Construction Of Primary Hybridization Pool For Soybean By Ac/Ds Transposon And The Application Of TAIL-PCR In Soybean Study

Soybean as the important economic food crop is one of the main sources of plant oil and protein for mankind. Although great progress has been achieved in soybean breeding in the recent years, improving soybean quantity and quality will still be the main task with the increasing population. Searching for interesting genes for breeding by identification and appraisal of plant gene function is of great importance. Studying the mutants to find out the function of related genes is one of the most effective methods in explore the genes.So it is urgent to study the gene function of soybean. The construction of soybean mutant library can not only provide us the useful materials in studying the gene function but also bring us soybeans with new phenotype and thus provide important materials and germplasm for genetic breeding.More than 9 hundred plants from T₄ of transformed soybean were checked for searching of the homozygous line. One mutant line was found and its quality was appraised. Different reagents were tested for screening the transformed plants. TAIL-PCR was used and the reaction conditions of this system were optimized as well. It will be of significance for producing mutants, screening the transformed plants and for gene cloning with TAIL-RCR.Research results are as follows :1. The sensitivity of different screening agents in Soybean germination and trifoliate expanding stage were systematically explored. The results indicated that: (1) Kanamycin have no effect in soybean germination stage, and it do not fit in transgenic soybean screening agent. The optimal screening concentrations of Hygromycin, chlorsulfuron and NAM on the transgenic soybean seed germination were 11㎎/L, 25p.p.m and 40 uM, respectively. (2) By leaf-smear screening in field condition at large scale, optimal concentration kanamycin, hygromycin and NAM on soybean were 700㎎/L, 110㎎/L and 600 uM, and chlorsulfuron is not appropriate for the field screening.2, Over 900 strains of the identified and suspected Ac, Ds homozygous T₃-generation descendants were detected by PCR. One homozygous Ac and one Ds pure line were initially determined. 3, Ac31-1 mutant strain was found in transgenic offspring. in the process of reproduction of T₃ generation, a large number of mutants were found, These mutants are as following, changes of seed coat color for 38, leave-shape alteration for 11, flower color change for 23 plants, flower time and maturity alteration for 23, alteration of plant heights for 7.4, Protein and oil content of the 73 groups mutants of Ac31 T₃ generation were tested. Protein content of mutation groups was in the range of 34.55-43.15%. Oil content was in the range of 15.99-19.97%. In the comparison of the protein and fatty acid content in Jilin No. 1, significant difference was discovered between the decedents and control.5, Soybean hybrid groups were constructed. Hybridization of 38 pairs were crossed, and more than 120 hybrid seeds were harvested.6,The feasibility TAIL-PCR technology in soybean application was explored, optimized application conditions in Jilin soybean varieties on the 1st. The feasibility of TAIL-PCR in a soybean gene cloning was confirmed.