Preliminary Study On Germplasm Resources Of Cyathula Officinalis Kuan.

Cyathula Officinalis Kuan. is a kind of plantation belongs to Amaranthaceae , Cyathula Blume; it is commonly called TianXi,which is one of the perennial plant species. Cyathula Officinalis is mainly distributed in Sichuan, Yunnan, and Guizhou Provinces, either wild or cultivated. Cyathula Officinalis grows in areas which is beyond 1500 meters altitude. Take the example of the productions in the resourceful districts such as Tianquan, Baoxing, Hanyuan and Leshan in Ya'an, Sichuan Province, are all ranking the biggest production rates. Among those districts, the most famous quality of Cyathula Officinalis is from Tianquan. The main components of Cyathula Officinalis are cyasterone,isocyasterone and capitasterone,etc. The major pharmacological functions of Cyathula Officinalis are activating blood to promote menstruation and curing rheumatism. In Tianquan production area, Cyathula Officinalis is facing the problems in species downgrading, quality decreasing, and even fake productions, because of the long term process in cultivation, naturally hybridize, which is leading the variance in this species.This research based on Cyathula Officinalis' variety in a wild range of resource in Cyathula Officinalis production areas. It aims to protect and fully utilize Cyathula Officinalis,to find higher production and better quality Cyathula Officinalis. Meanwhile, this research results in several useful methods in distinguishing authentic and fake Cyathula Officinalis.1.Variance analysis was used in the research according to the morphological index of 16 different soureces Cyathula Officinalis Kuan., the result showed that the morphological index according to plant height, stem diameter, branch number, root length, root diameter, number of split-root, 2nd leaf length, 3^rd leaf length, 2^nd leaf width, 3^rd leaf width, root weight are fit for botany characters observation and recordation of Cyathula Officinalis Kuan.. The study of comparative morphology indicated that the numeric taxonomy basically revealed the heritable differences among the varieties, and the research method canbe operation and the result is dependable, these studies have a important instruction meaning to seed selecting of Cyathula Officinalis Kuan..2.The study shows that the collection of germplasm material Cyathula Officinalis Kuan. chromosome numbers are 34, diploid, The karyotype formulas are diverse, the most long arm:the shortest arm (L:S) ratio in the 1.49-4.31, karyotype asymmetry coefficient between 61.89-67.59, a rich diversity of chromosome morphology. Study found that the long arm:the shortest arm (L:S) ratio is significant difference in two ecotypes(Bai NiuXi and Hong NiuXi), According to the significant difference, the 11 collected materials are divided into three categories: No. 1,2,3,4,5,7,8,9,11 (Bai NiuXi) between the range of 1.10-3.00; and No. 6, 10 (Hong NiuXi) (L:S) ratio are greater than 3.01.3. According to the PCR technical specificity, 19 rDNA ITS sequences of Cyathula Officinalis were under determination. The results of the sequences were compared to two categories. By doing the research in homology in Genbank, we found that No. 17 and 18 materials have the same sequence as the ITS of Achyranthes bidentata. The other 17 materials have the same or similar sequence as ITS sequences of Cyathula Officinalis. The comparisons of those sequence structure also indicated that ITS1-5.8S-ITS2 sequence length of Cyathula Officinalis and Achyranthes bidentata is 586-590bp, by deducting 5.8S's 162bp sequence length, ITSI+ITS2's length is 424-428bp after the right sequencing. ITS sequences of Cyathula Officinalis and Achyranthes bidentata have very obvious difference. However, based on different habitat field, different cultivation methods, different species, The germplasm resources of Cyathula Officinalis have no obvious difference in their sequences. This fact demonstrated that we can distinguish Cyathula Officinalis and imitated Achyranthes bidentata,but we cannot effectively distinguish Cyathula Officinalis (Bai NiuXi) and Hong NiuXi.17 Cyathula Officinalis marker distinguishing Chuan niuxi and Red niuxi. genetic differentiation is minor, it might result in the analyzable system classify of ITS sequence.4. Applied Mark RAPD analyzed 19 genetic diversity of Cyathula Officinalis. The results indicated that 23 10-mer arbitrary primers expanded 419 RAPD locus, among which there are 377 (90%) diversified locus; UPGMA analysis indicated 19 materials' analysis. 19 materials could be attributed to two main categories and two minor categories. RAPD mark indicated that the germplasm resources of Cyathula Officinalis did have genetic diversity in molecular level. By utilizing RAPD mark, not only we can solve problems in Cyathula Officinalis cultivated miscellany ,but also can use as an effective tool to build DNA fingerprint of Cyathula Officinalis.5.Based on the preparation of RAPD fingerprints for Cyathula Officinalis and its adulterant species, we can find an unique RAPD polymorphic band F300,F500 in order to sequence its order. Then we can design the specificity by sequencing and conduct unique DNA fragment's PCR expanding. By recycling, cloning, sequencing, a more steady SCAR mark can be converted, and the obtained SC-320 SCAR mark will distinguish the merchandised Cyathula Officinalis (Bai NiuXi and Hong NiuXi) in Cyathula Officinalis and Achyranthes bidentata, more quickly and more steadily. In this way, a new method to distinguish Cyathula Officinalis and its confusions by using SCAR mark was established.According to what we discussed above, the research of Cyathula Officinalis, The germplasm resources of Cyathula Officinalis in cytology and molecular level will help to understand its genetic diversity and authentic mechanism. This method established a solid foundation in cultivation of improved variety of seed, new product seed selecting, purifying fakes. Meanwhile, The results of this study in the morphology provides the main characteristics of indicators for the future breeding ,cytology study found that the ratio of the long arm of the nuclear type: the shortest arm,it can be used as the basis for the distinction between red and white bidentata identification;we can utilize RAPD molecular mark to detect higher level diversity, in order to build an useful tool in establishing the germplasm resources of Cyathula Officinalis DNA fingerprints. Also there is a new method to distinguish Cyathula Officinalis and its confusions by using conservatism of ITS sequence and transformed SCAR mark.