Study On BIO ORGANS And Its Mutant Ene In Lotus Japonicus Transforming Tobacco And Petunia Hybrida

BIO ORGANS (BIO) in Lotus Japonicus regulates floral organ symmetry and organ size. The mutant gene bio was obtained in large scale mutagenesis of EMS. This two genes have not been deeply studied. In order to study the regulatory mechanism, used Agrobacterium mediated transformation of tobacco and Petunia hybrida with BIO and the mutant gene bio. We observed the life cycles of tobacco and Petunia hybrida, in order to provide experiment basis for the study of regulatory mechanism of BIO gene in the next stage. The main results were as following:1. Used Agrobacterium mediated transformation of tobacco with BIO and the mutant gene bio.18BIO trans genie plants and21bio transgenic plants were obtained. The transgenic plants with BIO and bio had some similar phenotypes:the growth cycle was shorten significantly; incision appeared in almost half of the blades, and a tendency appeared that a leaf divided into two leaves; the color of corollas became shallow; the size, number and the morphology of floral organ all showed a certain change. Besides, some different phenotypes were also existed:compared with the wild type, BIO transgenic plants appeared obvious dwarfism and the organ size was reduced; bio transgenic plants were higher and the organ size was largen.2. Establishment of genetic transformation through different combination of hormones about Petunia hybrida, and confirm6-BA is beneficial to the proliferation of seedling, as well as NAA is beneficial to the induction of callus and adventitious shoot. MS medium was fit for the seed of Petunia hybrida; MS+O.lmg/L6-BA was suitable for the Rapid propagation; and MS+1.0mg/L6-BA+0.1mg/L NAA was suitable for the induction of callus and adventitious shoot, and could be use from preincubation to preliminary screening.3. Using Agrobacterium mediated transformation of Petunia hybrida with BIO and the mutant gene bio. Initial confirm the success of transformation by PCR and RT-PCR detection..The data displayed that both BIO and bio transgenic plants had abnormal leaves with irregular margins., even some of them showed a tendency that one leaf would divide into two. In BIO transgenic plants certain blades had smaller size and became thinner than wild-type with few of them had only main veins left.Phenotypic of tobacco and Petunia hybrida transgenic plants observations showed that BIO may participate in the morphogenesis of plant height, leaf shape and floral organ morphology, etc. And the lose sequence of bio may play an important role in the control of organ size. Sequenced the BIO and bio gene, the gene and protein sequences comparison show the similarity were above90%with a certain sequences in Glycine max and Medicago truncatula, and show a conserved locus may among legume genomes. The regulatory mechanism of BIO and the gene regulatory network should be study in the next stage and the results above provided experiment basis for it.