Identification Of Molecular Markers Linked To Pollen Sterility Gene And Convertion To SCAR Marker In Peach

Pollen sterility character of peach takes as a significant value which weighs fruit output, in search of molecular markers which tightly linked to pollen sterility gene,as it has significance of peach breeding.In this research,molecular markers linked to Pollen sterility character of peach were studied with RAPD and AFLP technology in 32 F₁ polulation hybrided from Chong yanghong peach and Da jiubao peach.The main results were as follows:Through detecting ,genome DNA was extracted by improved CTAB method which was suitable for analysis of RAPD, AFLP. RAPD reaction system of peach was Optimized:Total volume was 25 μl,including of 40ng template DNA, 2.0mM MgCl₂, 150μM dNTP, 2.5μl 10× buffer, 1U Taq DNA polymerase, 20pmol Primer.the rest was supplied with ddH₂O.Adopting BSA method,sample DNA of 5 individuals were selected from F₁ polulation mixed equally and consisted of gene pool of pollen fertility and sterility.Utilizing 180 RAPD primers, two linked RAPD markers (OPW03-900 and OPW03-1300) were screened, the genetic distance which linked to pollen sterility of peach were 6.46cM and 15.02cM respectively.16 primer combinations which represented polymorphism in pollen steriliy were screened from 96 AFLP primer combinations,through identification of F₁ polulation,the results indicate: there were no linkage relation between most fragments and Ps/ps,fragment was segregate from Ps/s which only primer E+AT/M+CTT amplified (400bp) .But recombination was too high(28.1%) and the linkage distance was 32.04cM.Conversion OPW03-900 of RAPD which linked to pollen sterility gene to SCW03-906 of SCAR marker.This sequence was adopted by Genebank,and the logging number is DQ659676. PCR anneal temperature of SCAR was gropped which linked to Ps/ps,the amplification was carried out: 94℃ pre-denaturing 5 min; 94℃ denaturing 45s, 68℃ annealing 45s, 72℃ extension 1 min, 35cycles; 72℃ final extension 5 min; 4℃ terminating. Through identification of F₁ polulation and varieties,SCAR marker which linked to pollen sterility has been transformed successfully. It can be used as an identification marker of pollen sterility in peach.