Radish Physiological And Biochemical Characterization Of Male Sterility Line And Its Maintainer Line And SSR Markers

Radish hybrids have obvious advantages,using hybrid generation of F1as production canimprove radish quality,disease resistance and yield.In order to improve hybrids purity,malesterle line breeding hybrids is the most econmically and effectively way to sove artificialemasculation.In order to expand the scope of application of radish male sterile line in theproduction,using the male sterility of new germ plasm of radish as test materilals,measuring thephysiological and biochemical indices of leaves,flowering buds and flowers in differentperiods,so that we can further explore the physiological and biochemical mechanisms of radishmale sterile and provide a theoretical basis for the breeding of radish male sterile.EST-SSR expressed sequence tag microsatellites is a new type of molecular markers basedon the design of EST simple sequence repeats. It solve the cumbersome problems of needing tobuild a genomic DNA library during the development process of genomic SSR and have beenwidely used in plant genetic mapping, evaluation of genetic diversity, comparative mappingand the identification of the genetic relationship. As the origin place of radish （Raphanussativus L.）, radish germ plasm resources are abundant in China. Lacking the radishbiotechnology research and the domestic application of SSR markers in radish has not yet beenreported, many important traits could not be effectively developed. Studying the factorsaffecting radish SSR amplification reaction to establish the best reaction system and screeningthe sterile line for SSR can mark important traits of radish for the SSR, and thus lay thefoundation for further research and utilization of Chinese radish germ plasm resources.In this experiment, using radish male sterile line and its maintainer line as test materials,determination the content of free proline, soluble protein, soluble sugar, malondialdehyde andsuperoxide anion, as well as the activity of SOD, POD and CAT in leaves, flowers and flowerbuds of different developmental stages. The results showed the contents of proline, solubleprotein and soluble sugar in the buds and flowers of male sterile lines were lower than themaintainer lines during flowering period, malondialdehyde and superoxide anion is higherthan the maintainer lines and the enzymatic activities of SOD, POD and CAT were relatedwith radish male sterile; using the DNA of the male sterile radish lines and fertile lines as testmaterial and orthogonal design L16to optimize the five factors （Taq DNA polymerase, Mg2+, the template DNA, dNTP, primers） of SSR-PCR reaction system of radish on4level andget the best of the reaction system, in a20ul reaction system,0.10mmol/L of dNTP,3. 0mmol/LMg²⁺, the50ng templates,0.500μmol/L,Primers,1the.0UTaq enzyme. Using theoptimal reaction system, the amplified bands were clear, can meet the material requirementsof different primer combinations and radish. This study lay the foundation for the use of SSRtechnology to classify radish germ plasm resources, construct genetic map and map gene inthe future. SSR analysis at the same time, the total of300SSR primers were screened, ofwhich five pairs of primers have difference in performance between the two pools and can befurther sequencing and screening.