CDNA-AFLP Analysis Of Restorer Genes Of K-cytoplasmic Male Sterile(CMS) In Wheat

Utilization of heterosis is one of the important ways to further explore thegenetic potential of wheat yield in order to break the bottleneck of wheat yield.Wheat K-type sterile line with small side effect of cytoplasm, a wide range ofmaintainer and restorer resources and other characteristics, is a nucleo-cytoplasmicinteraction male sterile type which extremely has the application prospect. But it alsohas a defect with lower and unstable fertility restoration, which also directly restrictsthe application in production of the K-type sterile line. In this study,cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis wasemployed to identify the difference expression of restorer gene between K-typesterile line Yumai3and NILs of restorer genes on2BL with the background ofYumai3during their dicaryophase of anther development. We investigated themolecular mechanism of fertility restoration in the cell signal transduction andtransmission, gene transcription and regulation, and receive the following mainresearch results:1. Optimized cDNA-AFLP analysis was employed to identify gene expressionprofile of K-type sterile line Yumai3and its maintainer line and two NILs of restorergenes BC9F2during their dicaryophase of anther development. After the120pairs ofprimer screened, the results showed that10757transcript-derived fragments (TDFs)existed between the materials, and53(4.9%) discrepant TDFs was detected by26pairs of primer. After large scale cloning and sequencing, eventually35differenceTDFs obtained, and all of them were extended by electronic method. By means ofBlastx comparison and functional classification analysis,23TDFs of them havesequence homology with NCBI, and their function mainly relates to cell and tissuemetabolism(37.1%), transcription factor(8.6%), Stress corresponding(5.7%),transcription promoting(5.7%), regulation of gene expression(5.7%) and mixedfunction (29%);3(8.6%)TDFs are highly homology with unknown function protein while9TDFs(25.7%) do not show significant matches to any genes in theGenedatabase. they may be new genes.2. The function of the35difference TDFs relates to many aspects. The results notonly showed the availability of cDNA-AFLP technology in researching target genes,but also indicated that the recovery mechanism of wheat K-type male sterile was acomprehensive, complex process controlled by a variety of physiological andbiochemical process.2difference TDFs (P1M3-1and P1M3-3) are involved in thesynthesis of indole acetic acid and proline metabolism, which can make the antherdevelopment proceeded normally. They most likely play a key role in fertilityrestoration, and have great application potential. In addition, some TDFs are relatedto cytochrome, calmodulin, and wheat mitochondrial DNA, and their roles need tobe further studied.