Studuty On Pollen Viability, Pollen Morphology And Cell Culture Of Forsythia Suspensa

Identification and screening of germplasm resources are not only the base of breed cultivate, but also the key to the modernization and the sustainable development of traditional Chinese medicine. Cell culture of modern biological technology is an effective mean for reasonable protection, exploitation, utilization of medicinal plants. Forsythiaside A was the main active ingredient and quality evaluation indicators of Forsythia suspensa.In this experiment, the traditional Chinese medicine was employed as the research object. Three staining methods (Riphenyl tetrazolium chloride staining method, iodide dyeing method, aceto carmine dyeing method) were applied to determine the pollen viability of Forsythia suspensa. Pollen morphology of 8 kinds of Forsythia suspensa were observed by using scanning electron microscope (SEM). Forsythia fruit, the medicine organ of Forsythia suspensa, was taken as explants to conduct cell suspension culture indicated by the biosynthesis of forsythiaside A and the growth level of cell.Our experiment results showed that:1% aceto carmine dyeing method was the best method for Forsythia suspensa’s pollen viability detection. The pollen viability of two stamens type forsythia suspensa was higher than three stamens type. The pollen viability of short style type Forsythia suspensa was higher than long style type when the number of stamens was same.The shape of 8 kinds of Forsythia suspensa pollen were subellipsoid, ellipsoidal or long ellipsoid. The size of reticulate sculpture on pollen exine was similar, and there were three colpate, with symmetrical shape also. The 8 kinds pollen above belong to smaller pollen, and there were some differences on pollen morphology(size and mesh ridge), which would provide an evidence for analyzing the phylogenetic relationship of Forsythia suspensa.The optimum scheme of explant sterilization of the green fruit of Forsythia suspensa was in 0.3% HgCl2 for 10 Minute. The optimum medium for the green fruit of Forsythia suspensa callus was MS culture medium, added 2.0 mg/L 2,4-D,0.5 mg/L 6-BA,0.2 mg/L KT and 3.0% sucrose. The optimum medium for cell suspension culture of Forsythia suspensa was B5 culture medium, added 2.0 mg/L 2,4-D,0.5 mg/L 6-BA,0.5 mg/L KT and 3.0% sucrose. The medium which the relative proportions of NH4+ and NO3- was 0/60 was conducive to the growth of suspension cell. The optimal medium for forsythiaside A accumulation was B5 culture medium added 0.7 mg/L KT,1.5 mg/L 2,4-D and 3.0% sucrose. The medium was conducive to the growth of suspension cell when the pH of medium ranged from 4.8 to 5.3. The kinetic analysis of cell growth and nutrient consumption during Forsythia suspensa suspension culture was performed, and an S-shape cell growth curve was showed. The pH of the medium was 4.1～7.0 during the cell culture process, and the cell growth curve was in accordance with the consumption of carbon, nitrogen and phosphorus.The experimental results provide an experimental basis for the application of cell culture technique in producing useful components of Forsythia suspensa.