Analysis Of Genetic Diversity Of Pyrola Calliantha By AFLP And SRAP Markers

Pyrola calliantha belongs to Pyrolaceae family, it is a wild and evergreen plant in the north. Pyrola calliantha hasthe characteristics of the evergreen, cold resistance, shade resistance and flowering in summer, so it is a goodgardening and greening material in the northern area. The distribution ranges of Pyrola calliantha are from GreaterKhingan to Lesser Khingan in the province of Heilongjiang. It has a wide range of applications in the medicine area,food area and chemical area. In order to use and protect the wide resource of Pyrola calliantha preferably,54materialswere collected and used as experiment materials from11areas of Heilongjiang province.Two molecular markers: AFLP and SRAP were used to study the genetic diversity, the genetic distance among thepopulations and clustering analysis and to discuss the genetic relationship among the materials from differentgeographical sources. It lays the foundations of introduction cultivation, landscaping and the protection of wildresource. The main results are as follows:1. There are plentiful phenols and polysaccharose. The improved CTAB method can extract high-quality DNA,the high-quality DNAaccord with AFLP and SRAP.2.The SRAP reaction system of Pyrola calliantha were optimized by the method of orthogonal experimentaldesign.The reaction system were showed:1.2U Taq DNA polymerase,1.5mmol·L^-1Mg²⁺,0.4umol·L^-1primer,0.4mmol·L^-1dNTPs,75ng template DNAand2.0μl10×PCR buffer, the reaction system was20ul.3.The8pairs of primers were screened out from64pairs of primers in the AFLP.228bands were detected by the8pairs of primers, and175bands were polymorphism bands.28.5bands were detected by average each pair ofprimers, the polymorphism ratio was76.75%. The genetic background was complicated among different genotype ofPyrola calliantha populations. The primer combination of E-ACG/M-CTG had the higher detection ratio. It wasshowed that the genetic distance range among the populations was0.0372⁰.3053by the analysis of AFLP molecularmarker.4. The3pairs of primers were screened out from16pairs of primers in the SRAP.75bands were detected by the3pairs of primers, and61bands were polymorphism bands.25bands were detected by average each pair of primers,the polymorphism ratio was81.33%.The primer combination of Me2Em3detected the most bands.28bands weredetected and23band were polymorphism bands, the polymorphism ratio was82.14%. It was showed that the geneticdistance range among the populations was0.0134⁰.2978by the analysis of SRAP molecular marker.5.The genetic diversity parameters values in different populations were showed that the AFLP analysis: theaverage value of Nei’s gene diversity index H=0.1384and the average value of Shannon information index I=0.2025;the SRAP analysis: the average value of Nei’s gene diversity index H=0.1239and the average value of Shannoninformation index I=0.1634, the values tended to accordance from the two molecular marker. Mantel test was used to detect the relativity between AFLP and SRAP. The result showed that the two molecular markers had prominentrelativity（r=0.8425, p=0.0120<0.05）.And the most value of the Nei’s gene diversity index and Shannon informationindex is Huzhong population in theAFLP and SRAP.6.The comprehensive analysis of AFLP and SRAP showed that the range of genetic distance was0.0329⁰.2664, the range of genetic similarity was0.7661⁰.9677in the Pyrola calliantha populations. Theclustering analysis results showed that there was further genetic relationship between Maoer mountain polulation andother populations.The clustered populations had the nearer geographical distribution. These datas shows that therewere relativity between genetic distance and geographical distribution. In order to analyze the genetic diversity ofPyrola calliantha populations better, the method of comprehensive datas of the two molecular markers to analyzecould obtain more detection locus and responsed the genetic relationship of Pyrola calliantha more comprehensive.