Construction Of Fingerprinting For Armeniaca Cathayana Based On SSR Molecular Markers

Armeniaca cathayana D. L. Fu et al., one of grain and oil woody plant in China, is a newspecies published in2010, with high ecological, economic and social value. The status ofhybrid varieties and homonym is widespread, and morphological marker is difficult todistinguish the abundant Armeniaca cathayana germplasm resources. Plant DNA molecularmarker technology could detect the plant genome directly without affection and restriction byenvironment and developmental stages during plant growth, therefore, the identificationmethod by DNA molecular markers is more scientific, accurate and convenient thanmorphological markers. In this study, the SSR molecular marker was used to develop theprimers of EST-SSR Marker of Armeniaca cathayana, to construct the DNA fingerprinting,and to analyze the genetic diversity. The results were as follows:1. In this paper, we used Armeniaca cathayana resources to optimize the microsatelliteDNA gelred fluorescence imaging technology. Results showed that bands of PG in AGE wereclearer and brighter and its background was cleaner, which suggested that PG was the bestmethod in AGE. In addition, NaCl could enhance banding brightness. In PAGE, IG was betterthan SS and PG. Results of the orthogonal design experiment indicated that immersion timesignificantly affected banding effect, the longer the brighter in this experiment. The bestdispose of combination was2X Gelred,10%NaCl and dyeing for40min.2. We used EST sequences from the database of dbEST to develop EST-SSR markers ofArmeniaca cathayana by using SSRHunter1.3software. The results showed that22527SSRsfrom8610non-redundancy sequences were obtained, which the frequency of occurrence was4.26%. In the search process, there were only two repeat types which was dinucleotide andtrinucleotide. Of the61designed EST-SSRs primers,24yielded clear amplified products withthe effective rate of39.34%, and10of them revealed polymorphism, indicating a polymorphicrate of16.39%. 3. Totally138alleles were detected and a DNA fingerprinting of Armeniaca cathayanaresources was constructed with the amplified data from63samples by using31pairs ofselected primers, Through analysing31primers’ polymorphism with heterozygosity, PIC,Shannon et al., BPPCT030and aprigms18were the best primers of identification ability, whilePchgms2was the worst.4. The genetic diversity of63Armeniaca resources were analysed by UPGMA. Theresults showed that:(1) All the tested samples could be distinguished at similarity coefficient of0.95.(2)63samples of Armeniaca were divided into four groups at similarity coefficient of0.65, in which group2was composed most of Armeniaca cathayana. Genetic relationship of’Xinsihao’ and ’Youyi’ was closer,’N34’ and ’N35’ was closer. Group1included Armeniacadasycarpa (Ehrh.)Borkh,’Yuxing’ and four samples of Armeniaca cathayana, group3included Armeniaca sibirica (L.)Lam and ’11L09’, group4was ’11X01’.