The AFLP And SCAR Detection On Mitochondrial DNA Of The Male Sterile Lines With Aegilops Kotschyi Cytoplasm In Wheat

Wheat is considered one of the important food crops in China, the research andutilization of heterosis is of great significance to the improvement of wheat quality and yield.Cytoplasmic male sterility (CMS) is a key way of crop heterosis utilization and also its mainpollination control system. Therefore, research on molecular mechanism of cytoplasm malesterility (CMS) for creating, identification and utilization of cytoplasmic male sterility (CMS)system has important guiding significance to cultivate the super-high-yield hybrid new crops.Wheat cytoplasmic male sterility (CMS) is a genetic phenomenon of nuclear andcytoplasmic interactions, while the present study showed that mitochondrial genome is themain carrier of cytoplasmic male sterile factor. Thus, wheat cytoplasmic male sterility (CMS)and mitochondria has a very close relationship, the variability of the mitochondrial genome islikely to involve the change in the nature of the male sterility. The analysis of mitochondrialgenome is of great significance in revealing the molecular mechanism of cytoplasm malesterility (CMS).This research utilized three kinds of wheat cytoplasmic male sterile lines which with thecytoplasm of Aegilops kotschyi, Ae. ventricosa and Triticum spelta as materials, studied theAFLP markers and convert it into SCAR marker on mitochondrial DNA in several kinds ofwheat male sterile lines which obtained the following important results:(1)Using the differences between the prolamine protein in these three types of theAegilops kotschyi cytoplasmic male-sterile lines to identify the material consistency ofgenetic background. Result shows that the genetic similarity coefficient is1in three types ofcytoplasmic male sterile lines of wheat varieties which repeated between individual，andgenetic similarity coefficient between sterility type is also1. That means the savedexperimental materials are highly homozygous in seed purity which through more than25generation backcross, while the genetic similarity is high between them and between parents,which nuclear genetic background is quite consistent.(2)This experiment used the combination of64pairs of primers to do the amplification intested materials which contained isonuclear alloplasmic male sterile line and its maintainerline in wheat, of which13primers showed polymorphism amplification.682bands were amplified out in total, including113polymorphic bands.(3)AFLP primers combination E8/M5only be amplified specific bands in Aegilopskotschyi types of wheat cytoplasmic male sterile lines (Kots)-90-110ms and ms (Kots)-8222, which the fragment size is about300bp.(4)Based on the result of AFLP markers, specific fragments were collected andsequenced and a SCAR marker YW2was successfully designed and validated. One specificfragment was amplified by YW2in ms (Kots)-90-110and ms(Kots)-8222which showedsequence similarity as high as99%. This SCAR marker can be used in molecularmarker-assisted breeding for identifying wheat male sterile lines with Aegilops kotschyicytoplasm and and provided theoretical and technical support for identifying cytoplasmictypes of the wheat.