Alisol,23-acetyl Alisol, but different samples are varied in their number of each elements. The fine variety breeding does not affect their chemical elements and the rate of each element does not show obvious similar pattern. Even if the seeds are from the same province, planted in the same location with exactly the same method, they will also show difference in their single chemical elements container and the rate of each element.(3) The total content determination of the active ingredients shows that the samples from Sichuan province attain the highest total content of the effective ingredients per unit area, which is obviously higher than the samples from other origins. It is likely that the seeds of Alisma Orientalis from Sichuan Province have their own advantages in product itself and in better ecological adaptability. After the F1generation seeds are planted, the effective elements of each sample per hectare increase apparently(P<0.05). The most marked sample has increased2.1times than the previous seeds, and the less remarkable one is about1.2times. In conclusion, the screening and breeding of the superior seeds has had great impact on the enhancing the quality of Alisma Orientalis. (4) The screening of superior Alisma Orientalis reveals that there are great differences between three prismatic plants and the non-three prismatic plants in their average size of the stems(P<0.01). The three prismatic plants obtain better advantage among other varieties. It is evident that the total chemical contents are distinct between before-screening Alisma Orientalis and after ones (P<0.01). The superior plants is inclined to have1.2times more total contents in active chemical ingredients. And the second year plants’quality will improve markedly than the previous year, making it clear that the superior plants have the capability to be improved.3. The Standards Building for the Quality of Seeds and Sprouts(1) The determining factors of seeds quality shows that net weight, thousand-grain weight, germination percentage and seed viability difference of the collected seeds is larger, and the F1seeds are smaller. The F1generation seeds possess higher thousand-grain weight, germination percentage and seed viability than those seeds collected from the market (P<0.01). The late-maturing seeds is better than the early-maturing seeds (P<0.01). The linear-regression analysis shows that the weight of one-thousand seeds, the germination percentage and vitality share a positive linear relationship with each other (r>r0.01). The weight of one-thousand seeds is an important evaluation indictor of the assessment of the quality of Alisma Orientalis seeds. So, The standards for evaluating the Alisma Orientalis seeds are as follows:the water percentage≤13%, seed purity≥90%, the weight of one-thousand seeds≥0.3700g, the germination percentage≥80.0%, vitality≥86.0%.(2) The standard for assessing the quality of cultivars indicates that if people breed the cultivars according to the cultivation techniques, then the health rate, the height, the number of the leaves and the length of the root of the cultivars is relatively stable. The total content of effective ingredients, the health rate, the height of a single Alisma Orientalis present positive relationships with each other (r>r0.01). The health percentage and the height of cultivars are of great significance to the assessment of the quality of Alisma Orientalis cultivars. So, The Standard for judging the quality of a Alisma Orientalis cultivar are as the following:the health rate of the cultivars≥93%;the height of the cultivars≥16cm;6p≤the number of the leaves on the cultivars≤8p. |
PDF Full Text Request