Study On The Identification Of Germplasm Resources By Molecular Marker And The Breeding Of Hybridization In Hypsizigus Marmoreus

Hypsizigus marmoreus is one of the rare Edible-medicinal fungus in China andforeign countries, and it is also an important Factory promotion cultivation product.With the rapid development of Hypsizigus marmoreus industry, the demand forbreeds which have better productive efficiency has became more and more urgent. Inthis dissertation, molecular marker technology and protoplasted monokaryontechnique were deployed to select hybrid breeds. Using fruiting experiments andmolecular marker validation, the genetic differences between hybrid strains andparental strains were identified. Hybrid breeds with excellent productive efficiencywere successful selected.The experimental results are as follows:1,Characterization of Hypsizigus marmoreus germplasm resources andhybrid parents.15RAPD primers,5ISSR primers and10SRAP primer werescreened and used to analyze16Hypsizigus marmoreus strains by RAPD, ISSR,SRAP technology. Using NTSYSpc2.1, the width of genetic similarity of theseHypsizigus marmoreus was calculated as0.566to0.922. The genetic width of anytwo of these strain was0.078to0.434and the average genetic similarity was0.738.The result of Cluster analysis indicated16strains had different degree inpolymerization when the GS was between0.68and0.93. When GS was0.73, thesestrains could be divided into6Groups: Group1: SM bai yu,Bai yu,19Cï¼›Group2:2A,H9600,18C,â…¢M1,Hai xian,Xie wei,H9602; Group3: H01; Group4: I4CF1,I2CF1,H515; Group5: Hui xie; Group6: Hui bai. Based on Cluster analysis andconsideration of the productive efficiency of tested strains, it was showed that Bai yuand Hui xie had potential for scaled production in factory. Therefore, Bai yu and Huixie was selected as the parents in the hybrid experiment.2,The breeding of Hybridization.2monokaryotic strains of Baiyu and13monokaryotic strain of Hui xie were regenerated by protoplast fusion and culture, andthe monocyte obtaining rate was relatively4.1%and9.56%.12strains of hybridstrains were obtained by Hybrid breeding. Obvious mushroom body difference between Bai13-49,Bai55-25,Bai55-4,Bai55-51,Bai55-6and their parentalstrains were found during the fruiting experiments. The pileus was Light yellow, anddiameter was intermediate between their parents. The stipe was yellow, and the stipelength was longer than their parents, but the diameter was intermediate between theirparents. The cultivation period (73d) was shorter than their parents (80d). And higherfruitbody yield indicated a marked hybrid successful. All these experiments suggestedusing Protoplast Monokaryogenesis Method in Hybrid breeding has a wideapplication prospects.3,The validation of the hybrid strains using molecular marker.5HybridStrains with fine productive efficiency were tested with RAPD in order to assess theirperformance in molecular level. The result of PCR amplification map indicated thesestrains were hybrid strains, and they have genetic differences between their parentalstrains. The similarity between these5strains and Bai yu was80.3%, and thesimilarity between these5strains and Hui xie was relatively83.5%,79.3%,81.0%,81.0%,79.4%. It also indicated that Bai55-4,Bai55-51,Bai55-6were the samestrain. After all, this study obtained3new strains with excellent productive efficiency.