Research On The Selection Of Primary Trisomics From Antherculture Of Wild Compatible Tetraploid Rice Of Pedigree Seed And Identification Of Primary Trisomics Via Molecular Marker

Five pedigree seed varieties of wild compatible tetraploid rice were used in this study of anther culture. In order to improve the regeneration efficiency, some factors such as sugar, agar concentration, basic media, and hormone on callus inducement were studied. Pollen plants were selected as primary trisomics by cytological karyotype analysis of the pachtene. The difficulties in karyotype analysis are commoning recognized due to the small size of somatic chromosome of rice. So in present study, a new method for identifying primary trisomics by RFLP marker was tried. The results are as follows:1.The difference of genotypes in varieties is one of the important factors which influence the rate of callus inducement and plant production. The average rate of callus inducement of wild compatible tetraploid rice is higher than that of diploid rice.2.Agar and sugar concentration are other influencing factors in dedifferentiation. In the rate of callus inducement of sucrose, 60g/L is higher than 8OgIL, sucrose 60g/L and high agar concentration of 1% is the best combination among the mediums from I to 12. As for the rate of callus inducement of maltose , concentration of 8OgIL is higher than 6OgIL, the effect is significant in combination of 0.5% agar.3.Kinetin or 2iP with 2mg/L greatly improved the rate of callus inducement. For example the rate of callus inducement from medium 22 with KT can reach 84%; and the medium 24 with 2iPcan reach 63%.4.The time of cold pretreatment also influences the rate of callus inducement.8 days is the proper cold pretreatment time for japonica 02428.5.N6, NB culture mediums are fitable for callus inducement, and MS, MB culture medium are fitable for plant regeneration.6.Among the callus inducement culture mediums, No. 16~. 22and 24 are the best ones for inducing callus; the callus obtained from the mediums of No.5 17~ 21~ 23 are high possible to regenerate the pollen plants.7.Cytological studiey demonstrated that 9983007-2-8, 9983009-2-4,998040-2-5, 998040-3-4 were all primary trisomics, and their chromosome number are 25.The kayotype analysis result of pachtene showed that 9983 007-2-8 is triplo-8, 998040-2-5 is triplo-5. And the dentification result by molecular marker is the same as that by kayotype analysis.