| Radix Polygalae is the dried root of Polygala tenuifolia Willd. or P. sibirica L It has a wide clinical use for its effection of calm the nerves, beneficial intellect, expectorant, detumescence, antiinflammatory and antimutagenesis. This essay contains four research sections:channels of high yield, quality assessment, genetic diversity, gene cloning of squalene epoxidase partial cds which is the key enzyme of biological synthesis of polygalacic acid.1. Ways of high yieldStudy out the standard of storage period of polygala tenuifolia seed and thousand kernel weight according to 1000-grain weight and germination rate:for the firsr-class seeds, length of storage≤2 years,1000-grain weight≥2.711g; for the second-class seeds, length of storage≤4 years,1000-grain weight≥2.411g; the third-class seeds are in poor quality.For the first time, establish the yield regression model with D-saturation design between the yield and application amount of N, P2O5, K2O:Y=5121+36.113X1+ 150.557X2+71.157X3-439.15X12-321.447X22-168.478X32-91.306X1X2-28.206X1X3+8.238X2X3. The sequence of yield effection of nitrogen, phosphor, kalium is:N (X1)> P(X2)> K(X3). The sequence of interaction effection is:NP> PK > NK.117.128kg nitrogen,120.333 kg phosphor, and 136.257 kg/hm2 kalium are in need if the prospect polygala tenuifolia yield is 5146.172 kg/hm2. and the proportion is 1:1.03:1.16.Carry out the water and fertilizer coupling experiment with pot culture in rainprotection shed, under the low water condition with less than 14% water in soil, early fertilizer (anthesis) is comparesively better than late fertilizer, and low nitrogen is better than high nitrogen; under the high water condition with 24% water in soil, add nitrogen during the period of seedling establishment and anthesis, the yield is very significant difference for other groups. Polygala tenuifolia has a narrow tolerance range of nitrogen, and has a better tolerance in early growth, and it is in great demanding of water and nitrogen during anthesis.Study on water demanding rule and pest contorl shows polygala tenuifolia is in highest yield if immgrated during bloom stage. The control effect of rogor and matrine are significant difference from that of derris alkali in aphid control, while rogor and matrine are no significant differences. Joint use of biological pesticide trichoderma preparation and chemical pesticide of tuzet has more significant effection than respective use in prevention and cure of root rot disease.For the first time, carry out the systematic study on growth dynamics of Polygala tenuifolia in 2007～2008, and the results indicates that the diameter of main steam or fresh root, fresh root/dried root proportion, weight of fresh root, weight of dry root, weight of dry steam and leave, biological yield increased along with the prolong of growth period. Branch number or rootlet number in 2007 was bigger than that in 2008. The height of plant changed in parabola, and higher in 2008 than in 2007. The fastest monthly increasing period of dry root weight was between 20th July and 20th August 2007, and between 27th June to 27th July 2008. The total accumulation of dry matters in root in 2008 was higher than in 2007.2 Quality assessment of Radix PolygalaeThe experimental results of HPLC fingerprint on Polygala tenuifolia shows: fingerprints are in high similarity among 23 batch radixes of Polygala tenuifolia, and only slight differences are observed between wild and cultivated species; There is similar structure in HPLC fingerprints of cultivated species for different grow periods and collection month, growth time has few effects on the types of active matters; the fingerprints of root xylem are similar to those of Radix Polygalae, while the peak areas are comparatively low, so the content is low, and the fingerprint similarity of root xylem is above 0.807 between the cultivated and the wild.The experimental results of effect matters of polygalic acid shows:producing area influences the content of polygalic acid significantly. Altidude and dimensionality have some effect on the content of polygalic acid. There are higher content of polygala tenuifolia in the different month collected polygala tenuifolia in 2008 than that in the same month in 2007. Polygalic acid is highly interrelated with growth years and the collected period, but may be not interrelated with the developmental stages. It is considered better to collection in June or July after polygala tenuifolia reaches its growth years. Polygalic acid can be found in root heart of polygala tenuifolia, and its accumulation may be negatively correlated with drugs. Leaves of polygala tenuifolia contain poor quantity of polygalic acid except P. sibirical L has slightly higher.3. Study on genetic diversity of germplasm resourcesFor the first time, with AFLP, capillary electrophoresis technique and allozyme Analysis, genetic diversity is studied in the ways of DNA, protein, allozyme on Polygala tenuifolia Willd. or P. sibirica L.Carry out amplified fragment length polymorphism (AFLP) method to study on the DNA genetic polymorphism,310 clear and repeatable gene bands are amplified. Among those bands,261 are polymorphism bands, accounting for 84.19%, and 21.75 allelic variation are found in each locus, while effective ones account for 1.49. The genetic identity of 24 polygala populations is between 0.4000～0.8645, and the genetic distance is between 0.1456～0.9163, which indicates that the genentic diversity is abundant. The result of AFLP experiment is in accordance with classical morphology result, and also the geographical distribution.The experimental results of protein genetic diversity by capillary electrophoresis show that the similarity coefficient of germplasm resources for polygala tenuifolia is between 0.316～0.895, and the genetic diversity is full. The protein genetic diversity reflected that the resources similarity is higher and genetic relationship is closer for species in closer geographical position. The cultivation species, however, the protein traits are in greater difference. Cultivated polygala tenuifolia in Shanxi has higher similarity with wild ones in Sichuan Maoxian and Gansu. The result of protein genetic diversity research is not all in accordance with the result of genetic diversity study.The outcome of soluble protein and allozyme shows the bands of peroxidase with mobility of 0.136 and the bands of esterase with mobility of 0.818 can identify Polygala tenuifolia Willd. and P. sibirica L., while the bands of peroxidase with mobility of 0.492 can identify polygala tenuifolia origins from Shanxi Qinyuan and other eight species. The electrophoresis band polymorphisms are between 0.198～0.444, and the genetic identity of 9 species is between 0.3-0.9. The genetic identity between Polygala tenuifolia Willd. and P. sibirica L. is lower than that among the populations of Polygala tenuifolia Willd, and the genetic distance is greater. The similarity of soluble protein and allozyme is accordance with peographical distribution in certain level.4. Study on cloning of squalene epoxidase geneFirstly clone the DNA gene of squalene epoxidase with 5 degenerate primer, and obtain a nucleotide with 517bp. After identification of Blastx, the fragment of the nucleotide belongs to SE gene fragment. According to the conjecture of Zea mays with full cDNA gene sequence (Accession:BT042800), the cloned SE fragment has 1426bp distance to 5'end, while 110bp to 3'end. At cloned part, the length of Zea mays is 23bp longer than SE gene fragment.
|
PDF Full Text Request