Studies On Innovative Technique Of Germplasm Resources In Saposhnikovia Divaricata

Saposhnikovia divaricata (Miq.) Schischk. is a perennial herbaceous plant of Saposhnikovia in Umbelliferae, a common traditional Chinese herb distributed in northeast, northwest and north of China and officially listed in the pharmacopoeia of the People's Republic of China and their unbolting roots can be used as medicines. The amount of natural S. divaricata decreased quickly due to non-plan and over-diggings, which has further aggravated the destruction of S. divaricata resources. Therefore,the study that the polyploid breeding of Saposhnikovia divaricate and the creation of the Germplasm Resources of autotetraploid lay the material and technical foundation for the further selection and breeding of improved varieties which are of high yields and contents rich in medicinal ingredients.With the employment of modern biological technology, with the amphiploid seeds of Radix Saposhnikoviae, the stem segments and callus of the test-tube seedlings as experimental materials,with the colchicine as the mutagen to double the chromosome,the changes of variant strains on morphology, cytology and other aspects can be investigated,the technology system which were appropriate for the doubling, purification and identification of chromosome can be establised so that we can artificially induce the autotetraploid plants of Radix Saposhnikoviae in vitro.1.The callus of Saposhnikovia divaricate was induced by the petioles, stems and leaves of Saposhnikovia divaricate. Saposhnikovia divaricate had the highest induction rate, which reached 98.00%.The tissues of Saposhnikovia divaricate grew rapidly, their density was high, their colour was green,their volume was large. This may be because their physical state and growth stages they stay at were different, and their concentration and ratio of endogenous hormones were different.2.By orthogonal test design,the most appropriate conditions for the induction of the callus of Saposhnikovia divaricate were investigated, taking the leaves of Saposhnikovia divaricate as explants and taking the kinds of culture medium, sucrose concentration, pH, plant hormone 2,4-D,6-BA concentration as factors. The results showed that the effects of sucrose concentration on the callus induction rate of Saposhnikovia divaricate were barely obvious, that the kinds of culture medium had the greatest impact on the callus induction rate of Saposhnikovia divaricate, and that several other factors also had rather obvious effects. The effects of the primary and secondary factors on the callus induction rate of Saposhnikovia divaricate were A (culture mediums),D (2,4-D),C (pH),E (6-BA),B(sucrose).The results were that the explants were the leaves of Saposhnikovia divaricate, the culture mediums were MS, the concentratin of sucrose was 25g/L,Ph 6.0, the concentratin of 2,4-D was 1.0 mg/L, the concentratin of 6-BA was 0.5mg/L.3. Using different concentrations of colchicine and treatment time, we studied the mutation rate of the seeds, stems and callus of Saposhnikovia divaricate. The smear method had no significant effects on the mutation rate of the different explants. It didn't work when smearing colchicine on the explants because of the poor absorption. Although we could gain some polyploids by soaking, the dosage of colchicine was too much, its toxic effects were most direct, and at the same time some explants died because of the lack of oxygen.—had a high requirement on the culture medium and colchicine because it had a heavy workload, and its absorption was poor. The injection had the best effects, it may be because the explants were in contact with colchicine fully, it had the strongest effect, so it reached the highest mutation rate. Using injection, the dosage of colchicine was less, the method was simple, the workload was small. Under the same condition, the callus induction had the highest survive rate and mutation rate among these three kinds of explants. Maybe this was related to their growing and physiological stages. When inducing the polyploidy, it was best to use 0.2% colchicine injecting the callus, and its mutation rate was 47%.4. The effects of NAA compared with riboflavin on the rooting of Saposhnikovia divaricate. The results were that with the rising concentration of NAA and riboflavin the rooting rate was significantly increased. But the concentration of NAA was higher than 0.5mg/L, the concentration of riboflavin was higher than 1.0mg/ L, the rooting rate was significantly decreased. It explained that NAA and riboflavin had an inhibited role on the rooting of Saposhnikovia divaricate. When using 1/2MS and 1.0mg/L riboflavin to induce the rooting of Saposhnikovia divaricate, the rooting rate was highest that reached 88.89%.5. According to the comparasion with the diploid of Saposhnikovia divaricate, after inducing its root system turned bold, its leaf color turned deeper, the leaves turned larger, the stem diameter turned thicker. After the determination of the chromosome number of the root tip cells,2n=4X=32, we can determine that it is tetraploid Saposhnikovia divaricate.