AFLP Detection On MtDNA Difference Of3Alloplasmic Cms Lines With Isonuclear And Its Maintainer In Wheat

With the rapid growth of the population, food crisis become more and more serious. Inorder to resolve this situation, crop heterosis utilization becomes more and more important,cytoplasmic male sterility (CMS) is the basis of heterosis in crops, people use this advantage,when they do the hybrid male sterility work, can save a lot of manpower and time so that theycan crop the combination with strong heterosis breeding. At the same time,people has done alot of research on the male sterile mechanism, the results show that, the mitochondrial DNA isclosely related with CMS.But in the cytoplasmic male sterility, it is difficult to determine the cytoplasmic genomedifferences are caused by the cytoplasmic genome itself or the nucleus cytoplasm effects.dueto the presence of nuclear-cytoplasmic interaction phenomena, Therefore, it is difficult todetermine the origin and nature of male sterile cytoplasm. if people does not select specializedresearch. In order to solve this problem, our team cultivate a number of isonuclear alloplasmicmale sterile wheat materials, these materials cross dozens of generations, they have the samenuclear genetic backgrounds. Therefore, it can be completely ruled out nuclear effects whendo the research on male sterility, The difference can be considered the essential differencefrom the cytoplasmic genome.if people use this as the tested material, This study is based onthis, we select three cases of isonuclear alloplasmic male sterile wheat and its correspondingmaintainer, and find the difference analysis on the mtDNA by way of the AFLP molecularmarkers.We can identify the different types and different male sterile cytoplasm sterilecytoplasm restoring and maintaining characteristics, in order to improve the nucleoplasminteraction type male sterile types of actual effectiveness, the important results are as follows:(1) The experiment use the method of density gradient centrifugation for mtDNAextraction,and the purity is very high,it can achieve the experiment requirements of AFLP.(2) We select5pairs of primer combinations from64primer combinations: E7/M21,E10/M16, E12/M18, E13/M18, E13/M20,they have the better polymorphism; we do it againand get the same result.(3) In the E7/M21primer combinations, there is an obvious difference in the location of 200bp; in the E12/M18primer combinations, there is an obvious difference in the location of250bp; in the E13/M20primer combinations, there is an obvious difference in the location of400bp,with these bands; we can easily identify these Isonuclear alloplasmic wheat malesmaterial.(4) The3cases of isonuclear alloplasmic male sterile wheat has many differencesthrough the primers,and its maintainer of mitochondrial mtDNA exist obvious polymorphism,these bands also illustrated tested Isonuclear alloplasmic wheat mitochondrial genome existsdifference, it can be used as a study and utilization of Heterosis in wheat used to distinguishdifferent cytoplasmic male sterile type of stable molecular marker.