General Application Analysis Of Nuclear And Chloroplast Microsatellite Derived From Prunus Mume On Stone Fruit Trees And Application On Genetic Diversity Of Prunus Mume

Stone fruit trees are important fruit trees among China and world,which include peach plum,mume,apricot and cherry.Prunus mume originated from China,it is one of important export fruit tree in the south of China.This paper studied orthogonal design in order to establish SSR reaction system in Fruiting-mei.Some nSSR and cpSSR markers from Prunus mume were screened for the transferability to stone fruit trees.Nssr and cpSSR markers were studied for genetic diversity of Prunus mume.1.Using Fruiting-mei 'hong-nong'as material,this paper studied orthogonal design for five factors including Mg²⁺,Taq polymerase,primer,template DNA and dNTPs at four content levels in order to establish SSR reaction system in Japanese apricot.The results showed that 1.875 mmol·L-1 Mg²⁺,0.375μmol·L^-1 primer,0.125 mmol·L^-1 dNTPs,0.75 U Taq polymerase,15 to 30 ng template DNA are the optimal conditions in the reaction of 20μL volume.The optimal annealing temperature of primer pair D-07 is 50.0℃.2.17 nSSR and 13 cpSSR Markers from Prunus mume were screened for the transferability to 24 cultivars of stone fruit trees.There were 13 nSSR and 13 cpSSR primer pairs with effective amplification.The useful rate of nSSR and cpSSR markers were 76.5%and100%.The average polymer phism information content(PIC) of nSSR and cpSSR markers were 0.591 and 0.654.They can be as effective molecular markers for studying on genome,plant genetics and corp breeding of stone fruit trees.These results showed that Prunus mume nSSR and cpSSR primers can be used well in stone fruit trees.3.The genetic diversity of 29 cultivars of Prunus mume were evaluated by 9 pairs of nSSR primers and 9 pairs of cpSSR primers.93 alleles were amplified,of which average 5.2 alleles per pair of primer were amplified and polymorphic percentage of amplified band was 83.3%-100%.Based on the SSR results,calculation of the similarity coefficient and also UPGMA cluster analysis showed that the Prunus mume cultivars could be classified into the three groups.Genetic relationship of some cultivars was close to which came nearly or bade the same type.