| Tobacco(Nicotiana tabacum L) is a solanaceous plant of the genus Nicotiana, who is a quite important economic crop over the world. Meanwhile, tobacco is widely used as a model plant in several related filed in molecular biology and life science. SNP (single nucleotide polymorphism) is a emerging molecular marker method with the advantages of high density, good accuracy and simple experiment, which make it turn into a hotspot in current research. This experiment, aiming at the drawback of the existing SNP prediction software, have developed a plant SNP predicting platform. Through the platform, there have researched the known tobacco EST sequences and developed a large number of SNP loci. Also, it have done the experimental verification by choosing SNP site and designing primers, as well as done the phylogeny of species identification. The main results are as follows:1. Plant SNP prediction platform CsSNP(http://biodb.sdau.edu.cn/cssnp/index.html.) was developed by using 60 plants as contrast.2. Based on 10 kinds of tobacco EST sequences,20,607,973 potential SNP loci were developed. Their abundance was between 67 SNPs/KB (N. tabacum) and 256 SNPs/KB (N. suaveolens x tabacum), with an average abundance of 87.57 SNPs/KB. statistics of 12 types of SNP showed that SNP is widely present in the genome. Statistical results indicated that the percentage of substitutions(59.22%) was greater than that of the transversion (40.78%) between bases.3. Designing primers based on SNP site, and predicting after amplification in 15 tobacco types. Alignment and cluster analysis of predicted sequences can distinguish related cultivars of tobacco.4. The first tobacco database NSNP(http://biodb.sdau.edu.cn/nsnp/) was built based on the acquired information in tobacco. |
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