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Mapping the Male Sterility Trait Obtained from Space Flight in Maize with Molecular Markers

Author: LiuHeYang
Tutor: CaoMoJu
School: Sichuan Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: maize space flight genetic male sterility(GMS) molecular marker gene mapping
CLC: S513
Type: Master's thesis
Year: 2013
Downloads: 37
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Abstract


It is well known that maize male sterility could be applied in hybrid seed production. So more and more attention were paid to this character. A male sterile materials was found by Maize Research Institute of Sichuan Agricultural University from the offspring of ChuanDan NO.9hybrid seeds flighted in space in1996,. The primary research indicated that the male sterile tait was governed by a single recessive gene. Based on F2segregation population, the male sterile gene was located within SSR markers umc2076and umcl674with3.7cM and4.4cM respectively on maize chromosome NO.3.In this experiment,the F2population was constructed by the cross of male sterile mutant and Mo17.The SSR markers were used for locating the male sterile gene, then built genetic linkage map on the region of GMS gene. The main results were concluded as follows:1. The F2population of A×Mo17were planted with totally14400, in which the number of sterile plants is3481, and the number of fertile plants is10919. Chi-square analysis showed that:the Xc2=5.20was small thanX(0.01,1)2=6.63. It demonstated that the fertility segregation fit to3:1Mendelian ratio, which suggested the GMS gene was controlled by a single recessive gene.2. According to the preliminary gene mapping results, five fertile plants DNA and five sterile plants DNA were selected randomly from the F2population as a template,and the molecular markers umcl674and umc2076were verified. The results showed that no polymorphism were found between male fertile plants and male sterile plants with the primers umc1674and umc2076.3. According to the BAC sequences between umc2076and umc1674which published by maize genetics database (http://www.maizegdb.org), microsatellite search software SSR Hunter was applied to search SSR loci of BAC and the sequences were compared with NCBI database sequence alignment to search for the single copy SSR loci, then use the software primer5.0to design28pairs of primers.3fertile plants DNA and3sterile plants DNA were selected randomly from F2population as a template, PCR was performed on28pairs of primers, and only two pairs of primers were not amplified bands, the efficiency of primer design is92.8%. Four of the rest26pairs of primers have differences between fertile and sterile materials. They were primer AC195817.3-3, primer AC194425.2-12, primer AC204515.3-10and primer AC204626.2-1. It was found that four pairs of primers were not closely linked to genic male sterility gene in further linkage genetic analysis.4. Owing to no SSR primers were found to closely link to the GMS gene. The F2population of A×Mo17was used as mapping population. According to the sterility investigation in field, fertile plants and sterile plants were selected randomly to screen170pairs of SSR primers in maize chromosome3and to conduct male sterility gene mapping.22of170pairs of SSR primers were found to be polymorphic primers. It was found that the SSR markers umc1974、 umc2050、umc1148、mmc0071、umc2276and umc1594were linked with the GMS gene in linkage analysis. The male sterile gene was located between SSR markers mmc0071and umc2276in maize chromosome3L with7.1cM and4.7cM respectively.5. Through screening for the plant which recombination took place between mmc0071and object GMS gene or between umc2276and object GMS gene, then bulked for resequencing, based on the bioninformation blast analysis to explore new molecular markers for fine mapping.In this study, The F2population of A×Mo17was treated as location population. The F2population included3481sterile plants. The results show that the SSR markers umc1974、 umc2050、umc1148、mmc0071、umc2276and umc1594were linked with the GMS gene. The male sterile gene was located between SSR markers mmc0071and umc2276with7.1cM and4.7cM genetic distance respectively in maize chromosome3L. With the two mapping population from A×S37or from A×Mo17, the target GMS gene were both located on maize chromosome

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