| In this experiment the adult stem nodes and seed embryos were used as materials for micropropagation and further callus culture and suspension culture, and the related anatomic obersavation was carried out in Ardisia crenata in Fujian; besides, RAPD was used for genetic diversity analysis and cluster analysis on the samples from Gushan Mountain's wild population and Wuping's seedling population.The main results were described as follows:1．In vitro propagation in Ardisia crenata Sims.(AG), Ardisia crenata Sims var. bicolor(AP) and the intermediary type (AGR) in FujianThe results showed that the appropriate medium for initiating culture was 1/2 MS or WPM medium supplemented with 1～5mg·L-1 BA, 0．1 mg·L-1 NAA and 0．2～0．5mg·L-1 GA for AG, 1/2MS medium supplemented with 1～5mg·L-1 BA, 0．1 mg·L-1 NAA and 0．2 mg·L-1 GA for AP, and 1/2 MS medium supplemented with 1mg·L-1 BA, 0．1 mg·L-1 NAA and 0．2～0．5mg·L-1 GA for AGR. The shoots proliferated on the MS medium supplemented with 3mg·L-1 KT, 0．01mg·L-1 NAA and 0．5mg·L-1 GA, and grew stronger and rooted on the multiplication media with the reduced mineral concentration. Raising auxin concentrations and reducing cytokinins concentrations properly rooted better.2．Embryo culture from seeds of two maturity degrees and callus culture from different parts of the tube piantiets derived from embryo cultureThe results showed that the callus was easier induced from more mature embryos then less ones. The callus could be induced on the MS medium supplemented with 0．5mg·L-1 2,4-D and 0．01mg·L-1 KT from every part of the piantiets derived from embryo culture. Among them, the radicle had the highest callus inducing rate, and the callus induced from the radicle was white and gibraltar-like, which could be well subcultured and maintained on the MS medium with 0．5mg·L-1 2,4-D and 0．01 mg·L-1 KT, supplemented with 5 mg·L -1 AgNO3 every two generation. After long-time culture of the callus on the different media, the other four kinds of calli,i.e., 1) the white, wet and soft callus , 2) the yellow and granular callus , 3) the kelly and nubbly callus , 4) the green and nubbly callus, were obtained.3．Root differentiation from the callusThe results showed that the fascicular root could be induced form the callus cultured on the 1/2 MS medium supplemented with 3% sugar without growth regulators. On the MS medium supplemented with 0．1mg·L-1 2,4-D and 5．0mg·L-1 KT, adding 100mg·L-1 inositol,4%sugar,3% mannitol and 5%CM, the highest root differentiation rate occurred, but it took a relatively long time. The weak light was beneficial to rooting by comparison. 4．Cell suspension culture from the callusFive kinds of calli (â… -the white, wet and soft callus,â…¡-the white gibraltar-like callus,â…¢-the yellow and granular callus,â…£-the kelly and nubbly callus,â…¤-the green and nubbly callus) were used to establish cell suspensions in the MS liquid medium supplemented with 0．5mg·L-1 2,4-D and 0．01mg·L-1 KT. The results showed that the calli ofâ…¡andâ…¢were more suitable for the suspension culture. Sucrose was the best carbon source for the suspension culture from the callus in Ardisia crcnata.5．Observation on paraffin sections of the abnormal plantlets with swelling shoot apical meristem (SAM) and the different kinds of calli in Ardisia crenataThe results showed that the structure of SAM in abnormal plantlets was different from the normal ones. In the natural environment, the periphery of strumae SAM was make up of many layers of suberized cells, and the mitosis of the cell inner SAM was frequent, the plantlet didn't grow. Under the in vitro condition, the cells of strumae SAM mitosed fleetly and abnormally, but there were not any suberized cells in the periphery of strumae SAM, so there was no structure differentiation. Besides, the different kinds of calli had the characteristics of gradual changes on their cell structure, and the root differentiation from calli belonged to exogenesis.6．RAPD analysis of the genetic diversity of 23 accessions of Ardisia crenata germplasm from Gushan Mountain's wild populationThe results indicated that in Gushan Mountain population, the number of effective alleles (Ne) was 1．4233, the Nei's gene diversity (H) was 0．2524, the average number of alleles (Na) was 1．8060, Shannon's index of diversity(I) was 0．3852．These data showed that the genetic diversity was hgih in the GuShan Mountain wild population. Besides, the genetic proximity was high in 23 accessions of Ardisia crenata germplasm, which suggested that they should be hgihly stable in heredity.7．Cluster analysis of 15 accessions of Ardisia crenata germplasm in the seedling population in Wuping ConutyThe results showed that when the combination distance was 0．598, the 15 accessions of Ardisia crcnata germplasm could be divided into 3 types: 1) the green leaves, 2) the green leaves with red sprout, and 3) the leaves with the purple side. Among the 3 types, the green leaves with red sprout was more close to the green leaves than to the leaves with the purple side. When the combination distance was 0．668, the 15 accessions of Ardisia crenata germplasm could be divided into 2 types: the green leaves and the leaves with the purple side. The above results were identical with the result of morphological classification. |
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