Assessing genetic diversity of native lily in Matsu using RAPD and rDNA ITS molecular markers
Date Issued
2010
Date
2010
Author(s)
Chien, Wan-Ting
Abstract
In order to establish genetic relationship of native lily in Matsu, Random Amplified Polymorphic DNA（RAPD）markers and rDNA Internal Transcribed Spacer（ITS）were analyzed. In RAPD analysis, 10 of 60 primers screened to provide polymorphic and reproducible bands. A total of 35 strong polymorphic bands were scored for 25 individuals of native lily collected in Matsu (21), Kinmen (3) and Guangxi China (1), and 15 individuals of outgroup. Principle coordinate analysis and unweighted pair group method with arithmetic mean（UPGMA）cluster analysis based on these RAPD profiles were performed. A clear distinction was found between the native lily in Matsu and outgroups. The results of analysis of molecular variance（AMOVA）of native lily in Matsu and Kinmen population indicated that 13％ of the total variation was attributable to the differences among population while 87％ was due to variation among individuals within population.
The results of direct sequencing of PCR products and sequencing by cloning PCR products of the ITS and the 5.8S coding regions of nuclear ribosomal DNA in native lily in Matsu, L. longiflorum and L. formosanum were compared and phylogenetically analyzed. The length of ITS region was 626 to 627 bp in native lily in Matsu, including 229 bp of ITS1, 164 bp of 5.8S rRNA gene, and 233 to 234 bp of ITS2. The GC content was about 55.49%~66.09%. Variable sites mainly occurred in the ITS1 and ITS2. A dendrogram generated by neighbor-joining（NJ）and UPGMA cluster analysis. According to the dendrogram, two main clusters were generated: (1) native lily in Matsu and Kinmen, (2) L. longiflorum and L. formosanum. In ITS sequence analysis, direct sequencing data analyzed by UPGMA method would be more reasonable than NJ method, while the NJ method could get a better interpretation with cloning sequencing data.
The results showed an explicit line of demarcation between native lily in Matsu and outgroup. Because the native lily in Matsu and other Lilium brownii were classified in the same group, we concluded that native lily in Matsu should be L. brownii. Owing to the sample number of L. brownii were too few, it needs further study to confirm that the native lily in Matsu is really unique.
Subjects
Matsu
Lilium brownii
genetic diversity
Molecular marker
RAPD (Random amplified polymorphic DNA)
ITS (Internal transcribed spacer)
Type
thesis
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