Identification And Analysis Of 8981 Genetically Upland Rice Cultivars

This topic selects 8981 genes as the target gene,through Agrobacterium transformation method to transfer the target gene into upland rice's genome,and this topic mainly carries on the later identification,using the herbicide,the PCR,the physiology screening,Southern hybridization and the TAIL-PCR.Finally we obtain a new upland rice system which is steady and hereditary.The specific findings are as follows:(1)Candidate Homozygous Strain Screening.In the experimental plots,through sprayed the concentration of 0.8mg/ml ammonium phosphate,97 resistant strains with herbicide resistance were screened for PCR identification,then 93 candidates have been screened out and 4 strains are mixed.(2)Identification of Resistance Strains and Selection of Mutants.After NaCl stress,8of the 93 candidate homozygous strains have stronger salt tolerance,comparing with wild type.After PEG stress,7 of the 93 candidate homozygous strains have stronger drought tolerance comparing with wild type.We screen out 93 strains with high herbicide resistance and the highest herbicide tolerance concentration is 8mg/ml.In addition,the 6strains with special agronomic characters and 33 plants with full grain are obtained.(3)Analysis of Southern Hybridization.In this section,the southern hybridization was carried out by DIG high prime DNA labeling and detection starter kit,and 2 strains of single copy transgenic strains,which heredity is stable,were screened out from 25 lines.The number of two copies of transgenic strains is 12;Multiple copies of transgenic strains are 11.(4)TAIL-PCR Analysis of Target Gene Insertion Sites.The 2 single copy transgenic strains,the 2-9-5 and the 2-19-5,were identified by TAIL-PCR,and the target gene's insertion site of the 2-9-5 is in chromosome 10 th Os10g0351500 gene,which encodes Tyrosine protein;The target gene insertion site of the 2-19-5 is in chromosome 1th Os01g0884300 gene which encodes No apical meristem protein.