| To study on phylogenetic relationship of wild species and cultived varieties in Chrysanthemum, some chrysanthemum cultivars and wild species were collected from China and the Japan. The methods of morphological analysis , ISSR and RAPD markers were used.The main results are as follows:1. In morphological analysis, the 48 morphological characteristics of the experimental materials were studied. The coefficient of variation of data consistency and variability of the quantitative traits were tested, and the contribution rate was detected by principal component analysis. And then, 40 taxonomic characters were selected by character R-clustering, and Q-type cluster analysis was studied.The results showed that C. chanetii had closest relationship with one small flower cultivar'Hongxuancao'(ground covered chrysanthemum), and wild species were closer relationship with the small flower chrysanthemum, and the clustering resultes were directly related to the petal type in large flower cultivars .2. In ISSR analysis, we have comparatively analyzed eleven wild species and twelve cultivars by using ISSR-PCR technology in order to make clear the genetic relationships and diversities among the wild species and cultivars of Chrysanthemum. We acquired 14 ISSR primers screened from tested 75 ISSR primers to identify the polymorphism among the tested materials. The results showed that 142 distinguishable bands were amplified that accounted for 95.1% polymorphism. The effective numbers of alleles (Ne), Nei's gene diversity (H) and shannon's Information index (I) in wild species were higher than that of tested cultivars, which illustrated that there were obvious gene differences in wild species, and higher polymorphism than that of cultivars. UPGMA cluster analysis indicated that the wild species had a evolutional tendencies from low ploidy to high ploidy and also had a complicated genetic relationships among the cultivars. Based on the evolutional relationships, we may have a conclusion that the flat petal type of Chrysanthemum might be the basic type. The germplasm of C. nankingense has the closest genetic relationships with all tested cultivars, and the three of C. chanetii, C. japonicum, or C. japonicum var. wakasaense have secondly close relatives with tested cultivars, C. indicum var. aromaticum has genetic relationship far from other tested varieties.3. In RAPD analysis, 10 wild species and 12 cultivars of Chrysanthemum. were used for analyzing genetic diversity and relationship by RAPD-PCR technology. The fourteen primers screened from 50 random primers, could generate 169 fragments, of which 163 ones (96.4%) were polymorphic, showed high polymorphism in Chrysanthemum. As analyzed by POPgene32, the similarity coefficient between cultivars ranged from 0.195 4 to 0.565 6. The average value of effective number of alleles, Nei's gene diversity and Shannon's informa-tion index were 1.515 8, 0.321 6 and 0.479 4. The number of polymorphic bands, the average value of effective number of alleles, Nei's gene diversity and Shannon's information index in the wild species were higher than the cultivars, showed that genetic diversity in the wild species was larger than the cultivars.The data were analyzed and clustered by using UPGMA method by NTsys 2.0. The results indicated that, the cultivars were most relatively close to C. japonicum var. wakasaense, secondly, C.indicum and C. chanetii. In the cultivars, the genetic relationship were basically related to their inflorescence size.4. The above results showed that culvited chrysanthemum was a very close relationship with the wild species of C. nankingense, C. japonicum var. wakasaense, C. chanetii, C. lavandulifolium, especially the farest to C. indicum var. aromaticum. The small flower chrysanthemum was closer relationship with the wild species, and then, the petal type and the inflorescence size could be used for classification of cultived chrysanthemum.These results indicated several wild species could be involved in origin of cultivated chrysanthemum. |
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