RAPD Analysis On Cultivars Or Lines Of Loquat And Offspring Group Of "DaWuxing"

In this study , variety identification and pedigree analysis were researched on 8 loquat cultivars or lines which cultured in Sichuan. Genetic diversity of 60 individuals of two groups was also analysised. The results were as follows:1. Genomic DNA of 68 individuals was successfully extracted by improved CTAB method. DNA quality was high, and 0D260/0D280 of DNA was around 1.80. The DNA without getting rid of RNA can be used for RAPD amplification.2.6 primers which could be used for stable amplification were selected from 80 random primers.3. The suitable PCR system for loquat were found: 25μl reaction volume with 2.5μl 10×PCR buffer, 2.5pμlMg²⁺, 0.25μl Taq polymerase, 0.5μl dNTP, 2.5μl primer, 15.75μl ddH₂0, 1ul template DNA.4.PCR thermo-cycle procedure used as follow: 94℃ 5min,94℃ 1 min, 38 ℃ 1 min, 72℃ 1 min, 40 cycling, final extension at 72℃ 5 min.5. 6 primers could be successfully used for amplifying the DNA of 8 loquat cultivars or lines and showing polymorphism. 54 bands were obtained, among which 26 bands showed polymorphism, amounting to 48. 15%. The genetic distance between Zaozhong No. 6 and Jiefangzhong was smallest, only 0.042, similar index was 95. 8%; the genetic distance between Chuannong No. 1 and Zaozhong No. 6 was biggest, 0. 217, similar index was only 78.3%.6. The DNA fingerprints of 8 cultivars or lines were established. The specific bands of some cultivars or lines were found out.7. All of 8 cultivars or lines could be identified by DNA fingerprints and the specific bands. Genetic distances and cluster analysis were also...