Apple Pear Hybrid Offspring Per Plant By Rapd Analysis

In this study, Hongjinqiu,Yanguangli, some mainly cultivars of pears and 21 F₁ hybrids ofHongjinqiu×Pingguoli (Hongjinqiu×Pingguoli'21 F₁ hybrids shorted NO.1～NO.21) were used asmaterials, analysis 27 samples by RAPD technique, researched variety identification and classifystation of them. The result indicated that:An improved CTAB method could extracted high-quality DNA from gene group ofPingguoli and the degree of purity is the highest, there are many degrdn fragments in the methodSDS-CTAB; pick-up rate of DNA is not high, through significance variance analysis, useingipv-SDS method can get more DNA from each kind of pear, but the purity is very low; themethod which high-salt and low-pH get lowest yield and purity, integrality is not good. Animproved CTAB method could extracted high-quality assemble DNA from Pingguoli filialgeneration genome in this experiment. It is suitable for analysis of RAPD amplification, and totalseventeen 10-mer random primer which polymorphism is more than 76.07 percent, defmited themost optimal RAPD-PCR reaction system of Pingguoli filial generation: DNA follow board 135ng/μL, in 25ul reaction system, primer concentration 15pmol,dNTP concentration 1.5 mmol/L,Mg²⁺ concentration 1.5 mmol/L and Taq DNA polymerase concentration 1.0 U. the rest wassupplied with ddH₂O.Amplification procedure: 94℃pre-denaturing 5min, 94℃denaturing 1min, 40℃annealtemperature 1min, 72℃extension 1.5min,35cycles; 72℃final extension 5 min.Establishment twenty varieties' finger printing. Get four samples characteristic marker.twenty samples in twenty-seven samples can use duplicate bands of one primer to bediscriminated from others.The progeny hybridized by Apple-pear and other varieties were proceeded in RAPDamplification. after cluster analyses, all researched samples were categorized 5 kinds in the termof origins, which was taken the genetic distance(0.30) as genetic distance, according to thegenetic distance. The genetic distance of 27 samples was between 0.10～0.46.The first group istotal 19 Progeny hybridized of Pingguoli: No.2,No.3,No.5～No.21; Hong jinqiu,No.1,No.4,Pingguoli as second group; it shows that the genetic relationship of No.1,No.4 andPingguoli is the closest. They inherit more gene from Pingguoli. Other progeny hybridized inheritmore gene from Hong jinqiu; Hong jinqiu is the progeny hybridized of big floral water andPingguoli, the genetic relationship of big floral water and the first breed is more closet, show thatHong jinqiu'gene from more Pingguoli. the third group is big floral water; small floral water asone group; the fifth group include Yanguang pear and Dongning No5.